Regeneration of skin burn in rat using adipose-derived stem cells (ADSCs) seeded into ovine small intestinal submucosa (OSIS)

Background and aim In the past 30 years, researchers have focused on skin regeneration, by using tissue engineering methods. Decellularized ECM-based scaffolds have been recently used as structures for repairing tissue defects, especially skin grafts for burns and cosmetic surgeries. Investigations have been mainly focused on the decellularized porcine SIS; however, there has rarely been report on OSIS. The aim of present study, investigation the capacity of regeneration of burn wound with a skin substitute which was fabricated of decellularized sheep small intestinal submucosal layer loaded with adipose-derived stem cells and was transplanted in rat full-thickness skin burn.MethodsIn the present study, OSIS was decellularized using two phase (chemical and physical), and their biomechanical and histological and biocompatibility properties were evaluated. In order to determine decellularized scaffold morphology and the degree of decellularization, hematoxylin & eosin (H&E), 6-diamidino-2-phenylindole (DAPI) and masson’s trichrome staining was applied. ADSCs were seeded on the scaffolds and adhesion of the cells to the scaffolds was assessed through Field emission scanning electron microscopy (FE-SEM). In addition, inducing the proliferation of ADSCs on the scaffolds was evaluated using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. At the second part of study, applications derived of in vitro phase were investigated in in vivo. In this phase, the effectiveness of the decellularized OSIS (DOSIS) with or without adipose mesenchymal stem cells (ADSCs) in full thickness burn wound was experimentally studied in a rat model. The experimental groups included; group I (untreated wound), group II (DOSIS scaffold) and group III (ADSCs seeded on DOSIS). Wound healing of all groups was examined and compared clinically, histopathologically and immunohistologically on days 5, 14 and 21 post-operations.ResultsResults revealed satisfactory decellularization of OSIS which could be due to its thin thickness. Mechanical properties, structural form and glycosaminoglycan contents were preserved in the decellularized groups. In decellularized groups, further cells and DNA residues were removed compared to the control groups. Importantly, the proliferation capacity of ADSCs on scaffolds significantly increased after 48 h incubation time (P<0.001). Our results represented ADSCs-seeded on DOSIS scaffolds accelerated wound contraction and healing compared to the other groups. Although we used allo and xeno origin of mesenchymal stem cells and scaffold respectively, no signs of inflammation, necrosis or rejection were observed in the experimental groups. Reepithelization was completed 21 days’ post-operation in trial groups. Conclusion The results of this study introduced DOSIS as an ECM based scaffold as novel candidate and alternative source for tissue engineering applications and accelerated healing of rat skin wound defects.

کلیدواژه ها:

decellularization ، small intestine sub-mucosa ، sheep ، mesenchymal stem cell