注射用聚左旋乳酸微球体内可促胶原再生  

In vivo neocollagenesis of injectable poly(L-lactic acid) microspheres

作　　者：张译心 罗倩 梁瀚文 陈建林 赵楠 何斌[4] Zhang Yixin;Luo Qian;Liang Hanwen;Chen Jianlin;Zhao Nan;He Bin(School of Smart Health,Chongqing College of Electronic Engineering,Chongqing 401331,China;School of Laboratory Medicine,Chengdu Medical College,Chengdu 610500,Sichuan Province,China;Puliyan(Nanjin)Medical Technology Co.,Ltd.,Nanjing 211500,Jiangsu Province,China;National Engineering Research Center for Biomaterials,Sichuan University,Chengdu 610064,Sichuan Province,China)

机构地区：[1]重庆电子工程职业学院智慧健康学院,重庆市401331 [2]成都医学院检验医学院,四川省成都市610500 [3]普丽妍(南京)医疗科技有限公司,江苏省南京市211500 [4]四川大学国家生物医学材料工程技术研究中心,四川省成都市610064

出　　处：《中国组织工程研究》2022年第34期5448-5453,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(51773130),项目负责人:何斌;四川省科技计划资助(2019JDRC0098),项目负责人:张译心;四川大学-企业横向合作项目(18H0350),项目负责人:何斌;四川省卫生和计划生育委员会科研课题(18PJ553),项目负责人:陈建林。

摘　　要：背景:聚左旋乳酸皮下填充剂是美国FDA批准可作为修复皮下软组织胶原流失的医美产品,由于其形态为不规则颗粒,容易产生过度炎症反应。目的:观察辐照条件对聚左旋乳酸微球分子质量和粒径形貌的影响,以及聚左旋乳酸微球皮下填充剂植入兔皮下的异物反应程度和刺激胶原再生情况。方法:采用乳液-溶剂挥发法制备聚左旋乳酸微球,参考Sculptra的配方配制成可注射皮下填充剂,分别经25,50 kGy辐照灭菌,分析辐照灭菌对微球分子质量和粒径形貌的影响。采用MTT法检测不同质量浓度(50,100,250,500 mg/L,以微球的浓度计)可注射皮下填充剂对小鼠成纤维细胞增殖的影响。将25 kGy辐照灭菌的可注射皮下填充剂(实验组)、生理盐水(对照组)分别注射至兔背部皮下,于设定的时间点进行注射部位背部组织苏木精-伊红、马松三色染色及CD68、Ⅰ型胶原和Ⅲ型胶原免疫荧光染色。结果与结论:(1)随着辐照剂量的增加,微球的黏度、黏均分子质量和数均分子质量降低明显,但辐照灭菌并未导致微球的粒径和形貌发生明显改变。(2)MTT检测结果显示,当填充剂的质量浓度低于250 mg/L时细胞存活率均>90%,即使材料质量浓度高达500 mg/L时细胞存活率也仍高于80%,并且长时间的孵育(72 h)也并未产生显著的细胞毒性。(3)动物实验苏木精-伊红染色与CD68免疫荧光染色显示,填充剂植入后的第0.5-4个月只引起轻微炎性反应,植入后第6个月时炎性反应程度达到最高,且部分微球表面出现孔洞结构或不规则形状;植入后第9个月微球完全降解。(4)动物实验马松三色染色与Ⅰ型胶原、Ⅲ型胶原免疫荧光染色显示,植入后第4个月,微球周围主要为Ⅰ型胶原,外围主要以Ⅲ型胶原为主;植入后第6个月,微球附近的I型胶原增多,外围的Ⅲ型胶原增多;植入后第9个月,纤维包囊内部主要以Ⅰ型胶原为主,外围则Ⅰ型胶原�BACKGROUND:Poly(L-lactic acid)(PLLA)microparticle(Sculptra?)has been approved by FDA as the dermal filler to regenerate collagen for anti-aging purpose.Because of its irregular particle shape,it is prone to excessive inflammation.OBJECTIVE:To investigate the effects of irradiation on the molecular weight,size and morphology of the poly(L-lactic acid)microspheres,as well as the foreign body reaction and the neocollagenesis stimulation of the poly(L-lactic acid)microspheres dermal filler implanted subcutaneously into back of rabbits.METHODS:Microspheres were prepared by the emulsion-solvent evaporation method and utilized as the dermal filler with the same Sculptra’s formula.The filler was sterilized by irradiation at 25 and 50 kGy,separately and the molecular weight,size and morphology of the microspheres were analyzed.The MTT assay was used to detect the effects of the microspheres at different concentrations(50,100,250,and 500 mg/L calculated by the concentration of microspheres)on the proliferation of mouse fibroblasts.The 25 kGy irradiated dermal filler(experimental group)and saline(control group)were injected into the back of the rabbit subcutaneously.Hematoxylin-eosin staining,Masson staining,and immunofluorescence staining of CD68,typeⅠcollagen and typeⅢcollagen were performed on the back tissue of the injection site at the set time points.RESULTS AND CONCLUSION:(1)With the increase of the radiation dose,the viscosity,viscosity-average molecular weight and number-average molecular weight of the microspheres decreased significantly,but the irradiation sterilization did not cause significant changes in the particle size and morphology of the microspheres.(2)MTT assay results demonstrated that when the mass concentration of the filler was lower than 250 mg/L,the cell survival rate was all>90%.Even when the material mass concentration reached the highest level 500 mg/L,the cell survival rate was still higher than 80%.Incubation for 72 hours also did not produce significant cytotoxicity.(3)In animal experi

关 键 词：聚左旋乳酸 高分子微球 皮下填充剂 炎性反应 胶原再生 Ⅰ型胶原 Ⅲ型胶原 体内降解 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]