CGF与脂肪干细胞联合应用修复大鼠全层皮肤缺损的研究

Biology and Role of ADSCs Induced by CGF in Regenerative Medicine of Rat Full-thickness Skin Wounds

目的:探讨以明胶海绵为支架材料,复合浓缩生长因子(concentrated growth factor,CGF)诱导的大鼠脂肪干细胞(adipose-derived stem cells,ADSCs)得到的组织工程皮肤替代物,对大鼠全层皮肤缺损愈合的作用。方法:取健康雄性大鼠脂肪组织分离、培养并鉴定脂肪干细胞,观察CGF对其增殖、分化的影响。在健康雌性大鼠背部制造一直径3 cm全层皮肤缺损,其上覆盖复合脂肪干细胞的明胶海绵(A组)、无细胞的明胶海绵(B组)或不作处理单纯包扎(C组)。术后观察各组缺损的愈合情况,并于7、14、21 d及完全愈合后,取背部全层皮肤作组织学观察。结果:CGF能促进体外培养的ADSCs增殖;术后伤口渗出物、红肿等炎症反应A组<B组<C组;伤口完全愈合所需时间A组<B组<C组。术后1周组织学观察见:A组创区细胞与创伤愈合相关的生长因子受体表达量相对B组、C组增加;A组的肉芽组织内新生毛细血管数目多于B组、C组。完全愈合后雌性大鼠新生皮肤存在雄性大鼠来源的细胞。结论:适当浓度的CGF能够促进ADSCs在体外的增殖;复合CGF与ADSCs的组织工程皮肤替代物利于大鼠皮肤创伤愈合;移植的ADSCs能够存活,并分化为上皮细胞。

Objective： To observe the effect of tissue-engineered skin consisting of adipose tissue-derived stem cells （ADSCs） induced by concentrated growth factor （CGF） as seed cells and gelatin sponge as scaffold on the healing process of rat full thickness skin defect. Methods： Rat ADSCs were isolated and cultured in vitro. The effect of CGF on ADSCs was observed. The tissue-engineered skin, made by gelatin sponge and ADSCs induced by CGF, was transferred to female rats＆#39; full thickness skin defect to observe the healing effect （group A）. To be contrast, two other groups were set：gelatin sponge with no cells （group B）, and none special treatment （group C）. The formation of new skin was investigated by immunohistochemical examination every 7 days until the wounds completely healed. Results：ADSCs showed a higher rate of proliferation when cultured with CGF. A slight inflammatory response could be observed in group A rats; it took shorter time to close up the wound for rats in group A. Microvascular and receptors of growth factors which related to wound healing were significantly more in group A than in group B and C. Cells originated from male rat could be found in female rats new regenerative skin in group A. Conclusion：Our findings demonstrate that the application of proper concentration of CGF can promote the proliferation of ADSCs in vitro. The tissue-engineered skin made of gelatin sponge and ADSCs induced by CGF can promote wound healing. The ADSCs can survive and differentiate into epithelial cells.