Immunohistochemical and biochemical assay of MMP-3 in human dentine

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Mazzoni, Annalisa ^{[1, 2, 3]} ; Papa, Veronica ^[4] ; Nato, Fernando ^[5] ; Carrilho, Marcela ^{[6, 7]} ; Tjaderhane, Leo ^{[8, 9]} ; Ruggeri, Jr., Alessandra ^[1] ; Gobbi, Pietro ^[5] ; Mazzotti, Giovanni ^[1] ; Tay, Franklin R. ^[10] ; Pashley, David H. ^[10] ; Breschi, Lorenzo ^{[11, 12]}

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^[1] Univ Bologna, Dept SAU&FAL, Bologna - Italy

^[2] Rizzoli Orthopaed Inst, Cell Biol Lab, Bologna - Italy

^[3] Rizzoli Orthopaed Inst, Lab Immunorheumatol & Tissue Regenerat Ramses Lab, Bologna - Italy

^[4] Univ Cassino, Dept Sport & Hlth Sci, I-03043 Cassino - Italy

^[5] Univ Carlo Bo, Dept STeVA, Urbino - Italy

^[6] Bandeirante Univ Sao Paulo UNIBAN, Sao Paulo - Brazil

^[7] Univ Estadual Campinas, Piracicaba Sch Dent, Dept Restorat Dent, Piracicaba - Brazil

^[8] Univ Oulu, Inst Dent, Oulu - Finland

^[9] Oulu Univ Hosp, Oulu - Finland

^[10] Med Coll Georgia, Sch Dent, Dept Oral Biol & Maxillofacial Pathol, Augusta, GA 30912 - USA

^[11] IOR, Unit Bologna, IGM CNR, Bologna - Italy

^[12] Univ Trieste, Dept Biomed, Unit Dent Sci & Biomat, I-34129 Trieste - Italy

• Tipo de documento: Artigo Científico
• Fonte: Journal of Dentistry; v. 39, n. 3, p. 231-237, MAR 2011.
• Citações Web of Science: 43

Resumo

Objective: The function of endogenous MMP-3 and its distribution within the human dentine is unclear. Thus, the aim of the present study was to assay the presence and distribution of MMP-3 within human sound dentine by means of biochemical and immunohistochemical assays. Methods: Powdered dentine from extracted human teeth was prepared and (1) partially demineralised with 1% H(3)PO(4) for 10 min or (2) untreated (control). The presence of MMP-3 was measured using a colorimetric assay system (QuantiSir (TM), Epigentek, USA). Additional cryo-fractured dentine fragments were processed for immunohistochemical identification of MMP-3 under FEI-SEM. Casein-zymography was used to investigate MMP-3 activity. Results: MMP-3 detected level was 2.732 ng/mu L in partially demineralised dentine powder, whilst it increased to 3.280 ng/mu L in mineralised dentine. The FEI-SEM analysis revealed positive immunolabelling patterns for MMP-3, predominantly localized on the intertubular collagen fibrillar network showing MMP-3 directly or indirectly bound to the collagen fibrils. Casein-zymograms showed positive proteolytic activity for MMP-3 in demineralised dentine powder. Conclusion: The results of the study clearly revealed the presence and distribution of MMP3 in human sound dentine. Whilst the presence was verified, its role is still unclear. Future studies are needed to investigate the possible involvement of MMP-3 in physiological and pathological condition of the dentine-pulp complex. (C) 2011 Elsevier Ltd. All rights reserved. (AU)

• Processo FAPESP: 07/54618-4 - Avaliação da atividade de colagenases na degradação da dentina humana tratada por procedimentos restauradores adesivos
• Beneficiário: Marcela Rocha de Oliveira Carrilho
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores