Tissue dynamics and regenerative outcome in two resorbable non-cross-linked collagen membranes for guided bone regeneration: A preclinical molecular and histological study in vivo

Clin Oral Implants Res. 2018 Jan;29(1):7-19. doi: 10.1111/clr.13032. Epub 2017 Jul 13.


Objectives: To investigate the molecular and structural patterns of bone healing during guided bone regeneration (GBR), comparing two resorbable non-cross-linked collagen membranes.

Materials and methods: Trabecular bone defects in rat femurs were filled with deproteinized bovine bone (DBB) and covered with either a membrane comprising collagen and elastin (CXP) or collagen (BG). Samples were harvested after 3 and 21 days for histology/histomorphometry and gene expression analysis. Gene expression analysis was performed on the membrane (at 3 days) and the underlying defect compartment (at 3 and 21 days).

Results: At the total defect level, no differences in bone area percentage were found between the CXP and BG. When evaluating the central area of the defect, a higher percentage of de novo bone formation was seen for the CXP membrane (34.9%) compared to BG (15.5%) at 21 days (p = .01). Gene expression analysis revealed higher expression of bone morphogenetic protein-2 (Bmp2) in the membrane compartment at 3 days in the BG group. By contrast, higher Bmp2 expression was found in the defect compartment treated with the CXP membrane, both at 3 and 21 days. A significant temporal increase (from 3 to 21 days) in the remodeling activity, cathepsin K (Catk) and calcitonin receptor (Calcr), was found in the CXP group. Molecular analysis demonstrated expression of several growth factors and cytokines in the membrane compartment irrespective of the membrane type. Bmp2 expression in the membrane correlated positively with Bmp2 expression in the defect, whereas fibroblast growth factor-2 (Fgf2) expression in the membrane correlated positively with inflammatory cytokines, tumor necrosis factor-alpha (Tnfa) and interleukin-6 (Il6) in the defect.

Conclusions: The results provide histological and molecular evidence that different resorbable collagen membranes contribute differently to the GBR healing process. In the BG group, bone formation was primarily localized to the peripheral part of the defect. By contrast, the CXP group demonstrated significantly higher de novo bone formation in the central portion of the defect. This increase in bone formation was reflected by triggered expression of potent osteogenic growth factor, Bmp2, in the defect. These findings suggest that the CXP membrane may have a more active role in regulating the bone healing dynamics.

Keywords: animal experiments; biomaterials; bone substitutes; genetics; growth factors; guided bone regeneration; morphometric analysis.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Bone Morphogenetic Proteins / genetics
  • Bone Morphogenetic Proteins / metabolism
  • Bone Regeneration / physiology*
  • Cathepsin K / genetics
  • Cathepsin K / metabolism
  • Collagen*
  • Cytokines / genetics
  • Cytokines / metabolism
  • Elastin
  • Femur / anatomy & histology
  • Femur / metabolism
  • Femur / physiology*
  • Gene Expression Profiling
  • Guided Tissue Regeneration*
  • Male
  • Models, Animal
  • Osteocalcin / genetics
  • Osteocalcin / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Calcitonin / genetics
  • Receptors, Calcitonin / metabolism


  • Bone Morphogenetic Proteins
  • Cytokines
  • Receptors, Calcitonin
  • Osteocalcin
  • Collagen
  • Elastin
  • Cathepsin K
  • Ctsk protein, rat