Chitosan scaffolds containing calcium phosphate salts and rhBMP-2: in vitro and in vivo testing for bone tissue regeneration

PLoS One. 2014 Feb 4;9(2):e87149. doi: 10.1371/journal.pone.0087149. eCollection 2014.

Abstract

Numerous strategies that are currently used to regenerate bone depend on employing biocompatible materials exhibiting a scaffold structure. These scaffolds can be manufactured containing particular active compounds, such as hydroxyapatite precursors and/or different growth factors to enhance bone regeneration process. Herein, we have immobilized calcium phosphate salts (CPS) and bone morphogenetic protein 2 (BMP-2)--combined or alone--into chitosan scaffolds using ISISA process. We have analyzed whether the immobilized bone morphogenetic protein preserved its osteoinductive capability after manufacturing process as well as BMP-2 in vitro release kinetic. We have also studied both the in vitro and in vivo biocompatibility of the resulting scaffolds using a rabbit model. Results indicated that rhBMP-2 remained active in the scaffolds after the manufacturing process and that its release kinetic was different depending on the presence of CPS. In vitro and in vivo findings showed that cells grew more in scaffolds with both CPS and rhBMP-2 and that these scaffolds induced more bone formation in rabbit tibia. Thus chitosan scaffolds containing both CPS and rhBMP-2 were more osteoinductive than their counterparts alone indicating that could be useful for bone regeneration purposes, such as some applications in dentistry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 2 / pharmacology
  • Bone Regeneration / drug effects*
  • Bone and Bones / diagnostic imaging
  • Bone and Bones / drug effects
  • Bone and Bones / physiology
  • Calcium Phosphates / pharmacology*
  • Cell Line
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Chitosan / pharmacology*
  • Elastic Modulus / drug effects
  • Humans
  • Kinetics
  • Male
  • Materials Testing / methods*
  • Mice
  • Rabbits
  • Recombinant Proteins / pharmacology
  • Rheology / drug effects
  • Salts / pharmacology*
  • Tensile Strength / drug effects
  • Tissue Scaffolds / chemistry*
  • Transforming Growth Factor beta / pharmacology
  • X-Ray Microtomography

Substances

  • Bone Morphogenetic Protein 2
  • Calcium Phosphates
  • Recombinant Proteins
  • Salts
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2
  • Chitosan
  • calcium phosphate

Grants and funding

This work was supported by Spanish Ministry of Economy and Competitiveness (MINECO) (CONSOLIDER project CSD2009-00088), Ministerio de Ciencia e Innovación (MAT2009-10214, MAT2011-25329, MAT2012-34811, SAF2011-27863) and Seventh Framework Programme of the European Commission (EuroNanomed ERA-NET initiative, REBONE project; PI10/02985FIS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.