chondroitin-sulfates and Inflammation

Chondroitin sulfate (CS) belongs to a class of molecules called glycosaminoglycans (GAGs). These are long, linear chains of polysaccharides comprising alternating amino sugars and hexuronic acid. Similar to other GAGs, CS is important in a multitude of biological activities. Alteration of CS levels has been implicated in several pathological conditions, including osteoarthritis (OA) and other inflammatory diseases, as well as physiological conditions, such as aging. Therefore, devising replenishment strategies for this molecule is an important area of research. In this review, we discuss the nature of CS, its function in different organs, and its implications in health and disease. We also describe different methods for the exogenous administration of CS.

Topics: Aging; Animals; Chondroitin Sulfates; Humans; Inflammation; Osteoarthritis

The remarkable structural heterogeneity of chondroitin sulfate (CS) and dermatan sulfate (DS) generates biological information that can be unique to each of these glycosaminoglycans (GAGs), and changes in their composition are translated into alterations in the binding profiles of these molecules. CS/DS can bind to various cytokines and growth factors, cell surface receptors, adhesion molecules, enzymes and fibrillar glycoproteins of the extracellular matrix, thereby influencing both cell behavior and the biomechanical and biochemical properties of the matrix. In this review, we summarize the current knowledge concerning CS/DS metabolism in the human cancer stroma. The remodeling of the GAG profile in the tumor niche is manifested as a substantial increase in the CS content and a gradual decrease in the proportion between DS and CS. Furthermore, the composition of CS and DS is also affected, which results in a substantial increase in the 6-O-sulfated and/or unsulfated disaccharide content, which is concomitant with a decrease in the 4-O-sulfation level. Here, we discuss the possible impact of alterations in the CS/DS sulfation pattern on the binding capacity and specificity of these GAGs. Moreover, we propose potential consequences of the stromal accumulation of chondroitin-6-sulfate for the progression and metastasis of cancer.

Topics: Animals; Chondroitin Sulfates; Dermatan Sulfate; Humans; Inflammation; Neoplasms; Stromal Cells; Tumor Microenvironment

The temporary anchorage devices (TADs) which include miniscrew implants (MSIs) have evolved as useful armamentarium in the management of severe malocclusions and assist in complex tooth movements. Although a multitude of factors is responsible for the primary and secondary stability of miniscrew implants, contemporary research highlights the importance of biological interface of MSI with bone and soft tissue in augmenting the success of implants. The inflammation and remodeling associated with MSI insertion or loading are reflected through biomarkers in peri-miniscrew implant crevicular fluid (PMICF) which is analogous to the gingival crevicular fluid. Analysis of biomarkers in PMICF provides indicators of inflammation at the implant site, osteoclast differentiation and activation, bone resorption activity and bone turnover. The PMICF for assessment of these biomarkers can be collected non-invasively via paper strips, periopaper or micro capillary pipettes and analysed by enzyme-linked immunosorbent assay (ELISA) or immunoassays. The markers and mediators of inflammation have been previously studied in relation to orthodontic tooth movement include interleukins (IL-1β, IL-2, IL-6 and IL-8), growth factors and other proteins like tumour necrosis factor (TNF-α), receptor activator of nuclear factor kappa-B ligand (RANKL), chondroitin sulphate (CS) and osteoprotegerin (OPG). Studies have indicated that successful and failed MSIs have different concentrations of biomarkers in PMICF. However, there is a lack of comprehensive information on this aspect of MSIs. Therefore, a detailed review was conducted on the subject.. A literature search revealed six relevant studies: two on IL-1β; one on IL-2, IL-6 and IL-8; one on TNF-α; one on CS; and one on RANKL/OPG ratio. One study showed an increase in IL-1β levels upon MSI loading, peak in 24 hours (h), followed by a decrease in 21 days to reach baseline in 300 days. A 6.87% decrease in IL-2 levels was seen before loading and a 5.97% increase post-loading. IL-8 showed a 6.31% increase after loading and IL-6 increased by 3.08% before MSI loading and 15.06% after loading. RANKL/OPG ratio increased in loaded compared to unloaded MSIs.. Cytokines (mainly ILs and TNF-α) and RANKL/OPG ratio showed alteration in PMICF levels upon loading of MSIs as direct or indirect anchorage.

Topics: Biomarkers; Bone Remodeling; Bone Screws; Chondroitin Sulfates; Cytokines; Gingival Crevicular Fluid; Humans; Inflammation; Interleukins; Orthodontic Anchorage Procedures; Osteoprotegerin; RANK Ligand; Tooth Movement Techniques; Tumor Necrosis Factor-alpha

Chondroitin sulphate (CS) is recommended by the European League Against Rheumatism as a symptomatic slow-acting drug for the treatment of osteoarthritis on the basis of numerous clinical trials and meta-analyses. Furthermore, recent clinical trials have also demonstrated the possible structure-modifying effects of CS. This review focuses on recent experimental results and data available in the scientific literature regarding the anti-inflammatory properties of CS with a view to understanding the molecular basis responsible for its activity. Several animal studies have demonstrated that orally administered CS significantly inhibited hind paw oedema, synovitis and destruction of the articular cartilage in a dose-dependent manner. Furthermore, CS proved to have a beneficial effect in slowing down the development of adjuvant arthritis and in reducing disease markers, findings which support its beneficial activity in humans as a chondroprotective drug. Finally, several in vitro studies have focused on the hypothesis that CS may reduce inflammatory processes by acting on the nuclear translocation of NF-κB, which is closely associated with the blood biomarkers of inflammation, primarily IL-1, IL-6 and C-reactive protein.

Topics: Animals; Anti-Inflammatory Agents; Chondroitin Sulfates; Clinical Trials as Topic; Humans; Inflammation; Meta-Analysis as Topic; Osteoarthritis

To evaluate the immune-modulator effect of chondroitin sulfate (CS) by means of the review of the literature.. Inflammatory reactions are primarily originated by infectious agents, immune reactions and by sterile tissue lesions that activate membrane receptors by means of pathogen-associated molecular patterns, tissue breakdown products and cytokines. The activation of membrane receptors triggers the phosphorylation of mitogen activated protein kinases and of the nuclear factor kappaB (NF-kappaB). The binding of NF-kappaB to the promoter of target genes enhances the expression of pro-inflammatory cytokines, inducible nitric oxide synthase, cyclooxygenase 2, phospholipase A2, and matrix metalloproteases, proteins that contribute to tissue damage and to the inflammatory reaction. The activation of NF-kappaB has a key role in the immune homeostasis and the inflammatory response and therefore, in the pathogenesis of numerous diseases. Chondroitin sulfate (CS) is able to diminish NF-kappaB activation and nuclear translocation in chondrocytes and synovial membrane, effects that may explain the benefits of CS in osteoarthritis. In addition, systemic CS reduces NF-kappaB nuclear translocation in macrophages and hepatocytes, raising the hypothesis that CS might be of benefit to treat other diseases with a strong inflammatory component. There is preliminary evidence in humans that CS improves moderate to severe psoriasis. Moreover, experimental and clinical data suggest that CS might be a useful therapeutic agent in diseases such as inflammatory bowel diseases, atherosclerosis, Parkinson's and Alzheimer's diseases, multiple sclerosis, amyotrophic lateral sclerosis, rheumatoid arthritis and systemic lupus erythematosus.. These results urge for double blinded placebo-controlled trials to confirm the utility of CS in diseases with immune and inflammatory components.

Topics: Anti-Inflammatory Agents; Atherosclerosis; Chondroitin Sulfates; Humans; Inflammation; Inflammatory Bowel Diseases; Osteoarthritis; Psoriasis

Osteoarthritis is a disabling disease of the aging generation, which results in loss of quality of life and increased healthcare costs. Cytokines appear to play an important role in the cartilaginous degeneration characterizing the pathological process. Increasing experience is being gained with cytokine-modulating therapies aimed at interfering with effects of chondrodegradative cytokines in the synovial fluid. Although in vitro and in vivo effectiveness of several of these therapies has been demonstrated, clinical effectiveness remains disputable, which may be related to the low levels of inflammatory cytokines found in osteoarthritic joints. By contrast, directly after joint trauma, which has been shown to predispose to early osteoarthritis, synovial fluid cytokine levels are strongly increased. Cytokine-modulating therapies, however, have hardly been considered for this indication. Increased knowledge of intra-articular soluble mediators correlating with cartilage pathology will lead to further development of cytokine-modulating products and, eventually, to effective inhibition of cartilage degeneration, in both the osteoarthritic as well as injured joints.

Topics: Animals; Cartilage; Chondroitin Sulfates; Cytokines; Humans; Immunotherapy; Inflammation; Knee Injuries; Knee Joint; Osteoarthritis; Risk Factors; Synovial Fluid

Topics: Amino Acid Sequence; Animals; Chondroitin Sulfates; Cytokines; Heparitin Sulfate; Humans; Inflammation; Midkine; Molecular Sequence Data; Neoplasms; Nerve Growth Factor; Protein Binding; Protein Conformation; Protein Structure, Tertiary

Approximately 50% of the weight of a mature mast cell (MC) consists of varied neutral proteases stored in the cell's secretory granules ionically bound to serglycin proteoglycans that contain heparin and/or chondroitin sulfate E/diB chains. Mouse MCs express the exopeptidase carboxypeptidase A3 and at least 15 serine proteases [designated as mouse MC protease (mMCP) 1-11, transmembrane tryptase/tryptase gamma/protease serine member S (Prss) 31, cathepsin G, granzyme B, and neuropsin/Prss19]. mMCP-6, mMCP-7, mMCP-11/Prss34, and Prss31 are the four members of the chromosome 17A3.3 family of tryptases that are preferentially expressed in MCs. One of the challenges ahead is to understand why MCs express so many different protease-proteoglycan macromolecular complexes. MC-like cells that contain tryptase-heparin complexes in their secretory granules have been identified in the Ciona intestinalis and Styela plicata urochordates that appeared approximately 500 million years ago. Because sea squirts lack B cells and T cells, it is likely that MCs and their tryptase-proteoglycan granule mediators initially appeared in lower organisms as part of their innate immune system. The conservation of MCs throughout evolution suggests that some of these protease-proteoglycan complexes are essential to our survival. In support of this conclusion, no human has been identified that lacks MCs. Moreover, transgenic mice lacking the beta-tryptase mMCP-6 are unable to combat a Klebsiella pneumoniae infection effectively. Here we summarize the nature and function of some of the tryptase-serglycin proteoglycan complexes found in mouse and human MCs.

