Kyu-Nam Kim; Jung-Eun Yang; Jea-Won Jang; Balaraman Sasikala; Wang Beng; Il-Kyu Kim

doi:10.5125/jkaoms.2010.36.3.161

Journal List > J Korean Assoc Oral Maxillofac Surg > v.36(3) > 1032382

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Kim, Yang, Jang, Sasikala, Beng, and Kim: Morphometric analysis on bone formation effect of β-TCP and rhBMP-2 in rabbit mandible

Original Article

J Korean Assoc Oral Maxillofac Surg 2010;36(3):161-171.

Published online: 10 January 2010

DOI: https://doi.org/10.5125/jkaoms.2010.36.3.161

Morphometric analysis on bone formation effect of β-TCP and rhBMP-2 in rabbit mandible

Kyu-Nam Kim, Jung-Eun Yang, Jea-Won Jang, Balaraman Sasikala, Wang Beng, Il-Kyu Kim

Department of Oral and Maxillofacial surgery, College of Medicine, Inha University, Incheon, Korea

Il-kyu Kim Department of Oral and Maxillofacial Surgery, Department of Dentistry, College of Medicine, Inha University #7-206, 3rd St. Shinheung-Dong, Choong-Gu, Incheon, 400-711, Korea TEL: +82-32-890-2470 FAX: +82-32-890-2475 E-mail: kik@inha.ac.kr

Received 10 March 2010 Revised 22 April 2010 Accepted 4 May 2010

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Introduction

This study was to assess the effectiveness of new bone formation and regeneration by using a rhBMP-2 and β-TCP as a carrier in rabbits’ mandible.

Materials and Methods

The mandibles of 36 rabbits were exposed and cortical bone was penetrated for this study. The experimental subjects were divided into 3 groups each 12 rabbits; control group, experimental group 1, and experimental group 2. Control group had the defect itself without any treatment, in the experimental group 1, β-TCP only was grafted, and in the experimental group 2, rhBMP-2 soaked in β-TCP was grafted. The rabbits were sacrificed after 1, 2, 3, 4, 6, and 8weeks, and new bone formation area was examined and measured for bone quantitative and qualitative analysis with light, fluorescent and polarized microscopy.

Results

In the experimental group 1, new bone formation from the adjacent host bone was made by osteoconduction, and in the experimental group 2, direct new bone formation by osteoinduction of rhBMP-2 as well as new bone formation by osteoconduction of β-TCP were observed.

Conclusion

rhBMP-2 of experimental group 2 is very effective in the bone formation in early 2weeks and bone remodelling from 3weeks.

Keywords: Bone morphogenetic protein, Tricalcium Phosphate (TCP), Osteoconduction, Osteoinduction

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Fig. 1.

Circular defect form in rabbit's mandible.

Fig. 2.

Photomicrographs of the histological section seen by light microscopy.(original magnification x10) (TCP: β-tricalcium phosphate)

Fig. 3.

Photomicrographs of decalcified specimen.(H&E staining) A. Experimental group 1, 2 weeks.(original magnification x200) B. Experimental group 2, 2 weeks.(original magnification x200) (TCP: β-tricalcium phosphate)

Fig. 4.

Photomicrographs of decalcified specimen.(H&E staining) A. Experimental group 1, 6 weeks.(original magnification x200) B. Experimental group 2, 6 weeks.(original magnification x200) (TCP: β-tricalcium phosphate)

Fig. 5.

Quantitative analysis of bone formation at TCP/rhBMP-2 grafts. (H&E staining, original magnification x100, 4 weeks) Boundary of new bone formation were marked and calculated with Image Pro Plus 4.0.(A1-A10) (TCP: β-tricalcium phosphate, rhBMP-2: recombinant human bone morphogenetic protein-2)

Fig. 6.

Quantitative analysis of bone formation at control group (defect only), TCP grafts only and TCP/rhBMP-2 grafts. (H&E staining, original magnification x100) (TCP: β-tricalcium phosphate, rhBMP-2: recombinant human bone morphogenetic protein-2)

Fig. 7.

Quantitative analysis of bone formation at control group (defect only), β-TCP grafts only and β-TCP/rhBMP-2 grafts. (TCP: β-tricalcium phosphate, rhBMP-2: recombinant human bone morphogenetic protein-2)

Fig. 8.

Photomicrographs of undecalcified specimen.(original magnification x100) A. Experimental group 2, 8 weeks. (Villanueva bone stain) B. Experimental group 2, 8 weeks.(fluorescence light) (TCP: β-tricalcium phosphate)

Fig. 9.

Comparison of Villanueva bone stain and fluorescence light microscopic photographs of undecalcified specimen at control group (defect only), TCP grafts only and TCP/rhBMP-2 grafts. (TCP: β-tricalcium phosphate, rhBMP-2: recombinant human bone morphogenetic protein-2)

Fig. 10.

Photomicrographs of undecalcified specimen.(original magnification x100) A. Experimental group 2, 8 weeks.(Villanueva bone stain) B. Experimental group 2, 8 weeks.(polarizing light) (TCP: β-tricalcium phosphate)

Fig. 11.

Comparison of Villanueva bone stain and polarizing light microscopic photographs of undecalcified specimen at control group (defect only), TCP grafts only and TCP/rhBMP-2 grafts. (TCP: β-tricalcium phosphate, rhBMP-2: recombinant human bone morphogenetic protein-2)

Table 1.

List of used graft materials

Material	Manufacturer
β-TCP	Cowellmedi Co. Korea
rhBMP-2	Cowellmedi Co. Korea

(β-TCP; β-tricalcium phosphate, rhBMP-2; recombinant human bone morphogenetic protein-2)

Table 2.

Number of experimental animals in this study

Experimental groups	Number of Rabbits	Site
Control group (defect only)	12	24
Experimental group 1 (β-TCP only)	12	24
Experimental group 2 (rhBMP-2/β-TCP)	12	24

(β-TCP; β-tricalcium phosphate, rhBMP-2; recombinant human bone morphogenetic protein-2)

Table 3.

Quantitative analysis of bone formation area (Unit; %, 1/1.0208 mm²)

	Control group (n=16)	Experimental group 1 (n=16)	Experimental group 2 (n=16)
1 Wks	0	0	0
2 Wks	0	1.29±0.9%	11.29±1.04%
		(0.01321±0.00915)	(0.11525±0.01065)
4 Wks	15.04±2.18%	19.28±2.95%	28.20±3.79%
	(0.15357±0.02223)	(0.19683±0.03009)	(0.28788±0.03869)
6 Wks	22.73±2.38%	29.87±3.52%	42.48±3.18%
	(0.23205±0.02433)	(0.30494±0.03590)	(0.43368±0.03247)

Table 4.

Comparative analysis of inter-grou up differences (Unit: times)

	Cont. vs Exp.1	Cont. vs Exp.2	Exp.1 vs Exp.2
2 Wks	−	−	8.72
4 Wks	1.28	1.87	1.46
6 Wks	1.31	1.87	1.42

(Cont.; control group, Exp.1; experimental group 1, Exp.2; experimental group 2)

Table 5.

Quantitative analysis of inter-group bone formation area (Unit; mm², %)

	Cont. vs Exp.1	Cont. vs Exp.2	Exp.1 vs Exp.2
2 Wks	0.01321 (1.3)	0.11525 (11.3)	0.10204 (10.0)
4 Wks	0.04326 (4.3)	0.13431 (13.2)	0.09105 (8.9)
6 Wks	0.07289 (7.1)	0.20163 (19.8)	0.12874 (12.6)

(Cont.; control group, Exp.1; experimental group 1, Exp.2; experimental group 2)

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