Investigation of osteoblast-like cells cultured on nano-hydroxyapatite/chitosan based composite scaffold in the treatment of bone defects and limited mobility

Design and construction of biocompatible and biodegradable scaffolds are among the main goals of tissue engineering. Recently, use of nano-hydroxyapatite as a bioactive bioceramic agent with high similarity to the mineral phase of the human bone tissue, in combination with biodegradable polymers and implant coatings has attracted the attention of researchers in the field of biomaterial sciences. The present study aimed to assess the differentiation of bone marrow stromal cells (BMSCs) in osteoblast-like cells on the chitosan/polyethylene oxide (PEO)/nano-hydroxyapatite scaffold in mature rats. Chitosan and PEO solution with the weight ratio of 80:20 and 70:30 were prepared, and 2% weight of nano-hydroxyapatite was added. Nanofibers were prepared using the electrospinning method, and the morphology was studied using scanning electron microscopy (SEM). Afterwards, the BMSCs of mature rats were cultured on nanofibers and differentiated by adding a differentiation medium. The survival of the differentiated cells was evaluated at the end of the first, second, and third week using acridine orange staining, and the morphology of the differentiated cells exposed to nanofibers was assessed using SEM. The mean diameter of the nanofibers with the ratio of 80:20 was 150±17 nanometers. The differentiation of BMSCs into the osteoblast-like cells on nanofibers was confirmed using Alizarin red staining. The results indicated a significant decrease in the survival of the differentiated cells in the nanofiber groups by the end of the third week of differentiation compared to the control samples. According to the results, BMSCs could be differentiated into osteoblast-like cells in the presence of the chitosan/PEO nanofibers containing nano-hydroxyapatite.