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人脐带华通氏胶间充质干细胞与仿生 3D 支架的体外相互作用。

In vitro interaction of human Wharton's jelly mesenchymal stem cells with biomimetic 3D scaffold.

机构信息

Trauma Research Center, Aja University of Medical Sciences, Tehran, Iran.

Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.

出版信息

J Biomed Mater Res A. 2019 Jun;107(6):1166-1175. doi: 10.1002/jbm.a.36608. Epub 2019 Mar 18.

Abstract

Study of cell-biomaterial interaction is a crucial aspect of bone tissue engineering to find a state-of-the-art functional substitute. In present study, the Wharton's jelly mesenchymal stem cells (hWJ-MSCs) behavior on three-dimensional biomimetic nano-hydroxyapatite/chitosan/gelatin (nHA/CS/Gel) scaffolds was investigated. The outcome was assessed by histological, biochemical and morphological tests. Results indicated that hWJ-MSCs attached onto the scaffold surface through membrane filopodia, uniformly spread throughout the contacting surface. It only took 3 days for the seeded cells to appear deep inside the scaffold, reflecting proper hWJ-MSCs adhesion and migration, evidenced by both scanning electron microscope and hematoxilin and eosin assessments. Additionally, the present fabricated nHA/CS/Gel scaffold proved to be non-toxic as it supported cell proliferation measured by 3-(4,5-dimethylthiazoyl-2-yl)-2,5-diphenyltetrazolium bromide assay. Moreover, 3-week culture of hWJ-MSCs on scaffolds, immersed in osteogenic medium, rendered the microenvironment in favor of hWJ-MSCs differentiation into osteoblast cells and extracellular matrix secretion. Finally, osteoblasts were immunologically positive for various osteogenic markers including osteocalcin, osteopontin, osteonectin, and alkaline phosphatase. Present findings indicate that nHA/CS/Gel scaffold appropriately harbored hWJ-MSCs, stimulating their growth, migration, proliferation, and differentiation. hWJ-MSCs-loaded nHA/CS/gel substitute may therefore be considered as a suitable platform for the rising demand in in vivo bone repair studies. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1166-1175, 2019.

摘要

研究细胞-生物材料相互作用是骨组织工程的一个关键方面,旨在寻找最先进的功能替代品。在本研究中,研究了牙髓间充质干细胞(hWJ-MSCs)在三维仿生纳米羟基磷灰石/壳聚糖/明胶(nHA/CS/Gel)支架上的行为。通过组织学、生化和形态学测试来评估结果。结果表明,hWJ-MSCs 通过细胞膜丝状伪足附着在支架表面,均匀地分布在接触表面上。接种细胞仅需 3 天即可深入支架内部,这反映了适当的 hWJ-MSCs 黏附和迁移,扫描电子显微镜和苏木精-伊红评估也证明了这一点。此外,本研究中制备的 nHA/CS/Gel 支架被证明是无毒的,因为它支持细胞增殖,这是通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐测定法测量的。此外,在成骨培养基中培养 3 周的 hWJ-MSCs 支架,使微环境有利于 hWJ-MSCs 分化为成骨细胞和细胞外基质分泌。最后,成骨细胞对各种成骨标志物呈免疫阳性,包括骨钙素、骨桥蛋白、骨粘连蛋白和碱性磷酸酶。本研究结果表明,nHA/CS/Gel 支架适当地容纳了 hWJ-MSCs,刺激其生长、迁移、增殖和分化。负载 hWJ-MSCs 的 nHA/CS/gel 替代物因此可以被认为是满足体内骨修复研究需求的合适平台。 © 2019 Wiley Periodicals, Inc. J 生物材料 Res 部分 A:107A:1166-1175,2019。

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