脂肪干细胞通过旁分泌作用改善异种脱细胞真皮基质体内植入效果的机制研究

Mechanism of Adipose-Derived Stem Cells Improving the Efficacy of Heterologous Acellular Dermal Matrix Implantation by Paracrine Effect

目的探讨脂肪干细胞(ADSCs)在异种脱细胞真皮基质(HADM)微环境中能否有效发挥其旁分泌功能,从而改善HADM体内植入的效果。方法将ADSCs分别接种于HADM及培养皿上。分别收集两组细胞的总RNA,通过q RT-PCR检测ADSCs旁分泌基因的表达。将ADSCs预处理前后的HADM分别植入大鼠体内,4周后取材行组织学观察。结果在HADM上培养的ADSCs与在普通培养皿上培养的ADSCs相比,其促血管形成因子(VEGF、HGF、bFGF)、炎症调节因子(TGF-β、i NOS、TSG-6、COX-2)及干性因子(SOX-2)均有所上调。经ADSC体外预培养后的HADM与未经处理的HADM相比,体内植入后表现出更好的血管化能力且炎症反应也更低。结论 HADM能为ADSCs发挥旁分泌作用提供合适的微环境,且HADM经过ADSCs体外预培养,其血管化程度更高,而炎症反应则得以缓解及局限化。

Objective To explore whether adipose-derived stem cells(ADSCs)can effectively exert its paracrine function in heterologous acellular dermal matrix(HADM)microenvironment and consequently improve the efficacy of HADM after implantation in vivo.Methods ADSCs were seeded on HADM and microplate.Total RNA of ADSCs from both materials were collected respectively,and qRT-PCR was used to evaluate the levels of genes associated with ADSC paracrine.In addition,HADM with and without ADSC pre-culture were implanted on the back of rats.All samples were harvested 4 weeks after operation for histological staining.Results The expression of genes with respect to inflammatory regulation(TGF-β,iNOS,TSG-6,COX-2),pro-angiogenesis(VEGF,HGF,bFGF)and stemness(SOX-2)increased when ADSCs were seeded on HADM compared to ADSCs seeded on microplate.After pre-culture,in vivo implanted ADSCs-HADM exhibited improved vascularization and mitigated inflammatory response compared to untreated HADM.Conclusion HADM can provide a beneficial microenvironment for ADSCs to exert their paracrine function.After pre-culture with ADSCs in vitro,the vascularization rate of in vivo implanted HADM is accelerated and the inflammatory response is attenuated and localized.