人脂肪干细胞复合猪小肠黏膜下层脱细胞基质/壳聚糖温敏凝胶体内构建组织工程脂肪的研究  

HUMAN ADIPOSE-DERIVED STEM CELLS COMBINED WITH SMALL INTESNITAL SUBMUCOSA POWDER/CHITOSAN CHLORIDE-β-GLYCEROL PHOSPHATE DISODIUM-HYDROXYETHYL CELLULOSE HYBRID FOR ADIPOSE TISSUE ENGINEERING

作　　者：张舒[1,2] 罗静聪[1] 吕青[2] 邓雪琴[1] 熊炳钧 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室.干细胞与组织工程研究室,成都610041 [2]四川大学华西医院乳腺外科,成都610041

出　　处：《中国修复重建外科杂志》2015年第8期1028-1033,共6页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的探讨人脂肪干细胞(human adipose-derived stem cells,h ADSCs)与猪小肠黏膜下层脱细胞基质微粉(small intestinal submucosa powder,SISP)/壳聚糖温敏水凝胶复合构建组织工程脂肪的可行性。方法取乳癌患者自愿捐赠脂肪组织,Ⅰ型胶原酶消化分离h ADSCs。取第3代细胞与SISP/壳聚糖温敏水凝胶混匀,制成浓度为1×106个/m L的液态凝胶。将24只5周龄健康雌性裸鼠随机分为实验组及对照组(n=12),于颈背部皮下分别注射1 m L h ADSCs+SISP/壳聚糖温敏水凝胶或SISP/壳聚糖温敏水凝胶。注射后0、1、2、4、8周测量植入物体积,评价其降解情况;注射后1、2、4、8周两组各处死3只裸鼠,大体观察后取材行组织学及免疫组织化学染色观察,评价植入物成分构成变化(HE染色)、成脂能力(油红O染色)、血管形成情况(CD31标记)及材料内h ADSC存活分化(人波形蛋白标记)。结果注射后随时间延长,两组植入物均逐渐缩小;但各时间点实验组植入物体积均大于对照组,且8周时差异有统计学意义(t=3.348,P=0.029)。大体观察示各时间点两组植入物均边界清楚、无粘连,至8周时均可见黄色新生组织,且表面有毛细血管包裹。组织学观察示,注射后植入物结构逐渐变紧密,SISP逐渐降解,且实验组降解速度较对照组慢,脂肪逐渐形成,至8周时实验组中央出现少量成熟脂肪组织;各时间点实验组油红O染色阳性面积均大于对照组,且8周时差异有统计学意义(t=3.411,P=0.027)。免疫组织化学染色示,各时间点实验组均有h ADSCs存活;植入物有新生血管生成,2周时达峰值,各时间点两组CD31阳性面积比较差异无统计学意义(P>0.05),但实验组植入物中血管化更均匀。结论 SISP/壳聚糖温敏水凝胶可作为载体,复合h ADSCs植入裸鼠体内后可构建组织工程脂肪。[Abstract] Objective To study the feasibility of human adipose-derived stem cells （hADSCs） combined with small intestinal submucosa powder （SISP）/chitosan chloride （CSC1）-13-glycerol phosphate disodium （GP）- hydroxyethyl cellulose （HEC） for adipose tissue engineering. Methods hADSCs were isolated from human breast fat with collagenase type I digestion, and the third passage hADSCs were mixed with SISP/CSC1-GP-HEC at a density of 1×106 cells/mL. Twenty-four healthy female nude mice of 5 weeks old were randomly divided into experimental group （n=12） and control group （n=12）, and the mice were subcutaneously injected with 1 mL hADSCs＋SISP/CSC1-GP-HEC or SISP/CSC1-GP-HEC respectively at the neck. The degradation rate was evaluated by implant volume measurement at 0, 1, 2, 4, and 8 weeks. Three mice were euthanized at 1, 2, 4, and 8 weeks respectively for general, histological, and immunohistochemical observations. The ability of adipogenesis （Oil O staining）, angiopoiesis （CD31）, and localized the hADSCs （immunostaining for human Vimentin） were identified. Results The volume of implants of both groups decreased with time, but it was greater in experimental group than the control group, showing significant difference at 8 weeks （t=3.348,β-0.029）. The general observation showed that the border of implants was clear with no adhesion at each time point; fat-liked new tissues were observed with capillaries on the surface at 8 weeks in 2 groups. The histological examinations showed that the structure of implants got compact gradually after injection, and SISP gradually degraded with slower degradation speed in experimental group; adipose tissue began to form, and some mature adipose tissue was observed at 8 weeks in the experimental group. The Oil O staining positive area of experimental group was greater than that of the control group at each time point, showing significant difference at 8 weeks （t=3.411, P=0.027）. Immunohistochemical staining for Vemintin showed that hA

关 键 词：仿生脊髓支架 3-D打印 神经干细胞 生物相容性 

分 类 号：R622[医药卫生—整形外科] R318.08[医药卫生—外科学]