Evaluation of growth and differentiation of mouse osteoblast progenitor cells on synthesized nano-hydroxyapatite bone scaffolds
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sara sharifi ghimbavani,1,* hamidreza samadi khah,2 Soheila Zamanlui Benisi,3
1. Rafsanjani University Campus, Islamic Azad University Central Tehran Branch
2. Rafsanjani University Campus, Islamic Azad University Central Tehran Branch
3. Rafsanjani University Campus, Islamic Azad University Central Tehran Branch
- Introduction: There are promising tools today for bone tissue regeneration. However, it is difficult to trace cells and maintain them at the site of injury. A potential solution is to culture the cells on bone scaffolds and then examine the source of osteoblast cells and the molecular factors that differentiate them. We used RT-PCR test to design the scaffold and then study the proliferation and differentiation of these cells. Scaffolds should have features such as controlled degradation, biocompatibility, increased cell viability, differentiation, production of extracellular matrix, food and excretory production, adhesion and integration with natural bone tissue, three-dimensional structure with internal pores associated with Both the chemical level and the surface are suitable. Tissue engineering is a growing field and can be a permanent solution for tissue damage and analysis. In this approach, cells, scaffolds and messaging factors are used. Scaffolds are three-dimensional structures that provide the necessary conditions for cell proliferation and differentiation, and their structure determines the final shape of the tissue. The aim of this project was to investigate the effect of bone scaffolds on the growth and differentiation of mouse osteoblast progenitor cells into adult osteoblasts. Hydroxyapatite with the compound Ca10 (PO4) 6OH2 due to its biocompatibility and bioactivity, is widely used in bone and tooth treatment applications and as a hard tissue.
- Methods: After synthesis of bone hydroxyapatite and sodium alginate scaffolds and its characterization (tests: degradation, swelling, mechanical, porosity, (FTIR-ATR), in order to evaluate their quality, osteoblast cells were cultured on the scaffolds and In order to study the morphology of the cells on the scaffold, SEM test, MTT cell viability and expression of Runx2ALP, COL1 genes, Real-timePCR test were performed.
- Results: Bone scaffold with new formulation affects mouse osteoblast cell line. And increases the expression of genes involved in cell growth and differentiation. Bone scaffolds synthesized for safe and successful use in clinical settings, biomaterials for bone tissue engineering show several characteristics such as biocompatibility, biodegradability, bone formation and bone conduction, scaffold pore structure, grain size and surface topography. Real-timePCR differentiation factors were examined to evaluate the differentiation and growth of osteoblast cells, and this issue was confirmed by increasing the expression of these genes in the study of gene expression.
- Conclusion: Real-Time PCR results show increased expression of type 1 collagen genes and alkaline phosphatase and Runx2 compared to control cells. By examining the expression of genes influencing cell growth and differentiation, it can be concluded that synthetic scaffolds have the ability to optimize the substrate for the growth of mouse osteoblast cells.
- Keywords: Cell scaffolding, alkaline phosphatase, osteoblasts, cell differentiation, collagen 1
