transforming-growth-factor-beta and Burns

Wound-healing complications following body contouring for massive weight loss patients are significant, with rates exceeding 40 percent. To better understand aberrant healing in this population, the authors have performed a comparative analysis of the wound milieu literature for patient populations with similar complication rates.. PubMed and Ovid databases were reviewed from January of 1985 to January of 2009 for key terms, including wound healing, obesity, cancer, burn, transplant, and body contouring. Serum and wound levels of multiple factors, including matrix metalloproteinases (MMPs) and cytokines, were assessed.. Complication rates in body contouring surgery range from 31 to 66 percent. Sixty-five studies were reviewed, and wound-healing complication rates were identified for cancer (45.8 percent), burn (30.4 percent), posttransplant (36 percent), and obese (43 percent) populations. In these groups, matrix metalloproteinases and tissue inhibitors of metalloproteinase (TIMPs) help regulate wound repair. Matrix metalloproteinase levels were elevated in cancer (4-fold increase in MMP-2), burn (20- to 30-fold increase in MMP-9), transplant (1.4-fold increase in MMP-2), and obese/chronic (79-fold increase) populations. TIMPs were increased in cancer (1.9-fold increase in TIMP-2) and burn (1.4-fold increase in TIMP-1) patients but decreased in chronic wound (55-fold decrease in TIMP-1) populations. Alterations to these regulatory proteins lead to prolonged matrix degradation, up-regulation of inflammatory mediators, and decreased growth factors, delaying the wound-healing process.. Complications after body contouring surgery are likely multifactorial; however, molecular imbalances to the massive weight loss wound milieu may contribute to poor surgical outcomes. Examining wound regulatory proteins including transforming growth factor-beta, vascular endothelial growth factor, and matrix metalloproteinases could aid in understanding the healing difficulties observed clinically.

Topics: Bariatric Surgery; Biomarkers; Burns; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Neoplasms; Obesity; Organ Transplantation; Plastic Surgery Procedures; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-2; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Weight Loss; Wound Healing

To review the current condition of growth factors and their application to clinical treatment of acute and chronic wounds.. Data from the literature and Medline were analyzed according to their different uses in acute and chronic wounds. Their potential side-effects were studied.. All data showed that wound healing time in acute and chronic wounds was accelerated and wound healing quality was improved after treatment with growth factors. No side-effect was observed.. The efficacy and safety of growth factors in improving wound healing were confirmed. However, some reconsideration about potential problems of growth factors must be made to apply them clinically in the future.

Topics: Animals; Burns; Epidermal Growth Factor; Fibroblast Growth Factor 2; Growth Substances; Humans; Recombinant Proteins; Skin; Soft Tissue Injuries; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Wound Healing

The pathogenesis of hypertrophic scars following thermal injury remains a complex and incompletely understood process but recent investigations into the composition of the tissue itself, the activities of the scar fibroblasts, and the effects of various cytokines and growth factors, have all contributed to the emergence of an increasingly clear picture. Although it may be considered just one example of a broad range of fibroproliferative disorders that afflict many different organs, often in response to diverse environmental insults, the nature of the burn injury and the special properties of skin probably play important roles in promoting the development of this especially troublesome variety of excessive connective tissue. This knowledge has provided the rationale for a number of experimental therapies that, individually or in some combination, may augment or one day supplant the more commonly employed surgical or physical treatments.

Topics: Burns; Carrier Proteins; Cicatrix, Hypertrophic; Connective Tissue Growth Factor; Growth Substances; Humans; Immediate-Early Proteins; Insulin-Like Growth Factor I; Intercellular Signaling Peptides and Proteins; Interferons; Transforming Growth Factor beta

Scar formation and fibrosis often cause devastating disabilities in children suffering severe burn injury. In contrast to the child, the fetus has the ability to heal skin injury without scar formation, and instead with regeneration of epithelial and mesenchymal tissues and restoration of normal skin architecture. In this paper we review those unique features of the fetus and fetal wound healing that may contribute to the scarless repair process. It is hoped that an understanding of these remarkable reparative capabilities may lead to the development of new wound healing therapies that reduce or prevent scar formation and fibrosis in the management of children with burns.

Topics: Burns; Cell Adhesion Molecules; Child; Cicatrix; Collagen; Fetus; Fibroblast Growth Factor 2; Humans; Hyaluronic Acid; Platelet-Derived Growth Factor; Skin Transplantation; Transforming Growth Factor beta; Wound Healing

Because molecular memories of past inflammatory events can persist in epidermal cells, we evaluated the long-term epidermal protein expression landscapes after dermal regeneration and in psoriatic inflammation. We first characterized the effects of two dermal regeneration strategies on transplants of indicator split-thickness skin grafts (STSGs) in ten adult patients with deep burns covering more than 20% of their body surface area. After fascial excision, three adjacent areas within the wound were randomized to receive a permanent dermal matrix, a temporary granulation-tissue-inducing dressing or no dermal component as control. Control areas were covered with STSG immediately, and treated areas after two-weeks of dermis formation. Epidermis-dermis-targeted proteomics of one-year-follow-up samples were performed for protein expression profiling. Epidermal expression of axonemal dynein heavy chain 10 (DNAH10) was increased 20-fold in samples having had regenerating dermis vs control. Given the dermal inflammatory component found in our dermal regeneration samples as well as in early psoriatic lesions, we hypothesized that DNAH10 protein expression also would be affected in psoriatic skin samples. We discovered increased DNAH10 expression in inflammatory lesions when compared to unaffected skin. Our results associate DNAH10 expression with cell proliferation and inflammation as well as with the epidermal memory resulting from the previous regenerative signals of dermis. This study (ISRCTN14499986) was funded by the Finnish Ministry of Defense and by government subsidies for medical research.

Topics: Adult; Burns; Cell Proliferation; Dermis; Dyneins; Epidermis; Female; Humans; Immunohistochemistry; Inflammation; Keratinocytes; Male; Middle Aged; Psoriasis; Regeneration; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing; Young Adult

There have been several case reports of improvement in the appearance of mature burn scars following treatment with fractional CO(2) lasers. However, the biochemical mechanisms responsible for these improvements have not been elucidated.. Ten patients with mature, full-thickness, hypertrophic burn scars received initial treatment with a fractional CO(2) laser. Clinical improvement was measured with Vancouver Scar Scale as well as Patient and Observer Scar Assessment Scale. Fresh tissue samples were obtained before the initial treatment and 48 hours after the first treatment for TaqMan Real-time RT-PCR analyses. Expressions of several scar-related biological markers, including types I and III procollagen, matrix metalloproteinase (MMP)-1, -13, transforming growth factor (TGF)-β1, β2, β3, and basic fibroblast growth factor (bFGF), as well as microRNA miR-17-92 cluster, were investigated.. There were significant improvements in both observer and subject ratings in all scales. Both types I and III procollagen mRNA levels were dramatically down-regulated after treatment, but the ratio of types I/III procollagen mRNA was not different. The expression of MMP-1 was significantly up-regulated after treatment, while TGF-β2, -β3, and bFGF levels were significantly down-regulated. Expression of miR-18a and miR-19a were dramatically up-regulated (P < 0.05) after treatment.. Our study indicated that fractional CO(2) resulted in clinical improvement of mature burn scar. Alteration of types I and III procollagen, MMP-1, TGF-β2, -β3, bFGF, as well as miRNAs miR-18a and miR-19a expression may be responsible for the clinical improvement after treatment. Our finding may have implications for novel treatments and further our understanding of fractional CO(2) laser treatment.

Topics: Adult; Biomarkers; Burns; Cicatrix, Hypertrophic; Down-Regulation; Female; Fibroblast Growth Factor 2; Humans; Lasers, Gas; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 13; MicroRNAs; Middle Aged; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Transforming Growth Factor beta; Treatment Outcome; Up-Regulation

Hypertrophic scarring is a common dermal fibroproliferative disorder that leads to poor quality wound healing, prolongs rehabilitation, and increases morbidity following major thermal and other injuries to the deep dermis. Local and systemic transforming growth factor (TGF)-beta has been implicated as a fibrogenic cytokine in the pathogenesis of many fibrotic disorders, whereas interferon (IFN) alpha-2b may improve the pathologic features of dermal fibrosis directly or by antagonizing the effects of TGF-beta and histamine. Nine patients with severe hypertrophic scarring were evaluated for 8 weeks before treatment with subcutaneous recombinant IFN alpha-2b, 2 x 10(6) IU three times per week for 24 weeks. Clinical assessment was performed using standardized photography, a burn scar assessment tool, and serial scar volume measurements. Monthly measurements of serum TGF-beta and plasma Ntau-methylhistamine were made prior to, during, and after IFN alpha-2b therapy and compared with 27 age-matched controls. Serial biopsies of the hypertrophic scars and normal skin were performed for evaluation of mast cell numbers. Significant improvement in scar assessment occurred in 7 of 9 patients, and 3 of 9 demonstrated significant reductions in scar volume with interferon therapy beyond that occurring during the 8-week control period. For the entire group, mean rates of improvement were significantly better during interferon therapy with no recurrence following treatment. Before interferon therapy, serum TGF-beta was significantly higher in the burn patients with hypertrophic scarring than in a control population (123.04 +/- 36.48 vs. 56.85 +/- 8.38 ng/ml, p < 0.05). Within 3 months of IFN alpha-2b therapy, serum TGF-beta levels fell significantly and remained within the normal range during therapy and after interferon therapy was stopped. Plasma Ntau-methylhistamine levels were also significantly elevated in the hypertrophic scar patients as compared with age and sex-matched controls (153.6 +/- 92.07 vs. 48.3 +/- 28.9 pg/ml, p < 0.05), and significant reductions were achieved with interferon therapy and maintained after interferon was discontinued. Paired biopsies of hypertrophic scarring and normal tissue demonstrated increased numbers of mast cells in hypertrophic scars compared with normal uninjured skin from the same patients (2.65 +/- 1.63 vs. 1.04 +/- 0.62 cells/high power field, p < 0.001); however, no significant change in mast cell content of the hypertrophic

Topics: Adult; Burns; Child; Cicatrix, Hypertrophic; Female; Humans; Interferon alpha-2; Interferon-alpha; Male; Middle Aged; Recombinant Proteins; Transforming Growth Factor beta