Topics: Amino Acid Sequence; Animals; Chondroitin Sulfates; Heparin; Humans; Immunity, Innate; Inflammation; Mast Cells; Mice; Molecular Sequence Data; Peptide Hydrolases; Proteoglycans; Tryptases

CD44 is a major cell surface receptor for the glycosaminoglycan, hyaluronan (HA). CD44 binds HA specifically, although certain chondroitin-sulfate containing proteoglycans may also be recognized. CD44 binding of HA is regulated by the cells in which it is expressed. Thus, CD44 expression alone does not correlate with HA binding activity. CD44 is subject to a wide array of post-translational carbohydrate modifications, including N-linked, O-linked and glycosaminoglycan side chain additions. These modifications, which differ in different cell types and cell activation states, can have profound effects on HA binding function and are the main mechanism of regulating CD44 function that has been described to date. Some glycosaminoglycan modifications also affect ligand binding specificity, allowing CD44 to interact with proteins of the extracellular matrix, such as fibronectin and collagen, and to sequester heparin binding growth factors. It is not yet established whether the HA binding function of CD44 is responsible for its proposed involvement in inflammation. It has been shown, however, that CD44/HA interactions can mediate leukocyte rolling on endothelial and tissue substrates and that CD44-mediated recognition of HA can contribute to leukocyte activation. Changes in CD44 expression (mainly up-regulation, occasionally down-regulation, and frequently alteration in the pattern of isoforms expressed) are associated with a wide variety of cancers and the degree to which they spread; however, in other cancers, the CD44 pattern remains unchanged. Increased expression of CD44 is associated with increased binding to HA and increased metastatic potential in some experimental tumor systems; however, in other systems increased HA binding and metastatic potential are not correlated. CD44 may contribute to malignancy through changes in the regulation of HA recognition, the recognition of new ligands and/or other new biological functions of CD44 that remain to be discovered.

Topics: Animals; Antigens, CD; Chondroitin Sulfates; Genetic Variation; Humans; Hyaluronan Receptors; Hyaluronic Acid; Inflammation; Neoplasm Metastasis; Neoplasms; Proteoglycans

Topics: Cell Adhesion Molecules; Chondroitin Sulfates; Glycosaminoglycans; Granulocytes; Humans; Inflammation

Delayed onset of muscle soreness (DOMS) often results from unaccustomed exercise or exercise that involves heavy eccentric loading of skeletal muscle. Chondroitin sulfate (CS) has been reported to possess anti-inflammatory and analgesic properties and has been used widely in the treatment of osteoarthritis. Based on these findings, the investigators sought to determine whether CS supplementation might reduce DOMS symptoms associated with eccentric arm exercise.. Sixteen non-resistance-trained males participated in a double-blind protocol. Subjects consumed 3,600 mg x day-1 of encapsulated CS or placebo for 14 days preceding the exercise protocol and during 48 hours of follow-up measurements. Subjects performed a maximum of 50 eccentric contractions of the elbow flexors against a resistance that was 120% of the concentric one-repetition maximum.. Significant time effects were present for soreness, relaxed elbow extension (joint angle), and creatine kinase; all measures were significantly altered 48 hours after the exercise session. No effects were present for complement system fragment C3a and prostaglandin E2. No significant treatment or interaction effects were detected for any of the variables.. CS supplementation was not found to be effective in the prevention of DOMS, nor did it influence biochemical indices of inflammation and muscle damage following heavy eccentric loading of the arm flexors.

Topics: Adult; Arm; Chondroitin Sulfates; Creatine Kinase; Dietary Supplements; Double-Blind Method; Exercise; Humans; Inflammation; Male; Muscle Contraction; Muscle Fatigue; Muscle, Skeletal; Muscular Diseases; Premedication; Range of Motion, Articular; Time Factors; Treatment Outcome

Topics: Adolescent; Adult; Chondroitin Sulfates; Cystitis, Interstitial; Female; Humans; Inflammation; Instillation, Drug; Male; Neutralization Tests; Patient Compliance; Patient Selection; Potassium; Quality of Life; Research Design; Surveys and Questionnaires

Anticoagulants have gained increasing attention in the treatment of sepsis. This study used danaparoid to investigate the role of factor Xa in endotoxin-induced coagulation and inflammation and its effectiveness when coagulation activation has already occurred. Thirty healthy volunteers were enrolled in the randomized, placebo-controlled trial. Subjects received 2 ng/kg endotoxin and danaparoid 10 min or 3 h thereafter or placebo. Endotoxin increased prothrombin fragment 1+2 (F(1+2)) levels from 0.5 to 7.0 nmol/L at 5 h in the placebo group. Early danaparoid infusion inhibited endotoxin-induced thrombin formation: maximum F(1+2) levels reached only 1.8 nmol/L (P<.01, vs. baseline or placebo). Delayed danaparoid infusion effectively blocked further thrombin formation. However, danaparoid did not alter endotoxin-induced changes in the fibrinolytic system, cytokine levels, activation of leukocytes, or tissue factor expression on monocytes. Danaparoid therefore selectively attenuates endotoxin-induced coagulopathy, even with delayed administration when coagulation activation is well under way.

Topics: Adult; alpha-2-Antiplasmin; Anticoagulants; Blood Coagulation; Chemokine CCL2; Chondroitin Sulfates; Cytokines; Dermatan Sulfate; Drug Combinations; Endotoxins; Factor X; Heparitin Sulfate; Humans; Inflammation; Interleukin-6; Kinetics; Lipopolysaccharides; Pilot Projects; Reference Values

Topics: Animals; Antioxidants; Chondroitin Sulfates; Dopamine; Furans; Hyaluronic Acid; Hydrogels; Inflammation; Intervertebral Disc Degeneration; Nucleus Pulposus; Rats

The alteration of the extracellular matrix (ECM) homeostasis plays an important role in the development of osteoarthritis (OA). The pathological changes of OA are mainly manifested in the large reduction of components in ECM, like type II collagen and aggrecan, especially hyaluronic acid and chondroitin sulfate and often accompanied by inflammation. Rebuilding ECM and inhibiting inflammation may reverse OA progression. In this work, we developed new magnesium-containing glycosaminoglycans (Mg-GAGs), to create a positive ECM condition for promoting cartilage regeneration and alleviating OA. In vitro results suggested that the introduction of Mg-GAGs contributed to promoting chondrocyte proliferation and facilitated upregulating chondrogenic genes and suppressed inflammation-related factors. Moreover, Mg-GAGs exhibited positive effects on suppressing synovial inflammation, reducing chondrocyte apoptosis and preserving the subchondral bone in the ACLT-induced OA rabbit model. This study provides new insight into ECM-based therapeutic strategy and opens a new avenue for the development of novel OA treatment.

Topics: Aggrecans; Animals; Cartilage; Cartilage, Articular; Chondrocytes; Chondroitin Sulfates; Collagen Type II; Extracellular Matrix; Glycosaminoglycans; Hyaluronic Acid; Inflammation; Magnesium; Osteoarthritis; Rabbits

Growing evidence for the importance of the gut-brain axis in Parkinson's disease (PD) has attracted researchers' interest in the possible application of microbiota-based treatment approaches. Using a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model, we looked into the prospect of treating PD with fucosylated chondroitin sulfate obtained from sea cucumbers

Topics: 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; alpha-Synuclein; Animals; Chondroitin Sulfates; Disease Models, Animal; DNA, Bacterial; Dopamine; Dysbiosis; Inflammation; Intestines; Mice; Mice, Inbred C57BL; NF-kappa B; Parkinson Disease; Polysaccharides; RNA, Ribosomal, 16S; Sea Cucumbers

Ageing-related bone impairment due to exposure to hyperglycemic environment is scarcely researched. The aim was to confirm the improvement effects of undenatured type II collagen (UC II) on bone impairment in ageing db/db mice, and the ageing model was established by normal feeding for 48-week-old. Then, the ageing db/db mice were randomly assigned to UC II intervention, the ageing model, and the chondroitin sulfate + glucosamine hydrochloride control groups. After 12 weeks of treatment, femoral microarchitecture and biomechanical parameters were observed, biomarkers including bone metabolism, inflammatory cytokines, and oxidative stress were measured, and the gastrocnemius function and expressions of interleukin (IL) 1β, receptor activator of nuclear factor (NF)-κB ligand (RANKL), and tartrate-resistant acid phosphatase (TRAP) were analyzed. The results showed that the mice in the UC II intervention group showed significantly superior bone and gastrocnemius properties than those in the ageing model group, including bone mineral density (287.65 ± 72.77 vs. 186.97 ± 32.2 mg/cm

Topics: Aging; Animals; Bone Density; Bone Resorption; Chondroitin Sulfates; Collagen Type II; Diabetes Mellitus, Type 2; Gene Expression Regulation; Glucosamine; Humans; Inflammation; Interleukin-1beta; Mice; Mice, Inbred NOD; Oxidative Stress; RANK Ligand; Tartrate-Resistant Acid Phosphatase

One major aspect of the aging process is the onset of chronic, low-grade inflammation that is highly associated with age-related diseases. The molecular mechanisms that regulate these processes have not been fully elucidated. We have identified a spontaneous mutant mouse line, small with kinky tail (

Topics: Age Factors; Animals; Apoptosis; Chondroitin Sulfates; Female; Glucuronosyltransferase; Inflammation; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Multifunctional Enzymes; Mutation; N-Acetylgalactosaminyltransferases; Neurodegenerative Diseases; Neurons; Protein Processing, Post-Translational; Proteins; Retinal Degeneration

