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Biocompatibility and biomineralization assessment of experimental sealers Sealepox and Sealepox RP compared to Pro-Root MTA ®: histological and immunohistochemical study in the subcutaneous tissue of rats


Tissue reactions against new filling materials should be checked earlier in subcutaneous connective tissue (ISO 10993-6. 2007), and biocompatibility can be observed by staining hematoxila and eosin (HE) (Cintra et al., 2006, 2010, 2012, Gomes-Filho et al., 2008, 2009, 2011). Moreover, the use of immunohistochemical technique offers the possibility of understanding the cellular and molecular mechanisms involved in the inflammatory response more accurately. The markers TNF-±, IL-6 and CD5 are often used in studies aimed at a better understanding of the phenomena that involve an inflammatory tissue response. In addition, despite to biocompatibility, it is evident a concern when the possibility of filling material stimulate mineralization adjacent tissue forming the biological apical sealing. Assessment of the biomineralization ability requires specific methods, such as the Von Kossa technique and by polarized light (Gomes-Filho et al., 2008, 2009, 2011, Cintra et al. 2012). The stained by the Von Kossa is used to detect structures mineralized tissue which stained microscopic analysis appears in black. Polarized light can observe structures having birefringent crystals of calcium carbonate originating from the combination of calcium ions and carbon dioxide of the material of the fabric (Gomes-Filho et al. 2008). Moreover, the immunohistochemical technique is able to accurately detect cells that are involved with the production of mineralized tissues via detection immunomarkers related to the process of cell differentiation, as RUNX-2, or by identifying molecules involved in the biomineralization, such as osteocalcin (OC) and osteopontin (OP). Given the development of experimental cements Sealepox Sealepox and RP, which contain calcium hydroxide in its formulation, we propose to evaluate quantitatively and qualitatively the tissue response to these sealers, comparing them to the Mineral Aggregate Trióxidos, Pro-Root MTA ®. Will be used 140 polyethylene tubes and 35 rats (four tubes per rat) divided into 4 groups: G1 - control (empty tubes), G2 - tubes containing Sealepox; G3 - tubes containing Sealepox RP; G4 - tubes containing the ProRoot ® MTA. The evaluation times are 7, 15, 30, 60 and 90 days. After each postoperative period, seven animals are sacrificed and the polyethylene tubes together with the surrounding tissue will be removed, fixed in 10% formalin and processed for light microscopy analysis, with inclusion in glycol methacrylate, cut serial 3¼m and stained with hematoxylin and eosin, the immune markers IL-6, TNF- ± and CD5 will be used to evaluate the material biological compatibility. Will be employed Von Kossa staining, and without staining under polarized light for analysis and osteoblastic markers to verify the activity of the materials ability biomineralization. The tissue sections will be evaluated by pre-established criteria by assigning scores to the microscopic events observed. Data will be analyzed from a statistical viewpoint using the Kruskall Wallis and Dunn with a significance level of 5% (p <0.05). (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
HOLLAND, ROBERTO; GOMES FILHO, JOAO EDUARDO; ANGELO CINTRA, LUCIANO TAVARES; DE AZEVEDO QUEIROZ, INDIA OLINTA; ESTRELA, CARLOS. Factors affecting the periapical healing process of endodontically treated teeth. Journal of Applied Oral Science, v. 25, n. 5, p. 465-476, SEP-OCT 2017. Web of Science Citations: 11.
ROBERTO HOLLAND; JOÃO EDUARDO GOMES FILHO; LUCIANO TAVARES ANGELO CINTRA; ÍNDIA OLINTA DE AZEVEDO QUEIROZ; CARLOS ESTRELA. Factors affecting the periapical healing process of endodontically treated teeth. Journal of Applied Oral Science, v. 25, n. 5, p. -, Out. 2017.
PEDRO FELICIO ESTRADA BERNABE; MARIANE MAFFEI AZUMA; LUCIANA LOUZADA FERREIRA; ELOI DEZAN-JUNIOR; JOAO EDUARDO GOMES-FILHO; LUCIANO TAVARES ANGELO CINTRA. Root Reconstructed with Mineral Trioxide Aggregate and Guided Tissue Regeneration in Apical Surgery: A 5-year Follow-up. Brazilian Dental Journal, v. 24, n. 4, p. -, Jul. 2013.

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