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Microarray analysis reveals that lncRNA PWRN1-209 promotes human bone marrow mesenchymal stem cell osteogenic differentiation on microtopography titanium surface in vitro.
Journal of Biomedical Materials Research Part B: Applied Biomaterials ( IF 3.2 ) Pub Date : 2020-05-24 , DOI: 10.1002/jbm.b.34620
Mingyue Wang 1 , Xiyuan Ge 2 , Yan Zheng 1 , Chenxi Wang 1 , Yu Zhang 1 , Ye Lin 1
Affiliation  

Sandblasted, large‐grit, and acid‐etched (SLA) titanium (Ti) with microtopography is currently one of the most widely used implant materials to accelerate osseointegration. Numerous long noncoding RNAs (lncRNAs) have been involved in bone remodeling, with their role in osseointegration, and the underlying mechanisms remain largely unclear. Here, microarrays of human bone marrow mesenchymal stem cells (hBMSCs) were used to identify differentially expressed lncRNAs during early cell differentiation stages (0–7 days) on SLA Ti and polished Ti surfaces. The function of lncRNAs in the osteogenic differentiation of hBMSCs was identified by RNA silencing and overexpression assays. RT‐PCR and Western blot were used to detect RNA and protein expression. Alkaline phosphatase (ALP) protein activity was tested by ALP staining. Altogether, 4112 differentially expressed lncRNAs were identified from day 0 to day 7 on SLA Ti with a novel lncRNA, Prader‐willi region non‐coding RNA 1‐209 (PWRN1‐209) upregulated. We then proved that PWRN1‐209 promoted osteogenic differentiation in hBMSCs by genetic tools. The upregulation of PWRN1‐209 was further confirmed to be related to the surface topography of Ti by comparing SLA Ti and polished Ti. Interestingly, this trend seems to have a certain correlation with the mRNA expression level of integrins (α2, αV, β1, β2) and the phosphorylation of focal adhesion kinase (FAK). Taken together, the lncRNA PWRN1‐209 was upregulated by the SLA microtopography Ti surface, which may regulate osteogenic differentiation of hBMSCs through integrin‐FAK‐ALP signaling. Our results provide new insights into the relationship between surface topography and osseointergration.

中文翻译:

微阵列分析表明,lncRNA PWRN1-209 在体外促进人骨髓间充质干细胞成骨微地形钛表面的分化。

具有微形貌的喷砂、大粒度和酸蚀刻 (SLA) 钛 (Ti) 是目前最广泛使用的加速骨整合的植入材料之一。许多长链非编码 RNA (lncRNA) 已参与骨重塑,并在骨整合中发挥作用,但其潜在机制尚不清楚。在这里,人骨髓间充质干细胞 (hBMSCs) 的微阵列用于在 SLA Ti 和抛光的 Ti 表面的早期细胞分化阶段(0-7 天)识别差异表达的 lncRNA。lncRNAs 在 hBMSCs 成骨分化中的功能通过 RNA 沉默和过表达测定来鉴定。RT-PCR 和蛋白质印迹用于检测 RNA 和蛋白质表达。通过 ALP 染色测试碱性磷酸酶 (ALP) 蛋白活性。共,从第 0 天到第 7 天,在 SLA Ti 上鉴定了 4112 种差异表达的 lncRNA,其中一种新型 lncRNA、Prader-willi 区域非编码 RNA 1-209 (PWRN1-209) 上调。然后我们通过遗传工具证明了 PWRN1-209 促进了 hBMSCs 的成骨分化. 通过比较 SLA Ti 和抛光钛,进一步证实 PWRN1-209 的上调与 Ti 的表面形貌有关。有趣的是,这种趋势似乎与整合素(α2、αV、β1、β2)的mRNA表达水平和粘着斑激酶(FAK)的磷酸化有一定的相关性。总之,lncRNA PWRN1-209被SLA微地形Ti表面上调,这可能通过整合素-FAK-ALP信号调节hBMSCs的成骨分化。我们的结果为表面形貌和骨整合之间的关系提供了新的见解。
更新日期:2020-05-24
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