An extracellular matrix (ECM) mimicking architecture was introduced with gelatin glycosaminoglycans like hyaluronic acid and chondroitin sulfate and a triterpenoid using asiatic acid, possessing biodegradable and biocompatible properties that mark the functionality for the treatment of second-degree burn wounds. In the present work, a foam-based scaffold was fabricated and sterilized with gamma radiation at a 2.5 Mrad dose. The scaffolds were further characterized for morphology, swelling, degradation behaviour, release of bioactive components, ATR-FTIR, mechanical, thermal properties and compared with control. In vitro cytocompatibility of the developed scaffold was studied with L929 mouse fibroblast cells and human mesenchymal stem cells based on deoxyribonucleic acid and lactate dehydrogenase assay. Additionally, the developed scaffold was evaluated for its biocompatibility on the Wistar rat to assess any toxicity induced to the animal based on blood biochemistry and histopathology analysis. Finally, we assessed the efficacy of developed foam scaffolds on the second-degree burn wound-induced Wistar rat with a scaffold alone and a scaffold seeded with human bone-marrow-derived mesenchymal stem cells in a wound healing study for 28 d. The wound contraction assay, histopathology, immunohistochemistry analysis and pro-healing marker quantification using hexosamine, hydroxyproline, and pro-inflammatory markers like TNF-α and MMP-2 were carried out and compared with the commercially available wound dressing. The results revealed that foam-based ECM mimic was cytocompatible, biocompatible and biodegradable in 18 ± 3 d in in vivo conditions and the scaffold fostered the process of healing of second-degree burns within 28 d of treatment. The obtained result proved that the scaffold has a potential for clinical settings in second-degree burn wound treatment.

Topics: Animals; Biocompatible Materials; Bone Marrow Cells; Calorimetry, Differential Scanning; Cell Line; Chondroitin Sulfates; Gelatin; Humans; Hyaluronic Acid; Hydroxyproline; Inflammation; Mesenchymal Stem Cells; Mice; Pentacyclic Triterpenes; Polymers; Rats; Rats, Wistar; Spectroscopy, Fourier Transform Infrared; Swine; Tensile Strength; Thermogravimetry; Tissue Scaffolds; Wound Healing

Topics: Animals; Blood Glucose; Chondroitin Sulfates; Diabetes Complications; Diabetes Mellitus, Experimental; Female; Gene Expression Regulation; Inflammation; Osteoporosis; Osteoprotegerin; Oxidative Stress; RANK Ligand; Rats; Rats, Sprague-Dawley

Fucosylated chondroitin sulfate from Isostichopus badionotus (fCS-Ib) is a kind of sulfated polysaccharides with well-repeated structure. In our former publications, fCS-Ib has been reported to be a functional food ingredient with hypoglycemic and antilipemic activities. However, there is no systematic study to investigate the effects of fCS-Ib on metabolic syndromes. In the present study, C57BL/6 mice fed on a high-fat and high sucrose diet (HFSD) for 6 weeks was used to cause metabolic syndromes. The final results showed that fCS-Ib alleviated obesity, hyperlipidemia, hyperglycemia, inflammation, liver steatosis, and adipocyte hypertrophy caused by HFSD. Meanwhile, fCS-Ib showed powerful effects on moderating gut microbiota dysbiosis in the HFSD-fed mice. Supplement of fCS-Ib could reduce ratio of Firmicutes to Bacteroidetes by decreasing abundance of Lachnospiraceae and Allobaculum while increasing abundance of Porphyromonadaceae, Barnesiella, and Bacteroides. Our results showed that fCS-Ib could be further developed as a potential pharmaceutical agent to prevent metabolic syndromes and gut microbiota dysbiosis.

Topics: Adipocytes; Animals; Chondroitin Sulfates; Diet, High-Fat; Dysbiosis; Fatty Liver; Fructose; Gastrointestinal Microbiome; Humans; Hyperglycemia; Hyperlipidemias; Hypertrophy; Inflammation; Metabolic Syndrome; Mice; Obesity; Sea Cucumbers

Biological regeneration of articular cartilage continues to be a challenge at present. Functional engineered implants with patient-specific sizes are difficult to achieve. The aim of this study is to fabricate a biocompatible cell-laden hydrogel with a designable structure. Covalent hydrogels were prepared with water soluble hydroxybutyl chitosan (HBC) and oxidized chondroitin sulfate (OCS) via a Schiff-base reaction. With the aid of three-dimensional (3D) bioprinted sacrificial molds, HBC/OCS hydrogel with various structures were obtained. After the material constituent optimization process, an injectable hydrogel with a uniform porous structure of 100 μm average pore size was developed to form macroporous hydrogel. In vitro and in vivo biocompatibility of optimized HBC/OCS hydrogel were also carefully assessed. The results indicated that human adipose-derived mesenchymal stem cells could be 3D cultured in HBC/OCS hydrogel maintaining good viability. Moreover, the hydrogels were found to trigger the least amount of pro-inflammatory gene expression of macrophage and to inhibit acute immune responses in 7 d. These results demonstrate the potential of HBC/OCS hydrogels as a cell delivery system for cartilage tissue engineering.

Topics: Animals; Biocompatible Materials; Biopolymers; Bioprinting; Cartilage; Cartilage, Articular; Cell Survival; Chitosan; Chondroitin Sulfates; Cross-Linking Reagents; Humans; Hydrogels; Immune System; Inflammation; Macrophages; Mesenchymal Stem Cells; Mice; Oxygen; Porosity; Printing, Three-Dimensional; Regeneration; Regenerative Medicine; Rheology; Spectroscopy, Fourier Transform Infrared; Tissue Engineering; Tissue Scaffolds

Osteoarthritis (OA) affects the integrity of the entire joint including the synovium. The most abundant cells in the synovium are fibroblasts (SF). Excessive mechanical loading might contribute to OA pathogenesis. Here, we investigate the effects of mechanical loading on SF derived from non-OA (N-SF) and OA patients (OA-SF). We treated N-SF and OA-SF with or without mechanical loading for 48h after 24h of preincubation. Then we assessed gene and protein expression of proinflammatory factors (TNFα, COX-2, PG-E2, IL-6), extracellular matrix (ECM) components (COL1, FN1) and glycosaminoglycans (GAGs) via RT-qPCR, ELISA, DMMB assay and HPLC. Mechanical loading significantly increased TNFα and PG-E2 secretion by N-SF and OA-SF, whereas in OA-SF IL-6 secretion was reduced. COL1 and FN1 secretion were downregulated in N-SF during loading. OA-SF secreted less COL1 compared to N-SF under control conditions. In contrast, OA-SF in general expressed more FN1. GAG synthesis was upregulated in N-SF, but not in OA-SF during loading with OA-SF displaying a higher charge density than N-SF. Mechanical loading enhanced proinflammatory factor expression and GAG synthesis and decreased secretion of ECM components in N-SFs, indicating a contributing role of SF to OA development.

Topics: Adult; Aged; Chondroitin Sulfates; Female; Fibroblasts; Glycosaminoglycans; Humans; Inflammation; Male; Middle Aged; Osteoarthritis; Reverse Transcriptase Polymerase Chain Reaction; Stress, Mechanical; Synovial Membrane

This study evaluated the possible prebiotic effects of dietary fucosylated chondroitin sulfate from Acaudina molpadioides (Am-CHS) on the modulation of the gut microbiota and the improvement in the risk factors for chronic inflammation in high fat diet-fed mice. The results showed that the Am-CHS treatment greatly modified the gut microbiota, including the decrease in Bacteroidetes, increase in Firmicutes, elevation in Lactobacillus (intestinal barrier protector) and short chain fatty acid (SCFA)-producing bacteria (Lactobacillus, Bifidobacterium, and Lachnospiraceae NK4A136 group), and reduction in the lipopolysaccharide (LPS) producer (Escherichia coli). This modulation inhibited inflammatory response, manifesting the decreases in circulating proinflammatory cytokines and their mRNA expression, and the increases in interleukin-10. Dietary Am-CHS caused reductions in serum and fecal LPS concentrations and inhibition of transcription of toll-like receptor 4 (TLR4) and its downstream proteins. In addition, there were increases in the portal levels of fecal SCFAs, which probably contributed to an increase in the adenosine monophosphate-activated protein kinase (AMPK) protein in Am-CHS-treated mice. These results suggest that modulation of gut microbiota by Am-CHS can improve chronic inflammation by reducing LPS levels and TLR4 signaling. Modulation also appears to increase the levels of fecal SCFAs, which activates AMPK and finally leads to inflammation resistance.

Topics: Animals; Chondroitin Sulfates; Cytokines; DNA, Bacterial; Escherichia coli Proteins; Fatty Acids, Volatile; Gastrointestinal Microbiome; Gene Expression Regulation; Inflammation; Intestinal Mucosa; Intestines; Lipopolysaccharides; Male; Membrane Proteins; Mice; Mice, Inbred C57BL; Obesity; Phosphotransferases; RNA, Messenger; Sea Cucumbers

Interstitial cystitis and/or bladder pain syndrome (IC/BPS) are characterized by discomfort, abdominal pain, and pelvic pain, and they are often associated with chronic diseases. Pathological conditions related to IC/BPS can occur due to a defect in the integrity of the bladder lining. This defect has been ascribed to damage to the glycosaminoglycan (GAG) layer of the urinary epithelium. In addition, the incipient cascade of inflammation events might prompt extracellular matrix degradation. Several medical devices based on GAG instillation were proposed to re-establish epithelial integrity by GAGs binding to proteoglycans or interacting with structural urothelium. However, to date, only in vitro studies have investigated the GAG, hyaluronic acid (HA). In the present study, TNFα treatment was used to mimic IC/BPS-induced damage in bladder cells in an in vitro model. Highly purified fermentative HA and pharmaceutical grade bovine chondroitin sulfate (CSb), alone or in combination, were evaluated for the ability to counteract bladder cell damage. We evaluated NF-κB with western blots, and we analyzed interleukin 6 and 8 expression at the transcriptional and protein levels with quantitative RT-PCR, western blotting, and ELISA. We also evaluated the expression of an antibacterial peptide, human β-defensin-2. We confirmed our results in a 3D bladder epithelium model. Our results demonstrated that inflammatory status was reduced in the presence of HA, CSb, and the combination of both (HA/CSb 1.6%/2% w/v). This result suggested that these GAGs might be suitable for treating IC/BPS. All the assayed biomarkers showed that HA/CSb treatment modulated cells towards a more physiological status. Finally, we compared two commercial products suggested for the IC/BPS treatments and found that the product with more Ca++, showed enhanced anti-inflammatory activity and provided superior mucoadhesivity.

