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人牙髓干细胞和成纤维细胞暴露于羟基磷灰石纳米颗粒后的成骨潜能:一项体外比较研究。

Osteogenic potential of human dental pulp stem cells, and human dermal fibroblasts exposed hydroxyapatite nanoparticles: A comparative in vitro study.

作者信息

Naseri Mohsen, Sarfi Sepideh, Hanafi-Bojd Mohammad Yahya, Azaryan Ehsaneh

机构信息

Cellular and Molecular Research Center, School of Medicine, Birjand University of Medical Sciences, Birjand, Iran.

Student Research Committee, Birjand University of Medical Sciences, Birjand, Iran.

出版信息

Int J Mol Cell Med. 2025 Jul 1;14(2):656-665. doi: 10.22088/IJMCM.BUMS.14.2.656. eCollection 2025.

Abstract

This study aimed to evaluate the osteogenic response of human dermal fibroblasts (hDFs) and human dental pulp stem cells (hDPSCs) when exposed to hydroxyapatite nanoparticles (HA-NPs), which are commonly employed biomaterials in the field of bone regeneration. The sol-gel method was employed to synthesize HA-NPs. Cell viability was assessed using the MTT assay after 24 and 72 hours of exposure to HA-NPs. Osteogenic differentiation was assessed through Alizarin red S staining (ARS), alkaline phosphatase (ALP) activity assay, and quantitative real-time PCR (qPCR) to evaluate the expression levels of osteogenic markers, including BMP-2, VEGF-A, RUNX2, and IL-6. The HA-NPs had a nanorod shape, with dimensions of 17-29 nm in width and 62-89 nm in length. Both hDPSCs and hDFs demonstrated enhanced osteogenic potential when exposed to HA-NPs, as indicated by increased ARS staining for calcium deposition and elevated ALP activity. Gene expression analysis showed up-regulation of BMP-2 and VEGF-A and down-regulation of RUNX2 in both cell types. IL-6 expression markedly increased in hDFs but did not show significant changes in hDPSCs compared to the control group. The findings of this study suggest that HA-NPs may enhance the osteogenic capability of hDPSCs and hDFs. The results demonstrate that while fibroblasts exhibit some mineralization potential, hDPSCs are the most suitable cell type for bone regenerative applications.

摘要

本研究旨在评估人真皮成纤维细胞(hDFs)和人牙髓干细胞(hDPSCs)在暴露于羟基磷灰石纳米颗粒(HA-NPs)时的成骨反应,HA-NPs是骨再生领域常用的生物材料。采用溶胶-凝胶法合成HA-NPs。在暴露于HA-NPs 24小时和72小时后,使用MTT法评估细胞活力。通过茜素红S染色(ARS)、碱性磷酸酶(ALP)活性测定和定量实时PCR(qPCR)评估成骨分化,以评估包括骨形态发生蛋白-2(BMP-2)、血管内皮生长因子-A(VEGF-A)、Runx2和白细胞介素-6(IL-6)在内的成骨标志物的表达水平。HA-NPs呈纳米棒形状,宽度为17-29nm,长度为62-89nm。hDPSCs和hDFs在暴露于HA-NPs时均表现出增强的成骨潜力,钙沉积的ARS染色增加和ALP活性升高表明了这一点。基因表达分析显示,两种细胞类型中BMP-2和VEGF-A均上调,Runx2下调。与对照组相比,hDFs中IL-6表达明显增加,但hDPSCs中未显示出显著变化。本研究结果表明,HA-NPs可能增强hDPSCs和hDFs的成骨能力。结果表明,虽然成纤维细胞表现出一定的矿化潜力,但hDPSCs是最适合骨再生应用的细胞类型。

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