PAR-1 and PAR-2 Expression Is Enhanced in Inflamed Odontoblast Cells

Author(s)

Alvarez, M. M. P. ^[1] ; Moura, G. E. ^[1] ; Machado, M. F. M. ^[2] ; Viana, G. M. ^[1] ; de Souza Costa, C. A. ^[3] ; Tjaderhane, L. ^{[4, 5, 6, 7, 8]} ; Nader, H. B. ^[1] ; Tersariol, I. L. S. ^[1] ; Nascimento, F. D. ^[2]

Affiliation

^[1] Fed Univ Sao Paulo UNIFESP, Dept Biochem, Mol Biol Div, Rua Tres Maio, 100-4th Floor, BR-04044020 Sao Paulo, SP - Brazil

^[2] Univ Mogi Das Cruzes, Interdisciplinary Ctr Biochem Invest CIIB, Av Dr Candido Xavier Almeida & Souza 200 1S15, BR-08780911 Mogi Das Cruzes, SP - Brazil

^[3] Univ Estadual Paulista, Univ Estadual Paulista, Araraquara Sch Dent, Dept Physiol & Pathol, Sao Paulo, SP - Brazil

^[4] Oulu Univ Hosp, Med Res Ctr Oulu, Oulu - Finland

^[5] Univ Oulu, Oulu - Finland

^[6] Univ Helsinki, Dept Oral & Maxillofacial Dis, Helsinki - Finland

^[7] Helsinki Univ Hosp, Helsinki - Finland

^[8] Oulu Univ Hosp, Res Unit Oral Hlth Sci, Oulu - Finland

• Document type: Journal article
• Source: JOURNAL OF DENTAL RESEARCH; v. 96, n. 13, p. 1518-1525, DEC 2017.
• Web of Science Citations: 1

Abstract

Protease-activated receptors (PARs) are G protein-coupled receptors, which are activated by proteolytical cleavage of the amino-terminus and act as sensors for extracellular proteases. We hypothesized that PAR-1 and PAR-2 can be modulated by inflammatory stimulus in human dental pulp cells. PAR-1 and PAR-2 gene expression in human pulp tissue and MDPC-23 cells were analyzed by quantitative polymerase chain reaction. Monoclonal PAR-1 and PAR-2 antibodies were used to investigate the cellular expression of these receptors using Western blot, flow cytometry, and confocal microscopy in MDPC-23 cells. Immunofluorescence assays of human intact and carious teeth were performed to assess the presence of PAR-1 and PAR-2 in the dentin-pulp complex. The results show for the first time that human odontoblasts and MDPC-23 cells constitutively express PAR-1 and PAR-2. PAR-2 activation increased significantly the messenger RNA expression of matrix metalloproteinase (MMP)-2, MMP-9, MMP-13, and MMP-14 in MDPC-23 cells (P < 0.05), while the expression of these enzymes decreased significantly in the PAR-1 agonist group (P < 0.05). The high-performance liquid chromatography and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis showed the presence of MMP-13 activity cleaving PAR-1 at specific, noncanonical site TLDPRS42 down arrow(FLL)-L-43 in human dental pulp tissues. Also, we detected a presence of a trypsin-like activity cleaving PAR-2 at canonical site SKGR(20)down arrow S(21)LIGRL in pulp tissues. Confocal microscopy analysis of human dentin-pulp complex showed intense positive staining of PAR-1 and PAR-2 in the odontoblast processes in dentinal tubules of carious teeth compared to intact ones. The present results support the hypothesis of activation of the upregulated PAR-1 and PAR-2 by endogenous proteases abundant during the inflammatory response in dentin-pulp complex. (AU)

• FAPESP's process: 13/05822-9 - Proteinase-Activated Receptors (PARs) in the dentin-pulp complex: identification, modulation and signal tranduction in caries disease
• Grantee: Fábio Dupart Nascimento
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 15/03964-6 - Glycosaminoglycans and proteoglycans: interplay between structure and function
• Grantee: Helena Bonciani Nader
• Support Opportunities: Research Projects - Thematic Grants