ginsenoside-rg3 and Cicatrix--Hypertrophic

Hypertrophic scarring (HS) remains a great challenge in wound dressing. Although various bionic extracellular matrix (ECM) biomaterials have been designed towards HS treatment, not all biomaterials can synergize biological functions and application functions in wound repair. Bionic scar-inhibiting scaffolds, loaded with biomolecules or drugs, become promising strategies for scarless skin regeneration. In this work, inspired by the physicochemical environment of ECM, a versatile fabrication of poly(γ-glutamic acid) based on electrospun photocrosslinkable hydrogel fibrous scaffolds incorporated with ginsenoside Rg3 (GS-Rg3) is developed for tissue repair and wound therapy. Decorated with adhesive peptide, bionic fibrous scaffolds can accelerate fibroblasts to sprout and grow, forming organized space-filling basement that gradually fills a depression before wound close up in the early stage. Additionally, by sustained release of GS-Rg3 in late stage, fibrous scaffolds promote scarless wound healing in vivo as evidenced by the promotion of cell communication and skin regeneration, as well as the subsequent decrease of angiogenesis and collagen accumulation. These ECM-inspired fibrous scaffolds, therefore, offer new perspectives on accelerated wound healing and tissue regeneration.

Topics: Animals; Biocompatible Materials; Cell Line; Cell Proliferation; Cicatrix, Hypertrophic; Drug Liberation; Extracellular Matrix; Fibroblasts; Ginsenosides; Hydrogels; Male; Oligopeptides; Polyglutamic Acid; Rats; Rats, Sprague-Dawley; Skin; Tissue Scaffolds; Wound Healing

Incorporation of bioactive drugs and biofunctionalization of polyester fibrous scaffolds are essential means to improve their bio-functions and histocompatibility for regenerative medicine. However, it is still a challenge to biofunctionalize such drug carriers via traditional biochemical methods while maintaining their properties without changes in drug activity and loading ratio. Here, we demonstrated a facile approach for biofunctionalization of PLGA fibrous scaffolds with various molecules (i.e., PEG polymer, RGD peptide and bFGF growth factor for cell repellent, adhesion and proliferation, respectively) via mussel-Inspired poly(dopamine) (PDA) coating in aqueous solution. By virtue of the mild and efficient nature of this approach, the drug-loaded PLGA fibers could be easily biofunctionalized and showed negligible effects on the scaffold properties, especially drug activity and loading ratio. Further, in vivo study showed that, a ginsenoside-Rg3-loaded fibrous scaffold functionalized with bFGF growth factor could not only promote the early-stage wound healing in rabbit ear wounds (bio-signal from bFGF), but also inhibit later-stage hypertrophic scars formation (release of Rg3 drug). Therefore, the mussel-inspired method for bio-modification provides a facile and effective strategy to combine drug and bio-function in one system, thus facilitating a synergistic effect of drug-therapy and bio-signal when such biomaterial is used for regenerative medicine.

Topics: Animals; Biocompatible Materials; Cell Count; Cell Survival; Cicatrix, Hypertrophic; Collagen; Epithelium; Female; Fibroblasts; Ginsenosides; Humans; Indoles; Lactic Acid; Microvessels; Photoelectron Spectroscopy; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Polymers; Rabbits; Surface Properties; Tissue Engineering; Tissue Scaffolds; Wound Healing

Prevention of hypertrophic scar formation of the skin requires a complex treatment process, which mainly includes promoting skin regeneration in an early stage while inhibiting hypertrophic formation in a later stage. Electrospinning PLGA with the three-dimensional micro/nano-fibrous structure and as drugs carrier, could be used as an excellent skin repair scaffold. However, it is difficult to combine the advantage of nanofibrous membranes and drug carriers to achieve early and late treatment. In this study, Ginsenoside-Rg3 (Rg3) loaded hydrophilic poly(D,L-lactide-co-glycolide) (PLGA) electrospun fibrous membranes coated with chitosan (CS) were fabricated by combining electrospinning and pressure-driven permeation (PDP) technology. The PDP method was able to significantly improve the hydrophilicity of electrospun fibrous membranes through surface coating of the hydrophilic fibers with CS, while maintaining the Rg3 releasing rate of PLGA electrospun fibrous membranes. Experimental wounds of animal covered with PDP treated fibrous membranes completely re-epithelialized and healed 3-4 days earlier than the wounds in control groups. Scar elevation index (SEI) measurements and histologic characteristics revealed that Rg3 significantly inhibited scar formation 28 days post-surgery. Moreover, RT-PCR assays and western blot analysis revealed that at day 28 after wound induction the expression of VEGF, mRNA and Collagen Type I in the scars treated with Rg3 was decreased compared to control groups. Taken together PLGA-Rg3/CS electrospun fibrous membranes induced repair of tissue damage in the early stage and inhibited scar formation in the late stage of wound healing. These dual-functional membranes present a combined therapeutic approach for inhibiting hypertrophic scars of the skin.