The aim of this study was to study the curative effect and the relative mechanism of modified photodynamic therapy combined with Taohong Siwu Decoction in the treatment of hyperplastic scar after severe burn, in order to provide a stable, safe and satisfactory scheme for scar repair.. Forty cases with hyperplastic scars after severe burns admitted to the plastic surgery department from May 2021 to May 2022 were divided into a control group and an observation group by means of the random number table method. The control group was treated with ordinary laser therapy combined with Taohong Siwu Decoction, while the observation group was treated with modified photodynamic therapy combined with Taohong Siwu Decoction. The Vancouver Scar Scale (VSS) was assessed in both groups, and the clinical effectiveness of both groups was compared. HE-staining was performed on the scar tissue of the same patient before and after treatment to observe the changes in the arrangement of fibroblasts. The Vascular Endothelial Growth Factor (VEGF), β-Transforming Growth Factor (TGF-β), and Platelet-Derived Growth Factor (PDGF) in the tissue samples of both groups were detected by quantitative real-time PCR. The patients were followed up for 6 months, and their satisfaction, side effects, and scar recurrence were observed.. Compared with the control group, the VSS score in the observation group was lower (p < 0.05). The therapeutic effect of the observation group was superior to the control group after 3 months (p < 0.05). After 3-months of therapy, the arrangement of fibroblasts in the scar became looser in two groups, and the observation group was more looser. The VEGF, TGF-β and PDGF levels in tissue samples of the observation group were lower than those in the control group after 3 months of treatment (p < 0.05). The satisfaction of the observation group was higher than that of the control group (p < 0.05). The adverse reactions between the two groups showed no difference (p > 0.05), while the recurrence rate was lower in the observation group (p < 0.05).. Modified photodynamic therapy combined with Taohong Siwu Decoction shows remarkable efficacy in patients with hyperplastic scars after severe burns. It can improve the color, thickness, vascular distribution, and softness of the scar, and reduce the level of cytokines related to tissue repair. At the same time, it can improve patients' satisfaction with the aesthetic appearance and reduce the recurrence rate, providing a new comprehensive therapy that is safer and more effective, simple and quick, and easy to promote in the clinic.

Topics: Burns; Cicatrix, Hypertrophic; Humans; Photochemotherapy; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Management of burn wounds with diabetes and microbial infection is challenging in tissue engineering. The delayed wound healing further leads to scar formation in severe burn injury. Herein, a silver-catechin nanocomposite tethered collagen scaffold with angiogenic and antibacterial properties is developed to enable scarless healing in chronic wounds infected with Pseudomonas aeruginosa under diabetic conditions. Histological observations of the granulation tissues collected from an experimental rat model show characteristic structural organizations similar to normal skin, whereas the open wound and pristine collagen scaffold treated animals display elevated dermis with thick epidermal layer and lack of appendages. Epidermal thickness of the hybrid scaffold treated diabetic animals is lowered to 33 ± 2 µm compared to 90 ± 2 µm for pristine collagen scaffold treated groups. Further, the scar elevation index of 1.3 ± 0.1 estimated for the bioengineered scaffold treated diabetic animals is closer to the normal skin. Immunohistochemical analyses provide compelling evidence for the enhanced angiogenesis as well as downregulated transforming growth factor- β1 (TGF-β1) and upregulated TGF-β3 expressions in the hybrid scaffold treated animal groups. The insights from this study endorse the bioengineered collagen scaffolds for applications in tissue regeneration without scar in chronic burn wounds.

Topics: Animals; Burns; Catechin; Collagen; Nanocomposites; Rats; Silver; Skin; Transforming Growth Factor beta; Wound Healing

Topics: Animals; Burns; Cell Proliferation; Collagen; Extracellular Matrix; Fibroblasts; Fibrosis; Gene Expression Regulation; Humans; Mice; Muscle Fibers, Skeletal; Muscle, Skeletal; Myostatin; Signal Transduction; Transforming Growth Factor beta

Burn wounds are associated with a series of risks, such as infection and pathologic scar tissue formation, which significantly delay wound healing and lead to complications. In this study, we successfully fabricated a dextran-hyaluronic acid (Dex-HA) hydrogel enriched with sanguinarine (SA) incorporated into gelatin microspheres (GMs), which had high porosity, good swelling ratio, enhanced NIH-3T3 fibroblast cell proliferation, and sustained SA release profile. The in vitro degradation results indicate that the SA/GMs/Dex-HA hydrogel can be degraded. The in vitro antibacterial tests showed that the SA/GMs/Dex-HA hydrogel can inhibit methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli). We evaluated the wound-healing effects and antibacterial properties of SA/GMs/Dex-HA hydrogels in a rat full-thickness burn infection model. The hematoxylin-eosin (H&E) and Masson's trichrome staining results of the SA/GMs/Dex-HA hydrogel showed that it improved re-epithelialization and enhanced extracellular matrix remodeling, and immunohistochemistry results showed that the expression of TGF-β1 and TNF-α was decreased, while the TGF-β3 expression was increased. Our findings suggest that the SA/GMs/Dex-HA hydrogel provides a potential way for infected burn treatment with high-quality and efficient scar inhibition.

Topics: Animals; Anti-Bacterial Agents; Benzophenanthridines; Burns; Cell Survival; Dextrans; Escherichia coli; Gelatin; Hyaluronic Acid; Hydrogels; Isoquinolines; Male; Methicillin-Resistant Staphylococcus aureus; Mice; NIH 3T3 Cells; Rats, Sprague-Dawley; Skin; Staphylococcal Infections; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing; Wound Infection

T cells play a critical role in host defense against intestinal bacteria. We have shown that ethanol combined with burn injury suppresses Peyer's patch (PP) Th17 cytokines 1 d after injury. We assessed the mechanism of suppressed Th17 effector functions. Mice were gavaged with ethanol 4 h before burn injury and euthanized 1, 3, and 7 d after injury. Mesenteric lymph nodes (MLNs), PPs, and spleen Th1 and Th17 cytokines were assessed. A significant decrease in IL-17, IL-22, IL-2, and IFN-γ were observed in all 3 lymphoid organs 1 and 3 d after injury. We used splenic cells to study the role of IL-6, IL-23, TGF-β, and aryl hydrocarbon receptor (AHR) in suppressing Th17 cytokines. We also assessed whether the AHR agonist 6-formylindolo (3, 2-b) carbazole (FICZ) modulates Th17 cytokines. We found a significant decrease in IL-6 and TGF-β after ethanol and burn; IL-23 was undetectable. The reconstitution of IL-23 in culture medium increased IL-17 by 2-fold and IL-22 by 20-fold in cells from burn ethanol mice. The restoration of IL-6 and TGF-β combined did not influence the release of Th17 cytokines. We observed that AHR was necessary for IL-23 restoration of IL-22 after ethanol and burn injury. The AHR agonist FICZ enhanced IL-22, but not IL-17. None of these treatments influenced the release of Th1 cytokines. Together, these results suggest that IL-23 plays a critical role in regulation of Th17 cytokines. Furthermore, IL-6 and TGF-β do not appear to influence IL-23-mediated restoration of Th17 cytokines after ethanol and burn injury.

Topics: Alcohol-Induced Disorders; Animals; Burns; Disease Models, Animal; Interleukin-23; Interleukin-6; Male; Mice; Th1 Cells; Th17 Cells; Transforming Growth Factor beta

Unfortunately burns are a common occurrence, leading to scarring or death. Platelet-rich plasma (PRP) contains many growth factors that can accelerate wound healing. We analyzed the use of PRP in deep second-degree (dSD), deep second-degree associated with diabetes mellitus (dSDD), and third-degree (TD) burns in rats. Sixty syngeneic rats divided into three groups (dSD, dSDD, and TD) were burned, half receiving topical PRP and half being used as control; 10 additional rats per group were used for PRP preparation. On day 21, the animals were sacrificed and skin biopsies were collected. dSD and dSDD wounds treated with PRP showed faster wound closure, reduction in CD31-, CD68-, CD163-, MPO-, and in TGF-β-positive cells, and an increase in MMP2-positive cells. The neo-epidermis was thinner in the control of both the dSD and dSDD groups and granulation tissue was less reduced in the control of both the dSDD and TD groups. These results indicate that PRP can accelerate the healing process in dSD and dSDD, but not in TD burns.

Topics: Animals; Antigens, CD; Biomarkers; Burns; Cicatrix; Collagen; Cytokines; Diabetes Mellitus, Experimental; Disease Models, Animal; Granulation Tissue; Male; Platelet-Rich Plasma; Rats; Rats, Wistar; Transforming Growth Factor beta; Wound Healing

Wound healing is an incredibly complex biological process that often results in thickened collagen-enriched healed tissue called scar. Cutaneous scars lack many functional structures of the skin such as hair follicles, sweat glands, and papillae. The absence of these structures contributes to a number of the long-term morbidities of wound healing, including loss of function for tissues, increased risk of re-injury, and aesthetic complications. Scar formation is a pervasive factor in our daily lives; however, in the case of serious traumatic injury, scars can create long-lasting complications due to contraction and poor tissue remodeling. Within this report we target the expression of connective tissue growth factor (CTGF), a key mediator of TGFβ pro-fibrotic response in cutaneous wound healing, with controlled local delivery of RNA interference. Through this work we describe both a thorough in vitro analysis of nanolayer coated sutures for the controlled delivery of siRNA and its application to improve scar outcomes in a third-degree burn induced scar model in rats. We demonstrate that the knockdown of CTGF significantly altered the local expression of αSMA, TIMP1, and Col1a1, which are known to play roles in scar formation. The knockdown of CTGF within the healing burn wounds resulted in improved tissue remodeling, reduced scar contraction, and the regeneration of papillary structures within the healing tissue. This work adds support to a number of previous reports that indicate CTGF as a potential therapeutic target for fibrosis. Additionally, we believe that the controlled local delivery of siRNA from ultrathin polymer coatings described within this work is a promising approach in RNA interference that could be applied in developing improved cancer therapies, regenerative medicine, and fundamental scientific research.

