혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구

The Effect of Platelet Derived Growth Factor - BB Loaded Chitosan/Calcium Metaphosphate on Bone Regeneration

  • 이승열 (서울대학교 치과대학 치주과학교실 및 치학연구소) ;
  • 설양조 (서울대학교 치과대학 치주과학교실 및 치학연구소, 성균관대학교 의과대학 치과학교실) ;
  • 이용무 (서울대학교 치과대학 치주과학교실 및 치학연구소) ;
  • 이주연 (이화여자대학교 약학대학 물리약학교실) ;
  • 이승진 (이화여자대학교 약학대학 물리약학교실) ;
  • 김석영 (영남대학교 공과대학 재료금속공학부) ;
  • 구영 (서울대학교 치과대학 치주과학교실 및 치학연구소) ;
  • 류인철 (서울대학교 치과대학 치주과학교실 및 치학연구소) ;
  • 한수부 (서울대학교 치과대학 치주과학교실 및 치학연구소) ;
  • 최상묵 (서울대학교 치과대학 치주과학교실 및 치학연구소) ;
  • 정종평 (서울대학교 치과대학 치주과학교실 및 치학연구소)
  • Lee, Seung-Yeol (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University) ;
  • Seol, Yang-Jo (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University, Department of Dentistry, College of Medicine, Sungkyunkwan University) ;
  • Lee, Yong-Moo (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University) ;
  • Lee, Ju-Yeon (Department Industrial Pharmacy, College of Pharmacy, Ewha Womans University) ;
  • Lee, Seung-Jin (Department Industrial Pharmacy, College of Pharmacy, Ewha Womans University) ;
  • Kim, Suk-Young (School of Materials and Metal Engineering, College of Engineering Yeungnam University) ;
  • Ku, Young (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University) ;
  • Rhyu, In-Chul (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University) ;
  • Han, Soo-Boo (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University) ;
  • Choi, Sang-Mook (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University) ;
  • Chung, Chong-Pyoung (Department of Periodontology and Dental Research Institute, College of Dentistry, Seoul National University)
  • 발행 : 2001.03.30

초록

Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

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