Molecular Response of Pulp Fibroblasts after Stimulation with Pulp Capping Materials

Autor(es)

Karin Cristina da Silva Modena ^[1] ; Adriana Maria Calvo ^[2] ; Carla Renata Sipert ^[3] ; Bella Luna Colombini-Ishikiriama ^[4] ; Thiago José Dionísio ^[5] ; Maria Fidela de Lima Navarro ^[6] ; Maria Teresa Atta ^[7] ; Carlos Ferreira Santos ^[8]

Afiliação do(s) autor(es)

^[1] Universidade de São Paulo. Bauru School of Dentistry. Department of Restorative Dentistry, Endodontics and Dental Materials - Brasil

^[2] Universidade de São Paulo, Bauru. Bauru School of Dentistry. Department of Biological Sciences - Brasil

^[3] Universidade de São Paulo. School of Dentistry. Department of Restorative Dentistry - Brasil

^[4] Universidade de São Paulo. School of Dentistry, Bauru School of Dentistry. Department of Pediatric Dentistry, Orthodontics and Public Health - Brasil

^[5] Universidade de São Paulo, Bauru. Bauru School of Dentistry. Department of Biological Sciences - Brasil

^[6] Universidade de São Paulo. Bauru School of Dentistry. Department of Restorative Dentistry, Endodontics and Dental Materials - Brasil

^[7] Universidade de São Paulo. Bauru School of Dentistry. Department of Restorative Dentistry, Endodontics and Dental Materials - Brasil

^[8] Universidade de São Paulo, Bauru. Bauru School of Dentistry. Department of Biological Sciences - Brasil

• Tipo de documento: Artigo Científico
• Fonte: Brazilian Dental Journal; v. 31, n. 3, p. 244-251, 2020-07-13.

Resumo

Abstract This in vitro study evaluated cell viability and metabolism, nitric oxide release and production of two chemokines and one cytokine by cultured human dental pulp fibroblasts (HDPF) in contact with two glass ionomer cements (Ketac Molar-KM and Vitrebond-VB), Single Bond (SB) and calcium hydroxide (Dycal-DY). Cultures of HDPF were established by means of an explant technique. The specimens were prepared under sterile conditions and in disks measuring 5 mm x 2 mm obtained from a prefabricated mold and placed on a permeable membrane to avoid direct contact with the cells. Cytotoxicity was assessed by Trypan Blue exclusion method and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide release in cell supernatant was detected by the Griess Method whereas stromal derived factor-1 alpha (SDF-1α or CXCL12), chemokine (C-X-C motif) ligand 8 [Interleukin 8 (IL-8 or CXCL8)] and interleukin-6 (IL-6) were detected by ELISA. RT-qPCR was employed for gene expression analysis. Statistical analyses were performed by One-way ANOVA followed by Tukey’s post hoc test for materials independent of the time, and Two-way ANOVA followed by Bonferroni correction test for the comparisons between materials and experimental time (p<0.05). Cytotoxic tests showed significant differences only for DY. Protein levels and mRNA expression were significantly increased for IL-8 for both periods of time. IL-6 production increased when fibroblasts were stimulated by KM. SDF-1α protein production and mRNA expression were not affected by any of the materials. There was a decrease in nitrate/nitrite levels only for KM. Although DY caused intense cell death and did not stimulate the production of the inflammatory mediators evaluated in this work, it is known that this event seems to be fundamental for the process of repair of the pulp tissue and formation of mineralized barrier. KM and VB increased production of proteins related to the inflammatory process, thus favoring tissue repair. Therefore, although these glass ionomer cements did not lead to large cell death, they should be used with caution. (AU)

• Processo FAPESP: 10/01230-1 - Avaliação do papel dos receptores TLR2 e TLR4 na produção de citocinas por fibroblastos humanos periodontais deficientes desses receptores
• Beneficiário: Carlos Ferreira dos Santos
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/03965-2 - Papel do sistema renina-angiotensina em diferentes modelos inflamatórios orais: uma abordagem interdisciplinar experimental e clínica
• Beneficiário: Carlos Ferreira dos Santos
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 05/60167-0 - Expressão de pró-colágeno tipo 1, MIP-1± alfa e SDF-1 alfa por fibroblastos da polpa humana estimulados por ácido lipoteicóico de Streptococcus mutans
• Beneficiário: Carlos Ferreira dos Santos
• Modalidade de apoio: Auxílio à Pesquisa - Regular