Study of bone repair mediated by recombination BMP-2/ recombination CXC chemokine Ligand-13-loaded hollow hydroxyapatite microspheres/chitosan composite

Life Sci. 2019 Oct 1:234:116743. doi: 10.1016/j.lfs.2019.116743. Epub 2019 Aug 10.

Abstract

Aims: The present study aimed to investigate the mechanism of bone repair mediated by recombination BMP-2 (rhBMP-2)/recombination CXC chemokine ligand-13 (rhCXCL13)-loaded hollow hydroxyapatite (HA) microspheres/chitosan (CS) composite.

Materials and methods: Firstly, the biological activity of rhBMP-2 and rhCXCL13 released from the complex was investigated. Secondly, the effect of rhBMP-2 sustained release solution on ALP activity and rhCXCL13 sustained release solution on cell migration of rat bone marrow mesenchyme stem cells was tested. Thirdly, osteoblasts differentiation test, X-ray scoring and three-point bending test were performed. Finally, the mRNAs expression of osteogenic marker genes and the protein expression of Runx2 was tested by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting (WB), respectively.

Key findings: RhBMP-2 could significantly promote the proliferation and differentiation, and RhCXCL13 could promote the migration of rat bone marrow MSCs. Detection of ALP activity and calcium salt deposition showed that rhBMP-2 and rhCXCL13 could significantly improve the biological activity and promote cell differentiation ability. X-ray scoring of radius and flexural strength test showed that rhBMP-2 and rhCXCL13 could promote bone healing and improve the bending resistance of bone tissue. The in vitro molecular experiments including RT-PCR and WB further demonstrated the roles of rhBMP-2 and rhCXCL13 in bone formation and bone repair.

Significance: Our results indicated that the hollow HA microspheres/CS composite could be effective as a delivery vehicle for rhBMP-2 and rhCXCL13 in bone regeneration and bone repair. In this process, rhBMP-2 may promote bone regeneration by regulating bone marrow MSCs cells recruited by rhCXCL13.

Keywords: Bone graft; Bone regeneration; Bone repair; Hollow HA microspheres/CS composite; Recombination BMP-2; Recombination CXC chemokine ligand-13.

MeSH terms

  • Animals
  • Biocompatible Materials / chemistry
  • Bone Morphogenetic Protein 2 / administration & dosage*
  • Bone Morphogenetic Protein 2 / pharmacology
  • Bone Regeneration / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Chemokine CXCL13 / administration & dosage*
  • Chemokine CXCL13 / pharmacology
  • Chitosan / analogs & derivatives*
  • Delayed-Action Preparations / chemistry*
  • Durapatite / chemistry*
  • Humans
  • Male
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Osteogenesis / drug effects*
  • Rabbits
  • Rats
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / pharmacology
  • Tissue Scaffolds / chemistry*
  • Transforming Growth Factor beta / administration & dosage*
  • Transforming Growth Factor beta / pharmacology

Substances

  • Biocompatible Materials
  • Bone Morphogenetic Protein 2
  • CXCL13 protein, human
  • Chemokine CXCL13
  • Delayed-Action Preparations
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2
  • Chitosan
  • Durapatite