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

Document Type : Original Article(s)

Authors

1 Assistant Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

2 PhD Student, Department of Anatomical Sciences, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

3 Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

5 Associate Professor, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.

Abstract

Background: 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 calcium phosphate hydroxyapatite (HA-TC) scaffolds and the behavior of these two types of cells in different cultures were analyzed.Methods: During the craniotomy operation in Alzahra hospital in five individual, bone samples were obtained and in sterile condition 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. After 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 was cultured in HA-TC scaffold and other as monolayer culture. Van kossa staining, flow cytometry, alkaline phosphatase assay and MTT method were used to investigate the characteristics of osteoblasts.Finding: Osteoblasts in monolayer culture were observed as fibroblast- like cells. 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 in 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 marker.Conclusion: According to the results of this study, the hydroxyapatite scaffold is better than monolayer culture for osteoblast cells. This could be because of having more calcium and similar elements to bone matrix that facilitate feeding cultured cells.

Keywords

References

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Journal of Isfahan Medical School
Volume 28, Issue 119 - Serial Number 119
November and December 2011
Pages 1348-1358

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History

  • Receive Date: 10 February 2011
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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