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 Escherichia coli (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 E. coli 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 E. coli 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.
Highlights
It is well known that bone tissue is constantly subjected to the dynamic processes of remodeling
Susceptibility to platelet adhesion was very high for polished medical steel AISI 316L, while much lower for the other biomaterials and can be ranked from lowest to highest—polyether ether ketone (PEEK), Ti6Al4V ELI and Ti6Al4V ELI-electron beam technology (EBT)
The number of expressed genes in Saos-2 cells exposed to contact with the examined biomaterials reached 9463 genes in total
Summary
It is well known that bone tissue is constantly subjected to the dynamic processes of remodeling. This process is carried out by osteoblasts, which produce and secrete matrix proteins and transport minerals into the bone matrix, as well as osteoclasts that break down bone tissue. It cannot be ruled out that direct contact of an implant surface with bone tissue has an impact on the process of bone remodeling and osteointegration. This effect may possibly be manifested by changes in gene expression caused by exposure and direct interaction of osteoblasts with an implant surface. It is becoming increasingly obvious that a holistic approach must be applied in order to understand the varied processes that determine the successful use of a particular implant
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