Topics: Animals; Cattle; Cell Line; Chondroitin Sulfates; Gene Expression Regulation; Glycosaminoglycans; Humans; Hyaluronic Acid; Hydrodynamics; Inflammation; Interleukin-6; Interleukin-8; Models, Biological; NF-kappa B; Signal Transduction; Tumor Necrosis Factor-alpha; Urinary Bladder; Zonula Occludens-1 Protein

Chondroitin sulphate (CS) has long been used to treat osteoarthritis. Some investigations have also shown that the treatment with CS could reduce coronary events in patients with heart disease but no studies have identified the mechanistic role of these therapeutic effects. We aimed to investigate how the treatment with CS can interfere with the progress of atherosclerosis. The aortic arch, thoracic aorta and serum were obtained from apolipoprotein E (ApoE) knockout mice fed for 10 weeks with high-fat diet and then treated with CS (300 mg/kg,

Topics: Animals; Anti-Inflammatory Agents; Aorta, Thoracic; Aortic Diseases; Atherosclerosis; Blood Glucose; C-Reactive Protein; Chondroitin Sulfates; Diet, High-Fat; Disease Models, Animal; Endothelial Cells; Foam Cells; Humans; Inflammation; Inflammation Mediators; Lipids; Lipoproteins, LDL; Male; Mice, Knockout, ApoE; Monocytes; Plaque, Atherosclerotic; THP-1 Cells

Osteoarthritic patients treated with high doses of chondroitin sulfate (CS) have a lower incidence of coronary heart disease--but the mechanistic aspects of these beneficial effects of CS remain undefined. We examined how CS treatment affects the formation of atheroma via interaction with endothelial cells and monocytes.. We characterized arterial atheromatous plaques by multiphoton microscopy and serum pro-inflammatory cytokines by immunoenzymatic techniques in obese mice receiving CS (1 g/kg/day, i.p.) or vehicle for 6 days. Effects of CS on signaling pathways, cytokine secretion and macrophage migration were evaluated in cultures of human coronary endothelial cells and in a monocyte cell line stimulated with TNF-α by Western blot, immunoenzymatic techniques and transwell migration assays.. Treatment of obese mice with CS reduced the extension of foam cell coverage in atheromatous plaques of arterial bifurcations by 62.5%, the serum concentration of IL1β by 70%, TNF-α by 82% and selected chemokines by 25-35%. Cultures of coronary endothelial cells and monocytes stimulated with TNF-α secreted less pro-inflammatory cytokines in the presence of CS (P < 0.01). CS reduced the activation of the TNF-α signaling pathway in endothelial cells (pErk 36% of reduction, and NFκB 33% of reduction), and the migration of activated monocytes to inflamed endothelial cells in transwells (81 ± 6 vs. 13 ± 2, P < 0.001).. CS interferes with the pro-inflammatory activation of monocytes and endothelial cells driven by TNF-α thus reducing the propagation of inflammation and preventing the formation of atherosclerotic plaques.

Topics: Animals; Atherosclerosis; Cell Line; Chondroitin Sulfates; Disease Models, Animal; Endothelium, Vascular; Inflammation; Male; Mice; Mice, Obese; Obesity

The purpose of this study was to clarify fundamental mechanical properties and biological responses of the sodium hyaluronate-containing double network (HA-DN) gel and chondroitin sulfate-containing double network (CS-DN) gel, which were newly developed using the molecular stent method. This study discovered the following facts. First, these hydrogels had high mechanical performance comparable to the native cartilage tissue, and the mechanical properties were not affected by immersion in the saline solution for 12weeks. Secondly, the mechanical properties of the CS-DN gel were not significantly reduced at 12weeks in vivo, while the mechanical properties of the HA-DN gel were significantly deteriorated at 6weeks. Thirdly, the degree of inflammation around the HA-DN gel was the same as that around the negative control. The CS-DN gel showed a mild but significant foreign body reaction, which was significantly greater than the negative control and less than the positive control at 1week, while the inflammation was reduced to the same level as the negative control at 4 and 6weeks. Fourthly, these gels induced differentiation of the ATDC5 cells into chondrocytes in the culture with the insulin-free maintenance medium. These findings suggest that these tough hydrogels are potential biomaterials for future application to therapeutic implants such as artificial cartilage.. The present study reported fundamental biomaterial properties of the sodium hyaluronate-containing double network (HA-DN) gel and chondroitin sulfate-containing double network (CS-DN) gel, which were newly developed using the molecular stent method. Both the HA- and CS-DN gels had high mechanical properties comparable to the cartilage tissue and showed the ability to induce chondrogenic differentiation of ATDC5 cells in vitro. They are potential biomaterials that may meet the requirements of artificial cartilage concerning the material properties. Further, these DN gels can be also applied to the implantable inducer for cell-free cartilage regeneration therapy.

Topics: Animals; Biocompatible Materials; Buffers; Cell Differentiation; Cells, Cultured; Chondrogenesis; Chondroitin Sulfates; Collagen Type II; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Hydrogels; Implants, Experimental; Inflammation; Materials Testing; Mechanical Phenomena; Muscles; Rabbits; Sterilization; Subcutaneous Tissue; Water

The microfracture technique for cartilage repair has limited ability to regenerate hyaline cartilage.. The current study made a direct comparison between microfracture and an osteochondral approach with microsphere-based gradient plugs.. The PLGA-based scaffolds had opposing gradients of chondroitin sulfate and β-tricalcium phosphate. A 1-year repair study in sheep was conducted.. The repair tissues in the microfracture were mostly fibrous and had scattered fissures with degenerative changes. Cartilage regenerated with the gradient plugs had equal or superior mechanical properties; had lacunated cells and stable matrix as in hyaline cartilage.. This first report of gradient scaffolds in a long-term, large animal, osteochondral defect demonstrated potential for equal or better cartilage repair than microfracture.

Topics: Animals; Bone and Bones; Calcium Phosphates; Cartilage, Articular; Chondrocytes; Chondroitin Sulfates; Female; Femur; Finite Element Analysis; Humans; Hyaline Cartilage; Inflammation; Knee Joint; Lactic Acid; Magnetic Resonance Imaging; Microspheres; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Regeneration; Sheep; Stress, Mechanical; Tissue Engineering; Tissue Scaffolds; Transforming Growth Factor beta

Inflammation is a host protective response to noxious stimuli, and excessive production of pro-inflammatory mediators by macrophages (mφ) can lead to numerous pathological conditions. In this study, immunomodulatory effects of immobilized and soluble glycosaminoglycans (GAGs) on mouse-bone-marrow-derived mφ were compared by measuring nitric oxide (NO). We demonstrate here that all GAGs studied except for heparin were able to modulate interferon-γ/lipopolysaccharide (IFN-γ/LPS)-induced NO release by mφ to varying extents after 24h of incubation. In particular, the modulatory activities of soluble chondroitin-6-sulfate (C6S), hyaluronic acid and heparan sulfate altered markedly after covalent immobilization. Of these, soluble C6S exhibited the strongest NO inhibitory activity, and the inhibition was dose- and time-dependent. Moreover, C6S significantly reduced pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α production by IFN-γ/LPS- or LPS-activated mφ. Specifically, the C6S-mediated suppression of mφ pro-inflammatory phenotype was accompanied by an increase in the IL-10 level, suggesting a possible switch towards anti-inflammatory/wound healing M2 state. In addition, the highest magnitude of inhibitory effects was obtained when cells were pre-treated with C6S prior to IFN-γ/LPS or LPS challenge, suggesting an additional role for C6S in protection against microbial infection. Further investigations reveal that the anti-inflammatory effects of C6S on activated mφ may be ascribed at least in part to suppression of NF-κB nuclear translocation.

Topics: Animals; Biological Transport; Cell Nucleus; Chondroitin Sulfates; Inflammation; Macrophages; Male; Mice; Mice, Inbred BALB C; NF-kappa B

Chondroitin Sulphate (CS), a natural glycosaminoglycan of the extracellular matrix, has clinical benefit in symptomatic osteoarthritis but has never been tested in gout. In vitro, CS has anti-inflammatory and positive effects on osteoarthritic chondrocytes, synoviocytes and subchondral bone osteoblasts, but its effect on macrophages is unknown. The purpose of our study was to evaluate the in vitro effects of CS on monosodium urate (MSU)-stimulated cytokine production by macrophages.. THP-1 monocytes were differentiated into mature macrophages using a phorbol ester, pretreated for 4 hours with CS in a physiologically achievable range of concentrations (10-200 μg/ml) followed by MSU crystal stimulation for 24 hours. Cell culture media were analyzed by immunoassay for factors known to be upregulated during gouty inflammation including IL-1β, IL-8 and TNFα. The specificity of inflammasome activation by MSU crystals was tested with a caspase-1 inhibitor (0.01 μM-10 μM).. MSU crystals ≥10 mg/dl increased macrophage production of IL-1β, IL-8 and TNFα a mean 7-, 3- and 4-fold respectively. Induction of IL-1β by MSU was fully inhibited by a caspase-1 inhibitor confirming inflammasome activation as the mechanism for generating this cytokine. In a dose-dependent manner, CS significantly inhibited IL-1β (p = 0.003), and TNFα (p = 0.02) production from macrophages in response to MSU. A similar trend was observed for IL-8 but was not statistically significant (p = 0.41).. CS attenuated MSU crystal induced macrophage inflammation, suggesting a possible role for CS in gout prophylaxis.

