Effect of gentamicin-nigella sativa fusion emulsions on osteoblast cell line for use in osteo healing-applications

Abstract

An alternative osteo-healing formulation with osteo-healing properties was formulated by combining gentamicin and Nigella sativa oil (NSO) in a form of gentamicin-N. sativa fusion emulsion (GNFE). This work aims to formulate a stable emulsion and to study the effects of GNFE on UMR-106 osteoblast-like rat osteosarcoma cell line in vitro and its related mechanisms of bone healing and regeneration. Emulsion A, B, C and D had been formulated, with final concentration of gentamicin was made constant at 0.1%, whereas NSO concentration was varied at 32.5%, 35.0%, 40.2% and 46.4% in all formulations respectively. Stability studies of emulsion A, B, C and D were performed at different storage conditions (8°C, 25°C and 50°C), followed by in vitro study of MTT assay, Alizarin Red S (ARS) staining, von Kossa staining and quantification, alkaline phosphatase (ALP) quantification and quantitation of collagen type-1 and osteocalcin (qPCR). Results showed that all emulsions were stable at storage temperature of 8°C. In vitro results showed that emulsion D produced the highest cell viability (97.1%) at 72 hours of post-incubation. The highest mineral deposits (2.64 ± 0.05) and ALP activity (2.19 ± 0.3 nmol) was produced by emulsion D at day 21. Lastly, the highest expression of collagen type-1 (29.4 ± 1.01 folds) and osteocalcin (1.8 ± 0.51 folds) were expressed by the cells treated with emulsion C. Thus, stable GNFE may have the ability to promote bone formation.