Journal of the Korean Association of Oral and Maxillofacial Surgeons

The Korean Association of Oral and Maxillofacial Surgeons (대한구강악안면외과학회)
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The effect of silk fibroin and rhBMP-2 on bone regeneration in rat calvarial defect model

백서 두개골결손모델에서 실크단백과 골형성단백 이식체가 골재생에 미치는 영향

  • Nam, Jeong-Hun (Department of Oral and Maxillofacial Surgery, Kangnam Sacred Heart Hospital, College of Medicine) ;
  • Noh, Kyung-Lok (Department of Oral and Maxillofacial Surgery, Kangnam Sacred Heart Hospital, College of Medicine) ;
  • Pang, Eun-O (Department of Oral and Maxillofacial Surgery, Kangnam Sacred Heart Hospital, College of Medicine) ;
  • Yu, Woo-Geun (Department of Oral and Maxillofacial Surgery, Kangnam Sacred Heart Hospital, College of Medicine) ;
  • Kang, Eung-Sun (Department of Oral and Maxillofacial Surgery, Kangnam Sacred Heart Hospital, College of Medicine) ;
  • Kweon, Hae-Yong (National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Seong-Gon (Department of Oral and Maxillofacial Surgery, College of Dentistry, Gangneung-Wonju National University) ;
  • Park, Young-Ju (Department of Oral and Maxillofacial Surgery, Kangnam Sacred Heart Hospital, College of Medicine)
  • 남정훈 (한림대학교 의과대학 강남성심병원 구강악안면외과학교실) ;
  • 노경록 (한림대학교 의과대학 강남성심병원 구강악안면외과학교실) ;
  • 방은오 (한림대학교 의과대학 강남성심병원 구강악안면외과학교실) ;
  • 유우근 (한림대학교 의과대학 강남성심병원 구강악안면외과학교실) ;
  • 강응선 (한림대학교 의과대학 강남성심병원 강악안면외과학교실) ;
  • 권해용 (농촌진흥청) ;
  • 김성곤 (강릉원주대학교 치과대학 구강악안면외과학교실) ;
  • 박영주 (한림대학교 의과대학 강남성심병원 구강악안면외과학교실)
  • Received : 2010.06.07
  • Accepted : 2010.10.19
  • Published : 2010.10.29
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Abstract

Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

Keywords

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