MC3T3-E1 proliferation and differentiation on biphasic mixtures of Mg substituted β-tricalcium phosphate and amorphous calcium phosphate

Mater Sci Eng C Mater Biol Appl. 2014 Dec:45:589-98. doi: 10.1016/j.msec.2014.03.032. Epub 2014 Mar 25.

Abstract

A low temperature aqueous approach was used to synthesize nanocrystalline, high surface area Mg(2+) substituted β-tricalcium phosphate (β-TCMP) to assess its potential use as a synthetic bone graft substitute. X-ray diffraction indicated that β-TCMP was the predominant crystalline phase formed. However, thermal analysis revealed the presence of a secondary amorphous phase which increased with increasing Mg(2+) concentration. Further analysis by Rietveld refinement indicated that the level of ionic substitution of Ca(2+) by Mg(2+) was significantly lower than the amount of Mg(2+) measured using elemental analysis, confirming the formation of a Mg(2+) rich secondary amorphous phase. MC3T3-E1 proliferation on substrates prepared using β-TCMP was assessed using the MTT assay. In comparison to commercially available β-TCP, increased proliferation was observed on samples prepared with 50% Mg, despite elevated Mg(2+) and PO4(3-) concentrations in culture media. Alkaline phosphatase (ALP) activity and qRT-PCR were used to study the effect of varying Mg(2+) substitution on osteogenic differentiation. Cells cultured on β-TCMP substrates prepared with increased Mg(2+) concentrations expressed significantly increased levels of ALP activity and osteogenic genes such as, osteocalcin, collagen-1, and Runx2, in comparison to those cultured on commercially available β-TCP.

Keywords: Calcium phosphate; Magnesium; Osteogenic differentiation; Rietveld refinement.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adsorption
  • Animals
  • Bone Substitutes / chemistry*
  • Bone Substitutes / pharmacology
  • Calcium Phosphates / chemistry*
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Magnesium / chemistry*
  • Mice
  • Nanostructures / chemistry
  • Osteocalcin / genetics
  • Osteocalcin / metabolism
  • Porosity
  • Proteins / chemistry
  • Proteins / metabolism

Substances

  • Bone Substitutes
  • Calcium Phosphates
  • Collagen Type I
  • Core Binding Factor Alpha 1 Subunit
  • Proteins
  • amorphous calcium phosphate
  • beta-tricalcium phosphate
  • Osteocalcin
  • Magnesium