School of Mechanical Engineering, University of Leeds, Leeds, LS2 9JT, UK.
UCL Centre for Nerve Engineering, UCL School of Pharmacy, University College London, London, UK.
Eur Spine J. 2020 Nov;29(11):2701-2712. doi: 10.1007/s00586-019-06177-w. Epub 2019 Oct 29.
Total disc replacements, comprising all-metal articulations, are compromised by wear and particle production. Metallic wear debris and ions trigger a range of biological responses including inflammation, genotoxicity, cytotoxicity, hypersensitivity and pseudotumour formation, therefore we hypothesise that, due to proximity to the spinal cord, glial cells may be adversely affected.
Clinically relevant cobalt chrome (CoCr) and stainless steel (SS) wear particles were generated using a six-station pin-on-plate wear simulator. The effects of metallic particles (0.5-50 μm debris per cell) and metal ions on glial cell viability, cellular activity (glial fibrillary acidic protein (GFAP) expression) and DNA integrity were investigated in 2D and 3D culture using live/dead, immunocytochemistry and a comet assay, respectively.
CoCr wear particles and ions caused significant reductions in glial cell viability in both 2D and 3D culture systems. Stainless steel particles did not affect glial cell viability or astrocyte activation. In contrast, ions released from SS caused significant reductions in glial cell viability, an effect that was especially noticeable when astrocytes were cultured in isolation without microglia. DNA damage was observed in both cell types and with both biomaterials tested. CoCr wear particles had a dose-dependent effect on astrocyte activation, measured through expression of GFAP.
The results from this study suggest that microglia influence the effects that metal particles have on astrocytes, that SS ions and particles play a role in the adverse effects observed and that SS is a less toxic biomaterial than CoCr alloy for use in spinal devices. These slides can be retrieved under Electronic Supplementary Material.
全金属关节的全椎间盘置换物会因磨损和颗粒生成而受损。金属磨损颗粒和离子会引发一系列生物学反应,包括炎症、遗传毒性、细胞毒性、过敏反应和假瘤形成,因此我们假设,由于靠近脊髓,神经胶质细胞可能会受到不良影响。
使用六工位销盘式磨损模拟器生成临床相关的钴铬(CoCr)和不锈钢(SS)磨损颗粒。通过使用活/死染色、免疫细胞化学和彗星试验,分别在 2D 和 3D 培养物中研究金属颗粒(每个细胞 0.5-50μm 碎片)和金属离子对神经胶质细胞活力、细胞活性(胶质纤维酸性蛋白(GFAP)表达)和 DNA 完整性的影响。
CoCr 磨损颗粒和离子在 2D 和 3D 培养系统中均导致神经胶质细胞活力显著降低。不锈钢颗粒不会影响神经胶质细胞活力或星形胶质细胞激活。相比之下,从 SS 释放的离子会导致神经胶质细胞活力显著降低,当星形胶质细胞在没有小胶质细胞的情况下单独培养时,这种影响尤其明显。在两种细胞类型和两种测试的生物材料中都观察到了 DNA 损伤。CoCr 磨损颗粒对星形胶质细胞激活具有剂量依赖性影响,通过 GFAP 表达来衡量。
本研究结果表明,小胶质细胞影响金属颗粒对星形胶质细胞的影响,SS 离子和颗粒在观察到的不良反应中起作用,并且 SS 是用于脊柱装置的比 CoCr 合金毒性更小的生物材料。这些幻灯片可在电子补充材料中检索。