The Characteristics of Human Osteoblast Cells in Two Methods of Monolayer Culture in Hydroxyapatite-Tricalcium Phosphate Scafolds

Bone diseases and injuries, including bone fractures during accidents and trauma, in all communities are prevalent and in most cases, crushed bone is not able to repair; various treatment ways including tissue graft from an allograft or autograft have risks and lead to problems. One method for treatment of bone fractures is using outologous osteoblasts with culturing on suitable scaffolds and then implant of cells to defect region. In this study, we cultured osteoblast cells on monolayer and more calcium phosphate hydroxyapatite (HA-TC) scaffolds and the behavior of these two types of cells in different cultures were analyzed. During the craniotomy operation in Alzahra hospital in five individual, bone samples were obtained and in sterile technique were sent to the laboratory. Each sample after washing with PBS was cut into small pieces and plates containing the medium culture were transferred to the incubator. In 10-12 days, the cells have started to leave the bones covering the floor plate and after using the enzyme Trypsin_EDTA isolated from plate divided into two groups. One group as HA-TC scaffolding and at the other group as a single layer were cultured. Staining Van kossa, MTT methods, alkaline phosphatase staining and flow cytometry were used to investigate osteoblast cells. Cultivation monolayer cells with fibroblast morphology were observed or round. Van kossa staining determined the mineral deposits in the two groups. The cell proliferation increased in cellular scaffolding HA-TC more than monolayer (P < 0.001). Results of MTT showed more activity items on living cells scaffold HA-TC than monolayer (P < 0.001). In alkaline phosphatase staining, blue granules were observed; in the flow cytometry, 57 percent of primary cultured cells and 5 percent in the third passage cells showed alkaline phosphatase activity. According to the results of this study, the hydroxyapatite scaffold is better than monolayer scaffold for osteoblast cells culture. This could be because of having more calcium and similar elements to bone matrix facilitate feeding cultured the cells.