Biphasic calcium phosphate (BCP) has received much interest for making various bone substitutes since its physicochemical properties can be easily tailored by tuning its phase composition. Due to high temperature processing, it is hard to prepare BCP with nanoscale characteristics. In the present study, we have made an attempt to optimize the heat treatment parameters for the synthesis of BCP with nanoscale characteristics from eggshell derived hydroxyapatite (HA) through rapid thermal processing (RTP). To accomplish this, eggshell derived HA was prepared by wet precipitation method and subjected to RTP at 750°C and 1150°C for 3 and 10 minutes. For comparison we have also studied conventional calcination at 750°C and 1150°C for 3 hours. XRD, FTIR, SEM, EDX, HRTEM, and BET analyses were used to understand the effect of RTP and conventional calcination on eggshell derived HA. Our results indicate that eggshell derived HA on RTP at 1150°C for 3 minutes and 10 minutes can offer nanoscale BCP with good dissolution, bioactivity, cytocompatibility, and mesoporous nature. Hence, RTP can be a potential method to prepare BCP with nanoscale features for biomedical applications.

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Drug repurposing has been gaining increasing interest recently due to the reduction in development cost and reduced development timelines. Here, we report the antibacterial activity of the anticancer drug etoposide investigated in combination with the eggshell-derived hydroxyapatite (EHA). Hydroxyapatite (HA) is a well-known bioactive material with enhanced osteoconductivity and possesses superior drug delivery properties. In the present work, we have synthesized etoposide-loaded EHA by the wet precipitation method. The physicochemical characterization of the samples confirmed the composition and amount of drug encapsulation. Screening for antibacterial activity confirmed the antibacterial effect of etoposide against Staphylococcus aureus. Biofilm formation test on pristine and etoposide-loaded samples showed the inhibition of biofilm formation on etoposide loading, which was further studied by confocal laser scanning microscopy (CLSM) and colony forming units (CFUs). It has been found that etoposide-loaded HA exhibited a sustained release of the drug upto 168 h. Analysis of the inhibition mechanism of etoposide against S. aureus revealed damage to the cell membrane and has been quantified using flow cytometry by the uptake of propidium iodide. Etoposide-loaded eggshell-derived HA (EHA-ET) exhibited excellent bioactivity and cytocompatibility against mouse fibroblast cells (L929) and supressed the growth of osteosarcoma cells (MG-63). Our studies reveal that the EHA-ET has a great potential for treating osteosarcoma and osteomyelitis.