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TGF-β1 和 -β3 对聚(乙烯醇)-壳聚糖-聚(乙二醇)支架上间充质干细胞软骨分化的影响。

TGF-β1 and -β3 for Mesenchymal Stem Cells Chondrogenic Differentiation on Poly (Vinyl Alcohol)-Chitosan-Poly (Ethylene Glycol) Scaffold.

机构信息

Department of Orthopaedic Surgery, Faculty of Medicine, Tissue Engineering Group (TEG), National Orthopaedic Centre of Excellent Research and Learning (NOCERAL), University of Malaya, Kuala Lumpur, Malaysia.

Faculty of Medicine, Nursing & Health Sciences, SEGi University, Kota Damansara, Petaling Jaya, Malaysia.

出版信息

Tissue Eng Part C Methods. 2022 Oct;28(10):501-510. doi: 10.1089/ten.TEC.2022.0112.

Abstract

Transforming growth factor-beta 1 (TGF-β1) has been reported to promote chondrogenic differentiation and proliferation in the multipotent stromal cell (MSCs), and the transforming growth factor-beta 3 (TGF-β3) tends to be exclusively in promoting cell differentiation alone. The objective of this study was to determine the effect of TGF-β1 and -β3 on the MSCs chondrogenic differentiation on the poly (vinyl alcohol)-chitosan-poly (ethylene glycol) (PVA-NOCC-PEG) scaffold, compared with that of monolayer and pellet cultures. In this study, P2 rabbit bone marrow-derived MSCs were seeded either on the untreated six-well plate (for monolayer culture) or onto the PVA-NOCC-PEG scaffold or cultured as a pellet culture. The cultures were maintained in a chemically defined serum-free medium supplemented with 10 ng/mL of either TGF-β1 or TGF-β3. Cell viability assay, biochemical assay, and real-time polymerase chain reaction were performed to determine the net effect of cell proliferation and chondrogenic differentiation of each of the growth factors. The results showed that the PVA-NOCC-PEG scaffold enhanced MSCs cell proliferation from day 12 to 30 ( < 0.05); however, no significant differences were observed in the cell proliferation between the cultures supplemented with or without TGF-β1 and TGF-β3 ( > 0.05). In terms of chondrogenic differentiation, the PVA-NOCC-PEG scaffold augmented the GAGs secretion in MSCs and the mRNA expression levels of , , , and were elevated ( < 0.05). However, there was no significant difference between both the TGF-β1 and TGF-β3-treated groups ( > 0.05). In conclusion, TGF-β1 and TGF-β3 enhanced the chondrogenic differentiation of MSCs seeded on the PVA-NOCC-PEG scaffold; however, there was no significant difference between the effect of TGF-β1 and TGF-β3. Impact statement Transforming growth factor-beta (TGF-β) superfamily members is a key requirement for the chondrogenic differentiation of mesenchymal stem cells (MSCs). In this study, the effects of TGF-β1 and -β3 on MSC chondrogenic differentiation and proliferation on a novel three-dimensional scaffold, the poly(vinyl alcohol)-chitosan-poly(ethylene glycol) (PVA-NOCC-PEG) scaffold, was evaluated. In this study, the results showed both TGF-β1 and TGF-β3 can enhance the chondrogenic differentiation of MSCs seeded on the PVA-NOCC-PEG scaffold.

摘要

转化生长因子-β 1(TGF-β1)已被报道能促进多能基质细胞(MSCs)的软骨分化和增殖,而转化生长因子-β 3(TGF-β3)则倾向于单独促进细胞分化。本研究的目的是确定 TGF-β1 和 -β3 对 PVA-NOCC-PEG 支架上 MSCs 软骨分化的影响,与单层和微球培养相比。在这项研究中,将 P2 兔骨髓来源的 MSCs 接种在未经处理的六孔板上(用于单层培养)或 PVA-NOCC-PEG 支架上,或作为微球培养。将培养物在补充有 10ng/mL 以下任一种生长因子的无血清化学定义培养基中维持。进行细胞活力测定、生化测定和实时聚合酶链反应,以确定每种生长因子对细胞增殖和软骨分化的净效应。结果表明,PVA-NOCC-PEG 支架从第 12 天到第 30 天增强了 MSCs 的细胞增殖(<0.05);然而,在补充或不补充 TGF-β1 和 TGF-β3 的培养物之间,细胞增殖没有显著差异(>0.05)。就软骨分化而言,PVA-NOCC-PEG 支架增加了 MSCs 中 GAG 的分泌,并上调了 、 、 和 的 mRNA 表达水平(<0.05)。然而,TGF-β1 和 TGF-β3 处理组之间没有显著差异(>0.05)。总之,TGF-β1 和 TGF-β3 增强了接种在 PVA-NOCC-PEG 支架上的 MSCs 的软骨分化;然而,TGF-β1 和 TGF-β3 的作用没有显著差异。影响说明转化生长因子-β(TGF-β)超家族成员是间充质干细胞(MSCs)软骨分化的关键要求。在这项研究中,评估了 TGF-β1 和 -β3 对新型三维支架——聚乙烯醇-壳聚糖-聚乙二醇(PVA-NOCC-PEG)支架上 MSC 软骨分化和增殖的影响。在这项研究中,结果表明 TGF-β1 和 TGF-β3 均可增强接种在 PVA-NOCC-PEG 支架上的 MSCs 的软骨分化。

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