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치의학석사학위논문 NAC 함유 PMMA 계골시멘트에서유출되는성분의 화학적및생물학적분석 Chemical and Biological Analysis of Components Released from NAC-supplemented Poly(methylmethacrylate)-based Bone Cement
NAC 함유 PMMA 계골시멘트에서유출되는성분의 화학적및생물학적분석 Chemical and Biological Analysis of Components Released from NAC-supplemented Poly(methylmethacrylate)-based Bone Cement
국문요약 ( 초록 ) NAC 함유 PMMA 계골시멘트에서유출되는성분의 화학적및생물학적분석 박희철 Dept. of Dental Biomaterials Science The Graduate School of Seoul National University 자기경화형의 PMMA계골시멘트는골과매식재사이에가해지는압력을효과적으로분배하여환자의통증감소와빠른회복력을유도하고시술이간편하여골과매식재혹은골간의접착제로서가장널리사용되어지고있는생체접착물질중하나이다. 그러나 PMMA 계골시멘트는일정시간경과후의단량체의용출로인한세포독성의증가, 주위의염증반응유발등이단점으로지적되고있다. 본실험에서는 PMMA계골시멘트시술후유출되는단량체의성분 (methyl methacrylate(mma), benzoyl peroxide(bpo), dimethyl para toluidine(dmpt)) 을 High Performance Liquid Chromatography(HPLC) 를이용하여분석하고유출성분이세포독성에미치는영향을알아보았다. 직경 6 mm, 높이 3 mm disc 형태의테프론몰드를사용하여골시멘트시편을제작후 DW에서 0-0.5hr/0.5-1.5hr/1.5-3hr/3-6hr 간격으로시편을이동하며추출하였다. 2X ɑ-mem 배지와혼합한추출물을 MC3T3-E1 세포에처리하여세포독성을측정하였다. 또기존의 HPLC 실험조건을이용하여, 골시멘트성분중에 MMA, BPO, DMPT의유출량을측정하고, MC3T3-E1 세포에직접처리하였을경우의세포독성을측정하여추출물의세포독성결과와비교하였다.
실험결과 PMMA계골시멘트에서유출되는단량체가 MC3T3-E1 세포독성에영향을주는것으로확인되었다. 세포독성에영향을주는주된요인이 BPO임을세포실험과 HPLC를이용한성분분석을통해확인하였다. 세포에 BPO와 MMA를같이처리한실험에서의세포독성이 BPO 단독의경우보다낮았다. MMA가 BPO와반응하여세포독성을줄여주고 DMPT는세포독성에영향을미치지않는것으로확인되었다. 골시멘트에 BPO와 MMA 비율이골시멘트물성에미치는영향과세포독성영향평가에대한추가적인실험이필요하다. 주요어 : 골시멘트, BPO, MMA, DMPT, NAC, 세포독성, HPLC 학번 : 2007-22097
목 차 1. 서론 1 2. 재료및방법 5 2-1 5 2-2 5 2-3 6 2-4 6 2-5 7 2-6 8 3. 결과 9 3-1 9 3-2 3-2 9 3-3 12 3-4 12 3-5 3-2 3-6 3-2 17 17 3-7 17 4. 고찰 24 5. 참고문헌 26
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mouse 이식 1주차다형백혈구세포 (Luekocyte(Neurophil)) 임파구 (Lymphocyte) 혈장세포 (Plasma cell) 대식세포 (macrophage) 거대세포 (Giant cell) 괴사 (necrosis) mouse 이식 1주차다형백혈구세포 (Luekocyte(Neurophil)) 임파구 (Lymphocyte) 혈장세포 (Plasma cell) 대식세포 (macrophage) 거대세포 (Giant cell) 괴사 (necrosis) mouse 이식 1주차다형백혈구세포 (Luekocyte(Neurophil)) 임파구 (Lymphocyte) 혈장세포 (Plasma cell) 대식세포 (macrophage) 거대세포 (Giant cell) 괴사 (necrosis) Control(Teflon tube) Up Slide Down 6 4 3 4.3 1 2 2 2 2 1 5 6 2 4.3 1 2 1 1 1.3 1 3 3 0 2 1 2 2 1 1.2 1 PMMA Up Slide Down 7 10 12 9.7 2 5 2 1 2.7 1 1 5 2 2.7 1 4 7 3 4.7 1 2 0 0 0.7 1 2 8 2 4 1 PMMA+1%NAC Up Slide Down 10 6 5 7 2 4 2 3 3 1 8 5 6 6.3 2 0 1 3 1.3 1 3 1 1 1.7 1 0 1 5 2 1-21 -
mouse 이식 2주차다형백혈구세포 (Luekocyte(Neurophil)) 임파구 (Lymphocyte) 혈장세포 (Plasma cell) 대식세포 (macrophage) 거대세포 (Giant cell) 괴사 (necrosis) mouse 이식 2주차다형백혈구세포 (Luekocyte(Neurophil)) 임파구 (Lymphocyte) 혈장세포 (Plasma cell) 대식세포 (macrophage) 거대세포 (Giant cell) 괴사 (necrosis) mouse 이식 2주차다형백혈구세포 (Luekocyte(Neurophil)) 임파구 (Lymphocyte) 혈장세포 (Plasma cell) 대식세포 (macrophage) 거대세포 (Giant cell) 괴사 (necrosis) Control(Teflon tube) Up Slide Down 6 5 1 4.3 1 4 5 3 2 1 6 4 4 4.3 1 2 3 2 1.3 1 3 3 4 2 1 2 2 4 1.7 1 PMMA Up Slide Down 8 11 14 9.7 2 5 4 2 2.7 1 3 5 2 2.7 1 6 7 5 4.7 1 2 4 4 0.7 1 4 8 1 4 1 PMMA+1%NAC Up Slide Down 8 6 7 7 2 5 2 2 3 1 7 5 7 6.3 2 4 1 3 2.6 1 2 1 1 1.7 1 4 1 5 3.3 1-22 -
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Abstract Chemical and Biological Analysis of Components Released from NAC-supplemented Poly(methylmethacrylate)-based Bone Cement Hee Chul Park Dept. of Dental Biomaterials Science The Graduate School of Seoul National University Self-curing poly(methylmethacrylate) (PMMA)-based bone cement is the most widely used bone cement because of its excellent physical properties and convenience to handle. However, PMMA bone cement has been known to cause biological adverse effects due to the elevated temperature during polymerization and release of harmful components into surrounded tissues. In this study, we analyzed the chemical released from PMMA bone cement and assessed their cytotoxicity in order to reveal a component responsible for the toxicity of PMMA. We also observed cell and tissue compatibility of PMMA supplemented with N-acetylcysteine(NAC), an antioxidant. Among the extracts obtained at interval, cytotoxicity was exhibited only in the extract obtained at the first 30 min. The cytotoxicity was not observed when NAC was added into the bone cement. Based on HPLC analysis, we also found that benzoyl peroxide(bpo) was a determining factor to induce cytotoxicity, although concentration of BPO was much less that of methyl methacrylate. Interestingly, cytotoxicity of BPO was reduced
in the presence of MMA, suggesting that BPO and MMA reacted each other. Finally, there was no significant difference in tissue reaction between un-supplemented and NAC-supplemented PMMA, when the bone cements were implanted subcutaneously at dorsal area of mice for 2 weeks. keywords : Bone cement, BPO, MMA, DMPT, NAC, Cytotoxicity, HPLC Student Number : 2007-22097