Topics: Cell Line; Chondroitin Sulfates; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical; Gout; Humans; Inflammation; Macrophages; Monocytes; Uric Acid

This study aimed to assess whether chronic administration of chondroitin sulfate (CS) affects baseline expression of cytochrome P450 isoforms and impedes the decrease in expression and activity of CYP1A2 and CYP3A6 in rabbits with a turpentine-induced inflammatory reaction (TIIR). Seven groups of 5 rabbits, 3 control groups and 4 receiving 20 mg/kg/day of CS for 20 and 30 days, were used. The rabbits of 1 control group and 2 groups receiving CS had a TIIR; finally, the rabbits of one of the control groups remained in the animal facilities for 30 days to assess the effect of time and environment on cytochrome P450. In control rabbits, intake of CS for 20 and 30 days did not affect CYP3A6, CYP1A2 and NADPH cytochrome P450 reductase (CPR) mRNA, protein expression and activity. Compared with control rabbits, the TIIR not only reduced mRNA, protein expression and activity of CYP3A6 and CYP1A2 but also that of CPR. In rabbits with TIIR, CS prevented the decrease of CYP3A6 expression but not the reduction in activity. CS did not impede TIIR-induced down-regulation of CYP1A2. Hepatic NO() concentrations and NF-κB nuclear translocation were increased by the TIIR, effect reversed by CS. In vitro, in hepatocytes, CS did not alter the expression and activity of CYP3A6, CYP1A2, and CPR. In conclusion, oral CS elicits a systemic effect but does not affect CYP1A2, CYP3A6, and CPR in control rabbits, although in rabbits with TIIR, CS prevents CYP3A6 protein down-regulation but not that of CYP1A2.

Topics: Animals; Aryl Hydrocarbon Hydroxylases; Chondroitin Sulfates; Cytochrome P-450 CYP1A2; Down-Regulation; Enzyme Inhibitors; Inflammation; Male; Rabbits; RNA, Messenger; Structure-Activity Relationship; Turpentine

Heat stroke is a condition characterized by high body temperature that can lead to hemorrhage and necrosis in multiple organs. Anticoagulants, such as danaparoid sodium (DA), inhibit various types of inflammation; however, the anti-inflammatory mechanism of action is not well understood. Given that heat stroke is a severe inflammatory response disease, we hypothesized that DA could inhibit inflammation from heat stress and prevent acute heat stroke.. Male Wistar rats were given a bolus injection of saline or DA (50 U/kg body weight) into the tail vein just prior to heat stress (42 °C for 30 min). Markers of inflammation were then determined in serum and tissue samples.. In rats pretreated with DA, induction of cytokines (interleukin [IL]-1β, IL-6, and tumor necrosis factor [TNF]-α), nitric oxide (NO), and high mobility group box 1 (HMGB1) protein were reduced compared with saline-treated rats. Histologic changes observed in lung, liver, and small intestine tissue samples of saline-treated rats were attenuated in DA-treated rats. Moreover, DA pretreatment improved survival in our rat model of heat stress-induced acute inflammation.. Collectively, our findings demonstrate that DA pretreatment may have value as a new therapeutic tool for heat stroke.

Topics: Acute Disease; Animals; Anticoagulants; Antithrombin III; Chondroitin Sulfates; Cytokines; Dermatan Sulfate; Disease Models, Animal; Fibrin Fibrinogen Degradation Products; Heat Exhaustion; Heat Stroke; Heparitin Sulfate; HMGB1 Protein; Inflammation; Intestine, Small; Liver; Lung; Male; Nitrates; Nitric Oxide; Nitrites; Peptide Hydrolases; Rats; Rats, Wistar; Survival Rate

CD44 is a cell surface receptor for the extracellular matrix glycosaminoglycan hyaluronan and is involved in processes ranging from leukocyte recruitment to wound healing. In the immune system, the binding of hyaluronan to CD44 is tightly regulated, and exposure of human peripheral blood monocytes to inflammatory stimuli increases CD44 expression and induces hyaluronan binding. Here we sought to understand how mouse macrophages regulate hyaluronan binding upon inflammatory and anti-inflammatory stimuli. Mouse bone marrow-derived macrophages stimulated with tumor necrosis factor α or lipopolysaccharide and interferon-γ (LPS/IFNγ) induced hyaluronan binding by up-regulating CD44 and down-regulating chondroitin sulfation on CD44. Hyaluronan binding was induced to a lesser extent in interleukin-4 (IL-4)-activated macrophages despite increased CD44 expression, and this was attributable to increased chondroitin sulfation on CD44, as treatment with β-d-xyloside to prevent chondroitin sulfate addition significantly enhanced hyaluronan binding. These changes in the chondroitin sulfation of CD44 were associated with changes in mRNA expression of two chondroitin sulfotransferases, CHST3 and CHST7, which were decreased in LPS/IFNγ-stimulated macrophages and increased in IL-4-stimulated macrophages. Thus, inflammatory and anti-inflammatory stimuli differentially regulate the chondroitin sulfation of CD44, which is a dynamic physiological regulator of hyaluronan binding by CD44 in mouse macrophages.

Topics: Animals; Carbohydrate Sulfotransferases; Cell Line, Tumor; Chondroitin Sulfates; Gene Expression Regulation; Humans; Hyaluronan Receptors; Hyaluronic Acid; Inflammation; Interferon-gamma; Interleukin-4; Lipopolysaccharides; Macrophage Activation; Macrophages; Mice; Mice, Knockout; Sulfotransferases; Tumor Necrosis Factor-alpha

Disturbed alveolar fibrin turnover is a characteristic feature of pneumonia. Inhibitors of coagulation could exert lung-protective effects via anticoagulant (inhibiting fibrin deposition) and possibly anti-inflammatory pathways, but could also affect host defense. In this randomized controlled in vivo laboratory study, rats were challenged intratracheally with Pseudomonas aeruginosa, inducing pneumonia, and randomized to local treatment with normal saline (placebo), recombinant human activated protein C (rh-APC), plasma-derived antithrombin (AT), heparin, or danaparoid. Induction of P. aeruginosa pneumonia resulted in activation of pulmonary coagulation and inhibition of pulmonary fibrinolysis, as reflected by increased pulmonary levels of thrombin-AT complexes and fibrin degradation products and decreased pulmonary levels plasminogen activator activity. Pseudomonas aeruginosa pneumonia was accompanied by systemic coagulopathy, since systemic levels of thrombin-AT complexes increased, and systemic levels of plasminogen activator activity decreased. Although rh-APC and plasma-derived AT potently limited pulmonary coagulopathy, neither heparin nor danaparoid affected net pulmonary fibrin turnover. Recombinant human APC also displayed systemic anticoagulant effects. Neither bacterial clearance nor pulmonary inflammation was affected by anticoagulant therapy. Nebulization of rh-APC or plasma-derived AT attenuated pulmonary coagulopathy, but not bacterial clearance or inflammation, in a rat model of P. aeruginosa pneumonia.

Topics: Animals; Anticoagulants; Antithrombins; Blood Coagulation; Chondroitin Sulfates; Dermatan Sulfate; Fibrinolysis; Heparin; Heparitin Sulfate; Inflammation; Male; Pneumonia; Pseudomonas aeruginosa; Rats; Rats, Sprague-Dawley

The chemokine IL-8 is critically important in inflammatory processes in human tissues, and IL-8 interactions with sulfated glycosaminoglycans have been implicated in modification of inflammatory responses in bronchial epithelium. To determine the role of chondroitin-4-sulfate (C4S) in mediating effects of IL-8, we silenced the enzyme N-acetylgalactosamine-4-sulfatase (arylsulfatase B [ASB]) that removes the 4-sulfate group from C4S, in the IB3-1 and C38 bronchial epithelial cell lines and in normal primary bronchial epithelial cells. When ASB was silenced and IL-8 production stimulated by exposure to TNF-alpha, ASB activity declined by roughly 75%, cellular C4S content increased by over 7.5 microg/mg protein, cell-bound IL-8 increased by over 530 pg/mg protein, and secreted IL-8 declined by over 520 pg/mg protein in all cell lines (P < 0.001). When cell lysates were immunoprecipitated with C4S antibody after ASB silencing and TNF-alpha, the IL-8 content of the immunoprecipitate was approximately 500 pg/mg protein, indicating that most of the cell-bound IL-8 was associated with C4S. Cell fractionation demonstrated that the IL-8 content associated with the cell membranes was about twice that of the cytosolic fraction. Also, ASB appeared to localize in the cell membrane, as well as in lysosomes. Neutrophil attraction to the cell lysates increased after ASB silencing, consistent with increased attraction to the cell-bound IL-8. These findings provide evidence for the influential role of ASB and C4S in the regulation of IL-8 secretion, and suggest that changes in ASB activity and C4S content may have a significant impact on IL-8-mediated inflammatory responses.

Topics: Bronchi; Cell Line; Chondroitin Sulfates; Cystic Fibrosis; Epithelial Cells; Gene Silencing; Glycosaminoglycans; Humans; Immunohistochemistry; Inflammation; Interleukin-8; Lysosomes; Microscopy, Confocal; N-Acetylgalactosamine-4-Sulfatase; Subcellular Fractions; Tumor Necrosis Factor-alpha

The present study examines the effect of chondroitin-4-sulfate (C4S) on the immediate (non-inflammatory conditions) and the delayed (inflammatory conditions) prostaglandin E(2) (PGE(2)) release from rat calvarial osteoblasts. An immediate low release of PGE(2) was induced by PAF, phorbol ester and arachidonic acid but not by IL1beta, TNF-alpha and LPS whereas a delayed high release of PGE(2) was induced by the inflammatory agents IL1beta, TNF-alpha and LPS but not by PAF, phorbol ester and arachidonic acid. C4S had no effect on the immediate PGE(2) release but inhibited the delayed release of PGE(2). IL1beta, TNF-alpha and LPS enhanced the expression of COX-2 and mPGES1 whereas phorbol ester enhanced COX-2 expression only. PAF and arachidonic acid had no effect on the expression of COX-2 and mPGES1. C4S inhibited the enhanced expression of COX-2 and mPGES1 but had no effect on the IL1beta-induced decrease of I-kappaBalpha and nuclear translocation of NF-kappaB. These results indicate that the beneficial effects of C4S in bone inflammatory diseases might be due to a specific inhibition of the delayed high PGE(2) release from osteoblasts.

