J Periodontal Implant Sci.  2020 Feb;50(1):14-27. 10.5051/jpis.2020.50.1.14.

Distinctive bone regeneration of calvarial defects using biphasic calcium phosphate supplemented ultraviolet-crosslinked collagen membrane

• Hong I
• Khalid AW
• Pae HC
• Cha JK
• Lee JS
• Paik JW
• Jung UW
• Choi SH

• KMID: 2470827
• DOI: http://doi.org/10.5051/jpis.2020.50.1.14

Abstract

PURPOSE
To overcome several drawbacks of chemically-crosslinked collagen membranes, modification processes such as ultraviolet (UV) crosslinking and the addition of biphasic calcium phosphate (BCP) to collagen membranes have been introduced. This study evaluated the efficacy and biocompatibility of BCP-supplemented UV-crosslinked collagen membrane for guided bone regeneration (GBR) in a rabbit calvarial model.
METHODS
Four circular bone defects (diameter, 8 mm) were created in the calvarium of 10 rabbits. Each defect was randomly allocated to one of the following groups: 1) the sham control group (spontaneous healing); 2) the M group (defect coverage with a BCP-supplemented UV-crosslinked collagen membrane and no graft material); 3) the BG (defects filled with BCP particles without membrane coverage); and 4) the BG+M group (defects filled with BCP particles and covered with a BCP-supplemented UV-crosslinked collagen membrane in a conventional GBR procedure). At 2 and 8 weeks, rabbits were sacrificed, and experimental defects were investigated histologically and by micro-computed tomography (micro-CT).
RESULTS
In both micro-CT and histometric analyses, the BG and BG+M groups at both 2 and 8 weeks showed significantly higher new bone formation than the control group. On micro-CT, the new bone volume of the BG+M group (48.39±5.47 mm3) was larger than that of the BG group (38.71±2.24 mm3, P=0.032) at 8 weeks. Histologically, greater new bone area was observed in the BG+M group than in the BG or M groups. BCP-supplemented UV-crosslinked collagen membrane did not cause an abnormal cellular reaction and was stable until 8 weeks.
CONCLUSIONS
Enhanced new bone formation in GBR can be achieved by simultaneously using bone graft material and a BCP-supplemented UV-crosslinked collagen membrane, which showed high biocompatibility and resistance to degradation, making it a biocompatible alternative to chemically-crosslinked collagen membranes.

Keyword

Absorbable implants; Animals; Bone regeneration; Collagen; Hydroxyapatite-beta tricalcium phosphate; Ultraviolet rays

MeSH Terms

Absorbable Implants
Animals
Bone Regeneration*
Calcium*
Collagen*
Membranes*
Osteogenesis
Rabbits
Skull
Transplants
Ultraviolet Rays
Calcium
Collagen

Figure

• 

  Figure 1 Experimental design in the rabbit calvarium. (A) Four circular defects 8 mm in diameter were prepared using a trephine bur. (B) Each defect was randomly assigned to an experimental group. Clockwise from top left; M, sham control, BG, and BG+M. BCP: biphasic calcium phosphate, UV: ultraviolet, M: group of defects covered with an a BCP-supplemented UV-crosslinked collagen membrane without graft material, BG: group of defects filled with BCP particles without membrane coverage, BG+M: group of defects filled with BCP particles and covered with a UV-crosslinked collagen membrane with added BCP in a conventional guided bone regeneration procedure.

• 

  Figure 2 In micro-CT images taken at 8 weeks, the new bone volume of the BG+M group (48.39±5.47 mm3) showed a significant difference compared to the BG group (38.71±2.24 mm3). CT: computed tomography, BCP: biphasic calcium phosphate, UV: ultraviolet, M: group of defects covered with an a BCP-supplemented UV-crosslinked collagen membrane without graft material, BG: group of defects filled with BCP particles without membrane coverage, BG+M: group of defects filled with BCP particles and covered with a BCP-supplemented UV-crosslinked collagen membrane in a conventional guided bone regeneration procedure. a)P=0.032.

• 

  Figure 3 Micro-CT views of the defects after 2 week, and 8 weeks of healing. At 2 weeks in the control and M groups, the defects were rarely filled, and even at 8 weeks, the defects were partially filled with a thin new bone bridge. Compared to natural bone, the newly-formed bone can be observed as a grayish area, and the BCP particles are observed as a more whitish area. CT: computed tomography, BCP: biphasic calcium phosphate, UV: ultraviolet, M: group of defects covered with an a BCP-supplemented UV-crosslinked collagen membrane without graft material, BG: group of defects filled with BCP particles without membrane coverage, BG+M: group of defects filled with BCP particles and covered with a UV-crosslinked collagen membrane with added BCP in a conventional guided bone regeneration procedure.

• 

  Figure 4 Reconstructed images of micro-CT views at 2 weeks and 8 weeks. At both 2 and 8 weeks, the BG group (upper right; A) showed scattering of BCP particles outside of the defect, while the BG+M group (B) showed enclosed BCP particles in the defect area. CT: computed tomography, BCP: biphasic calcium phosphate, UV: ultraviolet, M: group of defects covered with an a BCP-supplemented UV-crosslinked collagen membrane without graft material, BG: group of defects filled with BCP particles without membrane coverage, BG+M: group of defects filled with BCP particles and covered with a BCP-supplemented UV-crosslinked collagen membrane in a conventional guided bone regeneration procedure.

• 

  Figure 5 Histological images obtained at 2 and 8 weeks. Low-magnification views of the sham control, BG, M, and BG+M groups, respectively (hematoxylin and eosin staining). The applied collagen membrane remained intact over the defects at 2 weeks in the M and BG+M groups. The remaining collagen membranes in the 2-week groups are marked with an asterisk (*). BCP: biphasic calcium phosphate, UV: ultraviolet, M: group of defects covered with an a BCP-supplemented UV-crosslinked collagen membrane without graft material, BG: group of defects filled with BCP particles without membrane coverage, BG+M: group of defects filled with BCP particles and covered with a BCP-supplemented UV-crosslinked collagen membrane in a conventional guided bone regeneration procedure.

• 

  Figure 6 A few membrane remnants were observed in the M group at 8 weeks. Both in hematoxylin and eosin and Masson trichrome stained slides, the remaining portion of the crosslinked collagen membrane is observed as a thin film shape. The remaining membrane is marked with an asterisk (*). BCP: biphasic calcium phosphate, UV: ultraviolet, M: group of defects covered with a BCP-supplemented UV-crosslinked collagen membrane without graft material.

• 

  Figure 7 At 8 weeks in the BG+M group, matured new bone was found above the graft material area where the membrane was located, while new bone formation in the BCP-augmented area was insufficient. New bone that substituted the crosslinked collagen membrane is marked with an asterisk (*). BCP: biphasic calcium phosphate, UV: ultraviolet, BG+M: group of defects filled with BCP particles and covered with an BCP-supplemented UV-crosslinked collagen membrane in a conventional guided bone regeneration procedure.

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