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The change of gene expression in Raw 264.7 mouse macrophage cells induced by liberated proteins from silkworm cocoon and its application to clinical trials as a barrier membrane

원문보기

주제(키워드) Silk membrane , Microarray , RT-PCR , Clinical study
발행기관 강릉원주대학교 대학원
지도교수 김성곤
발행년도 2017
학위수여년월 2017. 8
학위명 박사
학과 및 전공 일반대학원 치의학과
세부전공 구강악안면외과
원문페이지 38
파일정보 2007 word
실제URI http://www.dcollection.net/handler/kangnung/000000009765
본문언어 영어

초록/요약

The silk mat produced by the silkworm Bombyx mori has been broadly reported as a barrier membrane for guided bone regeneration (GBR). It has been shown to regenerate new bone when using the middle layer of the silkworm’s cocoon. In this study, the supernatant from each layer of silk mat was administered to Raw 264.7 mouse macrophage cells, and changes in gene expression were analyzed by cDNA microarray assay. Two genes related to inflammatory responses were selected, and the change in expression of these genes was confirmed by quantitative RT-PCR. Microarray analysis and quantitative RT-PCR results demonstrated that interleukin-1β and tumor necrosis factor-alpha expression levels were significantly lower in the supernatant from the middle layer of the cocoon. In the clinical study, 20 patients who had bilateral impacted mandibular third molars were included. After flap elevation and surgical extraction, the clinical attachment level and probing depth were evaluated at the distobuccal (DB), distal (D) and distolingual (DL) aspects of the mandibular second molar. Both extraction sites were randomly covered with either a silk mat from the middle layer of the silkworm cocoon or an e-PTFE membrane. At 3 and 6 months after the operation, the clinical attachment level and probing depth were evaluated and compared. The silk-mat-treated group showed a significantly reduced probing depth and a higher gain of the clinical attachment level. This study revealed that the silk mat from the middle layer of the silkworm cocoon showed excellent ability as a barrier membrane for GBR.

Ⅰ. Introduction · · · · · · · · · · · · · · · · · · · · · · · · · · ·1
Ⅱ. Materials and Methods · · · · · · · · · · · · · · · · · 3
1. Preparation of silk nets for cell culture
2. Cell culture
3. Microarray
4. Quantitative RT-PCR
5. Clinical Study
Ⅲ. Results · · · · · · · · · · · · · · · · · · · · · · · · · · · · 7
1. Gene expression analysis with DNA microarrays compared with each layer of the silk-worm cocoon
2. RT –PCR
3. Clinical Study
Ⅵ. Discussion · · · · · · · · · ·· · · · · · · · · · · · · ··10
Ⅴ. Conclusions · · · · · · · · · · · ·· · · · · · · · · · 13
Ⅵ. References · · · · · · · · · · · · · · · · · · · · · · 14
Figure Legends · · · · · · · · · · · · · · · · · · · · · · 20
Figures · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 21
Appendix · · ·· · · · · · · · · · · · · · · · · · · · · · · · · 25
Abstract in Korean · · · · · · · · · · · · · · · · · · · 31

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The change of gene expression in Raw 264.7 mouse macrophage cells induced by liberated proteins from silkworm cocoon and its application to clinical trials as a barrier membrane

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