Topics: Animals; Burns; Cell Line; Cicatrix; Connective Tissue Growth Factor; Drug Delivery Systems; Fibrosis; Gene Silencing; Humans; Mice; Nanostructures; Polymers; Rats, Sprague-Dawley; Regeneration; RNA, Small Interfering; Skin; Sutures; Transforming Growth Factor beta

The aim of this study was to investigate the effects of low-level red laser on tissue repair in rats submitted to second-degree burn, evaluating if the timing of laser treatment influences the healing process. The animals had their backs shaved and divided as follows: control group (n = 12)-rats burned and not irradiated, early laser group (n = 12)-rats burned and irradiated from day 1 after injury for five consecutive days, and late laser group (n = 14)-rats burned and irradiated from day 4 after injury for five consecutive days. Laser irradiation was according to a clinical protocol (20 J/cm(2), 100 mW, continuous wave emission mode, 660 nm) as recommended by the laser device manufacturer. Half of the animals were sacrificed 10 days after burn, and the other animals were sacrificed 21 days after burn. The late laser group accelerated wound contraction 10 and 21 days after burn. The late laser group accelerated reepithelialization 18 days after burn. The late laser group increases the granulation tissue 10 and 21 days after burn. Both irradiated groups increased type III collagen expression and TGF-β 21 days after burn. Both irradiated groups increased macrophage and myofibroblast numbers 10 days after burn and decreased 21 days after. Low-level red laser exposure contributes to the process of tissue repair of second-degree burns, but the intervention during proliferative phase is crucial in the final outcome of the repair process.

Topics: Animals; Burns; Cell Proliferation; Collagen Type I; Collagen Type III; Granulation Tissue; Lasers, Semiconductor; Low-Level Light Therapy; Male; Rats; Rats, Wistar; Re-Epithelialization; Transforming Growth Factor beta; Treatment Outcome

Burn patients requiring hospitalization are often treated for anxiety with benzodiazepines (BDZs). Benzodiazepines are reported to influence immune system function. Immune system alterations are a major cause of burn-induced mortality. We wanted to determine whether the BDZ, midazolam given daily at an anxiolytic dose, had any influence on the burn injury-induced inflammatory response in the blood and wound. Mice received a 15% total body surface area flame burn and received either midazolam 1 mg/kg i.p. or saline 0.1 ml daily. Blood and skin wounds were harvested 24 h after injection on post-burn day 2, 3, 7, or 8. Mice treated with midazolam had significantly lower serum IL-1β (p=0.002), TNF-α (p=0.002), IL-6 (p=0.016), IL-10 (p=0.009), and TGF-β (p=0.004) than saline-treated mice, with little impact on serum chemokine levels. In the wound, TNF-α and IL-10 were the only cytokines significantly influenced by the drug, being lower (p=0.018) and higher (p=0.006), respectively. The chemokines in the wound influenced significantly by midazolam were MIP-1α, MIP-1β, and MIP-2 while MCP-1 and KC were not. There were more inflammatory cells at the burn wound margin in midazolam-treated mice on post-burn day 3. Although serum nitrate/nitrite was significantly increased by midazolam (p=0.03), both eNOS and iNOS mRNA expression in the wound were similar to the saline group. We found that midazolam given daily after burn injury significantly influenced the inflammatory response. The clinical implications of these findings on wound healing and shock following burn injury, especially larger burns, deserve further investigation.

Topics: Animals; Burns; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CXCL2; Inflammation; Interleukin-10; Interleukin-1beta; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Midazolam; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; RNA, Messenger; Skin; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing

The high incidence of morbidity and mortality following major burn can be in part attributed to immune dysfunction and wound healing complications. Inflammation plays a major role in the complex process of wound repair. Recently, a novel class of T-helper cells, termed Th-17 cells, has been found to secrete the pro-inflammatory cytokines IL-17 and IL-22. The Th-17 response also involves other cytokines, such as IL-6 and TGF-β, which have been shown to be associated with burn-induced inflammation. Nonetheless, the relationships between the Th-17 response and post-burn inflammation are unknown.. C57BL/6 male mice (n = 5-6/group) were subjected to a major burn (25% TBSA) or sham procedure. Three hours thereafter, skin samples were collected (uninjured skin and burn skin) and processed for the determination of Th-17 cytokine (IL-6, IL-17, IL-22, IL-23, IL-27, and TGF-β) levels by ELISA.. At 3h after burn a significant (~3-fold) increase in tissue levels of IL-17 and IL-22 was observed at the burn site as compared to sham skin. The burn-induced Th-17 response was independent of statistically significant changes in other Th-17 cytokines (i.e., IL-6, IL-23, IL-27 and TGF-β).. These findings indicate the development of a robust Th-17 response at the burn site that may play an important role in subsequent immune and wound healing derangements.

Topics: Animals; Burns; Cytokines; Enzyme-Linked Immunosorbent Assay; Interleukin-17; Interleukins; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Th17 Cells; Transforming Growth Factor beta

Topics: Bariatric Surgery; Biomarkers; Burns; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 3; Matrix Metalloproteinase 9; Neoplasms; Obesity; Organ Transplantation; Plastic Surgery Procedures; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinase-2; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Weight Loss; Wound Healing

Hypertrophic scar (HTS) following thermal injury is a dermal fibroproliferative disorder that leads to considerable morbidity. Previous clinical studies from our laboratory have suggested that interferon-alpha2b (IFN-alpha2b) improves scar quality as a result of suppression of fibroblast functions. Fibrocytes, which constitute a unique cell population, have recently been reported to contribute to wound healing and to a variety of fibrotic conditions, including HTS. Therefore, we hypothesize that improvement of scar in HTS patients after IFN-alpha2b treatment may be associated with a decreased number of fibrocytes or altered fibrocyte function. Using flow cytometry, immunofluorescent staining, and confocal microscopy, we demonstrate here that the marker protein leukocyte-specific protein 1 (LSP1) is stably expressed by fibrocytes for at least 2 months, whereas other potential fibrocyte markers, such as CD34 and CD45, gradually disappear. Using dual staining immunohistochemistry for LSP1 and procollagen, we demonstrated a significant reduction in numbers of fibrocytes in HTS tissue from patients after treatment with systemic IFN-alpha2b. IFN-alpha2b was shown to abolish fibrocyte differentiation from peripheral blood mononuclear cells (PBMCs) in vitro in a dose-dependent fashion. In addition, IFN-alpha2b inhibits proliferation of fibrocytes and T lymphocytes and reduces transforming growth factor-beta (TGF-beta)-mediated alpha-smooth muscle actin (alpha-SMA) expression in fibrocytes. Taken together with our previous study in which we showed that fibrocytes could indirectly regulate dermal fibroblasts in burn patients, the present study suggests that the improvement of scar quality in HTS patients after IFN-alpha2b treatment is associated with decreased numbers and activities of fibrocytes.

Topics: Actins; Adolescent; Adult; Burns; Cell Differentiation; Cell Proliferation; Child; Cicatrix, Hypertrophic; Female; Humans; Interferon alpha-2; Interferon-alpha; Leukocytes; Male; Microfilament Proteins; Middle Aged; Procollagen; Recombinant Proteins; Transforming Growth Factor beta; Wound Healing

CD4+CD25+ regulatory T cells (Tregs) play a critical role in suppressing the development of autoimmune disease, in controlling potentially harmful inflammatory responses, and in maintaining immune homeostasis. Because severe injury triggers both excessive inflammation and suppressed adaptive immunity, we wished to test whether injury could influence Treg activity. Using a mouse burn injury model, we demonstrate that injury significantly enhances Treg function. This increase in Treg activity is apparent at 7 days after injury and is restricted to lymph node CD4+CD25+ T cells draining the injury site. Moreover, we show that this injury-induced increase in Treg activity is cell-contact dependent and is mediated in part by increased cell surface TGF-beta1 expression. To test the in vivo significance of these findings, mice were depleted of CD4+CD25+ T cells before sham or burn injury and then were immunized to follow the development of T cell-dependent Ag-specific immune reactivity. We observed that injured mice, which normally demonstrate suppressed Th1-type immunity, showed normal Th1 responses when depleted of CD4+CD25+ T cells. Taken together, these observations suggest that injury can induce or amplify CD4+CD25+ Treg function and that CD4+CD25+ T cells contribute to the development of postinjury immune suppression.

Topics: Animals; Burns; CD4 Antigens; Cell Communication; Cell Proliferation; Cytokines; Flow Cytometry; Immunophenotyping; Lymph Nodes; Lymphocyte Depletion; Male; Mice; Mice, Inbred BALB C; Receptors, Interleukin-2; Reverse Transcriptase Polymerase Chain Reaction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Transforming Growth Factor beta1

To investigate the effect of asiaticoside on the expressions of transforming growth factor (TGF)-beta mRNA, matrix metalloproteinases (MMPS) and tissue inhibitors of metalloproteinases (TIMPs) in postburn hypertrophic scars.. Nine specimens of postburn (5-8 months) hypertrophic scars with asiaticoside treatment and 9 without asiaticoside treatment were collected for testing the expressions of MMPS, TIMPs, type I and III collagen and TGF-beta mRNA by immunohistochemistry and in situ hybridization methods, followed by image analysis of the results.. The expressions of TGF-beta mRNA and MMPS/TIMPS were all detected in the fibroblast cytoplasm. The expression of TGF-beta(1) mRNA in asiaticoside-treated scars was significantly lower than that in scars without asiaticoside treatment (P<0.01). In contrast, the expression of TGF-beta(3) mRNA was significantly higher in asiaticoside group (P<0.05). The expression of TIMP1 in asiaticoside group was significantly lower than that in non-asiaticoside group (P<0.01), and the expression of type I collagen in asiaticoside-treated scars was lower than that in non-asiaticoside-treated specimens (P<0.05), but the expression of MMP(1), MMP(2) and TIMP(2) and type III collagen exhibited no significant differences between the two groups (P>0.05).. Asiaticoside can down-regulate TGF-beta(1) mRNA and TIMP(1) expressions and up-regulate TGF-beta(3) mRNA expression in postburn hypertrophic scars, and is also capable of decomposing the products of type I collagen, contributing to the reduction of hypertrophic scar formation.

