Indirect coating of RGD peptides using a poly-L-lysine spacer enhances jaw periosteal cell adhesion, proliferation, and differentiation into osteogenic tissue

J Biomed Mater Res A. 2012 Aug;100(8):2034-44. doi: 10.1002/jbm.a.34062. Epub 2012 May 12.

Abstract

The aim of our study was to generate a biofunctionalized, three-dimensional (3D) biomaterial to enhance jaw periosteal cell (JPC) adhesion and differentiation into osteogenic tissue. Therefore, open-cell polylactic acid (OPLA) scaffolds were coated covalently with different RGD peptides (a conserved recognition sequence of the most ECM proteins--arginine-glycine-asparagine) and different coating variants. The linear and cyclic RGD peptides were either applied directly or indirectly via a poly-L-lysine (PLL) spacer. JPCs were analyzed on coated constructs in 2D and 3D cultures and showed enhanced rates for indirectly coated scaffolds using the PLL spacer. By gene expression, we detected significantly increased levels of osteogenic marker genes, such as alkaline phosphatase, RUNX2, and AMELY in JPCs seeded onto PLL/linear RGD constructs compared to the otherwise-coated constructs. An analysis of the JPC mineralization capacity revealed the highest amounts of calcium-phosphate precipitates in cells growing within the PLL/linear scaffolds. Additionally, the JPC adhesion behavior on OPLA scaffolds seems to be mediated by ITGB3, ITGB1, and ITGAV, as shown by blocking assays. We concluded that coating of OPLA constructs with linear RGD peptides via PLL represents a suitable approach for functionalizing the polymer surface and enhancing adhesion, proliferation, and mineralization of JPCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism
  • Calcification, Physiologic / drug effects
  • Calcification, Physiologic / genetics
  • Cell Adhesion / drug effects
  • Cell Culture Techniques
  • Cell Differentiation / drug effects*
  • Cell Proliferation / drug effects
  • Coated Materials, Biocompatible / pharmacology*
  • Flow Cytometry
  • Gene Expression Regulation / drug effects
  • Humans
  • Immobilized Proteins / pharmacology
  • Integrins / metabolism
  • Jaw / cytology*
  • Lactic Acid / pharmacology
  • Oligopeptides / pharmacology*
  • Osteogenesis / drug effects*
  • Osteogenesis / genetics
  • Periosteum / cytology*
  • Polyesters
  • Polylysine / pharmacology*
  • Polymers / pharmacology
  • Tissue Scaffolds / chemistry

Substances

  • Biomarkers
  • Coated Materials, Biocompatible
  • Immobilized Proteins
  • Integrins
  • Oligopeptides
  • Polyesters
  • Polymers
  • Polylysine
  • Lactic Acid
  • poly(lactide)
  • arginyl-glycyl-aspartic acid