Gene expression analysis of osteoblasts seeded in coral scaffold

J Biomed Mater Res A. 2008 Oct;87(1):215-21. doi: 10.1002/jbm.a.31765.

Abstract

Coral matrix of Porites sp. has the suitable properties for bone cell growth. This study was aimed to study the gene expression levels of osteoblast specific genetic markers; RUNX2, osteopontin, alkaline phosphatase and osteocalcin from osteoblasts seeded in coral scaffold, which are important in determining the feasibility of osteoblasts. Human osteoblasts were inoculated onto the processed coral in Dulbecco's Minimum Essential Medium. The cells were trypsinized on day 1, 7, 14, 18, and 21 and added with RNALater for preservation of RNA in cells. The RNA was extracted using commercial RNA extraction kit and the respective genes were amplified using RT-PCR kit and analyzed qualitatively on 1.5% agarose gel. The expressions were evaluated with the Integrated Density Value based on the intensity of band for different periods of cell harvest. Increased expressions of the RUNX2, osteopontin, alkaline phosphatase and osteocalcin genes in the present study proved that coral is a favorable carrier for osteogenetically competent cells to attach and remain viable.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / genetics
  • Animals
  • Anthozoa*
  • Cell Line
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Electrophoresis, Agar Gel
  • Gene Expression*
  • Genetic Markers
  • Humans
  • Osteoblasts / metabolism*
  • Osteocalcin / genetics
  • Osteogenesis / genetics
  • Osteopontin / genetics
  • RNA / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tissue Scaffolds*

Substances

  • Core Binding Factor Alpha 1 Subunit
  • Genetic Markers
  • RUNX2 protein, human
  • Osteocalcin
  • Osteopontin
  • RNA
  • Alkaline Phosphatase