Oral and Maxillofacial Surgery

, Volume 22, Issue 2, pp 203–213 | Cite as

Guided bone regeneration in calvarial critical size bony defect using a double-layer resorbable collagen membrane covering a xenograft: a histological and histomorphometric study in rats

  • Roy Abou Fadel
  • Rawad Samarani
  • Carole Chakar
Original Article



The aim of the present study was to evaluate histologically and histomorphometrically the bone regeneration in critical size calvarial defects in rats grafted with either a deproteinized bovine bone mineral (DBBM) alone or in combination with a single or double layer of native bilayer collagen membrane (NBCM). The secondary objective was to evaluate histologically and histomorphometrically the residual DBBM in these defects.

Material and methods

Thirty-two Wistar rats were divided into two groups: a control group of 16 rats with two critical size calvarial defects (CSD) of 5 mm performed each on either side of the median sagittal suture, where the frontal defect remained without any filling (negative control), while the occipital defect (positive control) was filled with DBBM; and then a test group of 16 rats, with two CSD filled with DBBM and covered by either a single (SM) or a double layer (DM) of NBCM. The animals were sacrificed at 4 and 8 weeks.


At 1 month, the histological and histomorphometric analysis showed new bone formation (NBF) in the defects that received only DBBM, DBBM+DM, and DBBM+SM (11.5, 17.3, and 22.7%, respectively), while the negative control defects showed only 0.4% of new bone formation. At 2 months, the histological and histomorphometric analysis showed NBF in the defects that received only DBBM, DBBM+DM, and DBBM+SM (16.8, 24.5, and 37%, respectively), while the negative control defects showed only 0.9% of new bone formation. The residual xenogeneic material (RXM) was higher in defects covered by SM (30.2% at 1 month and 25.3% at 2 months) or DM (32.5% at 1 month and 28.5% at 2 months) compared with defects that were not covered by membranes (15.3% at 1 month and 9.4% at 2 months).


This study demonstrated that GBR with a xenogeneic material in rat calvarial (CSD) of 5 mm requires the application of resorbable collagen membranes in either single or double layer, and a single layer alone is sufficient to promote this regeneration.


GBR Resorbable collagen membrane New bone formation Critical size defects Xenograft Xenogeneic 



The authors wish to thank Mr. Charbel Mansour for his technical assistance in the preparation of the histological sections and Dr. Nada Osta for her statistical advice and analysis.


This study was funded by Geistlich Pharma AG (FMD94).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of the animals were followed. All procedures performed on animals were in accordance with the ethical standards of the research committee at Saint Joseph University.

Informed consent

This study does not involve human participants, consent not available.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Periodontology, Faculty of DentistryUniversité Saint JosephBeirutLebanon

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