Improvement of skin-graft survival after autologous transplantation of adipose-derived stem cells in rats

https://doi.org/10.1016/j.bjps.2011.07.009Get rights and content

Summary

Background

Skin grafts are frequently used for a variety of indications in plastic and reconstructive surgery. Their necrosis is a common complication, while different therapies have been proposed. Currently, adipose-derived stem cells (ASCs) hold great promise for their angiogenic potential and role during tissue repair. In this study, autologous transplantation of ASCs was used in skin grafts in rats to determine if it increases angiogenesis, skin-graft survival and wound healing.

Methods

ASCs were isolated, cultured, labelled with fluorescent dye and injected under full-thickness skin grafts in 10 rats (group 1), while 10 others served as controls (group 2). Skin grafts were analysed after 1 week. Collagen’s framework was assessed with Masson’s trichrome stain and angiogenesis with von Willebrand factor (vWF) immunohistochemistry. In addition, immunohistochemical staining intensity of vascular endothelial growth factor (VEGF) and transforming growth factor b3 (TGFb3) was assessed in all grafts.

Results

Mean area of graft necrosis was significantly less in group 1 than in group 2 (6.12% vs. 32.62%, p < 0.01). Statistically significant increase of microvessel density, collagen density, VEGF and TGFb3 expression was noted in group 1 compared with group 2 (all: p < 0.01).

Conclusions

These findings suggest that autologous ASCs transplantation increases full-thickness skin-graft survival and shows promise for use in skin-graft surgery. This might be both due to in situ differentiation of ASCs into endothelial cells and increased secretion by ASCs of growth factors, such as VEGF and TGFb3 that enhance angiogenesis and wound healing.

Section snippets

Materials and methods

Twenty male Sprague–Dawley rats, weighing between 350 and 450 g, were used in this study. The National Research Council’s guidelines for the care and use of laboratory animals were followed. The experiments were performed under general anaesthesia using ketamine hydrochloride (100 mg kg−1 intramuscularly) and xylazine (5 mg kg−1 intramuscularly). Antibiotic prophylaxis with cefamandole nafate (20 mg kg−1 intramuscularly) and analgesia with carprofen (0.08 ml kg−1 subcutaneously) were used prior

ASCs’ immunophenotype and differentiation

Flow cytometry analysis showed that cultured cells of passage 1 were positive for stromal markers CD29 (integrin unit) and CD44 (receptor for hyalouronic acid) and negative for haematopoietic markers CD45 (pan-leucocyte marker), CD31 (differentiated endothelial marker) and CD34 (marks primitive haematopoietic progenitors) with 10–20% CD34(+). ASCs differentiated into adipocytes, chondrocytes and osteocytes, as assessed by the specific staining (Figure 1).

Skin-graft survival assessment

At 7 days postoperatively, the regions

Discussion

Our study evaluated the effect of autologous transplantation of ASCs in survival and wound healing promotion of full-thickness skin grafts in rats, with a statistically significant increase in skin-graft survival area in ASC-treated animals compared with controls. Although mean necrosis in our control group (32.62%) was relatively high compared with humans, this is in accordance with skin-graft necrosis in control groups in rat models in other studies.5, 6

Full-thickness skin-graft survival in

Ethical approval of the study

This study had the ethical approval of the Institutional Ethical Committee of the University of Athens and the ethical approval of the Hellenic Veterinary Medicine Committee.

Conflict of interest statement

None of the authors has a financial interest in any of the products, devices or drugs mentioned in this article.

Acknowledgements

This study was partially supported by the ‘Kapodistrias’ research programme of Athens University and by the ‘Bodossakis Foundation’ grant.

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