Effects of vitamin C on osteoblast proliferation and osteosarcoma inhibition using plasma coated hydroxyapatite on titanium implants.

Plasma-sprayed hydroxyapatite (HAp) coated titanium (Ti) implants are being extensively used in orthopedic surgeries and post-tumor resection to repair load-bearing segmental bone defects. In this study, vitamin C, an abundantly available natural biomolecule, is loaded onto plasma-sprayed HAp-coated commercially pure titanium (cpTi) surface to evaluate its chemopreventive and osteogenic properties, suggesting its clinical significance as an alternative or adjunct therapy in the treatment for osteosarcoma bone resection. Controlled release of vitamin C from HAp coated cpTi implant is assessed by in vitro drug release study, where Korsmeyer-Peppas model was applied to understand the release kinetics. After 21 days, the implants loaded with 400 and 800 μg of vitamin C showed a cumulative release of 62.7 and 74.1% in acidic microenvironment, whereas, 50.9% and 53.1% of total vitamin C release were observed by the implants loaded with 400 and 800 μg of vitamin C in physiological pH, respectively. To observe the effects of in vitro vitamin C release on osteosarcoma and osteoblast cellular activity, MG-63 (human osteosarcoma) and hFOB (human fetal osteoblast) cells were cultured on the surface of the implant and MTT cell viability assay and FESEM were carried out at 3 and 7 days of culture. Presence of high dosages 25 mM vitamin C shows a statistically significant (p≤0.05) decrease in osteosarcoma cell viability after 3 days, while both 5 mM and 25mM vitamin C reduced cellular viability by 2.5 folds (p≤0.05) compared to the control after 7 days. Interestingly, the presence of vitamin C showed no obvious signs of cytotoxicity towards osteoblast cell-line at day 3 and day 7, as confirmed by the MTT assay. Additionally, the FESEM images depict layers of hFOB cellular morphology on the surface of the implants, suggesting excellent cytocompatibility towards the osteoblast cells. These results suggest that vitamin C loaded HAp coated cpTi implant with improved osteogenic and chemopreventive properties can be considered as a promising reconstructive option to repair the post-tumor resection defects in osteosarcoma.