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Expression of procollagen type I, MIP-1alpha and SDF-1alpha by human gingival fibroblasts and dental pulp stimulated by Enterococcus faecalis lipoteichoic acid and lipopolysaccharide from Escherichia coli and Porphyromonas gingivalis

Grant number: 07/00306-1
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): August 01, 2007
Effective date (End): July 31, 2011
Field of knowledge:Health Sciences - Dentistry
Principal researcher:Carlos Ferreira dos Santos
Grantee:Carla Renata Sipert
Home Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil


Dental pulp is a connective tissue structure constituted by many different cell types. Among them, fibroblasts are the most abundant. When challenged by different aggressive agents, these cells are able to release some substances like cytokines and chemokines, which are essential to trigger the inflammatory process. Nevertheless, these cells also produce extracellular matrix compounds, like collagen, essential for tissue repair. Therefore, the aims of this study are to: 1. evaluate type I procollagen expression; 2. evaluate chemokines expression (MIP-1alpha and SDF-1alpha) by human gingival and dental pulp fibroblasts challenged by E. faecalis lipoteichoic acid (LTA) as well as E. coli and P. gingivalis lipopolysaccharide (LPS); 3. compare type I procollagen and chemokines release by fibroblasts when challenged by LTA or both LPS; 4. compare the cellular behavior when originated from dental pulp or from gingiva from the same subject using the same stimuli. The dental pulp from non-erupted third molars and health gingival samples will be collected from the same patient (n=5). Explants will be made and stocked in culture medium for fibroblasts growth. The cells will be used since the fourth passage. In 24-well plates and after cellular confluence, culture medium containing 0.8-80ug/mL of E. faecalis LTA or 1-100ug/mL of E. coli or P. gingivalis LPS will be added. After 1, 6 and 24 hours, the supernatant and the cells will be collected for analysis. Reverse transcription followed by real time polymerase chain reaction will be employed to evaluate mRNA expression for type I procollagen, MIP-1alpha and SDF-1alpha. Protein detection of type I procollagen will be achieved by Western blot analysis, whereas detection of MIP-1alpha and SDF-1alpha will be carried out by ELISA. Statistical analysis will be performed by GraphPad Prism 3.0 software, ANOVA and Bonferroni correction tests. Significance levels of 5% will be adopted. (AU)

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Scientific publications (5)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
REZENDE DELGADO, RONAN JACQUES; GASPAROTO, THAIS HELENA; SIPERT, CARLA RENATA; PINHEIRO, CLAUDIA RAMOS; DE MORAES, IVALDO GOMES; GARCIA, ROBERTO BRANDAO; HUNGARO DUARTE, MARCO ANTONIO; BRAMANTE, CLOVIS MONTEIRO; TORRES, SERGIO APARECIDO; GARLET, GUSTAVO POMPERMAIER; et al. Antimicrobial activity of calcium hydroxide and chlorhexidine on intratubular Candida albicans. International Journal of Oral Science, v. 5, n. 1, p. 32-36, . (09/14127-7, 07/00306-1)
PATRÍCIA YOSHINO; CELSO KENJI NISHIYAMA; KARIN CRISTINA DA SILVA MODENA; CARLOS FERREIRA SANTOS; CARLA RENATA SIPERT. In Vitro Cytotoxicity of White MTA, MTA Fillapex® and Portland Cement on Human Periodontal Ligament Fibroblasts. Brazilian Dental Journal, v. 24, n. 2, p. 111-116, . (05/60167-0, 07/00306-1)
SIPERT, CARLA RENATA; MORANDINI, ANA CAROLINA; DIONISIO, THIAGO JOSE; ANDRADE MOREIRA MACHADO, MARIA APARECIDA; PENHA OLIVEIRA, SANDRA HELENA; CAMPANELLI, ANA PAULA; KUO, WINSTON PATRICK; SANTOS, CARLOS FERREIRA. In Vitro Regulation of CCL3 and CXCL12 by Bacterial By-products Is Dependent on Site of Origin of Human Oral Fibroblasts. JOURNAL OF ENDODONTICS, v. 40, n. 1, p. 95-100, . (05/60167-0, 09/53848-1, 07/00306-1)
SIPERT, CARLA R.; MORANDINI, ANA C.; DIONISIO, THIAGO J.; TRACHTENBERG, ALEXANDER J.; KUO, WINSTON P.; SANTOS, CARLOS F.. MicroRNA-146a and microRNA-155 show tissue-dependent expression in dental pulp, gingival and periodontal ligament fibroblasts in vitro. JOURNAL OF ORAL SCIENCE, v. 56, n. 2, p. 157-164, . (05/60167-0, 09/53848-1, 07/00306-1)
GASPAROTO, THAIS H.; SIPERT, CARLA R.; DE OLIVEIRA, CARINE E.; PORTO, VINICIUS C.; SANTOS, CARLOS F.; CAMPANELLI, ANA P.; LARA, VANESSA S.. Salivary immunity in elderly individuals presented with Candida-related denture stomatitis. GERODONTOLOGY, v. 29, n. 2, p. E331-E339, . (05/60668-9, 07/00306-1, 05/60167-0)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
SIPERT, Carla Renata. In vitro study of chemokines and procollagen I by human gingival, periodontal ligament and dental pulp fibroblasts. 2011. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Odontologia de Bauru (FOB/SDB) Bauru.

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