J Periodontal Implant Sci.  2016 Oct;46(5):350-359. 10.5051/jpis.2016.46.5.350.

Increased osteoinductivity and mineralization by minimal concentration of bone morphogenetic protein-2 loaded onto biphasic calcium phosphate in a rabbit sinus

• Kim JS
• Cha JK
• Lee JS
• Choi SH
• Cho KS

• KMID: 2354933
• DOI: http://doi.org/10.5051/jpis.2016.46.5.350

Abstract

PURPOSE
The purpose of the present study was to evaluate the effectiveness of a minimal concentration of bone morphogenetic protein-2 (BMP-2) in terms of quantitative and qualitative analyses of newly formed bone in a rabbit maxillary sinus model.
METHODS
In 7 rabbits, sinus windows were prepared bilaterally. Biphasic calcium phosphate (BCP) loaded with 0.05 mg/mL BMP-2 was grafted into one sinus (the BMP group) and saline-soaked BCP was placed into the other (the control group) in each animal. The animals were allowed an 8-week healing period before being sacrificed. Specimens including the augmented area and surrounding tissues were then removed and evaluated both radiographically and histologically.
RESULTS
There was a difference in the mineralization of new bone between the groups. In the BMP group, the greater part of the new bone consisted of mature lamellar bone with an evident trabecular pattern, whereas the control group showed mostly woven bone, consisting only partially of lamellar bone. Histometrically, the area of new bone was significantly greater (4.55±1.35 mm2 vs. 2.99±0.86 mm2) in the BMP group than in the control group (P<0.05); however, the total augmentation volumes were not significantly different between the groups.
CONCLUSIONS
Within the limitations of this study, it can be suggested that a minimal concentration of BMP-2 (0.05 mg/mL) had an osteoinductive effect with accelerated mineralization in a rabbit sinus model using a BCP carrier.

Keyword

Bone morphogenetic proteins; Physiologic calcification; Hydroxyapatite-beta tricalcium phosphate; Sinus floor augmentation

MeSH Terms

Animals
Bone Morphogenetic Proteins
Calcification, Physiologic
Calcium*
Maxillary Sinus
Miners*
Rabbits
Sinus Floor Augmentation
Transplants
Bone Morphogenetic Proteins
Calcium

Figure

• 

  Figure 1 Photographs showing the surgical procedure. (A) Bilateral sinus windows were cut using a 5.5-mm circular reamer and (B) graft materials were placed into those windows: the test material (BMP-2-loaded BCP) on one side and the control material (saline-soaked BCP) on the other. BMP, bone morphogenetic protein; BCP, biphasic calcium phosphates.

• 

  Figure 2 Representative micro-CT image. Dome-shaped augmented sinuses were observed. The red portion is the BMP-2-loaded graft and the blue portion is the saline-soaked BCP (control) graft. micro-CT, micro-computed tomography; BMP, bone morphogenetic protein; BCP, biphasic calcium phosphates.

• 

  Figure 3 Histological images. (A, B) Low magnification images of the BMP and control group, respectively (scale bar=1 mm); (C, D) high magnification images of the central portion of the augmented area, the BMP and control group, respectively (scale bar=100 μm); (E, F) high magnification images of the sinus membrane area, the BMP and control group, respectively (scale bar=100 μm). NB, new bone; RP, residual particles; FV, fibrovascular tissue; SM, Schneiderian membrane, Masson trichrome staining; BMP, bone morphogenetic protein.

• 

  Figure 4 Highly magnified histologic images of (A) the BMP group and (B) the control group. The NB of the BMP group showed more maturation than the NB of the control group (white asterisks; the Haversian canals, Masson trichrome stains; scale bar=100 μm). BMP, bone morphogenetic protein; NB, new bone; LB, lamellar bone; RP, residual particles; WB, woven bone.

• 

  Figure 5 The proportion of NB, RP, and FV. The proportion of RP in the BMP group was significantly less than the control group (P<0.05). NB, new bone; RP, residual particles; FV, fibrovascular tissue; BMP, bone morphogenetic protein. *Statistically significant difference between the two groups (P<0.05).

Reference

1. Kim JS, Cha JK, Cho AR, Kim MS, Lee JS, Hong JY, et al. Acceleration of bone regeneration by BMP-2-loaded collagenated biphasic calcium phosphate in rabbit sinus. Clin Implant Dent Relat Res. 2015; 17:1103–1113.

2. LeGeros RZ, Lin S, Rohanizadeh R, Mijares D, LeGeros JP. Biphasic calcium phosphate bioceramics: preparation, properties and applications. J Mater Sci Mater Med. 2003; 14:201–209.

3. Lim HC, Zhang ML, Lee JS, Jung UW, Choi SH. Effect of different hydroxyapatite:beta-tricalcium phosphate ratios on the osteoconductivity of biphasic calcium phosphate in the rabbit sinus model. Int J Oral Maxillofac Implants. 2015; 30:65–72.

4. Mangano C, Sinjari B, Shibli JA, Mangano F, Hamisch S, Piattelli A, et al. A human clinical, histological, histomorphometrical, and radiographical study on biphasic HA-beta-TCP 30/70 in maxillary sinus augmentation. Clin Implant Dent Relat Res. 2015; 17:610–618.

