Sustained release of melatonin from poly (lactic‐co‐glycolic acid) (PLGA) microspheres to induce osteogenesis of human mesenchymal stem cells in vitro
Sustained release of melatonin from poly (lactic‐co‐glycolic acid) (PLGA) microspheres to induce osteogenesis of human mesenchymal stem cells in vitro
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DOI：
10.1111/j.1600-079x.2012.01016.x
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发表时间：
2013-01
影响因子：
10.3
通讯作者：
Liangming Zhang;Jinling Zhang;You Ling;Changhua Chen;Anjing Liang;Yan Peng;Hong Chang;Peiqiang Su;D. Huang
中科院分区：
文献类型：
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作者：
Liangming Zhang;Jinling Zhang;You Ling;Changhua Chen;Anjing Liang;Yan Peng;Hong Chang;Peiqiang Su;D. Huang
褪黑素通过受体依赖或受体独立的作用促进骨形成并防止骨退化。本研究的目的是将褪黑素包埋在聚乳酸- co -乙醇酸（PLGA）微球（PLGA - MEL - MS）中，建立褪黑素缓释系统，并在体外评价其对人间充质干细胞（hMSCs）成骨的影响。采用单乳液溶剂蒸发技术制备PLGA - MEL - MS。扫描电镜显示，褪黑激素的掺入不会干扰PLGA微球的大小和形态。体外释药试验表明，PLGA - MEL - MS呈双相释药模式：低初始爆发释药，前3天释药量约为40%，相对缓慢连续释药，25天释药量约为85%。细胞增殖实验表明，PLGA‐MEL‐MS对人间充质干细胞的增殖无明显影响。在成骨实验中，与对照组相比，PLGA‐MEL‐MS明显增强了碱性磷酸酶（ALP） mRNA的表达和ALP活性。同时，runx2、骨桥蛋白、骨钙素等成骨细胞分化标志物也显著上调。此外，基于茜素红的定量分析表明，与对照组相比，PLGA - MEL - MS显著增强了hMSCs的钙沉积。因此，这种简单的褪黑素缓释系统可以控制褪黑素的释放，形成一段时间内褪黑素浓度相对稳定的微环境，支持hMSCs体外成骨分化。这表明该系统可作为骨生长刺激剂用于体内骨愈合。
Abstract: Melatonin promotes bone formation and prevents bone degradation via receptor‐dependent or receptor‐independent actions. The aim of this study is to encapsulate melatonin into poly (lactic‐co‐glycolic acid) (PLGA) microspheres (PLGA‐MEL‐MS) and create a melatonin sustained release system, then to evaluate its effect on the osteogenesis of human mesenchymal stem cells (hMSCs) in vitro. PLGA‐MEL‐MS were prepared by single emulsion solvent evaporation technique. Scanning electron microscopy demonstrated the incorporation of melatonin did not disturb the conventional generation of PLGA microspheres in size and morphology. In vitro drug release assay showed that PLGA‐MEL‐MS exhibited a biphasic drug release pattern: a low initial burst release effect with approximately 40% drug release at the first 3 days and a relatively retarded and continuous release with about 85% drug release over the 25 days. Cell proliferation assay demonstrated that PLGA‐MEL‐MS had no apparent effect on proliferation of human MSCs. In an osteogenesis assay, PLGA‐MEL‐MS obviously enhanced alkaline phosphatase (ALP) mRNA expression and increased ALP activity compared to that in the control group. Meanwhile, several markers of osteoblast differentiation were also significantly upregulated, including runx2, osteopontin, and osteocalcin. Furthermore, quantificational alizarin red‐based assay demonstrated that PLGA‐MEL‐MS significantly enhanced calcium deposit of hMSCs compared to the controls. Therefore, this simple melatonin sustained release system can control released melatonin to generate a microenvironment with a relatively stable concentration of melatonin for a period of time to support osteogenic differentiation of hMSCs in vitro. This suggests that this system may be used as bone growth stimulator in bone healing in vivo.