Estudo in vitro da produção de quimiocinas e pró-colágeno I por fibroblastos de gengiva, ligamento periodontal e polpa dental humanos / In vitro study of chemokines and procollagen I by human gingival, periodontal ligament and dental pulp fibroblasts

Fibroblastos são as células mais numerosas encontradas nos tecidos orais como gengiva, ligamento periodontal e polpa dental. Além de exercerem função estrutural, estas células também desempenham papel importante na resposta imune destes tecidos através do reconhecimento de antígenos e produção de mediadores inflamatórios e citocinas. Evidências apontam ainda para o fato de que fibroblastos não constituem um grupo único de células. Sendo assim, os objetivos deste estudo foram: (I) avaliar a produção diferencial de fibroblastos de gengiva, ligamento periodontal e polpa dental de dentes permanentes e decíduos quanto à produção das quimiocinas CCL3 e CXCL12; (II) avaliar a produção de pró-colágeno I pelas células de polpa e (III) avaliar a expressão diferencial dos fibroblastos quanto a microRNAs. Dentes recentemente extraídos (terceiros molares hígidos) e fragmentos de gengivas saudáveis de três pacientes adultos foram obtidos no Laboratório de Farmacologia e Fisiologia Clínica da Faculdade de Odontologia de Bauru. Caninos decíduos de dois pacientes com indicação para extração por motivos ortodônticos foram obtidos na Clínica de Odontopediatria da mesma unidade. Culturas primárias de fibroblastos de gengiva (n=3), ligamento periodontal (n=3) e polpa de dente permanente (n=3) e polpa dental de dente decíduo (n=2) foram estabelecidas a partir de tecidos humanos por meio de técnica de explant. Após a quarta passagem, a produção de CCL3 e de CXCL12 foi avaliada após estímulo com concentrações crescentes (0 10 µg/mL) de ácido lipoteicóico de Enterococcus faecalis (EfLTA), lipopolissacarídeo de Porphyromonas gingivalis (PgLPS) ou LPS de Escherichia coli (EcLPS) por ELISA após 1, 6 e 24 h. O RNAm para as quimiocinas no grupo estimulado com EcLPS por 24 h foi avaliado por transcrição reversa seguida de reação em cadeia da polimerase quantitativa. A produção de pró-colágeno I por células de polpa estimuladas com EfLTA e PgLPS foi avaliada por imunofluorescência. O perfil...

Fibroblasts are the dominant cells within oral tissues such as gingiva, periodontal ligament and dental pulp. Besides the architectural maintenance of the connective tissues, fibroblasts are also involved in connective tissue immune response through antigen recognition and production of inflammatory mediators and cytokines. Recent studies also demonstrated that fibroblasts do not constitute a unique group of cells. Taken this togeter, the objectives of the present study were: (I) to evaluate the production of the chemokines CCL3 and CXCL12 by human gingival, periodontal ligament as well as permanent and deciduous dental pulp fibroblasts; (II) to evaluate the production of procollagen I by dental pulp fibroblasts and (III) to evaluate the differential pattern of expression of microRNAs by the oral fibroblasts. Recently extracted teeth (non-carious third molars) and fragments of healthy gingiva from three adults were obtained at the Laboratory for Clinical Pharmacology and Physiology at Dental School of Bauru. Deciduous canines from two patients with orthodontic indication for extraction were obtained at Pediatrics Clinics of Dental School of Bauru. Primary cultures of fibroblasts from gingiva (n=3), periodontal ligament (n=3) as well as permanent pulp (n=3) and deciduous pulp (n=2) were established through an explant technique. After the fourth passage, fibroblasts were challenged with increasing concentrations (0 10 µg/mL) of Enterococcus faecalis lipoteichoic acid (EfLTA), Porphyromonas gingivalis lipopolysaccharide (PgLPS) or Escherichia coli LPS (EcLPS) for 1, 6 and 24 h. The chemokines were assessed through ELISA while the mRNA for CCL3 and CXCL12 (EcLPS at 24 h) were assessed through reverse transcription followed by quantitative polymerase chain reaction. The expression of microRNAs was screened through a microarray assay. The production of CCL3 on cell supernatants was detected in all cellular groups, with higher amounts at gingival fibroblasts...

Increased serum concentration of procollagn peptides in essential hypertension and relation to cardiac alterations

This study was designed to evaluate fibrogenic activity in patients with essential hypertension by measuring serum procollagen type amino terminal peptide [PIIIP] and procollagen type I [PIP]. The study was performed on 50 patients with never-treated essential hypertension and 30 normotensive control subjects. Measurements were repeated in 43 hypertensive patients after six months of treatment with angiotensin- converting enzyme inhibitor lisinopril. The serum concentrations of PIIIP and PIP were measured by specific radioimmunoassay. Doppler ultrasound recordings and ECG were obtained to determine left ventricle anatomy and function and arrhythmias. Baseline serum PIIIP and PIP were increased in hypertensive patients as compared with the normotensive subjects and they decreased significantly to normal values in patients treated with lisinopril

Serum aminoterminal propertied of type III procollagen in nonmalignant gynecologic tumors

The present study included a total of 60 females in the childbearing period. They were divided into 3 groups, each comprised 20 females. These groups are the ovarian group, the uterine fibroid group and the control group. The serum aminoterminal propeptide of type III procollagen by radioimmunoassay technique was measured. The mean +/- S.D. in the fibroid group was significantly higher than that of the control groups. However, the means of serum aminoterminal propeptide of type III procollagen in the three studied groups were within the reference interval. Thus, the metabolism of type III collagen was not significantly altered in these benign gynecologic tumors

Serum procollagen III amino-terminal peptide [P III NP]: a non-invasive procedure for assessment of liver fibrosis

Serum pro-collagen III and liver function tests were estimated in 20 healthy subjects and 71 patients with chronic liver disease in a trial to find out its correlation with clinical, laboratory and histological parameters as well as its diagnostic value. From the results obtained, it was concluded that serum P III NP may be a good marker of liver fibrosis and may be used in differentiating fibrotic liver diseases from those with no or minimal fibrosis and may be a good prognostic clinical marker in the assessment of clinical severity of the disease

Study of serum procollagen III peptide changes before and after some antifibrotics in hepatosplenic schistosomiasis

The antifibrotic effect of colchicine and captopril on schistosomal hepatic fibrosis was evaluated in fifty patients with hepatosplenic schistosomiasis taking serum procollagen III peptide [PIII NP] as a marker for degree of fibrosis. The duration of study was two months. Serum PIII NP levels was non significantly decreased in patients received 25 mg, captopril twice daily [20 patients] while those received placebo [fifteen patients] showed no change in its levels. Opposite to these findings 0.5mg colchicine twice daily [fifteen patients] led to nonsignificant rise in the level of serum PIII NP. As it is expected that colchicines leads to reduction of serum PIII NP level after a period of therapy for six months at least, this initial rise after two months of therapy [secondary to release of incorporated PIII NP molecules on the surface of collagen in its hydrolysis by colchicines] indicate the start of the therapeutic effects. Beside this antifibrotic effect of captopril and colchicine. it was found that a significant reduction in spleen size had occurred while in those received placebo no change in the size of spleen had occurred. It is concluded that the measurement of serum PIII NP after two months of colchicine therapy may give a clue for continuing [with presence of initial rise of serum PIII NP] of therapy or not