chondroitin-sulfates and asiatic-acid

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

In the present work, a hydrogel platform composed of biopolymer gelatin, and glycosaminoglycan's (Hyaluronic acid and Chondroitin sulfate) incorporated with Asiatic acid (a triterpenoid) and nanoparticles (Zinc oxide and Copper oxide) has been designed and developed to find out the efficacy of healing in second degree burn wounds in Wistar rats. The developed hydrogel composite has been characterized by physico-chemical methods such as; SEM, swelling, mechanical strength, degradation and drug release kinetics. Results showed that the morphology of composite scaffolds are porous with maximum water uptake capacity of 1068% and possessed tensile strength of ~0.196 MPa. Anti-microbial evaluation depicted increase in zone of inhibition with hydrogel containing gelatin + ZnO (5.3 ± 0.2 mm in E. coli and 4.9 ± 0.6 mm in S. aureus) and gelatin + CuO (4.8 ± 0.7 mm in E. coli, 3.8 ± 0.3 mm in S. aureus) in comparison to hydrogel composite scaffold. In-vitro cytocompatibility of developed hydrogel composite was assessed in terms of MTT and DNA quantification on L929 fibroblast cells. In-vivo studies for the composite scaffolds were evaluated on Wistar rats after second degree burn wounds were induced and studied for 28 days which showed the significant wound healing activity in comparison to the control (NeuSkin™ and Cotton guaze) in terms of DNA, total protein, hexosamine and hydroxyproline content. Histopathology studies showed the significant progress in re-epithelization, collagen fibers arrangement and angiogenesis in comparison to control. Additionally, a decrease of TNF-α and increase of MMP-2 expression on day 7 of animal experiment support healing. Furthermore, no toxicity was seen with the developed scaffolds suggesting their suitability to use as a wound dressing in second degree burns.

Topics: Animals; Biocompatible Materials; Burns; Cell Line; Chondroitin Sulfates; Copper; Gelatin; Hyaluronic Acid; Hydrogels; Matrix Metalloproteinase 2; Metal Nanoparticles; Mice; Pentacyclic Triterpenes; Rats; Rats, Wistar; Skin; Tensile Strength; Tumor Necrosis Factor-alpha; Wound Healing; Zinc Oxide