拉伸条件下聚二甲基硅氧烷表面粗糙度对骨髓间充质干细胞成骨分化的影响

Effect of surface roughness of polydimethylsiloxane on osteogenic differentiation of bone marrow mesenchymal stem cells under stretching conditions

胡泽遵 ¹杨反磊 ¹徐浩 ¹罗宗平¹

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作者信息

1. 苏州大学附属第一医院骨科研究所,江苏省苏州市 215000
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摘要

背景:研究表明,机械刺激对骨髓间充质干细胞的谱系特异性分化至关重要.然而,机械拉伸条件下不同粗糙度表面的骨髓间充质干细胞成骨分化情况尚不清楚.目的:探讨拉伸条件下聚二甲基硅氧烷(polydimethylsiloxane,PDMS)表面粗糙度对骨髓间充质干细胞成骨分化的影响及作用机制.方法:通过不同目砂纸(120目、1 000目和10 000目)在PDMS表面构建3种不同粗糙度的形貌(PDMS-120M、PDMS-1000M和PDMS-10000M),与空气接触的PDMS表面作为对照组.采用RT-qPCR检测静态条件下和拉伸条件下(0%,2%,4%,6%拉伸幅度)不同粗糙度PDMS表面的骨髓间充质干细胞中成骨相关基因表达.采用RT-qPCR、Western blot检测2%拉伸条件下不同粗糙度表面骨髓间充质干细胞中SIRT1、成骨相关基因和蛋白表达,采用碱性磷酸酶染色、茜素红染色观察2%拉伸条件下不同粗糙度PDMS表面骨髓间充质干细胞的成骨分化能力.结果与结论:①静态条件下粗糙度为(13.51±2.11)µm PDMS-1000M表面的骨髓间充质干细胞成骨基因表达最高;②相对光滑PDMS表面的骨髓间充质干细胞在4%拉伸幅度条件下有更好的成骨分化效果,而PDMS-1000M表面的骨髓间充质干细胞在2%拉伸幅度条件下有更好的成骨分化效果;③在2%拉伸幅度条件下,粗糙度为(13.51±2.11)µm PDMS-1000M表面的骨髓间充质干细胞有更好的成骨分化效果,可能与激活SIRT1信号通路有关.

Abstract

BACKGROUND:Numerous studies have shown that mechanical stimulation is essential for the lineage-specific differentiation of bone marrow mesenchymal stem cells.However,osteogenic differentiation of bone marrow mesenchymal stem cells on surfaces with different roughnesses under mechanical stretching conditions is unknown.OBJECTIVE:To investigate the effects and action mechanisms of different roughness surfaces of polydimethylsiloxane(PDMS)on osteogenic differentiation of bone marrow mesenchymal stem cells under stretching conditions.METHODS:Three morphologies with different roughnesses(PDMS-120M,PDMS-1000M,and PDMS-10000M)were constructed on PDMS surfaces by means of different grits of sandpaper(120 grits,1 000 grits and 10 000 grits),and PDMS surfaces in contact with air served as a control group.With different amplitudes of 0%,2%,4%,and 6%,osteogenesis-related gene expression of bone marrow mesenchymal stem cells on different PDMS surfaces under static and stretching conditions was detected by RT-qPCR.RT-qPCR and western blot assay were used to detect the expression of SIRT1 gene and protein as well as osteogenesis-related genes and proteins in bone marrow mesenchymal stem cells on different roughness surfaces under 2%stretching conditions.Alkaline phosphatase staining and alizarin red staining were further used to observe the osteogenic differentiation ability of bone marrow mesenchymal stem cells on different PDMS surfaces under 2%stretching conditions.RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells on the PDMS-1000M surface with a roughness of(13.51±2.11)µm had better osteogenic gene expression under static conditions.(2)Bone marrow mesenchymal stem cells on the PDMS surface in contact with air had better osteogenic differentiation under 4%stretching conditions,while bone marrow mesenchymal stem cells on the PDMS-1000M surface had better osteogenic differentiation under 2%stretching conditions.(3)Bone marrow mesenchymal stem cells on the PDMS-1000M surface with a roughness of(13.51±2.11)µm had better osteogenic differentiation under 2%stretching conditions,which may be related to activation of SIRT1 signaling pathway.

关键词

骨髓间充质干细胞/成骨分化/机械刺激/机械拉伸/粗糙度/信号通路/SIRT1

Key words

bone marrow mesenchymal stem cell/osteogenic differentiation/mechanical stimulation/mechanical stretching/roughness/signaling pathway/SIRT1

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基金项目

国家自然科学基金(32071307)

国家自然科学基金(11802191)

出版年

2025

中国组织工程研究

中国康复医学会,《中国组织工程研究与临床康复》杂志社

中国组织工程研究

北大核心

影响因子：1.387

ISSN：2095-4344

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