不同浓度人源性抗菌肽LL-37/PLGA/β-TCP复合支架对兔骨髓间充质干细胞增殖、分化的影响(1)
【摘要】 目的：研究不同濃度人源性抗菌肽LL-37/聚乳酸聚乙醇酸共聚物(PLGA)/β-磷酸三钙(β-TCP)复合支架对于兔骨髓间充质干细胞(rBMSCs)增殖、分化的影响。方法：制备LL-37/PLGA/β-TCP复合支架材料，按LL-37浓度分为0、1、5、10 μmol/L组，另设置空白rBMSCs培养基为N组，每组三个培养皿，细胞密度为5×105个/皿。扫描电镜下观察不同浓度LL-37/PLGA/β-TCP复合支架形态。采用CCK-8检查方法检测各组干预rBMSCs 0、24、48、72、96、120 h后rBMSCs细胞增殖活力。干预rBMSCs 24 h后，采用荧光定量聚合酶链反应(qPCR)检测各组成骨相关基因碱性磷酸酶(ALP)、骨形态发生蛋白-1(BMP-1)及骨形态发生蛋白-7(BMP-7)的mRNA相对表达量。干预rBMSCs 24 h后，采用ELISA检测各组ALP、BMP-1及BMP-7的蛋白水平。每个实验独立重复三次。结果：LL-37/PLGA/β-TCP复合支架材料随着LL-37浓度增加而呈现多孔、细胞吸附增多的趋势，促进细胞贴附生长。干预24 h后，10 μmol/L组细胞增殖能力明显高于N组(P<0.05);随着干预时间延长，1、5、10 μmol/L组均对rBMSCs有促进增殖作用，且呈浓度依赖作用。1、5、10 μmol/L组ALP、BMP-1及BMP-7的mRNA相对表达量与蛋白表达水平均高于N组(P<0.05);1、5、10 μmol/L组ALP、BMP-1及BMP-7的mRNA相对表达量与蛋白表达水平均呈高表达，且呈浓度依赖。结论：LL-37/PLGA/β-TCP复合支架可以显著促进rBMSCs增殖与分化，是一种潜在的骨组织工程支架。
【关键词】 人源性抗菌肽 PLGA/β-TCP复合支架 兔骨髓间充质干细胞 碱性磷酸酶 骨形态发生蛋白
[Abstract] Objective： To investigate the effects of concentrations of human antimicrobial peptide LL-37/polylactic acid-polyglycolic acid copolymer (PLGA)/β-tricalcium phosphate (β-TCP) composite scaffolds on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (rBMSCs). Method： LL-37/PLGA/β-TCP composite scaffolds were prepared. The scaffolds were divided into 0， 1， 5， 10 μmol/L group according to the concentration of LL-37. In addition， the blank rBMSCs medium was set as group N， three plates in each group and cell density of 5×105/plates. The morphology of LL-37/PLGA/β-TCP composite scaffolds with different concentrations were observed under scanning electron microscope. The proliferation activity of rBMSCs was detected by CCK-8 after 0， 24， 48， 72， 96 and 120 h of rBMSCs intervention. After 24 h of rBMSCs intervention， the relative mRNA expression of osteogenesis-related genes alkaline phosphatase (ALP)， bone morphogenetic protein-1 (BMP-1) and bone morphogenetic protein-7 (BMP-7) were detected by fluorescence quantitative polymerase chain reaction (qPCR). After 24 h of rBMSCs intervention， the protein levels of ALP， BMP-1 and BMP-7 were detected by ELISA. Each experiment was repeated three times independently. Result： With the increase of LL-37 concentration， LL-37/PLGA/β-TCP composite scaffolds showed a trend of porous and cell adsorption， which promoted cell adhesion and growth. After 24 h of intervention， the proliferation ability of 10 μmol/L group was significantly higher than that of N group (P<0.05). With the extension of intervention time， the 1， 5 and 10 moL/L group all had the effect of promoting rBMSCs proliferation， and the effect was concentration dependent. The relative mRNA expression and protein expression level of ALP， BMP-1 and BMP-7 in 5， 10 μmol/L group were higher than those in N group (P<0.05). The relative mRNA expression and protein expression level of ALP， BMP-1 and BMP-7 in 1， 5， 10 μmol/L group were all highly expressed and concentration dependent. Conclusion： The LL-37/PLGA/β-TCP composite scaffold can significantly promote rBMSCs proliferation and differentiation. It is a potential bone tissue engineering scaffold., http://www.100md.com(罗万荣 易伟宏 胡广询)
【关键词】 人源性抗菌肽 PLGA/β-TCP复合支架 兔骨髓间充质干细胞 碱性磷酸酶 骨形态发生蛋白
[Abstract] Objective： To investigate the effects of concentrations of human antimicrobial peptide LL-37/polylactic acid-polyglycolic acid copolymer (PLGA)/β-tricalcium phosphate (β-TCP) composite scaffolds on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (rBMSCs). Method： LL-37/PLGA/β-TCP composite scaffolds were prepared. The scaffolds were divided into 0， 1， 5， 10 μmol/L group according to the concentration of LL-37. In addition， the blank rBMSCs medium was set as group N， three plates in each group and cell density of 5×105/plates. The morphology of LL-37/PLGA/β-TCP composite scaffolds with different concentrations were observed under scanning electron microscope. The proliferation activity of rBMSCs was detected by CCK-8 after 0， 24， 48， 72， 96 and 120 h of rBMSCs intervention. After 24 h of rBMSCs intervention， the relative mRNA expression of osteogenesis-related genes alkaline phosphatase (ALP)， bone morphogenetic protein-1 (BMP-1) and bone morphogenetic protein-7 (BMP-7) were detected by fluorescence quantitative polymerase chain reaction (qPCR). After 24 h of rBMSCs intervention， the protein levels of ALP， BMP-1 and BMP-7 were detected by ELISA. Each experiment was repeated three times independently. Result： With the increase of LL-37 concentration， LL-37/PLGA/β-TCP composite scaffolds showed a trend of porous and cell adsorption， which promoted cell adhesion and growth. After 24 h of intervention， the proliferation ability of 10 μmol/L group was significantly higher than that of N group (P<0.05). With the extension of intervention time， the 1， 5 and 10 moL/L group all had the effect of promoting rBMSCs proliferation， and the effect was concentration dependent. The relative mRNA expression and protein expression level of ALP， BMP-1 and BMP-7 in 5， 10 μmol/L group were higher than those in N group (P<0.05). The relative mRNA expression and protein expression level of ALP， BMP-1 and BMP-7 in 1， 5， 10 μmol/L group were all highly expressed and concentration dependent. Conclusion： The LL-37/PLGA/β-TCP composite scaffold can significantly promote rBMSCs proliferation and differentiation. It is a potential bone tissue engineering scaffold., http://www.100md.com(罗万荣 易伟宏 胡广询)
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