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钛合金支架对酶促抗氧化应激防御及骨髓细胞分化的影响

Influence of Titanium Alloy Scaffolds on Enzymatic Defense against Oxidative Stress and Bone Marrow Cell Differentiation.

作者信息

Rodrigues Lais Morandini, Lima Zutin Elis Andrade, Sartori Elisa Mattias, Mendonça Daniela Baccelli Silveira, Mendonça Gustavo, Carvalho Yasmin Rodarte, Reis de Vasconcellos Luana Marotta

机构信息

Department of Biosciences and Oral Diagnosis, São Paulo State University (UNESP), Institute of Science and Technology, São José dos Campos, Brazil.

Department of Biological Sciences, Oakland University, Rochester Hills, MI, USA.

出版信息

Int J Biomater. 2020 Jul 29;2020:1708214. doi: 10.1155/2020/1708214. eCollection 2020.

Abstract

Studies have been directed towards the production of new titanium alloys, aiming for the replacement of Ti-6 Aluminium-4 Vanadium (TiAlV) alloy in the future. Many mechanisms related to biocompatibility and chemical characteristics have been studied in the field of implantology, but enzymatic defenses against oxidative stress remain underexplored. Bone marrow stromal cells have been explored as source of cells, which have the potential to differentiate into osteoblasts and therefore could be used as cells-based therapy. The objective of this study was to evaluate the activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) in porous scaffolds of Ti-6 Aluminium-4 Vanadium (TiAlV), Ti-35 Niobium (TiNb), and Ti-35 Niobium-7 Zirconium-5 Tantalum (TiNbZrTa) on mouse bone marrow stromal cells. Porous titanium alloy scaffolds were prepared by powder metallurgy. After 24 hours, cells plated on the scaffolds were analyzed by scanning electron microscopy (SEM). The antioxidant enzyme activity was measured 72 hours after cell plating. Quantitative real time PCR (qRT-PCR) was performed after 3, 7, and 14 days, and (Runt-related transcription factor2) expression was evaluated. The SEM images showed the presence of interconnected pores and growth, adhesion, and cell spreading in the 3 scaffolds. Although differences were noted for SOD and CAT activity for all scaffolds analyzed, no statistical differences were observed ( > 0.05). The osteogenic gene presented high expression levels for TiNbZrTa at day 7, compared to the control group (TiAlV day 3). At day 14, all scaffolds had more than 2-fold induction for mRNA levels, with statistically significant differences compared to the control group. Even though we were not able to confirm statistically significant differences to justify the replacement of TiAlV regarding antioxidant enzymes, TiNbZrTa was able to induce faster bone formation at early time points, making it a good choice for biomedical and tissue bioengineering applications.

摘要

研究一直致力于新型钛合金的生产,目标是在未来取代Ti-6铝-4钒(TiAlV)合金。在植入学领域,已经研究了许多与生物相容性和化学特性相关的机制,但针对氧化应激的酶防御机制仍未得到充分探索。骨髓基质细胞已被作为细胞来源进行研究,这些细胞有分化为成骨细胞的潜力,因此可用于基于细胞的治疗。本研究的目的是评估Ti-6铝-4钒(TiAlV)、Ti-35铌(TiNb)和Ti-35铌-7锆-5钽(TiNbZrTa)多孔支架对小鼠骨髓基质细胞中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)抗氧化酶活性的影响。通过粉末冶金制备多孔钛合金支架。24小时后,通过扫描电子显微镜(SEM)分析接种在支架上的细胞。细胞接种72小时后测量抗氧化酶活性。在3天、7天和14天后进行定量实时聚合酶链反应(qRT-PCR),并评估(与 runt 相关的转录因子2)表达。SEM图像显示在这3种支架中存在相互连接的孔隙以及细胞生长、黏附和铺展。尽管在所分析的所有支架的SOD和CAT活性方面存在差异,但未观察到统计学差异(>0.05)。与对照组(TiAlV第3天)相比,成骨基因在第7天TiNbZrTa呈现高表达水平。在第14天,所有支架的mRNA水平诱导倍数均超过2倍,并与对照组相比有统计学显著差异。尽管我们无法在统计学上确认有显著差异来证明用其他材料取代TiAlV在抗氧化酶方面的合理性,但TiNbZrTa能够在早期诱导更快的骨形成,使其成为生物医学和组织生物工程应用的良好选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/390d/7411454/039e58839465/IJBM2020-1708214.001.jpg

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