Topics: Animals; Cell Proliferation; Cicatrix, Hypertrophic; Collagen Type I; Female; Fibroblasts; Ginsenosides; Humans; Lactic Acid; Male; Membranes, Artificial; Microscopy, Electron, Scanning; Microvessels; Photoelectron Spectroscopy; Polyglycolic Acid; Polylactic Acid-Polyglycolic Acid Copolymer; Rabbits; RNA, Messenger; Skin; Tissue Scaffolds; Vascular Endothelial Growth Factor A; Wound Healing

Intra-lesional injections of corticosteroids, interferon, and chemotherapeutic drugs are currently the most popular treatments of hypertrophic scar formation. However, these drugs can only be used after HS is formed, and not during the inflammatory phase of wound healing, which regulates the HS forming process.. To investigate a new, effective, combining therapeutic and safe drug for early intervention and treatment for hypertrophic scars.. Cell viability assay and flow cytometric analysis were studied in vitro. Animal studies were done to investigate the combining therapeutic effects of 20(S)-ginsenoside Rg3 (Rg3) on the inflammatory phase of wound healing and HS formation.. In vitro studies showed that Rg3 can inhibit HS fibroblasts proliferation and induce HSF apoptosis in a concentration-dependent manner. In vivo studies demonstrated that Rg3 can limit the exaggerated inflammation, and do not delay the wound healing process, which indicates that Rg3 could be used as an early intervention to reduce HS formation. Topical injection of 4 mg/mL Rg3 can reduce HS formation by 34%. Histological and molecular studies revealed that Rg3 injection inhibits fibroblasts proliferation thus reduced the accumulation of collagen fibers, and down-regulates VEGF expression in the HS tissue.. Rg3 can be employed as an early intervention and a combining therapeutic drug to reduce inflammation and HS formation as well.

Topics: Administration, Topical; Animals; Cell Proliferation; Cell Survival; Cells, Cultured; Cicatrix, Hypertrophic; Disease Models, Animal; Down-Regulation; Early Medical Intervention; Female; Fibroblasts; Ginsenosides; Humans; In Vitro Techniques; Male; Otitis; Rabbits; Vascular Endothelial Growth Factor A; Wound Healing

Hypertrophic scarring (HS) has been considered as a great concern for patients and a challenging problem for clinicians as it can be cosmetically disfiguring and functionally debilitating. In this study, Ginsenoside Rg3/Poly(l-lactide) (G-Rg3/PLLA) electrospun fibrous scaffolds covering on the full-thickness skin excisions location was designed to suppress the hypertrophic scar formation in vivo. SEM and XRD results indicated that the crystal G-Rg3 carried in PLLA electrospun fibers was in amorphous state, which facilitates the solubility of G-Rg3 in the PLLA electrospun fibrous scaffolds, and solubility of G-Rg3 in PBS is increased from 3.2 µg/ml for pure G-Rg3 powders to 19.4 µg/ml for incorporated in PLLA-10% fibers. The released G-Rg3 content in the physiological medium could be further altered from 324 to 3445 µg in a 40-day release period by adjusting the G-Rg3 incorporation amount in PLLA electrospun fibers. In vitro results demonstrated that electrospun G-Rg3/PLLA fibrous scaffold could significantly inhibit fibroblast cell growth and proliferation. In vivo results confirmed that the G-Rg3/PLLA electrospun fibrous scaffold showed significant improvements in terms of dermis layer thickness, fibroblast proliferation, collagen fibers and microvessels, revealing that the incorporation of the G-Rg3 in the fibers prevented the HS formation. The above results demonstrate the potential use of G-Rg3/PLLA electrospun fibrous scaffolds to rapidly minimize fibroblast growth and restore the structural and functional properties of wounded skin for patients with deep trauma, severe burn injury, and surgical incision.

Topics: Analysis of Variance; Animals; Chromatography, High Pressure Liquid; Cicatrix, Hypertrophic; Collagen; Fibroblasts; Ginsenosides; Humans; Microscopy, Electron, Scanning; Polyesters; Rabbits; Solubility; Tissue Scaffolds; X-Ray Diffraction

Clinically, hypertrophic scarring (HS) is a major concern for patients and has been a challenge for surgeons, as there is a lack of treatments that can intervene early in the formation of HS. This study reports on a Chinese drug, 20(R)-ginsenoside Rg3 (GS-Rg3), which can inhibit in vivo the early formation of HS and later HS hyperplasia by inducing the apoptosis of fibroblasts, inhibiting inflammation and down-regulating VEGF expression. Implantable biodegradable GS-Rg3-loaded poly(l-lactide) (PLA) fibrous membranes were successfully fabricated using co-electrospinning technology to control drug release and improve drug utilization. The in vivo releasing time of GS-Rg3 lasts for 3 months, and the drug concentration released in rabbits can be controlled by varying the drug content of the electrospun fibers. Histological observations of HE staining indicate that GS-Rg3/PLA significantly inhibits the HS formation, with obvious improvements in terms of dermis layer thickness, epidermis layer thickness and fibroblast proliferation. The results of immunohistochemistry staining and Masson's trichrome staining demonstrate that GS-Rg3/PLA electrospun fibrous membranes significantly inhibit HS formation, with decreased expression of collagen fibers and microvessels. VEGF protein levels are much lower in the group treated with GS-Rg3/PLA eletrospun membranes compared with other groups. These results demonstrate that GS-Rg3 is a novel drug, capable of inhibiting the early formation of HS and later HS hyperplasia. GS-Rg3/PLA electrospun membrane is a very promising new treatment for early and long-term treatment of HS.

Topics: Animals; Cicatrix, Hypertrophic; Collagen Type I; Epidermis; Ginsenosides; Immunohistochemistry; Implants, Experimental; Lactic Acid; Microscopy, Electron, Scanning; Microvessels; Polyesters; Polymers; Rabbits; Staining and Labeling; Tissue Scaffolds; Vascular Endothelial Growth Factor A; Wound Healing