Microbial biomineralization of hydroxyapatite nanocrystals using Bacillus tequilensis

Abstract

In this research, biomimetic deposition of “bone-like” apatite by novel Gram-positive bacterium Bacillus tequilensis was investigated. Hydroxyapatite (HA) was produced by Bacillus tequilensis using defined biomineralization media and dried. Calcination was carried out at different temperatures (100 °C–900 °C) and HA nanopowder was analysed for its structural phase composition, crystallinity, crystallite size and functional groups. X-ray diffractometry (XRD) indicated that increasing temperatures increased the crystallinity of HA nanocrystals. The presence of carbonate groups was evidenced by Fourier transform infrared (FTIR) spectrum and the purity of synthesized apatite nanocrystals was validated by absence of secondary peaks in XRD studies. Scanning electron microscopy (SEM) images depicted those uniform spherical agglomerates of HA comprised of nanosized crystallites. Transmission electron microscope (TEM) results identified needle-like crystal morphologies with average dimensions of 30–60 nm length and 3–10 nm width. Rich trace ion deposition was illustrated by energy dispersive x-ray spectroscopy (EDS) and quantified using inductively coupled plasma optical emission spectrometer (ICP-OES). Overall, microbial biomineralization by Bacillus tequilensis produced nanocrystals of HA that mimicked “bone-like apatite” as evidenced by pure phase, B-type carbonated form, poor crystallinity and trace amounts of vital elements (Mg, Na, K, Zn, Sr, Cl). Moreover, in vitro cytotoxicity studies revealed more than 80% cell viability highlighting the biocompatible nature of synthesized nano HA. Thereby, Bacillus tequilensis biomineralized nano HA reflects as a suitable candidate for applications in biomedicine addressing bone injuries and aiding regeneration.