摘要
目的:探讨微观形貌仿生的聚乳酸纤维膜对大鼠BMSCs生长、铺展、黏附、增殖和成骨分化的影响。方法:取第3代rBMSCs,按1×105个/mL密度接种于覆盖不同直径聚乳酸纤维膜的细胞培养板上,分别为仿生组(A组)直径(357±32)nm、有形貌对照组(B、C组)直径分别为(164±13)nm、(1311±93)nm,无形貌空白对照组(Ctrl组),在成骨分化实验时增设成骨诱导阳性对照组(Positive组)。扫描电镜观察BMSCs生长、铺展;CCK-8试剂盒检测BMSCs增殖、黏附;实时荧光定量PCR检测Runx2、COLI、ALP、OSX的mRNA表达;Western Blot检测骨桥蛋白(OPN)、骨钙素(OC)的表达。结果:扫描电镜示:A、B二组上细胞具有明显聚集区域,相互联系紧密;单个细胞形态观察,A组较B、C及Ctrl组,细胞铺展面积更大、伪足更多。黏附实验示:在8h、16h,A、B组均高于C组(P<0.05)。增殖实验示:第3、5、7、9天,Ctrl组、A组、B组增殖情况均优于C组(P<0.05),三组间对比,Ctrl组优于A、B组(P<0.05);第7、9天A组增殖情况优于B组(P<0.05)。实时荧光定量PCR显示:在第3、10天,A、Positive组ALP、COLI、Runx2、OSX的mRNA表达量均明显高于B、C及Ctrl组(P<0.05);A组低于Positive组(P<0.05)。蛋白质印迹示:第10天,A、Positive组OPN、OC表达量均明显高于B、C、Ctrl组(P<0.05);A组OC表达量低于Positive组(P<0.05);B、C、Ctrl三组间差异无统计学意义(P>0.05)。结论:微观形貌仿生的聚乳酸纤维膜能促进rBMSCs黏附、增殖、成骨分化。
Objective:To investigate the effect of biomimetic polylactic acid fiber membrane on the growth,spreading,adhesion,proliferation and osteogenic differentiation of rat BMSCs.Methods:The third generation BMSCs rats were inoculated on a cell culture board with different diameters of polylactic acid fiber membrane by 1×105 pieces/mL density.Respectively bionic group(A group)diameter of(357±32)nm,control group with morphology(B,C group)diameter were(164±13)nm,(1311±93)nm,blank control(Ctrl group),in osteogenic differentiation experiments adding osteogenic induction positive the control group(Positive group).Scanning electron microscopic observation of BMSCs growth;CCK-8 assay BMSCs proliferation and adhesion;Real-time fluorescence quantitative PCR detection of Runx2,COLI,ALP and OSX expression of mRNA;Western blotting detection of osteopontin and osteocalcin expression.Results:Scanning electron microscopy showed that the cells in A and B groups had obvious aggregation areas,and they were closely related to each other.Single cell morphology observation,A compared with B,C and Ctrl group,cell spreading area is greater.The adhesion experiment showed:in 8h,16h,compare between the A,B,C groups,A and B group were higher than C group(P<0.05);Proliferation experiments showm:in day 3,5,7,9 the value of Ctrl,A,B group were better than C group(P<0.05),the comparison between the three groups,Ctrl group is better than A and B group(P<0.05);the comparison between the A and B group,in 7,9 day,A group is better than B group(P<0.05);Real-time fluorescent quantitative PCR showed:in the 3 and 10 day,the ALP,COLI,Runx2 and OSX mRNA expression of A and positive group,were significantly higher than B,C and Ctrl group(P<0.05);A group is lower than Positive group(P<0.05);Western blotting showed:in 10 day,the OPN and OC expression of A and Positive group was significantly higher than B,C,Ctrl group(P<0.05);the OC expression of A group was lower than the Positive group(P<0.05);there was no statistically significant difference compared between the B,C and Ctrl groups(P>0.05).Conclusion:The biomimetic polylactic acid fiber membrane can promote the adhesion,proliferation and osteogenic differentiation of BMSCs.
作者
陈林
王明俊
孙锦波
谭国兵
谭通夏
陈春
CHEN Lin;WANG Mingjun;SUN Jinbo(Xingyi People’s Hospital,Xingyi City,Guizhou Province 562400)
出处
《医学理论与实践》
2024年第20期3421-3426,3450,共7页
The Journal of Medical Theory and Practice
关键词
仿生
成骨分化
BMSCS
形貌
聚乳酸
Bionics
Osteogenetic differentiation
Bone marrow mesenchymal stem cells
Morphology
Polylactic acid
