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The present study reported the preparation of food industry biowaste buffalo teeth-derived hydroxyapatite (BT-HA)/chitosan-polyvinyl pyrrolidone (CS-PVP)/aloe vera (ALV) natural biopolymer as a biocomposite for improved mechanical, antimicrobial and cytocompatible characteristics. The strengthening of biopolymer has a major role to play in improving the mechanical properties of the biocomposite, while the addition of aloe vera increases antimicrobial and cytocompatibility. Chemical structure, crystalline phase, morphology and mechanical property biocomposite were studied by FTIR, XRD, SEM-EDX and micro-hardness tests, respectively. Additionally, the antimicrobial effectiveness of the biocomposites was evaluated using Gram-positive and Gram-negative bacteria. The cytocompatibility of osteoblast cells to the biocomposites was calculated by the MTT assay examination. The findings demonstrated that the prepared biocomposite has the necessary mechanical, antimicrobial and biocompatible characteristics. Generally, the mixture of mechanical characteristics of CS-PVP, the antimicrobial and cytocompatible property of aloe vera in BT-HA/CS-PVP/ALV, makes the biocomposite a prospective restorative substance for various clinical applications.