第三代超声辅助吸脂获取的hADSCs构建组织工程软骨的实验研究  

Isolation of Human Adipose-Derived Stromal Cells Using Suction-Assisted or Third-Generation Ultrasound-Assisted Lipoaspiration for Constructing Tissue Engineered Cartilage

作　　者：王健[1] 周栩 WANG Jian;ZHOU Xu(Head and Neck Cosmetic Surgery Center,Plastic Surgery Hospital,Chinese Academy of Medical Science and Peking Union Medical College,Beijing 100144,China.)

机构地区：[1]中国医学科学院整形外科医院面颈整形中心,北京市100144

出　　处：《组织工程与重建外科杂志》2020年第2期91-96,共6页Journal of Tissue Engineering and Reconstructive Surgery

基　　金：协和小规模特色办学经费资助青年教师培养项目。

摘　　要：目的观察应用第三代超声辅助吸脂技术获取的人脂肪干细胞(hADSCs)与残耳软骨细胞共培养在体内构建组织工程软骨的效果。方法分离培养先天性小耳畸形患者来源的残耳软骨细胞,通过第三代超声辅助吸脂和常规负压吸脂术获取脂肪,并提取hADSCs。实验分为3组:对照组1,PGA/PLA支架接种残耳软骨细胞;对照组2,PGA/PLA支架接种残耳软骨细胞+常规负压抽脂获取的hADSCs;实验组,PGA/PLA支架接种残耳软骨细胞+第三代超声辅助抽脂获取的hADSCs。所有细胞-支架复合物经体外培养4周后植入裸鼠体内,8周后取材。对体外培养4周和体内培养8周的各组标本分别进行大体观察、湿重测量、糖胺多糖含量测定、组织学及免疫组化染色和生物力学检测。结果应用第三代超声辅助吸脂和常规负压吸脂都可获得hADSCs。细胞-支架复合物体外培养4周,对照组1、2与实验组都形成软骨样组织,组织学观察显示软骨特异性ECM沉积,但结构疏松,仍有未降解的PGA支架材料。体内培养8周后取材,对照组1组织学观察显示软骨组织不均一,大部分区域形成成熟软骨组织,但有部分区域组织结构疏松,软骨特异性ECM分泌较少;对照组2和实验组则形成成熟的软骨组织,结构均一,可见大量的软骨陷窝和细胞外基质分泌,支架材料完全降解。湿重、糖胺多糖、生物力学检测结果都显示对照组2和实验组显著优于对照组1,对照组2与实验组无显著差异。结论应用第三代超声辅助吸脂可安全有效地获取hADSCs,与残耳软骨细胞共培养可形成成熟的组织工程软骨。Objective To investigate the formation of tissue engineered cartilage by culture microtia chondrocytes and ADSCs obtained by suction-assisted lipoaspiration(SAL)or third-generation ultrasound-assisted lipoaspiration(UAL).Methods Microtia chondrocytes were isolated from patients with congenital microtia.ADSCs obtained by suction-assisted lipoaspiration or third-generation ultrasound-assisted lipoaspiration were isolated in vitro.There are 3 groups:Ctrl group 1,microtia chondrocytes were seeded on the PGA/PLA scaffolds;Ctrl group 2,microtia chondrocytes and ADSCs obtained by suction-assisted lipoaspiration were seeded on the PGA/PLA scaffolds;Exp group,microtia chondrocytes and ADSCs obtained by third-generation ultrasound-assisted lipoaspiration were seeded on the PGA/PLA scaffolds.After in vitro culture for 4 weeks and subcutaneous implantation for 8 weeks,the constructs were harvested for gross observation,wet weight measurement,glycosaminoglycan(GAG)quantification,histological and immunohistochemical staining and biomechanical evaluation.Results ADSCs can be obtained by SAL or third-generation UAL.In vitro culture for 4 weeks,chondroid tissues were formed in ctrl group 1,ctrl group 2 and Exp group.Histological observation showed chondrogenic ECM deposition,loosened structure and undegraded scaffolds.In vivo culture for 8 weeks,histological observation of the ctrl group 1 showed that the cartilage tissue was not uniform,most of the regions formed mature cartilage tissue,but some regions had loose tissue structure and less cartilage specific ECM secretion.In ctrl group 2 and Exp group,very mature cartilage tissues were formed with uniform structure,and a large number of cartilage lacunae and extracellular matrix were secreted,and the scaffold materials had been completely degraded.The test result of wet weight,glycosaminoglycan and biomechanics all showed that the ctrl group 2 and Exp group were significantly better than the ctrl group 1,but ctrl group 2 and Exp group revealed no significant difference.Conclusion

关 键 词：第三代超声辅助吸脂技术 负压吸脂技术 脂肪干细胞 组织工程软骨 

分 类 号：R318.1[医药卫生—生物医学工程]