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比较诱导多能干细胞在 2D 和 3D 聚偏二氟乙烯支架上的成骨分化潜能。

Comparison of osteogenic differentiation potential of induced pluripotent stem cells on 2D and 3D polyvinylidene fluoride scaffolds.

机构信息

Cellular and Molecular Research Center, Yasuj University of Medical Sciences, Yasuj, Iran.

Medicinal Plants Research Center, Yasuj University of Medical Sciences, Yasuj, Iran.

出版信息

J Cell Physiol. 2019 Aug;234(10):17854-17862. doi: 10.1002/jcp.28415. Epub 2019 Mar 9.

DOI:10.1002/jcp.28415
PMID:30851069
Abstract

In recent decades, tissue engineering has been the most contributor for introducing 2D and 3D biocompatible osteoinductive scaffolds as bone implants. Polyvinylidene fluoride (PVDF), due to the unique mechanical strength and piezoelectric properties, can be a good choice for making a bone bioimplant. In the present study, PVDF nanofibers and film were fabricated as 3D and 2D scaffolds, and then, osteogenic differentiation potential of the human induced pluripotent stem cells (iPSCs) was investigated when grown on the scaffolds by evaluating the common osteogenic markers in comparison with tissue culture plate. Biocompatibility of the fabricated scaffolds was confirmed qualitatively and quantitatively by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and scanning electron microscopy assays. Human iPSCs cultured on PVDF nanofibers showed a significantly higher alkaline phosphate activity and calcium content compared with the iPSCs cultured on PVDF film. Osteogenic-related genes and proteins were also expressed in the iPSCs seeded on PVDF nanofibers significantly higher than iPSCs seeded on PVDF film, when investigated by real-time reverse transcription polymerase chain reaction and western blot analysis, respectively. According to the results, the PVDF nanofibrous scaffold showed a greater osteoinductive property compared with the PVDF film and due to the material similarity of the scaffolds, it could be concluded that the 3D structure could lead to better bone differentiation. Taken together, the obtained results demonstrated that human iPSC-seeded PVDF nanofibrous scaffold could be considered as a promising candidate for use in bone tissue engineering applications.

摘要

近几十年来,组织工程学在引入 2D 和 3D 生物相容性成骨支架作为骨植入物方面做出了最大的贡献。聚偏二氟乙烯(PVDF)由于具有独特的机械强度和压电特性,因此可以成为制造骨生物植入物的良好选择。在本研究中,制备了 PVDF 纳米纤维和薄膜作为 3D 和 2D 支架,然后通过评估常见的成骨标志物,研究了人诱导多能干细胞(iPSC)在支架上的成骨分化潜力,并与组织培养板进行了比较。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐和扫描电子显微镜分析,定性和定量地证实了所制备的支架的生物相容性。与在 PVDF 薄膜上培养的 iPSCs 相比,在 PVDF 纳米纤维上培养的人 iPSCs 的碱性磷酸酶活性和钙含量明显更高。通过实时逆转录聚合酶链反应和 Western blot 分析分别研究了在 PVDF 纳米纤维上接种的 iPSCs 中骨形成相关基因和蛋白的表达情况,结果表明,与 PVDF 薄膜相比,PVDF 纳米纤维支架具有更强的成骨诱导特性,并且由于支架的材料相似性,可以得出结论,3D 结构可以促进更好的骨分化。综上所述,研究结果表明,人 iPSC 接种的 PVDF 纳米纤维支架可作为骨组织工程应用的有前途的候选材料。

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