Topics: Anti-Infective Agents; Burns; Cicatrix, Hypertrophic; Collagen Type I; Female; Humans; Male; Matrix Metalloproteinases; RNA, Messenger; Skin; Tissue Inhibitor of Metalloproteinase-1; Transforming Growth Factor beta; Triterpenes

This study aimed to investigate the interaction between serum levels of TGF-beta and active-immune cell infiltration in burn wounds of various depths.. Thirty female Sprague-Dawley rats were divided into three groups: full-thickness burns (F), partial-thickness burns (P), and no burns (S). After burn-induction, blood samples were obtained only once from shams and at postburn 1, 48 h, and 7 days in burn groups. Serum levels of TGF-beta were measured by means of the ELISA. The proportions of neutrophils, fibroblasts, vascular proliferation, CD68-macrophages, and CD3-lymphocytes were studied immunohistochemically and graded semiquantitatively.. Serum TGF-beta levels in the F and P groups were lower than those in sham at 1h after burn (p<.05). No significant differences in TGF-beta were observed between groups F and P on days 2 and 7 after injury. No local accumulation of macrophages and fibroblasts was noted in either burn group, but the proportion of lymphocytes was higher in the P group at 1h after burn. Neutrophils were higher in the F group than the P on day 7 after burn.. Prolonged neutrophil infiltration in full-thickness burn wounds and suppressed lymphocyte proliferation in partial-thickness burn wounds seem to be related to an increase in serum TGF-beta levels.

Topics: Animals; Burns; Cell Proliferation; Female; Fibroblasts; HLA-DR Antigens; Immunohistochemistry; Lymphocytes; Macrophages; Neutrophil Infiltration; Random Allocation; Rats; Rats, Sprague-Dawley; Transforming Growth Factor beta

Knowledge of the pathophysiology of hypertrophic scarring following deep dermal injuries is minimal due to the lack of an animal model. We previously confirmed that thick scars in female, red Duroc pigs (FRDP) are similar to human hypertrophic scar. The purpose of this study was to evaluate TGFbeta1, IGF-1, decorin, and versican expression in FRDP wounds. Deep and shallow wounds on the backs of two FRDPs were studied over 5 months. Immunohistochemistry was performed for TGFbeta1, IGF-1, decorin, and versican. TGFbeta1 and IGF-1 mRNA were evaluated by in situ hybridization and RT-PCR. In shallow wounds (1) TGFbeta1 protein was not detectable and IGF-1 protein was seen at 10 days post-wounding. TGFbeta1 and IGF-1 mRNA were elevated for 30 days. (2) Decorin protein was not detected at 10th day, but returned to levels of uninjured skin. (3) Versican protein was not detectable at any time. In deep wounds, (1) TGFbeta1 and IGF-1 protein and mRNA were elevated early, (2) decorin protein was greatly reduced for the first 90 days, and (3) versican protein was present from 30 to 150 days. These findings correlate with findings reported in the literature for human hypertrophic scar and further validate the FRDP model of hypertrophic scarring.

Topics: Animals; Burns; Chondroitin Sulfate Proteoglycans; Cicatrix; Cicatrix, Hypertrophic; Decorin; Dermis; Extracellular Matrix Proteins; Female; Humans; Immunohistochemistry; In Situ Hybridization; Insulin-Like Growth Factor I; Lectins, C-Type; Proteoglycans; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Swine; Transforming Growth Factor beta; Versicans

Burn injury induces immunosuppression, which is associated with an increased susceptibility to infection. Our laboratory has demonstrated that burn injury also impairs humoral immunity. We reported that burn injury enhanced expression of transforming growth factor-beta (TGF-beta) mRNA and that exogenous TGF-beta further impaired humoral immunity. The objective of this study was to clarify the role of TGF-beta on humoral immunity after burn injury with a neutralizing experiment.. Twelve BALB/c mice were randomly divided into two groups: sham and burn. Anesthetized mice received a 20% full-thickness burn or sham injury. The murine splenocytes containing 1.5 x 10 cells/mL were cultured with 2.5 microg/mL of lipopolysaccharide with or without 0.5 ng/mL of TGF-beta or 1 microg/mL of anti-TGF-beta neutralizing antibody, if necessary. Concentrations of immunoglobulin (Ig) M in the cell culture supernatant were determined by enzyme-linked immunosorbent assay and the number of IgM-secreting cells in the culture was measured by enzyme-linked immunospot assay.. After 2-day culture, neutralization of TGF-beta dramatically restored IgM synthesis after burn injury. After 5-day culture, however, it restored IgM concentration but failed to restore a number of IgM-secreting cells.. This neutralizing experiment demonstrated that TGF-beta is one of the inhibitors of IgM synthesis after burn injury. However, neutralization of TGF-beta was not enough to completely restore humoral immunity after burn injury. Investigation of the mechanism of impaired IgM synthesis after burn injury should be continued.

Topics: Animals; Antibody Formation; Burns; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Immunoglobulin G; Immunoglobulin M; Male; Mice; Mice, Inbred BALB C; Neutralization Tests; Spleen; Transforming Growth Factor beta

Placental extracts have been used as Chinese folk medicines to accelerate wound healing. However, the molecular mechanism of placental extracts on wound healing has not been identified. It is known that fibroblast growth factors (FGF) and transforming growth factors (TGF) are two key factors involved in wound healing.. To determine the molecular mechanism of placental extracts on wound healing.. The protein levels of both growth factors in rat skins with thermal injury were therefore studied to explore the molecular mechanism of placental extracts on wound healing. As cell proliferation is essential for wound healing, effects of placental extracts on fibroblast proliferation were also determined.. As compared with the controls, the S phase of fibroblasts was significantly increased by 1.5-, 1.7- and 4.7-fold for 1, 10 and 30 mg mL(-1) of placental extracts, respectively. The increase of the S phase was not due to the minute amount of sex hormones in the placental extracts as the addition of equivalent amounts of hormones showed no increase of the S phase. In addition, a 2.5-fold increase of TGF-beta1 in wound skin biopsy was noticed with 30 mg mL(-1) of porcine placental extracts. The FGF levels in the wound skin receiving 30 mg mL(-1) of porcine placental extracts were also significantly increased compared with the controls.. These ex vivo data support the observation that the application of 30 mg mL(-1) of placental extracts reduced the wound healing time by about 50%. To the best of our knowledge, this is the first report to explore the molecular mechanisms of porcine placental extracts on wound healing. These results may provide the insight into the potential use of porcine placental extracts as an alternative medicine for accelerating wound healing.

Topics: 3T3 Cells; Animals; Burns; Cell Division; Enzyme-Linked Immunosorbent Assay; Estradiol; Fibroblast Growth Factors; Fibroblasts; Male; Mice; Placental Extracts; Progesterone; Rats; Rats, Sprague-Dawley; Swine; Transforming Growth Factor beta; Wound Healing

Hypertrophic scarring occurs after deep dermal wounds. Our understanding of the etiology is poor; one reason is the lack of an animal model. In 1972, Silverstein described scarring in the Duroc pig but the model was never confirmed nor disproved. Another reason, as we previously suggested, is that hypertrophic scarring only occurs within regions of human skin that contain cones and the cones have not been studied in relation to hypertrophic scarring. We, therefore (i) explored healing in the female, red Duroc model for similarities to human hypertrophic scarring, studying wound thickness, appearance, healing status at 3 weeks, histology, and immunocytochemical localization of decorin, versican, TGFbeta1 and IGF-1; and (ii) examined Duroc skin for cones. We found that healing after deep wounds in Duroc pigs is similar, but not identical, to human hypertrophic scarring. We also found that Duroc skin contains cones. Healing in the female, red Duroc pig is sufficiently similar to human hypertrophic scarring to warrant further study so that it can be accepted or rejected as a model of human hypertrophic scarring. In addition, the relationship of the cones to hypertrophic scarring needs further detail and can be studied in this model.

Topics: Animals; Burns; Chondroitin Sulfate Proteoglycans; Cicatrix, Hypertrophic; Decorin; Extracellular Matrix Proteins; Female; Immunoenzyme Techniques; Insulin-Like Growth Factor I; Lectins, C-Type; Models, Animal; Proteoglycans; Swine; Transforming Growth Factor beta; Transforming Growth Factor beta1; Versicans; Wound Healing

Wound healing consists of re-epithelialization, contraction and formation of granulation and scar tissue. TGF-b is involved in these events, but its exact roles are not well understood. Here we demonstrate that topical application of a synthetic TGF-b antagonist accelerates re-epithelialization in pig burn wounds (100% re-epithelialization in antagonist-treated wounds vs. approximately 70% re-epithelialization in control wounds on postburn day 26) and reduces wound contraction and scarring in standard pig skin burn, pig skin excision and rabbit skin excision wounds. These results support the distinct roles of TGF-b in the complex process of wound healing and demonstrate the feasibility of manipulating wound healing by TGF-b antagonist.

Topics: Animals; Burns; Cicatrix; Epithelial Cells; Kinetics; Models, Biological; Peptide Fragments; Rabbits; Skin; Swine; Transforming Growth Factor beta; Transforming Growth Factor beta1; Wound Healing

To investigate the effect of antisense TGF-beta 1 in inhibiting scar formation during wound healing.. SD rats were divided into three groups after skin burn: group one was treated with antisense TGF-beta 1 oligonucleotide; group two was treated with antisense TGF-beta 1 recombinant plasmid and the control group. In different time, RT-PCR and immunohistochemistry were used to verify the expression of TGB beta 1 mRNA and protein. Type I Collagen mRNA expression was determined by hybridization in situ. Inflammatory reaction and collagen distribution were observed pathologically.. In the groups received antisense ODN and recombinant plasmid, the expression of TGF-beta 1 mRNA and protein reduced during 14 days after burn. In the control group, type I collagen mRNA began to express at the second week and reached a peak at the fourth week, while the antisense groups kept low expression. The antisense group also showed mild inflammatory reaction and less synthesis of collagen.. Antisense TGF-beta 1 could prevent the scar formation during wound healing.

Topics: Animals; Burns; Cicatrix; Male; Oligonucleotides, Antisense; Rats; Rats, Sprague-Dawley; Transforming Growth Factor beta; Transforming Growth Factor beta1; Wound Healing

Peripheral blood fibrocytes are a newly identified leukocyte subpopulation that displays fibroblast-like properties. These blood-borne cells can rapidly enter the site of injury at the same time as circulating inflammatory cells. We hypothesize that circulating fibrocytes represent an important source of fibroblasts for healing of extensive burn wounds where it may be difficult for fibroblasts to migrate from the edges of uninjured tissue. In this study we identified and quantified fibrocytes among the adherent cells cultured from human peripheral blood mononuclear cells (PBMC) obtained from 18 burn patients and 12 normal individuals, based on their ability to express type I collagen. Our results showed that adherent cells cultured from PBMC of burn patients differentiated to fibrocytes more efficiently than did those from normal individuals. The percentage of type I collagen-positive fibrocytes was significantly higher for patients than for controls (89.7 +/- 7.9% versus 69.9 +/- 14.7%, p < 0.001). This percentage was consistently higher for patients with a >/=30% total body surface area burn until 1 year, with the highest percentage appearing within 3 weeks of injury. A positive correlation was found between the levels of serum transforming growth factor-beta1 (TGF-beta1) and the percentage of fibrocytes developing in the cultures of PBMC derived from these patients. We also demonstrated that fibrocytes were derived from CD14(+) cells but not CD14(-) cells. Conditioned medium from CD14(-) cells was, however, required for fibrocyte differentiation, whereas direct contact between CD14(-) and CD14(+) cells was not necessary. Treatment of the cell cultures with TGF-beta1 enhanced the development of collagen-positive cells, whereas the inclusion of neutralizing anti-TGF-beta1 antibodies in the CD14(-) conditioned medium suppressed fibrocyte differentiation. These data suggest that the development of fibrocytes is up-regulated systemically in burn patients. Increased TGF-beta in serum stimulates the differentiation of the CD14(+) cell population in PBMC into collagen-producing cells that may be important in wound healing and scarring.

