Pre-coating deproteinized bovine bone mineral (DBBM) with bone-conditioned medium (BCM) improves osteoblast migration, adhesion, and differentiation in vitro

Clin Oral Investig. 2016 Dec;20(9):2507-2513. doi: 10.1007/s00784-016-1747-x. Epub 2016 Feb 15.


Objectives: Autogenous bone grafting has remained the gold standard for bone augmentation procedures with ability to release growth factors to the surrounding microenvironment. Recent investigations have characterized these specific growth factors released by autogenous bone chips with further isolation into a "bone-conditioned medium" (BCM). The aim of the present investigation was to utilize autologous growth factors from bone chips (BCM) in combination with deproteinized bovine bone mineral (DBBM) and investigate the ability for BCM to enhance osteoblast behavior.

Materials and methods: Mouse ST2 cells were seeded on (1) DBBM particles alone or (2) DBBM + BCM. Thereafter, samples were compared for cell recruitment, adhesion, proliferation, and real-time PCR for osteoblast differentiation markers including Runx2, collagen 1 alpha 2 (COL1A2), alkaline phosphatase (ALP), and osteocalcin (OCN). Alizarin red staining was used to assess mineralization.

Results: Coating BCM on DBBM particles improved cell migration of ST2 cells and significantly enhanced a 2-fold increase in cell adhesion. While no significant increase in cell proliferation was observed, BCM significantly increased mRNA levels of COL1A2, ALP, and OCN at 3 days post seeding. Furthermore, a 3-fold increase in alizarin red staining was observed on DBBM particles pre-coated with BCM.

Conclusion: Pre-coating DBBM with BCM enhanced the osteoconductive properties of DBBM by mediating osteoblast recruitment, attachment, and differentiation towards bone-forming osteoblasts. Future animal study is necessary to further characterize the added benefit of BCM as an autogenous growth factor source for combination therapies.

Clinical relevance: The application of BCM in combination with biomaterials may serve as an autogenous growth factor source for bone regeneration.

Keywords: Barrier membranes; Bone grafting; Bone-conditioned media; Growth factors; Guided bone regeneration.

MeSH terms

  • Alkaline Phosphatase / analysis
  • Animals
  • Biomarkers / analysis
  • Bone Substitutes / pharmacology*
  • Bone Transplantation
  • Cattle
  • Cell Adhesion
  • Cell Differentiation
  • Cell Movement
  • Cell Proliferation
  • Coated Materials, Biocompatible
  • Collagen Type I / analysis
  • Core Binding Factor Alpha 1 Subunit / analysis
  • Culture Media, Conditioned / pharmacology*
  • Mice
  • Osteoblasts / cytology*
  • Osteoblasts / drug effects*
  • Osteocalcin / analysis
  • Real-Time Polymerase Chain Reaction
  • Swine


  • Biomarkers
  • Bone Substitutes
  • Coated Materials, Biocompatible
  • Collagen Type I
  • Core Binding Factor Alpha 1 Subunit
  • Culture Media, Conditioned
  • Runx2 protein, mouse
  • Osteocalcin
  • Alkaline Phosphatase