Bony engineering using time-release porous scaffolds to provide sustained growth factor delivery

J Craniofac Surg. 2012 May;23(3):638-44. doi: 10.1097/SCS.0b013e31824db8d4.

Abstract

Microporous scaffolds designed to improve bony repair have had limited success; therefore, we sought to evaluate whether time-released porous scaffolds with or without recombinant bone morphogenetic protein 2 (rhBMP-2) could enhance stem cell osteoinduction. Custom-made 15/85 hydroxyapatite/β-tricalcium phosphate scaffolds were left empty (E) or filled with rhBMP-2 (E+), calcium sulfate (CS), or CS and rhBMP-2 (CS+). All scaffolds were placed in media and weighed daily. Conditioned supernatant was analyzed for rhBMP-2 and then used to feed human adipose-derived mesenchymal stem cells (ASCs). Adipose-derived mesenchymal stem cell ALP activity, OSTERIX expression, and bone nodule formation were determined. E scaffolds retained 97% (SD, 2%) of the initial weight, whereas CS scaffolds had a near-linear 30% (SD, 3%) decrease over 60 days. E+ scaffolds released 155 (SD, 5) ng of rhBMP-2 (77%) by day 2. In contrast, CS+ scaffolds released only 30 (SD, 2) ng (10%) by day 2, and the remaining rhBMP-2 was released over 20 days. Conditioned media from E+ scaffolds stimulated the highest ALP activity and OSTERIX expression in ACSs on day 2. However, after day 6, media from CS+ scaffolds stimulated the highest ALP activity and OSTERIX expression in ASCs. Adipose-derived mesenchymal stem cells exposed to day 8 CS+-conditioned media produced significantly more bone nodules (10.1 [SD, 1.7] nodules per high-power field) than all other scaffolds. Interestingly, day 8 conditioned media from CS scaffolds simulated significantly more bone nodules than either E or E+ scaffold (P < 0.05 for both). Time-released hydroxyapatite/β-tricalcium phosphate porosity provides sustained growth factor release, enhances ASC osteoinduction, and may result in better in vivo bone formation.

MeSH terms

  • Adipose Tissue / cytology*
  • Alkaline Phosphatase / metabolism
  • Analysis of Variance
  • Bone Morphogenetic Protein 2 / pharmacology*
  • Bone Regeneration / physiology*
  • Calcium Phosphates / pharmacology*
  • Calcium Sulfate / pharmacology
  • Cell Culture Techniques
  • Cell Differentiation
  • Durapatite / pharmacology*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Mesenchymal Stem Cells / metabolism*
  • Porosity
  • Recombinant Proteins / pharmacology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sp7 Transcription Factor
  • Staining and Labeling
  • Tissue Engineering / instrumentation*
  • Tissue Scaffolds*
  • Transcription Factors / metabolism
  • Transforming Growth Factor beta / pharmacology*

Substances

  • Bone Morphogenetic Protein 2
  • Calcium Phosphates
  • Recombinant Proteins
  • Sp7 Transcription Factor
  • SP7 protein, human
  • Transcription Factors
  • Transforming Growth Factor beta
  • beta-tricalcium phosphate
  • recombinant human bone morphogenetic protein-2
  • Durapatite
  • Alkaline Phosphatase
  • Calcium Sulfate