Comparison of Two Porcine Collagen Membranes Combined with rhBMP-2 and rhBMP-9 on Osteoblast Behavior In Vitro

Int J Oral Maxillofac Implants. Jul/Aug 2017;32(4):e221-e230. doi: 10.11607/jomi.5652.


Purpose: To investigate the bone-inducing properties of two types of collagen membranes in combination with recombinant human bone morphogenetic protein (rhBMP)-2 and rhBMP-9 on osteoblast behavior.

Materials and methods: Porcine pericardium collagen membranes (PPCM) and porcine dermis-derived collagen membranes (PDCM) were coated with either rhBMP-2 or rhBMP-9. The adsorption and release abilities were first investigated via enzyme-linked immunosorbent assay up to 10 days. Moreover, murine bone stromal ST2 cell adhesion, proliferation, and osteoblast differentiation were assessed by MTS assay; real-time polymerase chain reaction for genes encoding runt-related transcription factor 2 (Runx2); alkaline phosphatase (ALP); and osteocalcin, ALP assay, and alizarin red staining.

Results: Both rhBMP-2 and rhBMP-9 adsorbed to collagen membranes and were gradually released over time up to 10 days. PPCM showed significantly less cell attachment, whereas PDCM demonstrated comparable cell attachment with the control tissue culture plastic at 8 hours. While both rhBMPs were shown not to affect cell proliferation, collagen membranes combined with rhBMP-9 significantly increased ALP activity at 7 days and ALP mRNA levels at either 3 or 14 days compared with the control tissue culture plastic. Furthermore, rhBMP-9 increased osteocalcin mRNA levels and alizarin red staining at 14 days compared with the control tissue culture plastic.

Conclusion: The results from this study suggest that both porcine-derived collagen membranes combined with rhBMP-9 accelerated the osteopromotive potential of ST2 cells. Interestingly, rhBMP-9 demonstrated additional osteogenic differentiation compared with rhBMP-2 and may serve as a suitable growth factor for future clinical use.

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Animals
  • Bone Morphogenetic Protein 2 / pharmacology*
  • Cell Differentiation
  • Cell Proliferation
  • Collagen / metabolism*
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Growth Differentiation Factor 2 / pharmacology*
  • Humans
  • Mice
  • Osteoblasts / drug effects*
  • Osteoblasts / metabolism
  • Osteocalcin / metabolism
  • Osteogenesis
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Recombinant Proteins / pharmacology
  • Stromal Cells
  • Swine
  • Transforming Growth Factor beta / pharmacology*


  • Bone Morphogenetic Protein 2
  • Core Binding Factor Alpha 1 Subunit
  • Growth Differentiation Factor 2
  • RNA, Messenger
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2
  • Osteocalcin
  • Collagen
  • Alkaline Phosphatase