聚己内酯/氧化镁/氧化石墨烯支架中从间充质干细胞分化而来的成骨细胞的形态学和分子分析
Morphological and Molecular Analysis of Osteoblasts Differentiated from Mesenchymal Stem Cells in Polycaprolactone/Magnesium Oxide/Graphene Oxide Scaffold.
作者信息
Niknam Z, Zali H, Mansouri V, Rezaei Tavirani M, Omidi M
机构信息
Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Proteomics Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
出版信息
Int J Organ Transplant Med. 2019;10(4):171-182.
BACKGROUND
The loss or dysfunction of bone tissue that observed after bone tumor resections and severe nonunion fractures afflicts 200 million people worldwide. Bone tissue engineering is a promising approach to repair osteoporotic fractures.
OBJECTIVE
In this paper, polycaprolactone (PCL)/magnesium oxide (MgO)/graphene oxide (GO) nanofibrous scaffold was fabricated by electrospining method, and its biocompatibility and osteogenic differentiation of adipose-derived mesenchymal stem cells (MSCs) on this scaffold were evaluated and compared with pure PCL nanofibrous scaffold.
METHODS
SEM analysis, DAPI staining and MTT assay were used to evaluation biocompatibility of PCL/MgO/GO composite scaffold. In addition by ALP assay and proteomic approach, osteostimulatory effect of electrospun composite scaffold was investigated and the expression level of osteogenic markers including Runt-related transcription factor cbfa1/runx2 (runx2), collagen type I (Col1a1) and osteopontin (OPN) in MSCs seeded on PCL/MgO/GO composite scaffold was determined and compared with pure PCL scaffold. Then, RT-PCR technique was used to validate the level expression of these genes.
RESULTS
The obtained results showed that adhesion, viability and ALP activity of MSCs on PCL/MgO/GO scaffold considerably enhanced compared with pure PCL. As well as proteomic and real-time analysis illustrated the expression of osteogenic markers including runx2, Col1a1 and OPN increased (>2-fold) in cells seeded on PCL/MgO/GO composite scaffold.
CONCLUSION
It was concluded that MgO and GO nanoparticles could improve the biocompatibility of PCL scaffold and enhance the osteogenic differentiation of MSCs.
背景
骨肿瘤切除术后和严重骨不连骨折后观察到的骨组织丢失或功能障碍困扰着全球2亿人。骨组织工程是修复骨质疏松性骨折的一种有前景的方法。
目的
本文采用静电纺丝法制备聚己内酯（PCL）/氧化镁（MgO）/氧化石墨烯（GO）纳米纤维支架，并评估其生物相容性以及脂肪来源间充质干细胞（MSCs）在该支架上的成骨分化情况，并与纯PCL纳米纤维支架进行比较。
方法
采用扫描电子显微镜（SEM）分析、4',6-二脒基-2-苯基吲哚（DAPI）染色和噻唑蓝（MTT）法评估PCL/MgO/GO复合支架的生物相容性。此外，通过碱性磷酸酶（ALP）测定和蛋白质组学方法，研究静电纺丝复合支架的骨刺激作用，并测定接种在PCL/MgO/GO复合支架上的MSCs中成骨标志物包括 runt相关转录因子cbfa1/runx2（runx2）、I型胶原（Col1a1）和骨桥蛋白（OPN）的表达水平，并与纯PCL支架进行比较。然后，采用逆转录聚合酶链反应（RT-PCR）技术验证这些基因的表达水平。
结果
所得结果表明，与纯PCL相比，MSCs在PCL/MgO/GO支架上的黏附、活力和碱性磷酸酶活性显著增强。蛋白质组学和实时分析也表明，接种在PCL/MgO/GO复合支架上的细胞中，包括runx2、Col1a1和OPN在内的成骨标志物表达增加（>2倍）。
结论
得出结论，MgO和GO纳米颗粒可改善PCL支架的生物相容性并增强MSCs的成骨分化。
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