논문 상세보기
pp.267-276
Cementum is a hard connective tissue, produced by cementoblasts during tooth root formation, which provides for the attachment of the periodontal ligament to the roots and surrounding alveolar bone. Establishment of this attachment is an important event in the regeneration of lost periodontal tissues. We examined whether or not odontoblast conditioned media(CM) have a regulatory influence on the differentiation and mineralization of cementoblasts(murine cementoblastic cell line, OCCM-30) in vitro. To identify the effect of odontoblast conditioned media and dentin non collagenous proteins (dNCPs) on cementoblast differentiation and mineralization, we treated CM and dNCPs to cementoblast then differentiated the cells for 14 days. To evaluate the formation of mineralized nodules alizarin-red S staining was performed at 0,4,7 and 14 days. Expression of cementum matrix genes was measured by RT-PCR. Mineralization of cementoblasts was accelerated with CM from odontoblastic MDPC-23 and OD-11. The expression of BSP, ALP, and OC mRNA in cementoblastic OCCM-30 cells was facilitated by the MDPC-23 and OD-11 cells. The extracted dNCPs had little influence on the proliferation, cell cycle modification, and chemotaxis of OCCM-30 cells. Although the dNCPs did not exhibit chemotactic activities for cementoblasts, the dNCPs promoted the differentiation and mineralization of cementoblasts. In conclusion, the dentin matrix protein, or the secreted products of odontoblast, facilitates cementoblast differentiation and mineralization. This represents a new approach and suggests another avenue for cementum regeneration.
II. 재료 및 방법
1. 세포주와 세포배양
2. 조건배지와 co-culture system
3. 비아교질성 상아질 단백질의 분리와 정제
4. 단백질 전기영동과 western 분석
5. MTT와 chemotaxis assay
6. Flow cytometry 분석을 통한 세포주기의 확인
7. Total RNA 분리와 reverse transcriptase
8. 형태학적 관찰과 Alizarin red-S 염색
9. 통계분석
III. 연구 결과
1. 비아교질성 상아질 단백질의 확인(SDS-PAGE,
2. 조건배지를 처리한 백악모세포의 석회화, 형태학적인 변화와 유전자 발현 변화 분석
3. 비아교질성 상아질 단백질을 처리한 백악모세포의석회화, 형태학적인 변화와 유전자 발현 변화 분석
IV. 총괄 및 고찰
V. 참고문헌
