Analysis of an experimental scaffold using osteoblasts through gene expression of markers for Osteogenesis.

Objectives: The aim of this in vitro gene expression study was to analyse an experimental scaffold composed of nano-Hydroxyapatite and E-glass fibres for application in bone repair and regeneration.
Methods: Materials were grouped according to the concentration of nano-Hydroxyapatite with E-glass fibre (E) .i.e. E-10, E-20, E-30, E-50 and E-0 (control). MC3T3 E-1 cells were seeded onto the scaffolds and mRNA was extracted on the fifth day of experiment. Gene Expression of bone proteins, Osteopontin (OPN) and Collagen (Col) Type 1 was measured through Reverse Transcription Polymerase Chain Reaction (RT-PCR) and comparison was made between all the groups.
Results: The results of the experiment were expressed as Relative Expression Ratio (R). All the materials were able to up regulate expression of both proteins. OPN gene expression was highest in E-0 followed by E-10, E-20, E-30 and E-50 (p < 0.001). Highest Col expression was also seen in E-0 followed by E-20, E-10, E-30 and E-50 (p < 0.001). Col and OPN gene expression decreased significantly with decreasing concentration of HA. Only for E-0, Col expression was equal to OPN whereas for all the other experimental materials, OPN expression was greater in comparison to Col.
Conclusions: It was concluded on the basis of these results that the novel scaffold was osteogenic in nature and supported the growth of MC3T3 E-1 cells. In the presence of the material, these cells were producing an extracellular matrix containing the bone proteins, OPN and Col as measured by PCR.