Differential analysis of the dentin soluble proteomic

Autor(es)

Carrilho, Marcela R. ; Scaffa, Polliana M. C. ; Dionizio, Aline ; Ventura, Talita M. O. ; Buzalaf, Marilia A. R. ; Vidal, Cristina M. P.

• Tipo de documento: Artigo Científico
• Fonte: Journal of Dentistry; v. 131, p. 10-pg., 2023-02-24.

Resumo

Objectives: To perform a differential analysis of the dentin soluble proteomic and assess the effects of tissue health state and protocol for protein extraction. We hypothesized the dentin soluble proteomic varies according to the tissue physiopathological state (intact vs. caries-affected) and protocol used to extract its proteins. Methods: Dentin from freshly extracted non-carious and carious teeth were randomly assigned for protein extraction using either guanidine-HCl/ethylenediaminetetraacetic acid (EDTA) or acetic acid. Protein extracts from intact and caries-affected dentin were processed and digested with trypsin for shotgun label-free proteomic analysis (nLC-ESI-MS/MS). Peptides identification was performed on a nanoACQUITY UPLC-Xevo Q-Tof MS system. Peptides identified with scores of confidence greater than 95% were included in the quantitative sta-tistical analysis embedded in the PLGS software. Differences between experimental conditions were calculated using Student test-t with significance pre-set at alpha=0.05. Results: A total of 158 human proteins were identified. Approximately one-sixth of proteins (24/158) were present in at least two different extracts. Conversely, the greatest number of proteins (134/158) was identified uniquely in only one of the extracts. Overall, a larger number of soluble proteins was retrieved from caries -affected than intact dentin (86/158). Likewise, a greater number of proteins was extracted by the guanidine-HCl/EDTA (106/158) in comparison to acetic acid protocol. Several proteins detected in dentin extracts, mainly those from caries-affected teeth, are biological and/or metabolically involved with tissue turnover/ remodeling. Conclusion: The identity/abundance of soluble proteins retrieved from and remained in dentin noticeably depend on this tissue physiopathological state and protocol used to remove its minerals. Clinical significance: The present findings brought new insight into the proteomic phenotype of human dentin and may provide targets for the development of novel caries disease-prevention therapies. (AU)

• Processo FAPESP: 16/07996-2 - Síntese e caracterização de um cimento odontológico experimental obtido a partir de extrato de algas marinhas vermelhas
• Beneficiário: Marcela Rocha de Oliveira Carrilho
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/12316-5 - Estudo do perfil e atividade enzimática do fluido dentinário humano
• Beneficiário: Polliana Mendes Candia Scaffa
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 11/12226-8 - Avaliação da atividade de proteases na degradação da dentina humana tratada por procedimentos restauradores adesivos
• Beneficiário: Marcela Rocha de Oliveira Carrilho
• Modalidade de apoio: Bolsas no Exterior - Pesquisa