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纳米结构二氧化钛表面促进人骨髓间充质干细胞向成骨细胞分化。

Nanostructured TiO₂ Surfaces Promote Human Bone Marrow Mesenchymal Stem Cells Differentiation to Osteoblasts.

作者信息

Vercellino Marco, Ceccarelli Gabriele, Cristofaro Francesco, Balli Martina, Bertoglio Federico, Bruni Gianna, Benedetti Laura, Avanzini Maria Antonietta, Imbriani Marcello, Visai Livia

机构信息

Department of Molecular Medicine, Center for Health Technologies (CHT), UdR INSTM, University of Pavia, Viale Taramelli 3/b, Pavia 27100, Italy.

Department of Public Health, Experimental Medicine and Forensic, Human Anatomy Unit, Center of Health Technologies (CHT), University of Pavia, Viale Forlanini 8, Pavia 27100, Italy.

出版信息

Nanomaterials (Basel). 2016 Jun 24;6(7):124. doi: 10.3390/nano6070124.

DOI:10.3390/nano6070124
PMID:28335251
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5224601/
Abstract

Micro- and nano-patterning/modification are emerging strategies to improve surfaces properties that may influence critically cells adherence and differentiation. Aim of this work was to study the in vitro biological reactivity of human bone marrow mesenchymal stem cells (hBMSCs) to a nanostructured titanium dioxide (TiO₂) surface in comparison to a coverglass (Glass) in two different culture conditions: with (osteogenic medium (OM)) and without (proliferative medium (PM)) osteogenic factors. To evaluate cell adhesion, hBMSCs phosphorylated focal adhesion kinase (pFAK) foci were analyzed by confocal laser scanning microscopy (CLSM) at 24 h: the TiO₂ surface showed a higher number of pFAK foci with respect to Glass. The hBMSCs differentiation to osteoblasts was evaluated in both PM and OM culture conditions by enzyme-linked immunosorbent assay (ELISA), CLSM and real-time quantitative reverse transcription PCR (qRT-PCR) at 28 days. In comparison with Glass, TiO₂ surface in combination with OM conditions increased the content of extracellular bone proteins, calcium deposition and alkaline phosphatase activity. The qRT-PCR analysis revealed, both in PM and OM, that TiO₂ surface increased at seven and 28 days the expression of osteogenic genes. All together, these results demonstrate the capability of TiO₂ nanostructured surface to promote hBMSCs osteoblast differentiation and its potentiality in biomedical applications.

摘要

微纳图案化/修饰是改善表面性质的新兴策略，这些表面性质可能对细胞黏附和分化产生关键影响。本研究的目的是在两种不同培养条件下，即添加（成骨培养基（OM））和不添加（增殖培养基（PM））成骨因子的情况下，研究人骨髓间充质干细胞（hBMSCs）对纳米结构二氧化钛（TiO₂）表面与盖玻片（玻璃）相比的体外生物反应性。为了评估细胞黏附，在24小时时通过共聚焦激光扫描显微镜（CLSM）分析hBMSCs磷酸化黏着斑激酶（pFAK）焦点：TiO₂表面相对于玻璃显示出更多的pFAK焦点。在28天时，通过酶联免疫吸附测定（ELISA）、CLSM和实时定量逆转录PCR（qRT-PCR）在PM和OM培养条件下评估hBMSCs向成骨细胞的分化。与玻璃相比，TiO₂表面在OM条件下增加了细胞外骨蛋白的含量、钙沉积和碱性磷酸酶活性。qRT-PCR分析显示，在PM和OM中，TiO₂表面在第7天和第28天均增加了成骨基因的表达。总之，这些结果证明了TiO₂纳米结构表面促进hBMSCs成骨细胞分化的能力及其在生物医学应用中的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/a08c65a96254/nanomaterials-06-00124-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/76ef1199ca5c/nanomaterials-06-00124-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/a08c65a96254/nanomaterials-06-00124-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/76ef1199ca5c/nanomaterials-06-00124-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/d907f85f12ce/nanomaterials-06-00124-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/4f3aee0f225d/nanomaterials-06-00124-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/5450f2c62b41/nanomaterials-06-00124-g004.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/11b8/5224601/a08c65a96254/nanomaterials-06-00124-g007.jpg

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