Co-distribution of cysteine cathepsins and matrix metalloproteases in human dentin |
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| Affiliation: | 1. Faculty of Dentistry at Piracicaba, University of Campinas, Piracicaba, 13414-903, Brazil;2. Department of Biological Science, Bauru School of Dentistry, University of Sao Paulo, Bauru, SP, 17012-901, Brazil;3. Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, 40126, Italy;4. Department of Medical Sciences, University of Trieste, Trieste, 34128, Italy;5. Department of Operative Dentistry, College of Dentistry, University of Iowa, 801 Newton Rd, Iowa City, IA,52246, USA;6. IGM-CNR, Unit of Bologna c/o IOR, Bologna, 40136, Italy;7. Department of Pharmacy, Dentistry and Nursing, Federal University of Ceara, Fortaleza, 60020-181, Brazil;8. Department of SteVA, University “Carlo Bo”, Urbino, 61029, Italy;9. Emeritus Professor of Oral Biology, Dental College of Georgia, Georgia Health Sciences University, Augusta, Georgia, 30912, USA;10. Institute of Dentistry, University of Oulu, and Medical Research Center Oulu, Oulu University Hospital and University of Oulu, Oulu FI-90014, Finland;11. Department of Oral and Maxillofacial Diseases, University of Helsinki, and Helsinki University Hospital, Helsinki FI-00014, Finland;12. Department of Biochemistry, Federal University of São Paulo, São Paulo, 04021-001, Brazil;13. Centro Interdisciplinar de Investigação Bioquímica, University of Mogi das Cruzes, Mogi das Cruzes, 08773-520, Brazil;14. Biomaterials in Dentistry Program, Anhanguera University São Paulo (UNIAN-SP), São Paulo, 05145-200, Brazil;15. Biotechnology and Innovation in Health Program, Anhanguera University São Paulo (UNIAN-SP), São Paulo, 05145-200, Brazil |
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| Abstract: | It has been hypothesized that cysteine cathepsins (CTs) along with matrix metalloproteases (MMPs) may work in conjunction in the proteolysis of mature dentin matrix. The aim of this study was to verify simultaneously the distribution and presence of cathepsins B (CT-B) and K (CT-K) in partially demineralized dentin; and further to evaluate the activity of CTs and MMPs in the same tissue. The distribution of CT-B and CT-K in sound human dentin was assessed by immunohistochemistry. A double-immunolabeling technique was used to identify, at once, the occurrence of those enzymes in dentin. Activities of CTs and MMPs in dentin extracts were evaluated spectrofluorometrically. In addition, in situ gelatinolytic activity of dentin was assayed by zymography. The results revealed the distribution of CT-B and CT-K along the dentin organic matrix and also indicated co-occurrence of MMPs and CTs in that tissue. The enzyme kinetics studies showed proteolytic activity in dentin extracts for both classes of proteases. Furthermore, it was observed that, at least for sound human dentin matrices, the activity of MMPs seems to be predominant over the CTs one. |
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| Keywords: | Cysteine cathepsins Matrix metalloproteases Proteolytic activity Dentin Immunohistochemistry |
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