Adipose stem cell-laden injectable thermosensitive hydrogel reconstructing depressed defects in rats: filler and scaffold

Abstract

Facial depressed defects are a common cosmetic problem. Temporary fillers need to be re-injected frequently to maintain the desired outcomes. Here, the feasibility of a novel type of injectable hydrogel for persistent effect is demonstrated. We first useed agmatine to synthesize a poly(amidoamine) (PAA) to form a cell-attachable crosslinker and then the crosslinker was co-polymerized with N-isopropylacrylamide to obtain an injectable and temperature sensitive hydrogel. ¹H NMR showed the successful synthesis of the crosslinker. In vitro tests, CCK-8 assay and live/dead viability test showed that the hydrogel was non-toxic to adipose-derived stem cells (ASCs). SEM images also confirmed that ASCs could adhere to the hydrogel. Then we constructed a novel depressed defect model in rats and injected four different fillers in the depressed defects: (1) the hydrogel with ASCs, (2) the hydrogel only, (3) hyaluronic acid, and (4) PBS. After 4 weeks, gross and histological analyses showed the defects in hydrogel, hydrogel + ASCs, and HA groups improved significantly and there were no significant differences among them. Significant differences in thickness from skin to muscle in the defect was found between the hydrogel + ASCs group and the other groups after 6 months. The hydrogels degraded completely in defects in both the hydrogel group and the hydrogel + ASCs group, and were filled with adipocytes and multilocular immature adipocytes. Immunohistochemical study using s-100 and perilipin staining revealed adipocyte differentiation in the defect sites. We also used green fluorescent protein (GFP)–ASCs for tracing and found that exogenous added ASCs were involved in adipogenesis. In conclusion, such a cell attachable thermosensitive hydrogel has definite potential not only as a filler but also as a scaffold, and has a persistent effect for small depressed defects. It might ultimately become a new material in plastic and reconstructive surgery.