CPO-PCL微粒在缺氧条件下对脂肪间充质干细胞体外增殖及成骨分化作用的影响  

Effects of CPO-PCL microparticles on the proliferation and osteogenic differentiation of adipose-derived mesenchymal stem cells in vitro under hypoxia

作　　者：罗浙 练雷栋 胡珂琦 顾仕荣[2] 周珂 干开丰 LUO Zhe;LIAN Leidong;HU Keqi;GU Shirong;ZHOU Ke;GAN Kaifeng(Medical Department of Ningbo University,Ningbo 315000;Department of Orthopedics,Li Huili Hospital of Ningbo Medical Cente(r Affiliated Li Huili Hospital of Ningbo University),Ningbo 315040,China)

机构地区：[1]宁波大学医学部,浙江宁波315000 [2]宁波市医疗中心李惠利医院(宁波大学附属李惠利医院)骨外科,浙江宁波315040

出　　处：《南京医科大学学报（自然科学版）》2024年第8期1051-1061,共11页Journal of Nanjing Medical University(Natural Sciences)

基　　金：浙江省自然科学基金(LQ21H060002);宁波市自然科学基金(2022J251);宁波市公益类科技计划(2021S105);宁波市卫生健康青年技术骨干人才培养计划([2021]106)。

摘　　要：目的:探究聚己内酯(polycaprolactone,PCL)包载过氧化钙(calcium peroxide,CPO)的CPO-PCL微粒在缺氧条件下对脂肪间充质干细胞(adipose-derived mesenchymal stem cell,ADMSC)体外增殖及成骨分化作用的影响。方法:提取大鼠ADMSC,并加入制备的CPO-PCL微粒,在缺氧/常氧环境下,正常培养或成骨分化培养细胞,分别于培养第7天和第14天MTT实验检测ADMSC增殖,碱性磷酸酶(alkaline phosphatase,ALP)试剂盒检测ALP水平;茜素红染色观察钙化结节情况,免疫荧光染色观察ADMSC核心转录因子-2(Runt-related transcription factor 2,RUNX2)、骨钙素(Osteocalcin)和骨桥蛋白(Osteopontin)的荧光表达量。结果:在缺氧成骨分化培养条件下,1.00%CPO-PCL微粒显著促进细胞增殖(P<0.05),0.50%和1.00%CPO-PCL微粒显著增加ADMSC ALP和钙结节生成量(P<0.001),促进RUNX2、Osteocalcin和Osteopontin蛋白表达(P<0.05)。在缺氧正常培养条件下,1.00%CPO-PCL微粒增加ADMSC ALP含量,促进RUNX2、Osteocalcin和Osteopontin蛋白表达(P<0.05)。在常氧成骨分化培养条件下,1.00%CPO-PCL微粒增加ALP和钙结节生成量,0.50%和1.00%CPO-PCL微粒促进RUNX2、Osteocalcin蛋白表达(P<0.001)。在常氧正常培养条件下,0.50%和1.00%CPO-PCL微粒促进RUNX2、Osteocalcin和Osteopontin蛋白表达(P<0.05),但对细胞成骨分化无显著影响。结论:1.00%CPO-PCL微粒在缺氧成骨分化培养条件下能够促进ADMSC体外增殖及成骨分化。Objective:To investigate the effects of calcium peroxide(CPO)-loaded polycaprolactone(PCL)microparticles(CPO-PCL)on the proliferation and bone differentiation of adipose-derived mesenchymal stem cells(ADMSCs)in vitro under hypoxia.Methods:Rat ADMSCs were extracted and added with the prepared CPO-PCL particles for normal or osteogenic differentiation culture under hypoxia/normoxia environment.On the 7th and 14th days,cell proliferation,alkaline phosphatase(ALP)level,the calcification of nodulation,and the fluorescence intensity of Runt-related transcription factor 2(RUNX2),Osteocalcin and Osteopontin were examined by the MTT assay,alkaline phosphatase(ALP)kit,alizarine red staining and immunofluorescence staining,respectively.Results:Under hypoxia and osteogenic differentiation culture conditions,1.00%CPO-PCL microparticles significantly promoted ADMSCs proliferation(P<0.05),and CPO-PCL microparticles of 0.50%and 1.00%concentrations notably increased the production of ALP and calcium nodules(P<0.001),while enhancing the expressions of RUNX2,Osteocalcin and Osteopontin proteins(P<0.05).Under hypoxia and normal culture conditions,1.00%CPO-PCL microparticles elevated ALP levels,increased the expression levels of RUNX2,Osteocalcin and Osteopontin(P<0.05).Under normoxia and differentiation culture conditions,1.00%CPO-PCL microparticles increased the production of ALP and calcium nodules,and CPO-PCL microparticles of 0.50%and 1.00%concentrations promoted the expression levels of RUNX2 and Osteocalcin proteins(P<0.001).Under normoxia and normal culture conditions,0.50%and 1.00%CPO-PCL microparticles up-regulated RUNX2,Osteocalcin and Osteopontin expression levels(P<0.05),but rarely affect osteoblast differentiation.Conclusion:CPO-PCL microparticles of 1.00%promote the proliferation and bone differentiation of ADMSCs in vitro under hypoxia and osteogenic differentiation culture.

关 键 词：CPO-PCL微粒 ADMSC 成骨分化 细胞增殖 缺氧 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]