Topics: Animals; Arachidonic Acid; Chondroitin Sulfates; Dinoprostone; Inflammation; NF-kappa B; Osteoblasts; Rats; Skull

The endothelial glycocalyx is well endowed with the glycosaminoglycans (GAGs) heparan sulfate, chondroitin sulfate and hyaluronan. The current studies aimed to assess the relative contributions of each of these GAGs to the thickness and permeability of the glycocalyx layer by direct enzymatic removal of each using micropipettes to infuse heparinase, chondroitinase and hyaluronidase into post-capillary venules of the intestinal mesentery of the rat. The relative losses of GAGs due to enzymatic removal were compared with stimulated shedding of glycans induced by superfusing the mesentery with 10(-)(7)M fMLP. Thickness of the glycocalyx was assessed by infiltration of the glycocalyx with circulating FITC labeled 70kDa dextran (Dx70) and measuring the distance from the dye front to the surface of the endothelium (EC), which averaged 463nm under control conditions. Reductions in thickness were 43.3%, 34.1% and 26.1% following heparinase, chondroitinase and hyaluronidase, respectively, and 89.7% with a mixture of all three enzymes. Diffusion coefficients of FITC in the glycocalyx were determined using a 1-D diffusion model. By comparison of measured transients in radial intensity of a bolus of FITC with that of a computational model a diffusion coefficient D was obtained. Values of D were obtained corresponding to the thickness of the layer demarcated by Dx70 (D(Dx70)), and a smaller sublayer 173nm above the EC surface (D(173)), prior to and following enzyme infusion and superfusion with fMLP. The magnitude of D(Dx70) was twice that of D(173) suggesting that the glycocalyx is more compact near the EC surface. Chondroitinase and hyaluronidase significantly increased both D(Dx70) and D(173). However, heparinase decreased D(Dx70), and did not induce any significant change for the D(173). These observations suggest that the three GAGs are not evenly distributed throughout the glycocalyx and that they each contribute to permeability of the glycocalyx to a differing extent. The fMLP-induced shedding caused a reduction in glycocalyx thickness (which may increase permeability) and as with heparinase, decreased the diffusion coefficient of solutes (which may decrease permeability). This behavior suggests that the removal of heparan sulfate may cause a collapse of the glycocalyx which counters decreases in thickness by compacting the layer to maintain a constant resistance to filtration.

Topics: Animals; Cell Membrane Permeability; Chondroitin Sulfates; Chondroitinases and Chondroitin Lyases; Computer Simulation; Dextrans; Diffusion; Endothelial Cells; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Glycocalyx; Glycosaminoglycans; Heparin Lyase; Heparitin Sulfate; Hyaluronic Acid; Hyaluronoglucosaminidase; Inflammation; Infusions, Intravenous; Male; Mesentery; Models, Cardiovascular; N-Formylmethionine Leucyl-Phenylalanine; Rats; Rats, Wistar; Venules

Glycosaminoglycans (long-chain polysaccharides) are major components of the extracellular matrix, glycocalyx, and synovial fluid. These materials provide strength and elasticity to tissues and play a key role in regulating cell behavior. Modifications to these materials have been linked to multiple human pathologies. Although modification may occur via both enzymatic and nonenzymatic mechanisms, there is considerable evidence for oxidant-mediated matrix damage. Peroxynitrite (ONOO(-)/ONOOH) is a potential mediator of such damage, as elevated levels of this oxidant are likely to be present at sites of inflammation. In this study we demonstrate that hyaluronan and chondroitin sulfate are extensively depolymerized by HO(.) and CO3(.-), but not NO2(.), which may be formed from peroxynitrite. Polymer fragmentation is shown to be dependent on the radical flux, to be O2-independent, and to occur in a site-selective manner as indicated by the detection of disaccharide fragments. EPR spin trapping experiments with polymers, oligomers, and component monosaccharides, including 13C-labeled materials, have provided evidence for the formation of specific carbon-centered sugar-derived radicals. The time course of formation of these radicals is consistent with these species being involved in polymer fragmentation.

Topics: Carbon Radioisotopes; Carbonates; Chondroitin Sulfates; Electron Spin Resonance Spectroscopy; Extracellular Matrix; Free Radicals; Glycocalyx; Glycosaminoglycans; Humans; Hyaluronic Acid; Hydroxyl Radical; Inflammation; Nitrogen Dioxide; Oxidative Stress; Peroxynitrous Acid; Polymers; Synovial Fluid

Among the agents employed to manage osteoarthritis, chondroitin sulphate (CS) is a natural glycosaminoglycan with an anti-inflammatory effect on joint cells. CS might also influence the inflammatory component of atherosclerosis. Our aim was to examine the effect of CS administration on vascular injury and on markers of systemic inflammation in a rabbit model of atherosclerosis aggravated by systemic inflammation provoked by chronic antigen-induced arthritis.. Atherosclerosis was induced in rabbits by maintaining them on a hyperlipidaemic diet after producing an endothelial lesion in the femoral arteries. Simultaneously, chronic arthritis was induced in these animals by repeated intraarticular injections of ovalbumin in previously immunized rabbits. A group of these rabbits were treated prophylactically with CS (100 mg kg(-1)day(-1)) and when the animals were killed, serum and peripheral blood mononuclear cells (PBMC) were isolated. Furthermore, femoral arteries and thoracic aorta were used for gene expression studies and histological examination.. CS administration reduced the concentration of the proinflammatory molecules C-reactive protein and IL-6 in serum. Likewise, CS inhibited the expression of CCL2/monocyte chemoattractant protein (MCP)-1 and cyclooxygenase (COX)-2 in PBMC, and reduced the nuclear translocation of nuclear factor-kappaB. In the femoral lesion, CS also diminished the expression of CCL2 and COX-2, as well as the ratio of the intima/media thickness. Moreover, CS decreased the percentage of rabbits with atherosclerosis and chronic arthritis that developed vascular lesions in the aorta.. These findings suggest that CS treatment may to some extent impede the progression of atherosclerosis.

Topics: Animals; Anti-Inflammatory Agents; Aorta; Arthritis, Experimental; Atherosclerosis; C-Reactive Protein; Chemokine CCL2; Chondroitin Sulfates; Cyclooxygenase 2; Disease Models, Animal; Disease Progression; Gene Expression Regulation; Inflammation; Interleukin-6; Male; NF-kappa B; Rabbits

Familial Amyloidotic Polyneuropathy (FAP) is a disorder characterized by the extracellular deposition of fibrillar Transthyretin (TTR) amyloid, with a special involvement of the peripheral nerve. Several extracellular matrix proteins have been found elevated in tissues from FAP patients, namely metalloproteinase-9 (MMP-9), neutrophil gelatinase associated lipocalin (NGAL) and biglycan. In this work we assessed the levels of MMP-9, tissue inhibitor of metalloproteinase 1 (TIMP-1), NGAL, biglycan and chondroitin sulphate (CSPG) in an FAP V30M TTR-related transgenic mouse model at different stages of TTR deposition and after two different treatment approaches to remove fibrillar deposits. Immunohistochemistry or RT-PCR analysis showed that biglycan was already increased in animals presenting TTR deposited in a non-fibrillar state, whereas MMP-9, TIMP-1, NGAL and CSPG were elevated only in mice with TTR amyloid deposits. Mice treated with doxycycline, a TTR fibril disrupter, presented lower levels of MMP-9, TIMP-1 and NGAL, suggestive of matrix recovery. Mice immunized with TTR Y78F to remove TTR deposition showed significantly lower levels of all the five tested markers, suggesting removal of fibrillar and non-fibrillar deposits. Cellular studies using oligomeric TTR showed induction of MMP-9 when compared to soluble TTR, large aggregates or fibrils. Furthermore, this induction was neutralized by an anti-receptor for advanced glycation end products (RAGE) antibody, indicating RAGE engagement in this process. Further studies in a larger number of tissue samples will indicate the application of these ECM markers in parallel with Congo Red staining in tissue characterization of pre-clinical and clinical stages in FAP and other amyloidoses.

Topics: Amyloid Neuropathies, Familial; Animals; Biglycan; Cell Line, Tumor; Chondroitin Sulfates; Disease Progression; Extracellular Matrix; Extracellular Matrix Proteins; Follow-Up Studies; Inflammation; Matrix Metalloproteinase 9; Mice; Mice, Transgenic; Models, Biological; Proteoglycans; Rats; Tissue Inhibitor of Metalloproteinase-1

Inflammatory factors are known to play a key role in promoting tumorigenesis; therefore, it is a promising strategy to inhibit the inflammation for cancer prevention. The current study was performed to investigate the potential effects of chondroitin sulfate (CS) extracted from ascidian tunic on the expression of inflammatory factors induced by treatment with 12- O-tetradecanoylphorbol-13-acetate (TPA) and to elucidate the underlying molecular mechanism of CS action in mouse skin inflammation. TPA was topically applied to the shaven backs of ICR mice with or without CS (1 or 2 mg) for 4 h. The results demonstrated that CS suppressed TPA-induced edema and reduced the expression of cyclooxygenase-2, vascular cell adhesion molecule-1, and Akt signaling in mouse skin. These studies suggest that CS from ascidian tunic may be developed as an effective natural anti-inflammatory agent.