Topics: Adolescent; Adult; Burns; Cell Adhesion; Cell Count; Cell Differentiation; Cell Division; Cells, Cultured; Collagen; Female; Humans; Leukocytes; Lipopolysaccharide Receptors; Male; Middle Aged; Transforming Growth Factor beta; Transforming Growth Factor beta1; Wound Healing

In order to understand the roles of pro-inflammatory and anti-inflammatory cytokines in burn injury and sepsis post-burn, serial changes in serum levels of transforming growth factor beta-1 (TGF-beta-1) were determined and compared to those of IL-6 and IL-10 in 15 burned patients. Among these 15 patients, 8 recovered without sepsis. The other seven, who were septic, expired. Our results showed that an initial peak serum TGF-beta-1 response was detected within 1 day post-burn. Peak serum IL-6 and IL-10 responses were also detected within 4 days after the burn injury of these patients. Significant differences in peak serum IL-6, IL-10 and TGF-beta-1 levels were not found between patients with total body surface area (TBSA) of greater or less than 50% and between patients who survived or expired from burn injury. Afterwards, levels of circulating IL-6 and IL-10 remained low in the survivors. However, a second peak response in serum TGF-beta-1 levels was observed in all burned patients analyzed. The second peak serum TGF-beta-1 levels post-burn of the eight survivors and the seven non-survivors were from 28,542 to 76,554 pg/ml (a mean value of 51,256+/-14,264 pg/ml) and from 8616 to 40,851 pg/ml (a mean value of 24,079+/-10,399 pg/ml), respectively. A significant difference (P<0.01) in mean values of the second peak TGF-beta-1 responses between groups of survivors and non-survivors was detected. Levels of circulating IL-6 in the septic non-surviving patients showed a tendency to increase 1-2 weeks post-burn and reached high levels before the expiration of these patients. After an initial peak response, the serum IL-10 level remained low in one of the seven non-survivors, while it increased in the other six non-survivors. However, marked increases in circulating IL-10 levels were observed only just before the death of these non-survivors. In conclusion, an initial increase in serum levels of IL-6, IL-10 and TGF-beta-1 was detected post-burn. A marked increase in serum levels of IL-6 before death suggests its role in the pathophysiology of sepsis in burned patients. In addition, a low secondary TGF-beta-1 response and a lack and/or delay in the increase of circulating IL-10 in the non-survivors may all contribute to the pathophysiology of septic death in burned patients.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Burns; Female; Follow-Up Studies; Humans; Interleukin-10; Interleukin-6; Male; Middle Aged; Prognosis; Sepsis; Survival Rate; Transforming Growth Factor beta; Transforming Growth Factor beta1

To observe the effect of basic fibroblast growth factor (bFGF) on the outcome of myofibroblasts in burn wounds, and to further explore the mechanism of bFGF on wound healing.. Seventy-two Wistar rats were anaesthetized and put into hot water at the temperature of 98 degrees C with their back hair cut so as to cause full-thickness scald injury with an area of 30% of the total body surface. Then the rats were randomly divided into two groups of 36 mice: pure thermal injury group (administered with sterilization and dressing every other day for three times) and bFGF treatment group (administered locally with bFGF every other day in addition to the routine dressing). Three hours, six hours, one day, three days, seven days, and fourteen days after scalding samples of skin wound was taken, six rats for each time-point. Six rats were put into water at the temperature of 37 degrees C as controls and their skin samples were taken 8s after. In situ hybridization and immunohistochemical techniques were employed to detect the expression of caspase mRNA and proteins. alpha-smooth muscle actin (ASMA), and transforming growth factor-beta1 (TGF-beta1) were detected by immunohistochemical staining at different time points after scalding.. No obvious difference of ASMA expression in dermal tissues was seen at the early stage of injury. The number of cells with positive ASMA expression began to increase 1 approximately 3 days after and reached the peak by the 7th day after scalding, and then decreased gradually. The ASMA expression in bFGF group was remarkably increased by the 7th day, significantly higher than that in the pure thermal injury group (P < 0.05). By the 14th day, the ASMA expression in the bFGF group was still significantly higher than that in pure thermal injury group (P < 0.01), however, it was much lower than that in the bFGF group by the 7th day. By the 14th days after scald injury. The number of TGF-beta1 positive cells began to increase since the 3rd hour after scald injury and began to decrease by the 14th day in both experimental groups. However, the TGF-beta1 expression in bFGF group was stronger than that in pure thermal injury group. The expressions of caspase-3 mRNA and protein in bFGF group changed in the same way as in the simple injured group. Three hours after injury, the expression of caspase-3 mRNA was lower in bFGF group than in pure injury group (P < 0.05). Then the expression decreased till the 3rd day. Six hours after injury, no difference in the expression of caspase-3 mRNA was found between the two experimental groups. The expression of caspase-3 reached its second peak by the 7th day and then decreased again. However, the first expression peak of the bFGF group was lower than that of the pure thermal injury group, however, the second peak of the bFGF group was higher.. Myofibroblasts may play a critical role in wound closure and healing. bFGF treatment may increase the expression of TGF-beta1 and have a potential synergistic effect with other growth factors to stimulate the apoptosis of myofibroblasts during wound healing.

Topics: Actins; Animals; Burns; Caspases; Fibroblast Growth Factor 2; Fibroblasts; Gene Expression Regulation, Enzymologic; Immunohistochemistry; In Situ Hybridization; Male; Muscle, Smooth; Rats; Rats, Wistar; RNA, Messenger; Transforming Growth Factor beta; Transforming Growth Factor beta1; Wound Healing

Immune suppression is a common complication of injury. Transforming growth factor-beta(1) (TGF-beta(1)), a cytokine with diverse anti-inflammatory and anti-apoptotic effects, may play an important role. Smad 2 and Smad 3 are transcription factors that mediate the effects of TGF-beta(1). We hypothesized that burn-induced immunosuppression would be accompanied by increased apoptosis in lymphoid organs, a change likely associated with changes in TGF-beta(1) and Smad 2/3 expression. Mice were subjected to 18% body surface area flame burn. Lymph nodes, spleen, and thymus were harvested at multiple time points after injury. TGF-beta(1) and Smad 2/3 expression and levels of apoptosis were determined in whole tissue and in sorted T-cells by flow cytometry, RT-PCR, ELISA, and Western blot. TGF-beta(1) protein expression in the thymus increased from 1 to 7 days. Smad 2/3 protein expression was decreased at the same time points. This down-regulation was more dramatic in the non-T-cells than in the T-cells themselves. RT-PCR confirmed down-regulation of Smad 3 mRNA in the thymus from 3 to 6 h. Apoptosis in the thymus doubled at 1 day (6.4% control vs 12.8% burned), which corresponded with a marked decrease in thymus mass on subjective assessment. No changes were observed in other lymphoid organs. Burn injury in mice increases TGF-beta(1) expression in the thymus, while suppressing expression of its intracellular mediator, Smad 2/3. This response is most pronounced in the non-T-cell tissue, which suggests the thymic epithelial cells may be responsible for the increased thymic apoptosis. This TGF-beta(1) and Smad 2/3 counterregulation in response to injury may represent a potential mechanism for postinjury immune suppression.

Topics: Animals; Apoptosis; Burns; DNA-Binding Proteins; Flow Cytometry; Gene Expression; Mice; Mice, Inbred C57BL; RNA, Messenger; Smad2 Protein; Smad3 Protein; Thymus Gland; Trans-Activators; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Animals; Burns; Corticosterone; Dexamethasone; Endotoxins; Escherichia coli; Gene Expression Regulation; Glucocorticoids; Insulin-Like Growth Factor I; Male; Muscle, Skeletal; Myostatin; Rats; RNA, Messenger; Sepsis; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

A severe thermal injury is commonly associated with immune suppression and increased susceptibility to sepsis, frequently leading to multiple organ failure. Transforming growth factor-beta (TGF-beta) is a potent immunosuppressive cytokine involved in complications associated with major trauma. Interleukin- 4 (IL-4) is thought to synergize the immunosuppressive activity of TGF-beta by promoting naive lymphocytes to differentiate and generate TGF-beta secreting cells. This study examines the alterations in serum levels of TGF-beta and IL-4 after a thermal injury. Male Sprague-Dawley rats (300-400 g) were anesthetized and received a 50% total body surface area full-thickness scald burn followed by fluid resuscitation and analgesia. Control rats were given the same treatment, but were immersed in water at room temperature. Rats were sacrificed from 1 h to 8 days after injury. Blood samples were collected aseptically from the inferior caval vein. Serum levels of TGF-beta and IL-4 were measured by enzyme linked immunosorbent assay. Rats in the control and thermal injury groups showed similar increases in serum TGF-beta 1 h after injury. A progressive increase in serum TGF-beta was observed in burned animals compared to control animals starting on day 3 and continued through day 8 (P < 0.01). Serum IL-4 levels in control and thermally injured animals remained undetectable (< 15.6 pg/mL) throughout the experiment. Thermal injury induces a significant increase in serum TGF-beta, which may contribute to post-burn immunosuppression with an increased susceptibility to sepsis.