5. Choi Y, Lee JS, Kim YJ, Kim MS, Choi SH, Cho KS, et al. Recombinant human bone morphogenetic protein-2 stimulates the osteogenic potential of the Schneiderian membrane: a histometric analysis in rabbits. Tissue Eng Part A. 2013; 19:1994–2004.

6. Asai S, Shimizu Y, Ooya K. Maxillary sinus augmentation model in rabbits: effect of occluded nasal ostium on new bone formation. Clin Oral Implants Res. 2002; 13:405–409.

7. Brunner E, Langer F. Nonparametric analysis of ordered categorical data in designs with longitudinal observations and small sample sizes. Biom J. 2000; 42:663–675.

8. Torrecillas-Martínez L, Galindo-Moreno P, Ávila-Ortiz G, Ortega-Oller I, Monje A, Hernández-Cortés P, et al. Significance of the immunohistochemical expression of bone morphogenetic protein-4 in bone maturation after maxillary sinus grafting in humans. Clin Implant Dent Relat Res. 2016; 18:717–724.

9. Hong JY, Kim MS, Lim HC, Lee JS, Choi SH, Jung UW. A high concentration of recombinant human bone morphogenetic protein-2 induces low-efficacy bone regeneration in sinus augmentation: a histomorphometric analysis in rabbits. Clin Oral Implants Res. Forthcoming. 2015.

10. Boden SD, Kang J, Sandhu H, Heller JG. Use of recombinant human bone morphogenetic protein-2 to achieve posterolateral lumbar spine fusion in humans: a prospective, randomized clinical pilot trial: 2002 Volvo Award in clinical studies. Spine. 2002; 27:2662–2673.

11. Yon J, Lee JS, Lim HC, Kim MS, Hong JY, Choi SH, et al. Pre-clinical evaluation of the osteogenic potential of bone morphogenetic protein-2 loaded onto a particulate porcine bone biomaterial. J Clin Periodontol. 2015; 42:81–88.

12. Kim MS, Kwon JY, Lee JS, Song JS, Choi SH, Jung UW. Low-dose recombinant human bone morphogenetic protein-2 to enhance the osteogenic potential of the Schneiderian membrane in the early healing phase: in vitro and in vivo studies. J Oral Maxillofac Surg. 2014; 72:1480–1494.

13. Kenley RA, Yim K, Abrams J, Ron E, Turek T, Marden LJ, et al. Biotechnology and bone graft substitutes. Pharm Res. 1993; 10:1393–1401.

14. Haidar ZS, Hamdy RC, Tabrizian M. Delivery of recombinant bone morphogenetic proteins for bone regeneration and repair. Part B: delivery systems for BMPs in orthopaedic and craniofacial tissue engineering. Biotechnol Lett. 2009; 31:1825–1835.

15. Jung RE, Weber FE, Thoma DS, Ehrbar M, Cochran DL, Hämmerle CH. Bone morphogenetic protein-2 enhances bone formation when delivered by a synthetic matrix containing hydroxyapatite/tricalciumphosphate. Clin Oral Implants Res. 2008; 19:188–195.

16. Kim JW, Jung IH, Lee KI, Jung UW, Kim CS, Choi SH, et al. Volumetric bone regenerative efficacy of biphasic calcium phosphate-collagen composite block loaded with rhBMP-2 in vertical bone augmentation model of a rabbit calvarium. J Biomed Mater Res A. 2012; 100:3304–3313.

17. Jang JW, Yun JH, Lee KI, Jang JW, Jung UW, Kim CS, et al. Osteoinductive activity of biphasic calcium phosphate with different rhBMP-2 doses in rats. Oral Surg Oral Med Oral Pathol Oral Radiol. 2012; 113:480–487.

18. Lim HC, Hong JY, Lee JS, Jung UW, Choi SH. Late-term healing in an augmented sinus with different ratios of biphasic calcium phosphate: a pilot study using a rabbit sinus model. J Periodontal Implant Sci. 2016; 46:57–69.

19. Kim MS, Lee JS, Shin HK, Kim JS, Yun JH, Cho KS. Prospective randomized, controlled trial of sinus grafting using Escherichia-coli-produced rhBMP-2 with a biphasic calcium phosphate carrier compared to deproteinized bovine bone. Clin Oral Implants Res. 2015; 26:1361–1368.

20. Park JC, Kim JC, Kim BK, Cho KS, Im GI, Kim BS, et al. Dose- and time-dependent effects of recombinant human bone morphogenetic protein-2 on the osteogenic and adipogenic potentials of alveolar bone-derived stromal CELLS. J Periodontal Res. 2012; 47:645–654.

21. Choi Y, Yun JH, Kim CS, Choi SH, Chai JK, Jung UW. Sinus augmentation using absorbable collagen sponge loaded with Escherichia coli-expressed recombinant human bone morphogenetic protein 2 in a standardized rabbit sinus model: a radiographic and histologic analysis. Clin Oral Implants Res. 2012; 23:682–689.