Histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis of tissue response to sealing materials after furcation perforation
Histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis of tissue response to sealing materials after furcation perforation
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DOI：
10.1111/iej.13145
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发表时间：
2019-10-01
影响因子：
5
通讯作者：
Silva, L. A. B.
中科院分区：
文献类型：
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作者：
Silva, R. A. B.;Borges, A. T. N.;Silva, L. A. B.
目的：通过组织病理学、组织酶学、免疫组织化学和免疫荧光分析，评价Biodentine（TM）、矿物三氧化物聚合物（MTA）和牙胶封闭狗根分叉穿孔后的体内组织反应。方法学：根管治疗后，使用圆形金刚石钻在髓室底部的中心区域创建穿孔，并填充一种或另一种材料。120天后将动物安乐死，并对牙齿（n = 30）进行新矿化组织形成和胶原纤维重新插入的组织病理学分析、骨桥蛋白（OPN）和碱性磷酸酶（ALP）的免疫组织化学分析以及骨形态发生蛋白（BMP-2）、牙骨质附着蛋白（CAP）、骨唾液酸蛋白（BSP）、骨钙素（OCN）和牙骨质蛋白1（CEMP 1）。组织酶学检测TRAP活性和破骨细胞计数。使用卡方和Kruskal-Wallis检验对数据进行统计学分析（α = 0.05）。结果：牙胶不诱导矿化组织形成。MTA和Biodentine（TM）分别有88%和92%的标本形成矿化组织，差异无统计学意义（P> 0.05）。牙胶与平行于穿孔的散在胶原纤维相关。用MTA或Biodentine（TM）处理的组具有垂直于新形成的矿化组织的部分纤维再插入。所有材料均诱导OPN和ALP表达，牙胶最弱，MTA最强（P <0.05）。只有MTA诱导BMP-2、BSP、OCN、CAP和CEMP 1的表达。各组破骨细胞计数相似（P = 0.97）。结论：矿物三氧化物聚集体和Biodentine（TM）具有生物相容性，形成矿化组织和部分胶原纤维重新插入。此外，观察到几种分子参与了硅酸钙基材料诱导矿化组织的形成，ALP和OPN矿化标志物表达，而不干扰破骨细胞的数量。只有MTA刺激与牙骨质样矿化组织形成相关的蛋白质的表达。
Aim: To evaluate in vivo tissue responses after sealing furcation perforations in dog's teeth with either Biodentine (TM), mineral trioxide aggregate (MTA) or gutta-percha, by means of histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis. Methodology: After root canal treatment, perforations were created in the central region of the pulp chamber floor using a round diamond bur and filled with one or other of the materials. The animals were euthanized after 120 days, and the teeth (n = 30) were processed for histopathological analysis of new mineralized tissue formation and collagen fibre reinsertion, immunohistochemical analysis of osteopontin (OPN) and alkaline phosphatase (ALP) and immunofluorescence analysis for bone morphogenetic protein (BMP-2), cementum attachment protein (CAP), bone sialoprotein (BSP), osteocalcin (OCN) and cementum protein1 (CEMP1). Histoenzymology was performed for TRAP activity and osteoclast count. Data were analysed statistically (alpha = 0.05) using chi-square and Kruskal-Wallis tests. Results: Gutta-percha did not induce mineralized tissue formation. MTA and Biodentine(TM) formed mineralized tissue in 88% and 92% of specimens, respectively, with no significant difference (P > 0.05). Gutta-percha was associated with scattered collagen fibres parallel to the perforations. Groups treated with MTA or Biodentine(TM) had partial fibre reinsertion perpendicular to the newly formed mineralized tissue. All materials induced OPN and ALP expression, weakest for gutta-percha and strongest for MTA (P < 0.05). Only MTA induced BMP-2, BSP, OCN, CAP and CEMP1 expression. Osteoclast counts were similar in all groups (P = 0.97). Conclusions: Mineral trioxide aggregate and Biodentine(TM) were biocompatible, with formation of mineralized tissue and partial reinsertion of collagen fibres. In addition, the participation of several molecules by which calcium silicate-based materials induce the formation of mineralized tissue were noted, with expression of ALP and OPN mineralization markers, without interference in the number of osteoclasts. Only MTA stimulated the expression of proteins associated with the formation of a cementum-like mineralized tissue.