Topics: Animals; Biomarkers; Burns; Enzyme-Linked Immunosorbent Assay; Male; Rats; Rats, Sprague-Dawley; Time Factors; Transforming Growth Factor beta

Transforming growth factor-beta (TGF-beta) isoforms are multifunctional cytokines that play an important role in wound healing. Transgenic mice overexpressing TGF-beta in the skin under control of epidermal-specific promoters have provided models to study the effects of increased TGF-beta on epidermal cell growth and cutaneous wound repair. To date, most of these studies used transgenic mice that overexpress active TGF-beta in the skin by modulating the latency-associated-peptide to prevent its association with active TGF-beta. The present study is the first to use transgenic mice that overexpress the natural form of latent TGF-beta 1 in the epidermis, driven by the keratin 14 gene promoter to investigate the effects of locally elevated TGF-beta 1 on the healing of partial-thickness burn wounds made on the back of the mice using a CO(2) laser. Using this model, we demonstrated activation of latent TGF-beta after wounding and determined the phenotypes of burn wound healing. We found that introduction of the latent TGF-beta1 gene into keratinocytes markedly increases the release and activation of TGF-beta after burn injury. Elevated local TGF-beta significantly inhibited wound re-epithelialization in heterozygous (42% closed versus 92% in controls, P < 0.05) and homozygous (25% versus 92%, P < 0.01) animals at day 12 after wounding. Interestingly, expression of type I collagen mRNA and hydroxyproline significantly increased in the wounds of transgenic mice, probably as a result of a paracrine effect of the transgene.

Topics: Animals; Burns; Cell Division; Collagen Type I; Epidermis; Gene Expression Regulation; Humans; Hydroxyproline; Immunohistochemistry; Keratin-14; Keratinocytes; Keratins; Lasers; Mice; Mice, Transgenic; Promoter Regions, Genetic; Recombinant Fusion Proteins; RNA, Messenger; Skin; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transgenes; Wound Healing

Pseudomonas aeruginosa infection, one of the major complications of burn wounds, may lead to sepsis and death. Using the Multi-Probe Template/RNase protection assay, we have compared the expression of different cytokine genes within the skin and livers of thermally injured mice infected with P. aeruginosa PAO1. Thermal injury alone enhanced or up-regulated certain cytokines, including macrophage colony-stimulating factor (M-CSF), interleukin 1 (IL-1)RI, IL-1 beta, macrophage inflammatory protein (MIP)-1 beta and MIP-2; while PAO1 challenge alone up-regulated tumour necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) expression. The combination of thermal injury plus PAO1 infection enhanced the expression of several pro-inflammatory and haematopoietic cytokines [stem cell factor (SCF), leukocyte inhibitory factor (LIF), IL-6 and TNF-alpha]; induced the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) and G-CSF by 5 h and the expression of additional cytokines, including TGF-beta, TNF-beta, lymphotoxin beta (LT-beta), interferon gamma (IFN-gamma), and IFN-beta by 40 h post-burn/infection. While the most intense cytokine expression occurred in the skin, the majority of cytokines tested were also expressed in the liver by 40 h post-burn/infection. These results suggest that in P. aeruginosa infection of burn wounds: (1) up-regulation of the expression of different cytokines, locally and within the livers of burned mice, is an indication of P. aeruginosa -induced sepsis; and (2) IL-6 and G-CSF play an important role in the host response mechanism.

Topics: Animals; Burns; Cytokines; Female; Gene Expression Profiling; Granulocyte Colony-Stimulating Factor; Interleukin-6; Interleukins; Liver; Macrophage Colony-Stimulating Factor; Mice; Mice, Inbred C57BL; Pseudomonas Infections; Sepsis; Skin; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Up-Regulation

To localize the distribution of basic fibroblast growth factor (bFGF) and transforming growth factor-beta(TGF-beta) in tissues from dermal chronic ulcer and hypertrophic scar and to explore their effects on tissue repair.. Twenty-one cases were detected to localize the distribution of bFGF and TGF-beta, among them, there were 8 cases with dermal chronic ulcers, 8 cases with hypertrophic scars, and 5 cases of normal skin.. Positive signal of bFGF and TGF-beta could be found in normal skin, mainly in the keratinocytes. In dermal chronic ulcers, positive signal of bFGF and TGF-beta could be found in granulation tissues. bFGF was localized mainly in fibroblasts cells and endothelial cells and TGF-beta mainly in inflammatory cells. In hypertrophic scar, the localization and signal density of bFGF was similar with those in granulation tissues, but the staining of TGF-beta was negative.. The different distribution of bFGF and TGF-beta in dermal chronic ulcer and hypertrophic scar may be the reason of different results of tissue repair. The pathogenesis of wound healing delay in a condition of high concentration of growth factors may come from the binding disorder of growth factors and their receptors. bFGF may be involved in all process of formation of hypertrophic scar, but TGF-beta may only play roles in the early stage.

Topics: Adult; Burns; Chronic Disease; Cicatrix, Hypertrophic; Female; Fibroblast Growth Factor 2; Humans; Male; Skin Ulcer; Transforming Growth Factor beta; Wound Healing

The liver plays a critical regulatory role in the acute inflammatory response to injury, although the mechanisms of this regulation are not well understood. transforming growth factor-beta1 (TGF-beta1) is induced after burn injury and may contribute to an inhibitory or fatal effect on hepatocytes. We investigated the association over time between plasma concentration of TGF-beta1, expression of TGF-beta1 m-RNA in liver tissue, and histologic analysis of liver apoptosis after burn injury.. Male BALB/c mice were anesthetized and randomized to receive 0% (sham), moderate (approximately 25%) (M), or large (approximately 50%) (L) body surface area full-thickness contact burn, followed by resuscitation and analgesia. Animals were killed over a time course from 15 minutes to 24 hours after burn injury, and liver tissue and peripheral blood were collected. Plasma levels of TGF-beta1 (nanograms per milliliter) were measured by enzyme-linked immunosorbent assay. TGF-beta1 m-RNA was extracted from liver and measured by reverse transcription-polymerase chain reaction. Histology of liver apoptosis was examined after fixation and staining with TdT-mediated dUTP nick-end labeling (TUNEL) method.. The plasma concentration of TGF-beta in burn group L was significantly increased at 4 hours after burn when compared with sham and M burn groups. This rise in plasma TGF-beta1 was preceded by an increase in hepatic TGF-beta1 m-RNA expression at 30 minutes, 1, 2, and 4 hours after burn in the L group. Histologic analysis found greater hepatocyte death in the L group than in the M group at 8 hours after burn.. The levels of induced TGF-beta1 and TGF-beta1 m-RNA after L burn injury are higher and peak earlier than after M burn injury. Elevated TGF-beta1 may be associated with cell death in hepatocytes. The TGF-beta1 rise may be associated with hepatocyte injury and systemic response to massive burn.

Topics: Acute Disease; Animals; Apoptosis; Body Surface Area; Burns; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; In Situ Nick-End Labeling; Inflammation; Injury Severity Score; Liver; Male; Mice; Mice, Inbred BALB C; Random Allocation; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Time Factors; Transforming Growth Factor beta

Keratinocytes are increasingly recognized as key regulators of skin inflammation and remodeling, as they are capable of producing growth factors and cytokines that are important mediators in the wound healing process. We investigated the expression and distribution of TGF-beta 1 mRNA by mRNA in situ hybridization and of TGF-beta 1, TGF-beta 2, TGF-beta 3, bFGF and VEGF protein expression using immunohistochemistry in spontaneously healed, partial-thickness burns and compared this with the expression of these markers in matched unburned skin. This was done to assess their role in the remodeling phase of burn wound healing. Punch biopsies were taken from both partial-thickness burns after re-epithelialization and from matched, unburned skin. At 4 and 7 months post-burn, biopsies were taken of normotrophic and hypertrophic scars that had developed in these wounds. We observed a higher expression of all mentioned growth factors in keratinocytes in scars at 1 month post-burn compared with matched unburned skin. At 4 months, keratinocytes still displayed a higher expression of TGF-beta 3 and bFGF, but the expression of TGF-beta 1, TGF-beta 2 and VEGF was normalized. The expression of TGF-beta 3 in the epidermis of hypertrophic scars was slightly higher than in normotrophic scars. At 7 months post-burn, all growth factors studied showed a normal expression on keratinocytes. Our results suggest that keratinocytes are not only involved in re-epithelialization, but also in the scar maturation. The data support the idea that keratinocytes not only respond to cytokines and growth factors in an autocrine fashion, but also exert regulatory paracrine effects on contiguous cells.

Topics: Adult; Aged; Burns; Cicatrix; Endothelial Growth Factors; Fibroblast Growth Factor 2; Gene Expression; Humans; Immunohistochemistry; In Situ Hybridization; Keratinocytes; Lymphokines; Middle Aged; RNA, Messenger; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors; Wound Healing

Hypertrophic scarring remains the most disabling sequela for burn survivors. Little is known about its pathogenesis. Collagen accumulation, however, has been consistently observed in burn hypertrophic scars (HS).. We have studied collagen production in the dermal fibroblasts derived from HS, which has developed for 9 months to 2 years. Reconstructive surgery was performed to remove HS from which the fibroblasts were cultured. Similarly, the normal cells were grown from the patient's donor site (DS), which provided autografting to the HS site. Collagen production in HS and DS fibroblasts was compared and analyzed in minimal essential amino acid medium containing 5% fetal bovine serum with inclusion of L-ascorbic acid (100 microg/mL) and beta-aminopropoinitrile (100 microg/mL) by monitoring a 20-h [3H]proline incorporation into bacterial collagenase III-digestible protein in the conditioned media.. We failed to detect any significant difference in collagen production in vitro between HS and DS. Irrespective of the fibroblasts from HS or DS, collagen production was substantially stimulated by human recombinant transforming growth factor beta-1 (TGF-beta1) (20 ng/mL) by approximately 250% after a 3-day pretreatment. In contrast, sodium nitroprusside (SNP) at 100 microM exhibited significant suppression (68%), which was rescued by hemoglobin (10 microM). TGF-beta1 significantly decreased nitric oxide (NO) production by 55%. In contrast, NO level drastically increased by 350% following SNP treatment. Epidermal growth factor showed no effect on either collagen production or NO level. The linear regression analysis shows a significant inverse correlation (r = 0.72; p < 0.05) of NO level with collagen production, suggesting the involvement of NO signaling in the modulation of collagen production. Consistent with the notion, we further showed that N-nitro-L-arginine methyl ester (100 microM) caused a synergistic stimulation and an arrested inhibition of collagen production in the presence of TGFbeta-1 and SNP, respectively. 8-BrcGMP (300 microM) mimicked the NO inhibitory action, while methylene blue (50 microM) restored the collagen production which was inhibited by SNP. Moreover, 8-BrcGMP offset the stimulation of collagen production.. The dermal fibroblasts derived from HS were not different from normals with respect to collagen production and their responses to regulations. The inhibition of collagen production was achieved by a cGMP-dependent NO action. TGFbeta-1 inhibited NO/cGMP signaling to ensure its stimulatory effect on collagen production in the dermal fibroblasts.

