Ultrasonic-assisted green synthesis and characterization of nano-hydroxyapatite from Cirrhinus molitorella fish scales bio-waste for biomedical applications

Abstract

Herein, we demonstrate a facile green procedure to produce natural nano-hydroxyapatite (nHAp) using an ultrasonic-assisted method from Cirrhinus molitorella fish scale bio-waste. Characterization results confirmed the successful preparation of nHAp without any impurities. Further, X-ray diffraction (XRD) peaks matched the hydroxyapatite standard JCPDS 00–009–0432, which also confirmed the hexagonal pattern of nHAp. Fourier-transform infrared (FT-IR) spectroscopy analysis exhibited the existence of phosphate, hydroxyl, and B-type carbonate groups. The energy dispersive X-ray spectroscopy (EDX) was utilized to determine the calcium/phosphorous (Ca/P) molar ratio, which is 1.67. Further, the elemental composition of nHAp was identified using X-ray photoelectron spectroscopy (XPS) and EDX analysis. The surface area (111.96 m² g⁻¹), pore size (14.81 nm in diameter), and pore volume (0.4167 cm³ g⁻¹) of prepared nHAp were confirmed by the Brunauer-Emmett-Teller (BET). Atomic force microscopy (AFM) was employed to measure the shape parameters and surface roughness, and it was confirmed that nHAp has a dominant spiky surface. Transmission electron microscopy (TEM) showed the agglomerated rod-like nanostructured morphology of the nHAp powder. Thermogravimetric analysis (TGA) results of nHAp exhibited excellent thermal stability and a low weight loss (9.7 %). The mechanical properties of nHAp were checked using the measurement of compressive strength value, which was 114 ± 4.70 MPa. The hemolysis rate of nHAp powders was less than 3 %; hence, nHAp powders can be considered non-hemolytic material. The MTT results indicate that the prepared nHAp is more biocompatible with human osteosarcoma (MG-63) cells, which shows excellent cell viability compared to commercial hydroxyapatite (cHAp). Moreover, nHAp encourages the development of MG-63 cells, which exhibit better Giemsa staining and alkaline phosphatase (ALP) activity than cHAp. The size, structure, functional groups, compressive strength, hemocompatibility, and superior biocompatibility of synthesized nHAp make it suitable for use as a biomaterial in the biomedical field.