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파골 전구세포에서 여러 마모입자에 의한 NF-κB와 c-Jun/AP-1 전사 인자의 활성화

Activation of Transcription Factor NF-κB and c-Jun/AP-1 by Various Compositions of Particles in Osteoclast Precursor Cells

권순용|권재영|최남용|한석구|김용식
대한고관절학회 (The Korean Hip Society)|2008.09|Hip and Pelvis|제20권 제3호|pp. 209-214

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키워드

NF-κBParticle마모 입자Transcription factorElectrophoretic mobility shift assay전사 인자c-Jun/AP-1파골 전구세포Osteoclast precursor cell

초록

목적: 파골 전구세포에서 다양한 조성과 입자의 크기를 갖는 여러 마모입자들이 NF-κB와 c-Jun/AP-1 활성화에 미치는 영향 을 관찰하고자 한다. 대상 및 방법: C57BL 쥐의 골수 대식세포로부터 파골 전구세포를 배양하여 PMMA (polymethylmethacrylate) 시멘트, polystyrene, titanium 입자 및 실패한 무시멘트형 인공 고관절 치환술 환자에서 채취한 마모입자에 대한 NF-κB와 c-Jun/AP- 1 전사 인자의 DNA binding activity를 electrophoretic mobility shift assays (EMSA)로 관찰하였다. 결과: 내독소 검사상 PMMA 입자와 polystyrene 입자는 내독소 (endotoxin) 음성이었으며, titanium 입자와 환자에서 채취한 마모입자는 양성이었다. PMMA 입자에 의해 NF-κB와 c-Jun/AP-1 활동도는 증가하였으며, 특히 자극 30분 후 0.6 mg/ml에 서 최적의 반응을 나타내었다. 또한 polystyrene, titanium 입자와 추출한 마모입자도 대조군에 비해 NF-κB와 c-Jun/AP-1의 활동도를 증가시켰으며 내독소 제거 시 약 40%의 활동도가 감소되었다. 결론: 파골 전구세포에서 다양한 조성과 크기의 PMMA 입자, titanium 입자, polystyrene 및 환자에서 추출한 마모입자 모두 파 골세포 분화 및 활성에 중요한 두 신호전달 체계인 NF-κB와 c-Jun/AP-1 전사 인자의 DNA binding activity를 증가시켜 인체 의 인공관절 주위 골용해 기전에 NF-κB와 c-Jun/AP-1 전사 인자는 중요한 역할을 할 것으로 사료된다.

Abstract

Purpose: This study was performed in order to investigate the effects of various particle preparations on NF-κB and c-Jun/AP-1 activity in osteoclast precursor cells. Materials and Methods: Osteoclast precursor cells isolated from C57BL mice were treated with PMMA (polymethylmethacrylate) spheres, polystyrene, titanium particles, and retrieved metal particles from failed cementless total hip replacements. NF-κB and c-Jun/AP-1 DNA binding activities were analyzed using electrophoretic mobility shift assays (EMSA). Results: Commercially available PMMA and polystyrene spheres routinely showed negativity on endotoxin assays, but titanium particles and retrieved metal particles consistently showed positivity. PMMA spheres, with a maximal response noted at 30 minutes with an optimal concentration of 0.6 mg/ml, were potent stimulator of NF-κB and c-Jun/AP-1 activity in osteoclast precursor cells. Other particles (polystyrene, titanium, metal retrievals) also activated transcription factor NF-κB and c-Jun/AP-1 compared to controls. Endotoxin removal from retrieved metal particles diminished the biologic effect by approximately 40%. Conclusion: Particles of various compositions and sizes (PMMA, polystyrene, titanium, and retrieved metal particles) activated the NF-κB and c-Jun/AP-1 signaling pathways. This suggests that NF-κB and c-Jun/AP-1 may have important roles in the pathogenesis of periprosthetic osteolysis.

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