Topics: Burns; Cells, Cultured; Cicatrix; Collagen; Cyclic GMP; Fibroblasts; Humans; Linear Models; Microscopy, Fluorescence; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitroprusside; Proline; Signal Transduction; Transforming Growth Factor beta; Up-Regulation

Cytokines, such as the transforming growth factor beta (TGF-beta) isoforms, have been linked to the formation of proliferative scars. This study examines the stimulating effects of exogenous TGF-beta2 on cultured keloid, burn hypertrophic scar, and normal skin fibroblasts and whether such effects can be suppressed with TGF-beta2 antibody.. In vitro, the fibroblast-populated collagen lattice (FPCL) is used in the evaluation of fibroblast activation by measuring contraction of the lattice over time. Primary cultures of fibroblasts were grown from keloids, burn hypertrophic scars, and normal skin using standard cell culture techniques. TGF-beta2 (10 ng/ml) was added to each of the three types of cell cultures and placed on prefabricated FPCLs. Each was tested against their normal control counterparts. TGF-beta2 antibody (100 ng/ml) was then placed on the TGF-beta2-treated FPCLs. All lattices were allowed to contract and areas were measured for 5 days.. Compared to controls, keloid fibroblasts were most affected by the addition of exogenous TGF-beta2. Normal skin fibroblasts did not show a significant increase in contraction early on, yet a significant difference was seen as time progressed. The addition of TGF-beta2 antibody inhibited the function of keloid and burn hypertrophic scar fibroblasts. It also reversed the increased contraction of the TFG-beta2-treated proliferative scar fibroblasts.. By utilizing an in vitro model, we have demonstrated that TGF-beta2 antibody reverses the increased contraction of FPCLs by proliferative scar fibroblasts treated with TGF-beta2. This points to a possible treatment modality in patients afflicted with this disfiguring problem.

Topics: Antibodies; Burns; Cell Division; Cells, Cultured; Cicatrix; Fibroblasts; Humans; Keloid; Reference Values; Transforming Growth Factor beta

Transforming growth factor-beta (TGF-beta) has been shown to be an inhibitor of immunoglobulin (Ig) synthesis and may contribute to decreased Ig synthesis after burn injury. This study investigated the relationship between TGF-beta and Ig synthesis after burn injury.. Twenty-four BALB/c mice received either a 30% body surface area full-thickness contact burn or no burn. Splenocytes were isolated 8 days after burn and were cultured with 0, 0.05 or 0.5 ng/mL TGF-beta. After culture, total IgG and total IgM were measured by enzyme-linked immunosorbent assay. The number of IgM-secreting cells per 10(5) cells was measured by enzyme-linked immunoabsorbent spot forming assay. Total IgM per IgM-secreting cell (pg/cell) was calculated.. Total IgG, IgM, IgM-secreting cells, and B-cell number after culture were decreased by burn injury, and the decrease was exacerbated by the presence of TGF-beta. The total IgM per IgM-secreting cells, however, was significantly increased by TGF-beta at 0.5 ng/mL.. These data demonstrates that TGF-beta does not specifically impair IgM secretion by committed IgM B cells but appears to decrease B-cell proliferation or clonal expansion.

Topics: Animals; B-Lymphocytes; Burns; Cell Count; Cell Division; Cells, Cultured; Enzyme-Linked Immunosorbent Assay; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Male; Mice; Mice, Inbred BALB C; Spleen; Transforming Growth Factor beta

Keloid and hypertrophic scars are fibrous dermal tumors characterized by overabundant collagen deposition. Previous studies demonstrated that exogenous transforming growth factor beta (TGF-beta) might increase collagen production in incisional wound models and in vitro. Using an in vivo model of human scar xenografts maintained in congenitally athymic, asplenic "nude" rats, we examined endogenous TGF-beta(2), collagen I, and collagen III levels in keloids and burn hypertrophic scars treated with TGF-beta(2) and TGF-beta(2) antibody.. Fresh keloid and burn hypertrophic scar specimens excised from human subjects were explanted to pedicled flaps based on the superficial inferior epigastric vessels in athymic "nude" rats. These flaps were allowed to mature for 3 weeks, after which the scar explants were directly perfused with 200 ng of TGF-beta(2) or 250 microg of TGF-beta(2) antibody daily for 5 consecutive days, then again on Days 10, 15, and 20. Biopsies were taken 30 and 120 days following the initiation of treatment. Immunohistochemical staining was then performed for TGF-beta(2), collagen I, and collagen III. The intensity of staining was quantified.. Our results demonstrated that treatment of human proliferative scars with exogenous TGF-beta(2) results in a significant increase in endogenous TGF-beta(2), collagen I, and collagen III production. By contrast, exogenous addition of anti-TGF-beta(2) antibody significantly decreased endogenous TGF-beta(2), collagen I, and collagen III production.. This study supports a causative role for TGF-beta(2) in the formation of proliferative scars and suggests that TGF-beta(2) antibody may be a new potential antiscarring agent.

Topics: Animals; Burns; Cell Division; Cicatrix; Collagen; Humans; Rats; Rats, Nude; Transforming Growth Factor beta; Transplantation, Heterologous

To investigate the gene expression of PCMV4-hTGF beta 1 as nucleic acid vaccine regulation and the effects on burn wound healing and scarring in rats and study the feasibility of gene therapy in burns.. Sixty wistar rats were divided into three groups: 1. burn + nucleic acid vaccine; 2. nucleic acid vaccine only(control); 3. burn only(control). ELISA was used to determine the dynamic changes in anti-TGF beta 1 neutralizing antibody in serum; determination of the quantities of DNA retained in muscles by Southern blot and in situ hybridization; observation of the ratio in collagen I/III during the wound healing by special stain method.. The level of anti-TGF beta 1 antibody in serum reached the peak at the third week after naked DNA was injected, and a slight drop in the 4th week. TGF beta 1 plasmid DNA could be detected 5 minute after injection, and lasted 3 hours. In situ hybridization showed a positive staining in muscle fibers 5 days after injection. During the day 0-day 9, the wound healing speed in vaccine group was faster than control, and after day 10, no significant difference was found between groups, but the ratio of collagen I/III was reduced remarkably in vaccine group.. Injection of PCMV4-hTGF beta 1 in rats can really cause general immune response reaction. It was showed that PCMV4-hTGF beta 1 was similar to TGF beta 1, and it had the effect of stimulating of epidermic cells to accelerate wound healing at early stage, and at later stage, it was similar to anti-TGF beta 1 neutralizing antibody, having the effect of inhibiting hyperplasia of scar. So it is confirmed that PCMV4-hTGF beta 1 as nucleic acid vaccine has the effect of promoting healing of burn wound and controlling the formation of scar.

Topics: Animals; Burns; Cicatrix, Hypertrophic; Cytomegalovirus; Male; Plasmids; Random Allocation; Rats; Rats, Wistar; Recombinant Proteins; Transforming Growth Factor beta; Vaccines, DNA; Wound Healing

1. Fibroblast cultures were established from biopsies of hypertrophic scar and normal dermis taken from nine patients recovering from second- and third-degree burns. The capacity of these fibroblasts to synthesize the small proteoglycan decorin was assessed by quantitative Western blot analysis of conditioned medium collected from confluent cultures. Levels of mRNA for decorin were assessed by quantitative Northern analysis. Since transforming growth factor-beta 1 is implicated in various fibrotic conditions, including post-burn hypertrophic scar, its effect on decorin synthesis by these paired fibroblast cell strains was assessed. 2. Production of decorin was lower in all cell strains of hypertrophic scar fibroblasts tested, compared with normal dermal fibroblasts cultured from the same patients (mean 49 +/- 23%; P < 0.001, n = 9). Levels of mRNA for decorin were also lower (mean 59 +/- 28%; P < 0.02, n = 7) but those for biglycan and versican were not significantly different. Four pairs of cell strains were examined at more than one passage and the differences in decorin protein were found to be phenotypically persistent. Treatment of confluent cultures with transforming growth factor-beta 1 for 3 days caused a reduction in both decorin protein and mRNA in all six strains of hypertrophic scar fibroblasts tested and in five of six strains of normal dermal fibroblasts. An increase in the length of the dermatan sulphate chain on decorin, a previously reported characteristic of this glycosaminoglycan in hypertrophic scar, was seen in all but two of the strains treated with transforming growth factor-beta 1. The depression of decorin synthesis by transforming growth factor-beta 1 was reversed on removal of the agent and passaging the fibroblasts. 3. The reduced capacity of fibroblasts in hypertrophic scar tissue to synthesize decorin may have implications for the development of the condition since this small proteoglycan is involved in tissue organization and may also play a role in modulating the activity in vivo of fibrogenic cytokines such as transforming growth factor-beta 1.

Topics: Adolescent; Adult; Blotting, Northern; Blotting, Western; Burns; Cells, Cultured; Child; Child, Preschool; Cicatrix, Hypertrophic; Decorin; Extracellular Matrix Proteins; Female; Fibroblasts; Humans; Male; Middle Aged; Proteoglycans; RNA, Messenger; Statistics, Nonparametric; Transforming Growth Factor beta

To study the application of exogenous TGF-beta to excision, incision and burn wounds accelerating wound healing.. The expression of endogenous TGF-beta during burn wound healing was observed by mRNA dot blot hybridization and immunohistochemistry, respectively.. Thermal injury could induce TGF-beta gene expression in cutaneous wounds. Expression level of TGF-beta mRNA increased over time to reach a peak around day 5 postburn, and declined at day 10 after thermal injury. Immunohistochemistry evidences indicated spatially and temporally that TGF-beta peptide mainly distributed at hair follicles and migrating epidermis at early and middle stages of repair process, and stronger immunoreactivity from days 5 to 10 after wound, which was directly proportional to epidermal cell migration and dermal fibroblast proliferation.. The expression models of TGF-beta protein and gene suggests that TGF-beta play a critical role in regulation of repair process. Whether the inhibition of expression and overexpression of TGF-beta is correlated to imparied repair and scar formation remains to be studied.