Topics: Animals; Carcinogens; Cell Extracts; Chondroitin Sulfates; Cyclooxygenase 2; Down-Regulation; Female; Gene Expression; Humans; Inflammation; Mice; Mice, Inbred ICR; NF-kappa B; Phorbol Esters; Phosphorylation; Random Allocation; Signal Transduction; Skin; Urochordata; Vascular Cell Adhesion Molecule-1

Inflammatory mediators are known to play a key role in tumorigenesis, therefore, it is a promising strategy to inhibit the inflammation for cancer prevention and/or treatment. Current study was performed to investigate the effects of chondroitin sulfate (CS) extracted from Styela clava tunic on TNF-alpha-induced inflammation and to elucidate the mechanism of CS on the regulation of inflammatory factors in JB6 cells. Our results showed that CS inhibited TNF-alpha-induced NF-kappaB activation and subsequent vascular cell adhesion molecule 1 and inducible nitric oxide synthase expressions by blocking Akt signals in JB6 cells. Our results suggest that CS may be developed as an effective anti-inflammatory agent in the future.

Topics: Animals; Anti-Inflammatory Agents; Blotting, Western; Cell Line; Cell Survival; Chondroitin Sulfates; Down-Regulation; Epidermis; Inflammation; Inflammation Mediators; Mice; NF-kappa B; Nitric Oxide Synthase Type II; Proto-Oncogene Proteins c-akt; Signal Transduction; Tumor Necrosis Factor-alpha; Urochordata; Vascular Cell Adhesion Molecule-1

This study was undertaken to examine the mechanism by which danaparoid sodium (DS), a heparinoid that contains mainly heparan sulfate, prevents reperfusion-induced hepatic damage in a rat model of ischemia/reperfusion (I/R)-induced liver injury. Administration of DS significantly reduced liver injury and inhibited the decrease in hepatic tissue blood flow in rats. DS attenuated hepatic I/R-induced increases in hepatic tissue levels of tumor necrosis factor (TNF) and myeloperoxidase (MPO) in vivo. In contrast, neither monocytic TNF production nor neutrophil activation was inhibited by DS in vitro. DS enhanced I/R-induced increases in levels of calcitonin-gene related peptide (CGRP), a neuropeptide released from sensory neurons, and of 6-ketoprostaglandin (PG) F (1a) , a stable metabolite of PGI (2) , in liver tissues. The therapeutic effects of DS were not seen in animals pretreated with capsazepine, an inhibitor of sensory neuron activation. The distribution of heparan sulfate in the perivascular area was significantly increased by DS administration in this rat model. DS significantly increased CGRP release from isolated rat dorsal root ganglion neurons (DRG) in vitro, while DX-9065a, a selective inhibitor of activated factor X, did not. DS enhanced anandamide-induced CGRP release from DRG in vitro. These observations strongly suggested that DS might reduce I/R-induced liver injury in rats by attenuating inflammatory responses. These therapeutic effects of DS might be at least partly explained by its enhancement of sensory neuron activation, leading to the increase the endothelial production of PGI (2) .

Topics: Animals; Calcitonin Gene-Related Peptide; Chondroitin Sulfates; Dermatan Sulfate; Epoprostenol; Ganglia, Spinal; Heparitin Sulfate; Inflammation; Liver; Male; Neurons, Afferent; Rats; Rats, Wistar; Regional Blood Flow; Reperfusion Injury

TSG-6 protein, up-regulated in inflammatory lesions and in the ovary during ovulation, shows anti-inflammatory activity and plays an essential role in female fertility. Studies in murine models of acute inflammation and experimental arthritis demonstrated that TSG-6 has a strong anti-inflammatory and chondroprotective effect. TSG-6 protein is composed of the N-terminal link module that binds hyaluronan and a C-terminal CUB domain, present in a variety of proteins. Interactions between the isolated link module and hyaluronan have been studied extensively, but little is known about the binding of full-length TSG-6 protein to hyaluronan and other glycosaminoglycans. We show that TSG-6 protein and hyaluronan, in a temperature-dependent fashion, form a stable complex that is resistant to dissociating agents. The formation of such stable complexes may underlie the activities of TSG-6 protein in inflammation and fertility, e.g. the TSG-6-dependent cross-linking of hyaluronan in the cumulus cell-oocyte complex during ovulation. Because adhesion to hyaluronan is involved in cell trafficking in inflammatory processes, we also studied the effect of TSG-6 on cell adhesion. TSG-6 binding to immobilized hyaluronan did not interfere with subsequent adhesion of lymphoid cells. In addition to immobilized hyaluronan, full-length TSG-6 also binds free hyaluronan and all chondroitin sulfate isoforms under physiological conditions. These interactions may contribute to the localization of TSG-6 in cartilage and to its chondroprotective and anti-inflammatory effects in models of arthritis.

Topics: Animals; Anti-Inflammatory Agents; Biotinylation; Cell Adhesion; Cell Adhesion Molecules; Cell Line; Cell Line, Tumor; Chickens; Chondroitin Sulfates; Dose-Response Relationship, Drug; Edetic Acid; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Hydrogen-Ion Concentration; Inflammation; Insecta; Ions; Kinetics; Mice; Oocytes; Ovary; Ovulation; Protein Binding; Protein Isoforms; Protein Structure, Tertiary; Temperature; Time Factors; Up-Regulation

This study examined the effects of chondroitin sulfate (CS) alone and CS plus glucosamine sulfate (GS) in a dietary bar formulation on inflammatory parameters of adjuvant-induced arthritis and on the synthesis of interleukin-1beta (IL-1beta) and matrix metalloprotease-9 (MMP-9). Following 25 days pretreatment with dietary bars containing either CS alone, CS plus GS, or neither CS nor GS, rats were either sham injected or injected with Freund's complete adjuvant into the tail vein. Rats were fed their respective bars for another 17 days after inoculation. Parameters of disease examined included clinical score (combination of joint temperature, edema, hyperalgesia, and standing and walking limb function), incidence of disease, levels of IL-1beta in the serum and paw joints, levels of MMP-9 in the paw joints, paw joint histology, and joint cartilage thickness. Treatment with CS plus GS, but not CS alone, significantly reduced clinical scores, incidences of disease, joint temperatures, and joint and serum IL-1beta levels. Treatment with CS alone and CS plus GS inhibited the production of edema and prevented raised levels of joint MMP-9 associated with arthritis. Similarly, CS alone and CS plus GS treatment also prevented the development of cartilage damage associated with arthritis. Combination CS plus GS treatment in a dietary bar formulation ameliorates clinical, inflammatory, and histologic parameters of adjuvant-induced arthritis. The benefits of CS and GS in combination are more pronounced than those of CS alone. The reduction of arthritic disease by CS plus GS is associated with a reduction of IL-1beta and MMP-9 synthesis.

Topics: Animals; Arthritis, Experimental; Cartilage, Articular; Chondroitin Sulfates; Drug Therapy, Combination; Freund's Adjuvant; Glucosamine; Hindlimb; Inflammation; Interleukin-1; Joints; Male; Matrix Metalloproteinase 9; Rats; Rats, Wistar; Time Factors

Histidine-rich glycoprotein (HRG) is an alpha2-glycoprotein found in mammalian plasma at high concentrations (approximately 150 microg/ml) and is distinguished by its high content of histidine and proline. Structurally, HRG is a modular protein consisting of an N-terminal cystatin-like domain (N1N2), a central histidine-rich region (HRR) flanked by proline-rich sequences, and a C-terminal domain. HRG binds to cell surfaces and numerous ligands such as plasminogen, fibrinogen, thrombospondin, C1q, heparin, and IgG, suggesting that it may act as an adaptor protein either by targeting ligands to cell surfaces or by cross-linking soluble ligands. Despite the suggested functional importance of HRG, the cell-binding characteristics of the molecule are poorly defined. In this study, HRG was shown to bind to most cell lines in a Zn(2+)-dependent manner, but failed to interact with the Chinese hamster ovary cell line pgsA-745, which lacks cell-surface glycosaminoglycans (GAGs). Subsequent treatment of GAG-positive Chinese hamster ovary cells with mammalian heparanase or bacterial heparinase III, but not chondroitinase ABC, abolished HRG binding. Furthermore, blocking studies with various GAG species indicated that only heparin was a potent inhibitor of HRG binding. These data suggest that heparan sulfate is the predominate cell-surface ligand for HRG and that mammalian heparanase is a potential regulator of HRG binding. Using recombinant forms of full-length HRG and the N-terminal N1N2 domain, it was shown that the N1N2 domain bound specifically to immobilized heparin and cell-surface heparan sulfate. In contrast, synthetic peptides corresponding to the Zn(2+)-binding HRR of HRG did not interact with cells. Furthermore, the binding of full-length HRG, but not the N1N2 domain, was greatly potentiated by physiological concentrations of Zn2+. Based on these data, we propose that the N1N2 domain binds to cell-surface heparan sulfate and that the interaction of Zn2+ with the HRR can indirectly enhance cell-surface binding.

Topics: Animals; Baculoviridae; Blotting, Western; Cell Membrane; Chelating Agents; CHO Cells; Chondroitin Sulfates; Complement C1q; COS Cells; Cricetinae; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Fibrinogen; Flow Cytometry; Glucuronidase; Glycosaminoglycans; Heparitin Sulfate; Histidine; Humans; Immunoglobulin G; Inflammation; Jurkat Cells; Ligands; Microscopy, Fluorescence; Neoplasm Metastasis; Peptides; Plasmids; Plasminogen; Proline; Protein Binding; Protein Structure, Tertiary; Proteins; Recombinant Proteins; Thrombospondins; Transfection; Zinc

Mixtures of hyaluronan (HA), chondroitin sulfate (CS)-A and CS-C oligosaccharides were generated through the enzymatic digestion of the polysaccharides with either mammalian hyaluronidase or bacterial HA lyase or chondroitinase. Compared to mammalian enzymes, bacterial enzymes hydrolyze the polysaccharides through a different mechanism yielding chemically distinct sets of oligosaccharides. Peripheral leukocytes and a human monocytic cell line were exposed to these oligosaccharides and the amount of interleukin-12 released by the cells was measured. For all types of oligosaccharide tested, we found that the amount of interleukin-12 induced by oligosaccharides generated with bacterial enzyme was significantly lower than the amount of interleukin-12 induced by oligosaccharides generated with mammalian enzyme. In addition, we observed that CS oligosaccharides generated with bacterial enzyme were capable of reducing the lipopolysaccharide-induced interleukin-12 production in macrophages. Our results indicate that HA or CS oligosaccharides generated with mammalian enzymes might possess pro-inflammatory potential, while HA or CS oligosaccharides generated with bacterial enzymes might possess non- or anti-inflammatory properties. The implications of our findings in view of the ongoing investigation of the potential therapeutic benefits of HA and CS in arthritis or other inflammatory pathologies are discussed.

