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