Evaluation of processed bovine cancellous bone matrix seeded with syngenic osteoblasts in a critical size calvarial defect rat model

J Cell Mol Med. 2006 Jul-Sep;10(3):695-707. doi: 10.1111/j.1582-4934.2006.tb00429.x.


Introduction: Biologic bone substitutes may offer alternatives to bone grafting procedures. The aim of this study was to evaluate a preformed bone substitute based on processed bovine cancellous bone (PBCB) with or without osteogenic cells in a critical size calvarial defect rat model.

Methods: Discs of PBCB (Tutobone) were seeded with second passage fibrin gel-immobilized syngenic osteoblasts (group A, n = 40). Cell-free matrices (group B, n = 28) and untreated defects (group C; n=28) served as controls. Specimens were explanted between day 0 and 4 months after implantation and were subjected to histological and morphometric evaluation.

Results: At 1 month, bone formation was limited to small peripheral areas. At 2 and 4 months, significant bone formation, matrix resorption as well as integration of the implants was evident in groups A and B. In group C no significant regeneration of the defects was observed. Morphometric analysis did not disclose differences in bone formation in matrices from groups A and B. Carboxyfluorescine-Diacetate-Succinimidylester (CFDA) labeling demonstrated low survival rates of transplanted cells.

Discussion: Osteoblasts seeded into PBCB matrix display a differentiated phenotype following a 14 days cell culture period. Lack of initial vascularization may explain the absence of added osteogenicity in constructs from group A in comparison to group B. PBCB is well integrated and represents even without osteogenic cells a promising biomaterial for reconstruction of critical size calvarial bone defects.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biocompatible Materials
  • Bone Substitutes / chemistry
  • Bone and Bones / pathology*
  • Cattle
  • Cell Transplantation
  • Fluorescent Dyes / pharmacology
  • Imaging, Three-Dimensional
  • Male
  • Models, Statistical
  • Osseointegration
  • Osteoblasts / cytology*
  • Osteoblasts / metabolism
  • Rats
  • Rats, Inbred WKY
  • Tissue Engineering / methods*


  • Biocompatible Materials
  • Bone Substitutes
  • Fluorescent Dyes