Biomimetic collagen scaffolds for human bone cell growth and differentiation

Tissue Eng. Jul-Aug 2004;10(7-8):1148-59. doi: 10.1089/ten.2004.10.1148.

Abstract

Type I collagen provides a structural framework for connective tissues and plays a central role in the temporal cascade of events leading to the formation of new bone from progenitors. The aim of this study was to examine the ability of the cell-binding domain of type I collagen (P-15 peptide) to promote human bone marrow stromal cell adhesion, proliferation, and differentiation on three-dimensional scaffolds. Human bone marrow stromal cells were selected, expanded, and cultured on particulate microporous ABM ("pure" hydroxyapatite) phase adsorbed with or without P-15 under basal or osteogenic conditions. Immobilized P-15 increased alkaline phosphatase activity and bone morphogenetic protein 2 (BMP-2) gene expression after 1 and 5 days as determined by real-time polymerase chain reaction. P-15 promoted human bone marrow stromal cell attachment, spreading, and alignment on ABM as well as alkaline phosphatase-specific activity in basal and osteogenic cultures. The presence of mineralized bone matrix, extensive cell ingrowth, and cellular bridging between three-dimensional matrices adsorbed with P-15 was confirmed by confocal microscopy, scanning electron microscopy, and alizarin red staining. Negligible cell growth was observed on ABM alone. In vivo diffusion chamber studies using MF1-nu/nu mice showed bone matrix formation and organized collagen formation after 6 weeks. These studies indicate the potential of P-15 to generate appropriate biomimetic microenvironments for osteoblasts and demonstrate the potential for the exploitation of extracellular matrix cues for osteogenesis and, ultimately, bone regeneration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Bioartificial Organs
  • Biomimetic Materials / chemistry
  • Biomimetic Materials / pharmacology*
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / physiology
  • Bone Matrix / physiology
  • Bone Substitutes*
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Cells, Cultured
  • Coated Materials, Biocompatible / chemistry
  • Coated Materials, Biocompatible / pharmacology
  • Collagen / chemistry
  • Collagen Type I / chemistry
  • Collagen Type I / pharmacology
  • Extracellular Matrix / physiology
  • Humans
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects*
  • Mesenchymal Stem Cells / physiology*
  • Osteoblasts / cytology*
  • Osteoblasts / drug effects
  • Osteoblasts / physiology*
  • Osteogenesis / drug effects
  • Osteogenesis / physiology
  • Peptide Fragments / chemistry
  • Stromal Cells / cytology
  • Stromal Cells / drug effects
  • Stromal Cells / physiology
  • Tissue Engineering / methods

Substances

  • Bone Substitutes
  • Coated Materials, Biocompatible
  • Collagen Type I
  • Peptide Fragments
  • cell-binding peptide P-15
  • Collagen