Decellularization of bovine small intestinal submucosa and its use for the healing of a critical-sized full-thickness skin defect, alone and in combination with stem cells, in a small rodent model

Abstract

In this study, we initially described an efficient decellularization protocol for bovine-derived small intestinal submucosa (bSIS), involving freeze-thaw cycles, acid/base treatment and alcohol and buffer systems. We compared the efficacy of our protocol to some previously established ones, based on DNA content and SEM and histochemical analyses. DNA content was reduced by ~89.4%, significantly higher than compared protocols. The sulphated GAG content of the remaining interconnected fibrous structure was 5.738 ± 0.207 µg/mg (55% retained). An in vitro study was performed to evaluate whether rat bone marrow mesenchymal stem cells (MSCs) could attach and survive on bSIS membranes. Our findings revealed that MSCs can preserve their viability and proliferate on bSIS for > 2 weeks in culture. We conducted in vivo applications for the treatment of an experimental rat model of critical sized (7 cm ) full-thickness skin defect. The wound models treated with either MSCs-seeded (1.5 × 10 cells/cm ) or non-seeded bSIS membranes were completely closed by week 7 without significant differences in closure time; on the other hand, the open wound control was closed at ~47% at this time point. Immunohistopathology results revealed that the group which received MSCs-seeded bSIS had less scarring at the end of the healing process and was in further stages of appendage formation in comparison with the non-seeded bSIS group. Copyright © 2015 John Wiley & Sons, Ltd.

A Gelatin-sulfonated Silk Composite Scaffold based on 3D Printing Technology Enhances Skin Regeneration by Stimulating Epidermal Growth and Dermal Neovascularization

“…However, these grafts are not satisfactory and effective in repairing large area skin damage 56 and chronic skin wounds due to lower granulation 57 , and lack of vascularization during the process of wound healing 35 . Our silk fibroin derivative has demonstrated great potential for such types of major cutaneous defects, and further investigations in big animal models will be performed in the near future.…”

Critical‐sized full‐thickness skin defect regeneration using ovine small intestinal submucosa with or without mesenchymal stem cells in rat model

“…In BM‐MSCs‐seeded DOSIS group, epithelization was faster and fully took place at the subsequent time points. This may be attributed to the capacity of BM‐MSCs to inhibit inflammation, modulate immune responses and differentiate into various skin cell types, such as fibroblasts and keratinocytes, therefore, improve the quality of wound healing …”

“…This may be attributed to the capacity of BM-MSCs to inhibit inflammation, modulate immune responses and differentiate into various skin cell types, such as fibroblasts and keratinocytes, therefore, improve the quality of wound healing. 22,48 CONCLUSION DOSIS layer can be prepared in considerably cell-free form, with low substantially DNA content using the decellularization protocol. Decellularized OSIS layer accelerated healing of rat skin wound defects that was created at critical-size and full-thickness.…”

“…They have been in the spotlight of the regenerative medicine because of their various remarkable properties, such as self‐renewal ability, immunomodulatory properties, strong secretory properties, and their ability to differentiate into various epidermal cell types, such as keratinocytes and fibroblasts as well as dermal appendixes including, apocrine glands or hair follicle. It has also been reported that increase in basic fibroblast growth factor (b‐FGF), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF) levels are induced by MSCs . Local transplantation of rat adipose‐derived MSCs to an excisional wound‐healing model revealed enhanced granulation tissue deposition and epithelialization …”

“…It has also been reported that increase in basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF) levels are induced by MSCs. [20][21][22][23] Local transplantation of rat adipose-derived MSCs to an excisional wound-healing model revealed enhanced granulation tissue deposition and epithelialization. 24 Since, in vivo application of decellularized ovine small intestinal submucosal (DOSIS) layer as, a scaffold is yet to be investigated.…”