| Objective:Through to the BMSCs and ADSCs respectively and combined in the constructed artificial composite skin in vitro,applied to rabbit full-thickness skin wounds,to observe and compare the wound healing ability in order to find a better kind of artificial composite skin. Methods:1.Take the bone marrow and neck adipose of 1 clean and healthy Japanese big ear rabbit to culture the BMSCs and ADSCs in vitro respectively and the cells were expanded to the third generation.Take the third generation ADSCs into osteoblast and adipocyte differentiation experiment.Use the green fluorescent protein and cell tracer CM-Di L to mark the BMSCs and ADSCs.Prepare the SIS through acellular method.The BMSCs and ADSCs are planted in the SIS to form the artificial composite skin.2.18 clean and healthy Japanese big ear rabbits,each takes 2*2cm full thickness skin wounds in the back,randomly divided into five groups: A(ADSCs+ SIS)、B(BMSCs+SIS)、C(ADSCs+BMSCs+SIS)、D(SIS)、E(blank control group), observe the local conditions and adverse reaction after 14、21、28 days after the surgry,measure the size of the wound,and sentenced to death for all grafts.HE staining to observe the tissue structure. Results:1.BMSCs cultured to the third generation were spindle shape,polygonal,into whorls,faster proliferation;the third generation of ADSCs were spindle shaped, triangular, strong proliferation.Adipogenic and osteogenic differentiation experiments showed that the isolated cells were ADSCs.BMSCs showed green fluorescence labeling, fluorescence intensity; ADSCs showed red fluorescence labeling, fluorescence intensity. BMSCs and ADSCs can be grown in SIS with strong proliferation ability.BMSCs and ADSCs could be cultured in SIS to form the artificial composite skin.2.Animal experiment:(1)general observation: 14 days after surgery saw the central wound covering the thin crust,darker color; 21 days after surgery A、B、C three groups wound healing,covering more dander,wound area reduced,light color,group D and E were not healed completely, covering less dander,deep wound color; 28 days after surgery A、B、C three groups covering more dander, wound area reduced significantly, wound color and the surrounding normal tissue are similar. The skin surface of A、B、C three groups was smooth after surgery,the healing effect is better than that of group D and E. 14、21、28 days after the surgery, A、B、C three groups of wound contraction rate are lower than group D and E(P<0.05), but there is no difference in group A、Band C; A、B、C three groups of wound healing time are less than group D and E(P<0.05), there is no difference in group A、Band C.(2) 14 days after surgery, A、B、C three groups saw fibroblast and inflammatory cells with new capillary vessels; group D and E saw less fibroblast and inflammatory cells without new capillary vessels; 21、28 days after surgery, A、B、C three groups see large amount of collagen fiber, neatly arranged,new capillary vessels uniformly distributed, group D and E see collagen fiber arranged in disorder,no sign of new capillary vessels. Conclusion:Using the composite skin made of BMSCs and ADSCs to repair full-thickness wound can promote the formation of small blood vessels and wound epithelialization, increasing wound healing.The combination of the two culture of wound and the single culture construction of artificial composite skin are no significant differences in wound healing quality and reducing the rate of wound contraction. |
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