Human Adipose Stem Cells Cell Sheet Constructs Impact Epidermal Morphogenesis in Full-Thickness Excisional Wounds

Abstract

Among the wide range of strategies to target skin repair/regeneration, tissue engineering (TE) with stem cells at the forefront, remains as the most promising route. Cell sheet (CS) engineering is herein proposed, taking advantage of particular cell-cell and cell-extracellular matrix (ECM) interactions and subsequent cellular milieu, to create 3D TE constructs to promote full-thickness skin wound regeneration. Human adipose derived stem cells (hASCs) CS were obtained within five days using both thermoresponsive and standard cell culture surfaces. hASCs-based constructs were then built by superimposing three CS and transplanted into full-thickness excisional mice skin wounds with delayed healing. Constructs obtained using thermoresponsive surfaces were more stable than the ones from standard cell culture surfaces due to the natural adhesive character of the respective CS. Both CS-generating strategies lead to prolonged hASCs engraftment, although no transdifferentiation phenomena were observed. Moreover, our findings suggest that the transplanted hASCs might be promoting neotissue vascularization and extensively influencing epidermal morphogenesis, mainly through paracrine actions with the resident cells. The thicker epidermis, with a higher degree of maturation characterized by the presence of rete ridges-like structures, as well as a significant number of hair follicles observed after transplantation of the constructs combining the CS obtained from the thermoresponsive surfaces, reinforced the assumptions of the influence of the transplanted hASCs and the importance of the higher stability of these constructs promoted by cohesive cell-cell and cell-ECM interactions. Overall, this study confirmed the potential of hASCs CS-based constructs to treat full-thickness excisional skin wounds and that their fabrication conditions impact different aspects of skin regeneration, such as neovascularisation, but mainly epidermal morphogenesis.

Allogeneic Transplantation of an Adipose-Derived Stem Cell Sheet Combined With Artificial Skin Accelerates Wound Healing in a Rat Wound Model of Type 2 Diabetes and Obesity

“…Lin et al (31) and McLaughlin and Marra (32) reported that transplantation of human ASC sheets with fibrin-coated membranes accelerates wound healing in a 12-mm-diameter full-thickness skin defect on the backs of 6-week-old athymic nude mice. Cerqueira et al (33) reported that human ASC sheet transplantation promotes wound regeneration in 10-mm-diameter fullthickness skin defects on the backs of 5-week-old normal mice. In contrast to previous studies, the current study created a wider full-thickness skin defect with exposed bone (wound area 15 mm 2 ) on the parietal region of 16-week-old ZDF rats.…”

Accelerated wound healing in a diabetic rat model using decellularized dermal matrix and human umbilical cord perivascular cells

“…It is known that there are some differences in the wound healing process in rodent and human skin. The main reason of wound contraction that normally happens in rodent skin is basically due to the existence of a layer situated directly below the skin (paniculosus carnosus layer), while the healing in humans only occurs via re-epithelization and granulation tissue formation [52]. Therefore, in our study, the wound model was created by harvesting full-thickness skin and superficial fascia, in order to prevent wound contraction.…”

Stem Cells in Skin Wound Healing: Are We There Yet?

“…Considering the different number of works using stem cells either injected at the wound borders 15,36 or randomly incorporated within biomaterial-based matrices, [37][38][39][40][41][42] there is skepticism regarding the need for 3D organization. Nonetheless, this discrepancy can be correlated with the limitations of MSC differentiation toward epithelial and endothelial lineages, 43 as well as to the deficient purity/stability of epidermal stem cells cultures.…”

“…43 Stem cells from distinct sources, including MSCs, have shown a strong effect in HF formation. 37,39,44,45 In our recently published work, 37 the enhanced HF formation appears to be associated with keratinocyte growth factor (KGF) overexpression resulting from a direct interaction between resident keratinocytes (KCs) and transplanted hASCs cell sheet constructs. MSCs mechanisms that improve HF formation are, however, still elusive.…”