Effects of rabbit adipose-derived mesenchymal stem cells on the healing of skin deep partial-thickness scald wound of rabbit

Abstract

To investigate the effects of local injection of rabbit adipose-derived mesenchymal stem cells (ADSCs) on the healing of skin deep partial-thickness scald wound of rabbit. ADSCs were isolated from adipose tissue of one New Zealand rabbit and then sub-cultured. ADSCs of the third passage were used in the following experiments. Twenty-four rabbits were divided into ADSCs group (n=12) and control group (n=12) according to the random number table, and one deep partial-thickness scald wound with diameter of 5 cm on the two sides of the back near the buttocks was made. From post injury day (PID) 2, 2 mL suspension of EdU-labeled ADSCs with the number of 5×10(5) per mL was subcutaneously injected in wounds of rabbits in ADSCs group, while the rabbits in control group were given 2 mL serum-free DMEM until the wounds were healed. Wound healing processes of rabbits in two groups were observed every day, and the healing time was recorded. On PID 7, 14, 21, and 28, areas of wound of three rabbits in two groups were measured and the healing rates were calculated, respectively. The healed wound tissue was harvested to observe the morphology by HE staining, and the expression of collagen fiber was observed by Masson staining. The distribution of EdU-labeled ADSCs in healed wound tissue on PID 28 was observed by inverted fluorescence microscope. The expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) of healed wound tissue on PID 7, 14, and 21 were detected by enzyme-linked immunosorbent assay. Data were processed with analysis of variance of factorial design and paired samples t test. (1) The wound healing time of rabbits in ADSCs group was (19.5±1.1) d post injury, which was significantly shorter than that in control group [(23.3±1.5) d, t=4.50, P<0.05]. On PID 7, wounds of rabbits in two groups were dry with no obvious exudation, and redness and swelling around wounds disappeared gradually, the wound healing rate of rabbits in ADSCs group was (15.1±2.4)%, which was close to that in control group [(13.7±3.1)%, t=1.20, P>0.05]. On PID 14, wounds of rabbits in ADSCs group were dry and scabbed obviously, and the wound healing rate was (73.1±5.7)%, while wounds of rabbits in control group were little scabbed with little exudation, and the wound healing rate was significantly lower than that in ADSCs group [(52.9±5.1)%, t=8.06, P<0.01]. On PID 21, wounds of rabbits in ADSCs group were generally healed, and the wound healing rate was (95.6±3.0)%, while a few wounds still existed in rabbits of control group, and the wound healing rate was significantly lower than that in ADSCs group [(78.6±3.7)%, t=9.73, P<0.01]. On PID 28, wounds of rabbits in two groups were totally healed with the healing rate of 100%, and texture and microvascular responses of healed wound tissue in ADSCs group were better than those in control group. (2) On PID 7, fibroblasts in healed wound tissue of rabbits in two groups were all increased, and there were little vascular and collagen fiber proliferation with no obvious differences. On PID 14, the number of fibroblasts in healed wound tissue of rabbits in ADSCs group was more than that in control group, and the collagen fibers in healed wound tissue of rabbits in ADSCs group were arranged in dense and uniform, while those in control group were sparse and irregular. On PID 21, skin layers were differentiated in healed wound tissue of rabbits in two groups, and collagen fibers in healed wound tissue of rabbits in ADSCs group were still denser than that in control group. On PID 28, newborn skin was well differentiated in healed wound tissue of rabbits in ADSCs group, which was better than that in control group. There were a lot of thick collagen fibers in healed wound tissue of rabbits in two groups, and EdU-labeled ADSCs were involved in skin texture of rabbits in ADSCs group. (3) The expressions of VEGF and EGF in healed wound tissue of rabbits in two groups were similar on PID 7 (with t values respectively 0.70 and 0.91, P values above 0.05), which in ADSCs group were significantly higher than those in control group on PID 14 and 21 (with t values from 2.85 to 4.81, P values below 0.01). The transplantation of ADSCs can promote the wound healing of skin deep partial-thickness scald wound of rabbit and shorten the wound healing time.