J Vet Sci.  2014 Jun;15(2):289-295. 10.4142/jvs.2014.15.2.289.

Comparable bone healing capacity of different bone graft matrices in a rabbit segmental defect model

• Kim JM
• Kim MH
• Kang SS
• Kim G
• Choi SH

• KMID: 1784651
• DOI: http://doi.org/10.4142/jvs.2014.15.2.289

Abstract

We compared the bone healing capacity of three different demineralized bone matrix (DBM) products applied using different carrier molecules (hyaluronic acid [HA] vs. carboxymethylcellulose [CMC]) or bone compositions (cortical bone vs. cortical bone and cancellous bone) in a rabbit segmental defect model. Overall, 15-mm segmental defects in the left and right radiuses were created in 36 New Zealand White rabbits and filled with HA-based demineralized cortical bone matrix (DBX), CMC-based demineralized cortical bone matrix (DB) or CMC-based demineralized cortical bone with cancellous bone (NDDB), and the wound area was evaluated at 4, 8, and 12 weeks post-implantation. DBX showed significantly lower radiopacity, bone volume fraction, and bone mineral density than DB and NDDB before implantation. However, bone healing score, bone volume fraction, bone mineral density, and residual bone area at 4, 8, and 12 weeks post-implantation revealed no significant differences in bone healing capacity. Overall, three DBM products with different carrier molecules or bone compositions showed similar bone healing capacity.

Keyword

bone graft substitutes; bone regeneration; carboxymethylcellulose; hyaluronic acid; rabbit

MeSH Terms

Animals
Bone Matrix/*physiology
Bone Transplantation
Carboxymethylcellulose Sodium/*pharmacology
Histology
Hyaluronic Acid/*pharmacology
Rabbits
*Wound Healing
X-Ray Microtomography
X-Rays
Carboxymethylcellulose Sodium
Hyaluronic Acid

Figure

• 

  Fig. 1 Micro-CT images of DBX (A), DB (B), and NDDB (C). Three bone graft substitutes were imaged using a micro-CT. DBX clearly shows lower radiopacity than DB and NDDB, while there are few radiopaque particles in DBX.

• 

  Fig. 2 Radiographic images of no treatment, DBX, DB, and NDDB at 0, 4, 8, and 12 weeks post-implantation. There was callus formation but no union in the no treatment group at 12 weeks after the surgery. DBX was similar to the no treatment at 0 weeks post-implantation due to its low radiopacity. There were increased new bone densities, but no difference in DBX, DB, and NDDB at 4, 8, and 12 weeks post-implantation.

• 

  Fig. 3 Bone healing scores measured by radiographic image analysis. Four rabbits from each group were euthanized at 4, 8, or 12 weeks after surgical procedures, respectively, and X-ray images were taken with an X-ray machine. Digital images were used to evaluate the degree of bone healing based on the criteria defined by Cook et al. [6]. Bone healing scores increased nearly linearly during the experimental period, and there were no significant differences in this trend among groups. The values shown are the mean ± SD (n = 8).

• 

  Fig. 4 Changes in bone volume fraction (%) after implantation. The samples from the euthanized rabbits were imaged using a micro-CT at 4, 8, and 12 weeks after implantation. The scanned data were reconstructed using software. Bone mineral density (BMD) and the ratio of bone volume to total volume (BV/TV) of three DBM products were calculated according to the program set by the software. The values are the mean ± SD (n = 8).

• 

  Fig. 5 Changes in bone mineral density after implantation. The samples from the euthanized rabbits were imaged using a micro-CT at 4, 8, and 12 weeks after implantation. The scanned data were reconstructed using the software. The BMD of the three DBM products were calculated according to program set by the software (CT-analyzer; Skyscan). The values are the mean ± SD (n = 8).

• 

  Fig. 6 Light micrograph images taken at 4, 8, and 12 weeks post-implantation. The samples were decalcified and embedded in paraffin. The tissue sections obtained in 4-µm thickness were stained with H&E. The arrow indicates the junction between normal bone and host bone. All experimental groups show considerable new bone formation at the defect sites and the grafted BDMs surrounded by these new bones at 4 weeks post-implantation. All groups show initial signs of marrow formation at 8 weeks. Bone remodeling was nearly complete in all groups at 12 weeks. The magnification was 12.5.

• 

  Fig. 7 Measurement of the residual area (mm2) of DBX, DB, and NDDB after implantation. The samples were decalcified and embedded in paraffin. The five tissue sections (100 µm away from each section) were 4-µm thick and stained with H&E, after which they were thoroughly observed under a microscope and the regions of proximal and distal host bone in the slides were photographed. Residual graft areas (mm2) were calculated using a digital image analyzer to evaluate the resorption rate of the grafts. The values are the mean ± SD (n = 8).

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