Probing Cytocompatibility, Hemocompatibility, and Quantitative Inflammatory Response in Mus musculus toward Oxide Bioceramic Wear Particulates and a Comparison with CoCr

Abstract

Peri-prosthetic bone resorption and loosening of artificial joints have been widely recognized to limit the performance of the load-bearing implants. Therefore, the present study probes into the cyto-, hemo-, and histocompatibility of the small sized wear particulates of ZrO2-toughened Al2O3 (ZTA). In order to develop a comprehensive, yet clinically relevant understanding, a comparison is made with two baseline ceramics (Al2O3 and ZrO2) and metallic material (CoCr), all in particulate forms. While in vitro cytotoxicity assessment was carried out with mouse osteoblast cells, preclinical testing of intra-articularly injected particulates up to the concentration 25 wt % in PBS over the period of 12 weeks was performed in mouse model. Interestingly, mouse osteoblast cells, cultured in media with ZTA, Al2O3, and ZrO2 particulates of three different concentrations (0.25, 2.5, and 25 mg/mL) exhibited uncompromised cell viability and considerable cell spreading, up to the time frame of 72 h. In contrast, similar experiments with CoCr particulates demonstrated significant decrease in cellular growth with drastic change in osteoblast proliferation behavior. Furthermore, the decrease in RBC damage after contact with ZTA, Al2O3, and ZrO2 particulates at 25 mg/mL of dose level illustrated 4.4, 5.8, and 1.25% hemolysis, respectively, confirming clinically acceptable hemocompatibility. However, hematic activity of CoCr particles was reflected with 15.6% of hemolysis. In vivo, the absence of any significant effect of intra-articularly injected ceramic (Al2O3, ZrO2, ZTA) as well as metallic (CoCr) particulates on complete hemogram and serum biochemistry of Balb/C mice was recorded at all the time points up to 12 weeks. The extensive histological analysis confirmed the absence of any signature of the tissue-level toxicity at all time points. The pro-inflammatory cytokine analysis using TNF-α and IL-1β markers provided complementary evidence toward nongranulomatous and nonimmunogenic response of synovial membrane of knee joint and other vital organs of mice that were exposed to Al2O3, ZrO2, ZTA, and CoCr particulates. Taken together, our results establish the nontoxic nature of oxide ceramic particulates to bone cells, in vitro, as well as to periprosthetic tissue, in vivo.