Endodontic pathogens possess collagenolytic properties that degrade human dentine collagen matrix
Endodontic pathogens possess collagenolytic properties that degrade human dentine collagen matrix
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DOI：
10.1111/iej.13018
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发表时间：
2019-04-01
影响因子：
5
通讯作者：
Finer, Y.
中科院分区：
文献类型：
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作者：
Marashdeh, M. Q.;Gitalis, R.;Finer, Y.
目的测定粪肠球菌和藤黄微球菌的胶原蛋白酶活性及其对牙本质胶原的降解能力。方法E. faecalis ATCC 29212、ATCC 47077和M.使用荧光测定法测量黄体对一般和特异性人基质金属蛋白酶（MMP）底物的反应。细菌降解牙本质胶原蛋白的能力进行了测试，通过定量的羟基脯氨酸释放到媒体的细菌或热灭活细菌（HIN）与脱矿物质的人牙本质样品孵育24小时后，通过扫描电子显微镜（SEM）的量。采用多因素方差分析和Tukey事后检验进行统计学处理（P < 0.05）。结果所有菌株均具有MMP样活性，但对底物的亲和力不同; faecalis ATCC 29212、ATCC 47077和M.黄体对MMP-8（7.75 +/- 0.88 μ mol L-1/3 × 10（6）CFU）、MMP-9（33.86 +/- 5.16 μ mol L-1/3 × 10（6）CFU）和一般MMP（26.08 +/- 4.48 μ mol L-1/3 × 10（6）CFU）具有最大的亲和力。三种菌株从脱矿牙本质释放的羟脯氨酸的量相似（P > 0.05）（范围为1.8 +/- 0.17至2.38 +/- 0.39 μ g 50 μ L-1），并且与它们的HIN对应物（0.61 +/- 0.22 μ g 50 μ L-1）相比显著更高（P < 0.001）。SEM显示与HIN相比，与细菌孵育后胶原网络降解增加。结论根管致病菌具有胶原蛋白酶的特性，使他们能够降解牙本质胶原，可能会损害牙齿和密封剂-牙齿界面。这些胶原蛋白酶的特性可以促进病原菌迁移到根管系统，并部分解释了它们在根管感染中的作用。
Aim To measure collagenolytic protease activity from Enterococcus faecalis and Micrococcus luteus and their ability to degrade human dentinal collagen. Methodology Proteases activity of E. faecalis ATCC 29212, ATCC 47077 and M. luteus towards generic and specific human matrix metalloproteinase (MMP) substrates was measured using a fluorimetric assay. The ability of the bacteria to degrade dentinal collagen was tested by quantifying the amount of hydroxyproline released into the media following incubation of the bacteria or heat-inactivated bacteria (HIN) with demineralized human dentine samples for 24 h and by scanning electron microscopy (SEM). Multifactorial anova and Tukey's post hoc test were used to analyse the data (P < 0.05). Results All strains had MMP-like activities, but with different substrate affinity; E. faecalis ATCC 29212, ATCC 47077 and M. luteus had the greatest affinity towards MMP-8 (7.75 +/- 0.88 mu mol L-1/3 x 10(6) CFU), MMP-9 (33.86 +/- 5.16 mu mol L-1/3 x 10(6) CFU) and generic MMP (26.08 +/- 4.48 mu mol L-1/3 x 10(6) CFU), respectively. The amount of hydroxyproline released from demineralized dentine was similar (P > 0.05) for the three strains (range 1.8 +/- 0.17 to 2.38 +/- 0.39 mu g 50 mu L-1) and was significantly higher (P < 0.001) compared to their HIN counterparts (0.61 +/- 0.22 mu g 50 mu L-1). SEM revealed increased collagen network degradation after incubation with bacteria versus HIN. Conclusions Endodontic pathogens possess collagenolytic protease properties that enable them to degrade dentinal collagen, potentially compromising the restoration-tooth and sealer-tooth interfaces. These collagenolytic protease properties could facilitate the migration of pathogenic bacteria into the root canal system and explain in part their role in root canal infections.