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수산화칼슘 처리된 Porphyromonas endodontalis Lipopolysaccharide가 다형핵백혈구의 IL-1과 TNF-α 생성에 미치는 영향에 관한 연구
박찬제,박동성,유현미,오태석,임성삼 大韓齒科保存學會 2002 Restorative Dentistry & Endodontics Vol.27 No.5
Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines , namely, interleukin 1 and tumor necrosis factor-α from immune cells, Although monocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-α. Calcium hydroxide has been shown to be an effective medicament in root canal infectionsm, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)_2 may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromanas endodonatalis LPS with calcium hydroxide alters its biological action as neasured by human PMN secretion of IL-1 and TNF-α, and it was compared with Escherichia coli LPS. P.endodonatalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified . 100 ㎍/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)_2 at 37℃ for 7 days. The supernatants were subjected to ultrafiltration. And the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4 × 10^6 cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10㎍/ml) for 24 hours at 37℃ in 5% CO_2 incubator. The supernatants of cells were collected and the levels of IL-1α, IL-1β and TNF-α were measured by enzyme-linked immunosorbent assay. The results were as follows; 1. The levels of IL-1α, IL-1β, TNF-α from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05). while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05). 3. The levels of secretion or all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with P.endodontalis LPS were significantly lower than those released from PMN stimulated with E. coli LPS (p<0.05).
곽경인,박동성,유현미,오태석 대한치과보존학회 2000 Restorative Dentistry & Endodontics Vol.25 No.4
임상에서 점차 Mineral Trioxide Aggregate (MTA) 에 대한 우수성이 소개되면서, apical matrix로의 사용은 주목할 만하다 할 수 있다. 본 연구의 목적은 개방형근관에서 MTA가 apical matrix로 사용될 때의 치근단 밀폐효과를 알아보고, 근관충전의 시기와 matrix의 두께가 치근단 밀폐에 미치는 영향을 치근단 미세누출의 측면과 matrix탈락빈도의 측면에서 관찰하고자 하는 것이다. 개방형 근관을 재현하고자 45개의 발거된 단근치에 #90크기로 근단공을 형성하였고, 투명레진등을 이용하여 치근단병소를 갖는 치조골을 재현한 후, 4개의 실험 군과 1개의 대조 군으로 분류하였다. A군: 2mm두께의 MTA matrix 형성후, 열연화된 Gutta-percha와AH26 sealer를 이용하여 즉시 근관충전. B군: 2mm두께의 MTA matrix 형성후, A군과 같은 방법으로 24시간 후 근관충전. C군: 4mm 두께의 MTA matrix 형성후, A군과 같은 방법으로 즉시 근관충전. D군: 4mm두께의 MTA matrix 형성후, A군과 같은 방법으로 24시간 후 근관충전. 대조군: matrix를 사용하지 않고 열연화된 Gutta-percha와 AH26 sealer를 이용하여 근관충전하였다. Matrix의 탈락이 있는 경우 기록하고 완전수세 후 다시 시행하였다. 색소침투정도의 평가를 위해 methylene blue 에 치아를 48시간 침수시킨 후 협설로 양분하였다. 각 시편들을 고배율 현미경 관찰 후 비디오촬영 하였고, digitalized image analysing program 을 이용하여 두명의 관찰자가 누출의 정도를 평가한 후, One-way ANOVA로 통계적 유의성을 검증하여 다음과 같은 결과를 얻었다. 1. Matrix를 사용한 실험 군이 matrix를 사용하지 않은 대조 군에 비해 유의성 있게 낮은 누출을 보였다(p<0.05) . 그러나 근관충전시기와 matrix의 두께를 달리한 실험군 내에서는 누출에 유의성 있는 차이가 나타나지 않았다. 2 모든 실험군에서 gutta-percha를 이용한 근관충전의 시기에 matrix탈락 및 gutta-percha 의 압출은 발생하지 않았다.