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脊柱和矫形外科用生物材料的综合生物学评价

Comprehensive Biological Evaluation of Biomaterials Used in Spinal and Orthopedic Surgery.

作者信息

Komorowski Piotr, Siatkowska Małgorzata, Kamińska Marta, Jakubowski Witold, Walczyńska Marta, Walkowiak-Przybyło Magdalena, Szymański Witold, Piersa Katarzyna, Wielowski Patryk, Sokołowska Paulina, Białkowska Kamila, Makowski Krzysztof, Elgalal Marcin, Kierzkowska Agnieszka, Ciupik Lechosław, Walkowiak Bogdan

机构信息

Molecular and Nanostructural Biophysics Laboratory, "Bionanopark" Ltd., Dubois 114/116, 93-465 Lodz, Poland.

Division of Biophysics, Institute of Materials Science, Lodz University of Technology, Stefanowskiego 1/15, 90-924 Lodz, Poland.

出版信息

Materials (Basel). 2020 Oct 26;13(21):4769. doi: 10.3390/ma13214769.

DOI:10.3390/ma13214769
PMID:33114571
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7672648/
Abstract

Biological acceptance is one of the most important aspects of a biomaterial and forms the basis for its clinical use. The aim of this study was a comprehensive biological evaluation (cytotoxicity test, bacterial colonization test, blood platelets adhesion test and transcriptome and proteome analysis of Saos-2 cells after contact with surface of the biomaterial) of biomaterials used in spinal and orthopedic surgery, namely, Ti6Al4V ELI (Extra Low Interstitials), its modified version obtained as a result of melting by electron beam technology (Ti6Al4V ELI-EBT), polyether ether ketone (PEEK) and polished medical steel American Iron and Steel Institute (AISI) 316L (the reference material). Biological tests were carried out using the osteoblasts-like cells (Saos-2, ATCC HTB-85) and bacteria (DH5α). Results showed lack of cytotoxicity of all materials and the surfaces of both Ti6Al4V ELI and PEEK exhibit a significantly higher resistance to colonization with cells, while the more porous surface of the same titanium alloy produced by electron beam technology (EBT) is more susceptible to microbial colonization than the control surface of polished medical steel. None of the tested materials showed high toxicity in relation to cells. Susceptibility to platelet adhesion was very high for polished medical steel AISI 316L, whilst much lower for the other biomaterials and can be ranked from the lowest to the highest as follows: PEEK < Ti6Al4V ELI < Ti6Al4V ELI-EBT. The number of expressed genes in Saos-2 cells exposed to contact with the examined biomaterials reached 9463 genes in total (ranging from 8455 genes expressed in cells exposed to ELI to 9160 genes in cells exposed to PEEK). Whereas the number of differentially expressed proteins detected on two-dimensional electrophoresis gels in Saos-2 cells after contact with the examined biomaterials was 141 for PEEK, 223 for Ti6Al4V ELI and 133 for Ti6Al4V ELI-EBT. Finally, 14 proteins with altered expression were identified by mass spectrometry. In conclusion, none of the tested biomaterials showed unsatisfactory levels of cytotoxicity. The gene and protein expression analysis, that represents a completely new approach towards characterization of these biomaterials, showed that the polymer PEEK causes much more intense changes in gene and protein expression and thus influences cell metabolism.

摘要

生物相容性是生物材料最重要的方面之一，也是其临床应用的基础。本研究的目的是对脊柱和矫形外科手术中使用的生物材料进行全面的生物学评估（细胞毒性试验、细菌定植试验、血小板黏附试验以及与生物材料表面接触后Saos-2细胞的转录组和蛋白质组分析），这些生物材料包括Ti6Al4V ELI（超低间隙）、通过电子束技术熔炼得到的其改性版本（Ti6Al4V ELI-EBT）、聚醚醚酮（PEEK）以及经过抛光的美国钢铁协会（AISI）316L医用钢（参考材料）。使用成骨样细胞（Saos-2，ATCC HTB-85）和细菌（DH5α）进行生物学试验。结果显示所有材料均无细胞毒性，Ti6Al4V ELI和PEEK的表面对细胞定植均表现出显著更高的抗性，而通过电子束技术（EBT）生产的同一钛合金的多孔表面比经过抛光的医用钢对照表面更容易被微生物定植。所有测试材料对细胞均未显示出高毒性。对于经过抛光 的AISI 316L医用钢，其对血小板黏附的敏感性非常高，而其他生物材料的敏感性则低得多，从最低到最高排序如下：PEEK＜Ti6Al4V ELI＜Ti6Al4V ELI-EBT。与所检测生物材料接触的Saos-2细胞中表达的基因总数达到9463个（范围从接触ELI的细胞中表达的8455个基因到接触PEEK的细胞中的9160个基因）。而在与所检测生物材料接触后的Saos-2细胞的二维电泳凝胶上检测到的差异表达蛋白质数量，PEEK为141个，Ti6Al4V ELI为223个，Ti6Al4V ELI-EBT为133个。最后，通过质谱鉴定出14种表达发生改变的蛋白质。总之，所有测试的生物材料均未显示出不令人满意的细胞毒性水平。基因和蛋白质表达分析代表了一种全新的生物材料表征方法，结果表明聚合物PEEK会引起基因和蛋白质表达更强烈的变化，从而影响细胞代谢。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/5db4e4a94a64/materials-13-04769-g007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/5db4e4a94a64/materials-13-04769-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/9e09119b24aa/materials-13-04769-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/80db091275f4/materials-13-04769-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/53a0222440f9/materials-13-04769-g003.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/9a414a0d8d31/materials-13-04769-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f908/7672648/5db4e4a94a64/materials-13-04769-g007.jpg

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