Topics: Chondroitin Sulfates; Humans; Hyaluronic Acid; Inflammation; Inflammation Mediators; Interleukin-12; Macrophages

Inter-alpha-inhibitor (IalphaI) and pre-alpha-inhibitor (PalphaI) are the main members of a set of multichain serine proteinase inhibitors. Present in human plasma, they may be involved in control of the inflammatory process. They are composed of homologous heavy chains (H1 and H2 for IalphaI; H3 for PalphaI) covalently linked by a protein-glycosaminoglycan-protein cross-link to bikunin, which is a chondroitin 4-sulfate proteoglycan. During the acute-phase response, biosynthesis of IalphaI and PalphaI is downregulated and upregulated, respectively. In this work, we provide evidence that, in inflammatory diseases, the chondroitin sulfate chain of bikunin increases in size proportionally to the severity of the inflammatory response. As a consequence, all IalphaI-related components that contain bikunin are structurally modified. Therefore, the changes in glycosylation of the acute-phase proteins are not restricted to N-linked glycans but also affect glycosaminoglycans. The implications of these findings are discussed with regard to biosynthesis and biological role, especially the anti-inflammatory effects of IalphaI-related proteinase inhibitors.

Topics: Acute-Phase Proteins; Acute-Phase Reaction; Alpha-Globulins; Chondroitin Sulfates; Glycosaminoglycans; Glycosylation; Humans; Inflammation; Membrane Glycoproteins; Molecular Weight; Polysaccharides; Serine Proteinase Inhibitors; Trypsin Inhibitor, Kunitz Soybean

Paclitaxel can inhibit vascular smooth muscle proliferation in vitro, and early studies suggest that paclitaxel may be useful in preventing restenosis. Early and late intimal growth and local vascular pathological changes associated with paclitaxel delivered via stents have not been fully explored.. Localized drug delivery was accomplished with balloon-expandable stainless steel stents coated with a cross-linked biodegradable polymer, chondroitin sulfate and gelatin (CSG), containing various doses of paclitaxel. CSG-coated stents with paclitaxel (42.0, 20.2, 8.6, or 1.5 microgram of paclitaxel per stent), CSG-coated stents without paclitaxel, and uncoated stents (without paclitaxel or CSG) were deployed in the iliac arteries of New Zealand White rabbits, which were killed 28 days after implant. Mean neointimal thickness at stent strut sites was reduced 49% (P<0.0003) and 36% (P<0.007) with stents containing 42.0 and 20.2 microgram of paclitaxel per stent, respectively, versus CSG-coated stents without paclitaxel. However, histological findings suggested incomplete healing in the higher-dose (42.0 and 20.2 microgram) paclitaxel-containing stents consisting of persistent intimal fibrin deposition, intraintimal hemorrhage, and increased intimal and adventitial inflammation. Stents coated with CSG alone (without paclitaxel) had similar neointimal growth as uncoated stents. In a separate group of rabbits killed at 90 days, neointimal growth was no longer suppressed by CSG-coated stents containing 42.0 or 21.0 microgram of paclitaxel. CSG coating appears to be a promising medium for localized drug delivery. Paclitaxel polymer-coated stents reduce neointima formation but are associated with evidence of incomplete healing at 28 days. However, neointimal suppression was not maintained at 90 days.

Topics: Angiogenesis Inhibitors; Animals; Cell Division; Chondroitin Sulfates; Dose-Response Relationship, Drug; Drug Delivery Systems; Fibrin; Gelatin; Hemorrhage; Iliac Artery; Inflammation; Male; Paclitaxel; Polymers; Rabbits; Stents; Time Factors; Tunica Intima

It has been reported that the disaccharide composition of dermatan sulphate shows transient changes after epicutaneous application of the hapten 2,4-dinitrofluorobenzene to mouse skin, and that these changes are most conspicuous in healing skin on day 15 after chemical insult [Kuwaba, Nomura, Irie and Koyama (1999) J. Dermatol. Sci. 19, 23-30]. In the present study it was found that the molecular size of dermatan sulphate was increased on day 15 after hapten application. The molecular size of decorin increased in healing skin, whereas the size of dermatan-sulphate-depleted core protein did not increase. The length and localization of decorin dermatan sulphate were investigated by electron microscopy. Dermatan sulphate filaments oriented orthogonally to collagen fibrils were longer in healing skin than in control skin. In control skin, dermatan sulphate filaments were found among tightly packed collagen fibrils. In contrast, the interfibrillar gaps between each collagen fibril were enlarged in healing skin; elongated dermatan sulphate filaments extended from the surface of collagen fibrils across the enlarged gap. These results suggest that the increase in molecular size of decorin dermatan sulphate is important in organizing collagen fibrils separated by enlarged interfibrillar gaps in healing skin.

Topics: Animals; Anticoagulants; Blotting, Western; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Collagen; Decorin; Dermatan Sulfate; Electrophoresis, Polyacrylamide Gel; Extracellular Matrix Proteins; Female; Inflammation; Mice; Mice, Inbred BALB C; Microscopy, Electron; Proteoglycans; Skin; Time Factors; Ultrasonography; Wound Healing

The concept that aging impairs wound healing is largely unsubstantiated, the literature being contradictory because of poor experimental design and a failure to adequately characterize animal models. This study tested the hypothesis that aging retards the rate of wound repair using standardized cutaneous incisional wounds in a well-characterized aging mouse colony. Against the background of age-related changes in normal dermal composition, marked differences in healing were observed. Immunostaining for fibronectin was decreased in the wounds of the old mice, with a delay in the inflammatory response, re-epithelialization, and the appearance of extracellular matrix components. Heparan sulfate and blood vessel staining were both unexpectedly increased in the wounds of the old animals at late time points. Despite an overall decrease in collagen I and III deposition in the wounds of old mice, the dermal organization was surprisingly similar to that of normal dermal basket-weave collagen architecture. By contrast, young animals developed abnormal, dense scars. Intriguingly, some of these age-related changes in scar quality and inflammatory cell profile are similar to those seen in fetal wound healing. The rate of healing in young animals appears to be increased at the expense of the scar quality, perhaps resulting from an altered inflammatory response.

Topics: Aging; Animals; Chondroitin Sulfates; Collagen; Elastin; Endothelium; Extracellular Matrix; Heparitin Sulfate; Immunohistochemistry; Inflammation; Male; Mice; Mice, Inbred C57BL; Neovascularization, Physiologic; Skin; Up-Regulation; Wound Healing

The biologic response to fibrillar collagen (collagen) and fibrillar collagen plus heparin (collagen/heparin) implants have been compared in the rat subcutaneous and guinea pig dermal wound models. The reconstituted bovine dermal collagen implants were injected subcutaneously in rats at concentrations ranging from 18 to 30 mg/ml and in volumes ranging from 0.5 to 1.0 ml. The biologic response to the collagen implants alone was characterized by a transient invasion of a modest number of inflammatory cells within the first three days of implantation that was followed by limited fibroblast invasion into the peripheral 1/3 of the implant during the course of the next three to four weeks. Occasionally, blood vessels were observed to invade the peripheral regions of the implant. The degree (number) and extent (depth) of cell invasion were inversely related to initial collagen implant concentration. Addition of heparin (0.3-20 micrograms/mg collagen) to these implants resulted in a significant dose-dependent increase in the degree and extent of fibroblast invasion. Radiolabeling studies showed that the collagen and collagen/heparin implants were cleared from the subcutis at identical rates. Implantation of these formulations in a guinea pig dermal wound model was also performed, using a semi-occlusive wound dressing (Opsite) to maintain the implant in the wound site. The fibrillar collagen implant alone was pushed upward by developing granulation tissue at the base of the wound and served as a support for epidermal cell migration, proliferation, and differentiation as wound closure proceeded. The implant was slowly invaded and turned over as granulation tissue developed from the base and margins of the wound bed. The inclusion of heparin in these implants resulted in a significantly different pattern of wound healing. The collagen/heparin implants histologically presented a more broken-up or porous appearance following implantation, which was associated with a greater degree of penetration of developing granulation tissue into the implant itself as compared to the collagen implants. Radiolabeling studies revealed that clearance rates of implants with and without heparin from wound sites were similar, as noted in the rat subcutis. Laser doppler flowmetry studies suggested that the heparin--containing implants were more vascular than control wound sites or sites treated with collagen alone.

Topics: Animals; Blood Flow Velocity; Chondroitin Sulfates; Collagen; Female; Guinea Pigs; Heparin; Inflammation; Lasers; Male; Prostheses and Implants; Rats; Rats, Inbred Strains; Time Factors; Wound Healing

Topics: Animals; Burns; Chondroitin; Chondroitin Sulfates; Collagen; Female; Inflammation; Injections, Intradermal; Male; Rats; Rats, Inbred Strains; Time Factors; Wound Healing

Topics: Antirheumatic Agents; Biochemical Phenomena; Chondroitin; Chondroitin Sulfates; Glycosaminoglycans; Humans; Inflammation; Phenylbutazone; Prednisone; Rheumatic Diseases

Topics: Cartilage; Chondroitin Sulfates; Humans; Inflammation