可注射人脱细胞脂肪组织基质匀浆的制备与应用  

Preparation and application of injectable human decellular adipose tissue matrix homogenate

作　　者：郑洋洋 刘毅[1] 王嘉 彤郭镇 Zheng Yangyang;Liu Yi;Wang Jiatong;Guo Zhen(Department of Burns and Plastic Surgery and Wound Repair Surgery,Second Hospital of Lanzhou University,Lanzhou 730030,China;Lan Zhou University Second Hospital,Lanzhou 730030,China;Department of Burns and Plastic Surgery,The 94th Hospital of the Joint Logistics Support Force,Lanzhou 730050,China)

机构地区：[1]兰州大学第二医院烧伤整形与创面修复外科,兰州730030 [2]兰州大学第二临床医学院,兰州730030 [3]解放军联勤保障部队第九四〇医院,兰州730050

出　　处：《中华医学美学美容杂志》2021年第5期368-373,共6页Chinese Journal of Medical Aesthetics and Cosmetology

摘　　要：目的自主研发可控温高速软组织匀浆机将人脱细胞脂肪组织基质(DAT)制备成可注射匀浆,探讨其细胞相容性及填充效果。方法2019年3月至2021年3月,于解放军联勤保障部队第九四〇医院对正常脂肪组织进行脱细胞处理后得到DAT,将DAT与生理盐水按照1∶12混匀,用可控温高速软组织匀浆机以803×g匀浆,使制备DAT匀浆可顺利通过27 G针头。扫描电镜观察DAT匀浆并检测其细胞相容性;将脂肪颗粒、DAT匀浆、DAT匀浆+脂肪干细胞(ADSC)分别注射入大鼠背部,比较填充效果、血管生成能力、炎症浸润情况等。结果脂肪组织经脱细胞处理成功获得DAT,经可控温高速软组织匀浆机制备DAT匀浆,粒径(749.91±136.79)nm;细胞与其复合培养后,黏附率(92.16±1.00)%,CCK-8检测显示随时间延长,吸光度(A)值不断升高,细胞生长情况良好,匀浆对细胞无毒性。体内试验显示,DAT匀浆、DAT匀浆+ADSC填充效果明显好于脂肪颗粒组(P<0.01)。脂肪颗粒组有大量脂肪细胞坏死融合形成油囊,而DAT匀浆、DAT匀浆+ADSC未见明显降解,并有部分脂肪细胞生成。CD31染色结果显示,DAT匀浆组及DAT匀浆+ADSC组微血管数量高于脂肪颗粒组(P<0.01);CD68染色结果显示,DAT匀浆组及DAT匀浆+ADSC组炎症浸润低于脂肪颗粒组(P<0.01)。结论自主研发的可控温高速软组织匀浆机可将DAT制备为能通过27 G针头的DAT匀浆,具有较好的细胞相容性以及填充效果,且注射过程简单,创伤小,可作为填充材料。Objective To prepare human decellular adipose tissue matrix(DAT)as injectable homogenate by a specially developed high-speed soft tissue homogenizer with controllable temperature,and to investigate its cellular compatibility and filling effect.Methods From March 2019 to March 2021,DAT was obtained in the 940th Hospital from normal adipose tissue after decellular treatment.The DAT was mixed with normal saline at the rate of 1:12.The specially developed high-speed soft tissue homogenizer with controllable temperature was used to conduct homogenization at 803×g for 10 min.The produced DAT homogenizer could pass through the 27G needle smoothly.DAT homogenate was observed under scanning electron microscope and its cytocompatibility was detected.Finally,granular fat,DAT homogenate and DAT homogenate+ADSCs were injected into the back of rats respectively,and the filling effect,angiogenesis ability and inflammatory infiltration were compared.Results After decellular treatment,adipose tissue changed into DAT without cellular residue.The particle size of DAT homogenate was about(749.91±136.79)nm,which was prepared by the specially developed temperature controlled high-speed soft tissue homogenater.The adhesion rate was(92.16±1.00)%,and the A value increased with time.The cell growth was good,and the homogenate had no toxicity to the cells.In vivo experiment,the filling effect of DAT homogenate and DAT homogenate+ADSCs was significantly better than that of granulated fat group(P<0.01).In the granulated fat group,a large number of adipocytes were necrotic and fused to form oil sacs,while DAT homogenate,DAT homogenate+ADSCs showed no obvious degradation,and some adipocytes were generated.The results of CD31 staining indicated that the number of microvessels in DAT homogenate group and DAT homogenate+ADSCs group was higher than that in granulated fat group(P<0.01).The results of CD68 staining indicated that the inflammatory infiltration in DAT homogenate group and DAT homogenate+ADSCs group was lower than that in granule fa

关 键 词：大鼠 填充剂 软组织填充 脂肪干细胞 脱细胞脂肪组织基质 

分 类 号：R318.08[医药卫生—生物医学工程]