Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Inhibitory Effect of Conditioned Medium of Silk Fibroin-Treated Osteoblasts in Osteoclast Differentiation

실크피브로인을 처리한 MC3T3-E1 조골세포 조건배양액의 파골세포 분화억제효과

  • Yeo, Joo-Hong (Dept. of Agricultural Biology, National Institute of Agricultural Science and Technology) ;
  • Park, Kyung-Ho (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Ju, Won-Chul (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Lee, Jin-Ah (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University) ;
  • Lee, Kwang-Gill (Dept. of Agricultural Biology, National Institute of Agricultural Science and Technology) ;
  • Woo, Soon-Ok (Dept. of Agricultural Biology, National Institute of Agricultural Science and Technology) ;
  • Han, Sang-Mi (Dept. of Agricultural Biology, National Institute of Agricultural Science and Technology) ;
  • Kweon, Hae-Yong (Dept. of Agricultural Biology, National Institute of Agricultural Science and Technology) ;
  • Kim, Sung-Su (Dept. of Anatomy, Chung-Ang University) ;
  • Cho, Yun-Hi (Dept. of Medical Nutrition, Graduate School of East-West Medical Science, Kyung Hee University)
  • 여주홍 (농촌진흥청 농업생물부) ;
  • 박경호 (경희대학교 동서의학대학원 의학영양학과) ;
  • 주원철 (경희대학교 동서의학대학원 의학영양학과) ;
  • 이진아 (경희대학교 동서의학대학원 의학영양학과) ;
  • 이광길 (농촌진흥청 농업생물부) ;
  • 우순옥 (농촌진흥청 농업생물부) ;
  • 한상미 (농촌진흥청 농업생물부) ;
  • 권해용 (농촌진흥청 농업생물부) ;
  • 김성수 (중앙대학교 의과대학 해부학교실) ;
  • 조윤희 (경희대학교 동서의학대학원 의학영양학과)
  • Published : 2008.08.30
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Abstract

In this study, we investigated the indirect effect of silk-fibroin on osteoclastic differentiation of RAW264.7 cells. The conditioned medium were collected from MC3T3-E1 osbeoblasts treated with $0.001\;mg/mL{\sim}0.1\;mg/mL$ silk fibroin for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of osteoclastic cytokines in the conditioned medium, the protein expression of osteoprotegerin (OPG) with silk-fibroin was not significantly different. However, the protein expression of interleukin (IL)-$1{\beta}$ was specifically lower in a dose dependent manner. In RAW264.7 cells, the conditioned medium with silk-fibroin inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts in a dose dependent manner. Taken together, we demonstrated that the conditioned medium of silk-fibroin treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with inhibiting selective expression of IL-$1{\beta}$.

본 연구는 MC3T3-E1 조골전구세포에 실크피브로인 처리 시 파골세포분화 관련 국소인자 중 파골세포로의 분화를 강력히 유도하는 IL-$1{\beta}$의 발현 및 파골세포로의 분화억제에 관여하는 국소인자인 OPG의 발현변화를 조골세포 조건 배양액에서 살펴보았으며, 이 조골세포 조건배양액을 RAW264.7 파골전구세포에 처리 시 파골세포로의 분화를 억제하는지를 알아보았다. 조골세포 조건배양액에서 OPG의 발현은 농도별($0.001\;mg/mL{\sim}0.1\;mg/mL$) 실크피브로인 처리 시 $86%{\sim}100%$로 양성대조군(+C)과 유의적 차이가 없었다. 그러나 IL-$1{\beta}$의 발현은 0.1 mg/mL 실크피브로인 처리 시 양성대조군(+C)의 1.8% 수준으로 그 발현을 현저히 억제 하였다. 조골세포조건배양액을 처리한 RAW264.7 세포의 파골세포로의 분화정도는 농도 의존적으로 현저히 감소 하였다. 특히, 0.1 mg/mL의 실크피브로인을 처리한 조골세포 조건배양액 처리 시 파골세포로 분화하지 않은 음성대조군(-RANKL) 수준으로 파골세포 분화를 현저히 억제한 것은 OPG의 발현 증가가 아닌 IL-$1{\beta}$의 발현 억제에 의한 것으로 사료되며, 추후 연구를 통해 관련 기전의 연구가 필요할 것으로 생각되어진다.

Keywords

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