Topics: Animals; Burns; Male; Rats; Rats, Wistar; RNA, Messenger; Transforming Growth Factor beta; Wound Healing

The purpose of this study was to try to elucidate a possible biobehavioral mechanism associated with decreased immune function in trauma patients by determining whether there is an interaction between the effects of ACTH, a stress hormone, and TGF beta, a cytokine, on peripheral blood lymphocyte proliferation. Peripheral mononuclear lymphocytes (PMLs) from healthy donors were preincubated with varying concentrations of ACTH for 24 hr, stimulated with concanavalin A and increasing concentrations of TGF beta, and incubated for 72 hr. Proliferation was assayed by tritiated thymidine incorporation. A parallel aliquot of PMLs were incubated in the presence of ACTH to determine the direct effect of ACTH on mononuclear cell TGF beta production. While harvested supernatant from cells incubated in the presence of ACTH did not contain any detectable TGF beta, ACTH as well as TGF beta were found to significantly decrease cellular proliferation independent of one another. An even greater decrease in cellular proliferation was found when both ACTH and TGF beta were used, compared to either ACTH or TGF beta alone. These results suggest a biobehavioral interaction between ACTH and TGF beta at the cellular level and that interactions to relieve stress may assist in improving function and recovery from trauma.

Topics: Adrenocorticotropic Hormone; Analysis of Variance; Burns; Cell Division; Cells, Cultured; Concanavalin A; Dose-Response Relationship, Drug; Drug Interactions; Humans; Monocytes; Psychoneuroimmunology; Reference Values; Transforming Growth Factor beta; Wounds and Injuries

The distributions of the small proteoglycans, decorin and biglycan and the large proteoglycan, versican, in normal skin and post-burn hypertrophic and mature scars, were compared using monoclonal and polyclonal antibodies to the core proteins. Biglycan and versican were virtually undetectable in normal dermis but readily seen in hypertrophic scars. Staining for decorin was strong throughout the dermis in normal skin but restricted to the deep dermis and a narrow zone under the epidermis in hypertrophic scar--areas which did not stain for versican. Decorin was absent or reduced in the nodules in these specimens. In mature post-burn scars, staining for all three proteoglycans demonstrated an intensity that was intermediate between that in normal dermis and that in the nodules of the hypertrophic scars. Transforming growth factor-beta was present in the nodules of hypertrophic scars but the deep dermis of these specimens stained most intensely for this cytokine which was also found in the dermis of mature scars but was not detectable in normal dermis. The apparent co-distribution of decorin and transforming growth factor-beta suggests that this proteoglycan may play an active role in the resolution of the scars. Changes in proteoglycan type and distribution could possibly account, at least in part, for the derangement of collagen and the altered physical properties of hypertrophic scar tissue.

Topics: Adult; Amino Acid Sequence; Antibody Specificity; Biglycan; Burns; Case-Control Studies; Child; Chondroitin Sulfate Proteoglycans; Cicatrix; Cicatrix, Hypertrophic; Decorin; Epitopes; Extracellular Matrix Proteins; Female; Humans; Immunohistochemistry; Lectins, C-Type; Male; Molecular Sequence Data; Proteoglycans; Transforming Growth Factor beta; Versicans

Hypertrophic scar is the result of abnormal healing that often follows thermal injury. Hypertrophic scar is characterized by excessive dermal fibrosis and scarring. Five cases of human hypertrophic scar were compared with normal skin using in situ hybridization to localize mRNAs for procollagen types I and III and transforming growth factor-beta 1. Expression of type I procollagen and TGF-beta 1 were also examined with immunohistochemistry. The results demonstrated a significant increase in the expression of mRNA for types I and III procollagen and type I procollagen protein by fibroblasts in hypertrophic scar compared with normal skin. In all cases of hypertrophic scar, significant numbers of cells expressed TGF-beta 1 mRNA or peptide. Neither TGF-beta 1 mRNA nor protein was detected in control tissues. These results suggest a profound increase in production and expression of types I and III collagen mRNA by the fibroblasts in hypertrophic scar. This may result from increased TGF-beta 1 production, through paracrine and autocrine pathways, as have been described for this fibrogenic cytokine.

Topics: Base Sequence; Burns; Cicatrix, Hypertrophic; Collagen; Fibroblasts; Gene Expression; Humans; Immunohistochemistry; In Situ Hybridization; Molecular Sequence Data; Procollagen; RNA, Messenger; Skin; Transforming Growth Factor beta

Following severe thermal injury and other injuries to the deep dermis of the skin, patients frequently develop hypertrophic scarring (HSc) which is characterized by an over-abundance of extracellular matrix (ECM) proteins including collagen. Our previous work revealed a synchronous elevation in expression of mRNA for transforming growth factor (TGF)-beta 1, type I and type III procollagen in human HSc tissue, suggesting a possible role of locally synthesized TGF-beta 1 in matrix production. In this study the immunoreactive sites of TGF-beta 1 protein in hypertrophic and normal dermal (ND) tissue obtained from the same patients have been determined by an immunoperoxidase staining system. We used two TGF-beta 1-specific rabbit polyclonal antibodies known as anti-LC and anti-CC which were prepared to different synthetic preparations of a peptide corresponding to the first 1-30 amino acids of the amino-terminus of mature TGF-beta 1. These antibodies have affinity for two distinct epitopes of TGF-beta 1. The anti-LC antibody localized TGF-beta 1 to non-proliferating/differentiated epidermal cells, suprabasal keratinocytes in both normal and HSc tissues. The intensity of this staining was significantly higher (150 +/- 26 sq. micron vs 77 +/- 7 sq. micron, n = 5, P < 0.05) in normal tissues compared to HSc tissues. When the anti-CC antibody was used as the primary antibody, intense staining of focal regions was observed in the dermis of HSc tissue, but not in ND tissue obtained from the same patient. These foci contained collagen which was nodular, distributed in whorl-like arrangements and highly enriched with microvessels.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Animals; Antibodies; Antibody Specificity; Biopsy; Burns; Cell Differentiation; Cicatrix, Hypertrophic; Female; Humans; Immunoblotting; Immunoenzyme Techniques; Immunohistochemistry; Male; Middle Aged; Rabbits; Reference Values; Skin; Transforming Growth Factor beta

Topics: Burns; Dinoprostone; Humans; Macrophages; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wounds and Injuries

To explore the possible role of locally synthesized transforming growth factor beta-1 (TGF-beta 1) and procollagen gene expression in postburn hypertrophic scars, we have compared mRNA levels for type I and type III procollagen and TGF-beta 1 in human hypertrophic scar tissue with normal dermis obtained from the same patients as a control. Northern blot analysis of total RNA extracted from hypertrophic scar tissue and normal skin demonstrated two transcripts for the pro-alpha 1(I) chain (5.8 kb and 4.8 kb) and for the pro-alpha 1(III) chain (5.4 kb and 4.8 kb) and one transcript (4.9 kb) for TGF-beta 1. Quantitative analysis of dot blot autoradiograms of mRNA from three samples of hypertrophic scar tissue and normal skin showed average increases of 102% (p < 0.05) for pro-alpha 1(I), 91% (p < 0.06) for pro-alpha 1(III), and 61% (p < 0.05) for TGF-beta 1. Three additional hypertrophic scar samples were quantitatively analyzed on Northern blots and showed increases of 246%, 102%, and 250% of the specific messages for pro-alpha 1(I), pro-alpha 1(III), and TGF-beta 1 relative to a normal skin control. Two transcripts (4.9 kb and 2.5 kb) for TGF-beta 1 were identified in cultured fibroblasts. In contrast to the results from tissue, the level of these transcripts in fibroblasts cultured from hypertrophic scar tissue and normal skin were not significantly different, suggesting that the synthesis of this growth factor is stimulated in tissue by a presently unknown mechanism.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adolescent; Adult; Blotting, Northern; Burns; Child; Cicatrix; Female; Fibroblasts; Gene Expression; Humans; Hypertrophy; Male; Middle Aged; Procollagen; RNA, Messenger; Skin; Transforming Growth Factor beta

We have analysed and compared, by in situ hybridisation, the effects of exogenously applied TGF-beta s on expression of endogenous TGF-beta mRNAs in partial thickness thermal wounds in old and young mice. Although injury induced the expression of TGF-beta 1 mRNA in the epidermis and dermis at the wound margins, expression of TGF-beta 2- or TGF-beta 3-mRNA was not detected. Biopsies taken 24 hours following injury revealed a focally clustered distribution of TGF-beta 1 hybridisation signals in the dermis, the number of positive cells and expression levels being reduced in old mice. Topical application of all three TGF-beta isoforms enhanced TGF-beta 1 mRNA expression in the dermis of old and young mice. In biopsies taken three days following injury, TGF-beta 1 hybridisation signals were most prominent in the regenerating epidermis although at this timepoint differences in expression levels between treated and non-treated animals were less pronounced.

Topics: Animals; Biopsy; Burns; Humans; In Situ Hybridization; Mice; Mice, Inbred C57BL; Recombinant Proteins; RNA, Messenger; Skin; Time Factors; Transforming Growth Factor beta; Wound Healing

We previously reported that increased production of prostaglandin E2 by monocytes is a pivotal mechanism in posttrauma immunopathology. Here we characterize monocyte levels of transforming growth factor beta and examine the effects of elevated transforming growth factor beta on prostaglandin E2 release by patients' monocytes. Trauma patients' and normals' monocyte supernates (+/- stimulation with muramyl dipeptide) were acid treated and assayed for transforming growth factor beta using the mink lung-cell bioassay. Alternatively, human transforming growth factor beta was added to patients' and normals' monocytes and prostaglandin E2 production assayed. Significantly elevated transforming growth factor beta levels (median = 181.7 pmol/10(6) monocytes) were detected in immunosuppressed patients' monocytes but not immunocompetent trauma patients' (median = 32.0 pM) or normals' (median = 20.4 pM) monocytes. Adding transforming growth factor beta to monocytes resulted in a significant elevation of prostaglandin E2 levels. Elevated monocyte transforming growth factor beta levels in trauma patients could be both suppressing T-lymphocyte functions and maintaining elevated monocyte prostaglandin E2 synthesis.

Topics: Acetylmuramyl-Alanyl-Isoglutamine; Adult; Aged; Burns; Dinoprostone; Female; Humans; Immune Tolerance; Interferon-gamma; Male; Middle Aged; Monocytes; Transforming Growth Factor beta; Wounds and Injuries