Effect of Surface Chemistry on Hemolysis, Thrombogenicity, and Toxicity of Carbon Nanotube Doped Thermally Sprayed Hydroxyapatite Implants.

Assessing blood compatibility is crucial before in vivo procedures and is considered more reliable than many in vitro tests. This study examines the physiochemical properties and blood compatibility of bioactive powders ((0.5-2 wt % carbon nanotube (CNT)/alumina)-20 wt %)) produced through a heterocoagulation colloidal technique followed by ball milling with hydroxyapatite (HAp). The 1 wt % CNT composite demonstrated a surface charge ∼5 times higher than HAp at pH 7.4, with a value of -11 mV compared to -2 mV. This increase in electrostatic charge is desirable for achieving hemocompatibility, as evidenced by a range of blood compatibility assessments, including hemolysis, blood clotting, platelet adhesion, platelet activation, and coagulation assays (prothrombin time (PT) and activated partial thrombin time (aPTT)). The 1 wt % CNT composite exhibited hemolysis ranging from 2 to 7%, indicating its hemocompatibility. In the blood clot investigation, the absorbance values for 1-2 wt % CNT samples were 0.927 ± 0.038 and 1.184 ± 0.128, respectively, indicating their nonthrombogenicity. Additionally, the percentage of platelet adhered on the 1 wt % CNT sample (∼5.67%) showed a ∼2.5-fold decrement compared to the clinically used negative control, polypropylene (∼13.73%). The PT and aPTT experiments showed no difference in the coagulation time for CNT samples even at higher concentrations, unlike HAC2 (80 mg). In conclusion, the 1 wt % CNT sample was nontoxic to human blood, making it more hemocompatible, nonhemolytic, and nonthrombogenic than other samples. This reliable study reduces the need for additional in vitro and in vivo studies before clinical trials, saving time and cost.

Thermal spray processes influencing surface chemistry and in-vitro hemocompatibility of hydroxyapatite-based orthopedic implants.

Orthopedic implants made from titanium are a popular choice in the medical field because of their remarkable strength-to-weight ratio. Nevertheless, they may not interact well with human blood, resulting in thrombosis and hemolysis. In fact, non-hemocompatibility is believed to be responsible for about 31 % of medical device failures in the US alone, requiring painful and expensive revision surgery. To address this issue, bioactive hydroxyapatite coatings are applied to Ti-6Al-4V implants using thermal spray techniques. However, the temperature used during thermal processing impacts the coating's surface properties, affecting the mechanical and biological properties. Furthermore, the effectiveness of HA coatings on titanium for orthopedic applications has not been validated by biocompatibility tests, particularly hemocompatibility. In this study, we aimed to investigate the relative efficacy of three thermal spray processes of different temperature ranges: Atmospheric plasma spray (APS) (high temperature), Flame spray (FS) (moderate temperature), and High-Velocity Oxy-Fuel spray (HVOF) (low temperature), and study their impact on coating's surface properties, affecting blood components and implant's strength. The crystallinity of the HA coating increased by 32 % with a decrease in the operating temperature (APS < FS < HVOF). HVOF coating exhibited a ~ 34 % and ~ 120 % improvement in adhesion strength and ~ 31 % and 59 % increment in hardness compared to APS and FS coating, respectively, attributed to its low porosity, low coating thickness (~55 µm), and high degree of crystallinity. The HVOF coating showcased a significant increase in non-hemolytic behavior, with hemolysis rates ~8 and ~ 11 times lower than APS and FS coatings, respectively, owing to its smooth texture and high degree of crystallinity (p < 0.05). Furthermore, the HVOF coating exhibited minimal blood clotting based on the whole blood clotting assay, again confirmed by PT and aPTT assays showing delayed clotting time, indicating its non-thrombogenic behavior. The number of platelets adhered to the three coatings showed no significant difference compared to Ti-6Al-4V. APS and FS coatings showed low platelet activation, unlike HVOF coating and titanium, which revealed round platelets, similar to the negative control. Neither titanium nor HA coatings exhibited antibacterial properties, which may be due to their high affinity for organic substances, which promotes bacterial adhesion and replication. Among the three thermal processes, HVOF coating displayed good apatite growth, non-hemolytic, and non-thrombogenicity with no platelet activation owing to its low processing temperature, high degree of crystallinity (89.7 %), hydrophilicity, smooth (~4 µm) and dense (~97 %) microstructural properties. The results demonstrated that the HVOF-HA coating presented in this work meets the hemocompatible requirements and shows promise for prospective application as an orthopedic implant. Furthermore, this study has the potential to significantly reduce the use of animals in in-vivo research and improve their welfare while also cutting costs.