Effect of ScLL and 15d-PGJ2 on viability and cytokine release in LPS-stimulated fibroblasts: an in vitro study.

This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFß-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 µg/ml and treated with 15d-PGJ2 1 and 2 µg/ml, and ScLL 2 and 5 µg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFß-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 µg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFß-1 release (p > 0.05). Results indicated that ScLL 5 µg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFß-1 release.

Effect of ScLL and 15d-PGJ2 on viability and cytokine release in LPS-stimulated fibroblasts: an in vitro study

Abstract This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFβ-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 μg/ml and treated with 15d-PGJ2 1 and 2 μg/ml, and ScLL 2 and 5 μg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFβ-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 μg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFβ-1 release (p > 0.05). Results indicated that ScLL 5 μg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFβ-1 release.

Replanted teeth stored in a newly developed powdered coconut water formula.

BACKGROUND/AIM: A newly developed powdered coconut water formulae (ACP-404) has been proposed to preserve periodontal ligament cell viability. The aim of this study was to investigate the periodontal healing of dog teeth replanted after storage in ACP-404 with 2 different osmolalities. MATERIALS AND METHODS: Sixty teeth of 10 adult beagle dogs were extracted and randomly subjected to the following protocols: immediate replantation after extraction (positive control); dry conditions (negative control); and storage in ACP-404 I (G1), ACP-404 II (G2), in coconut water long shelf life (G3) or in pH-neutralized coconut water long shelf life (G4) for 50 minutes before replantation. The animals were euthanized 28 days post-operatively and processed for histomorphometric analysis using an optical microscope. The groups were subjected to histomorphometric analysis compared with immediate replantation and dry conditions. The data were statistically analysed using the Kruskal-Wallis test with post-analysis using Dunnett's test. RESULTS: The normal periodontal area was significantly higher for the G1 than for the G2 and G3 groups (P < .001). The positive control group presented similar normal periodontal areas compared to the experimental groups, except for the G3 and negative control groups (P < .001). G1 presented the lowest level of external root replacement resorption compared to the other groups (P < .05). All storage solutions had similar periodontal conditions for repaired resorption areas, inflammatory root resorption, ankylosis and replacement resorption (P > .05). CONCLUSIONS: ACP-404 I (G1) was better overall and was able to preserve the normal periodontium and had less external root replacement resorption.

Colagem de fragmento dental, tratamento conservador ­ relato de caso / Bonding of dental fragment, conservative treatment ­ case report

O traumatismo dental acomete frequentemente crianças e adolescentes, comprometendo, principalmente, os dentes permanentes. Dentre as injúrias traumáticas, as fraturas coronárias não complicadas ocorrem mais comumente. Nesses casos, o tratamento ideal é a colagem do fragmento dental para restabelecer função e estética, devolvendo qualidade de vida ao paciente. Diante disso, o objetivo desse trabalho foi apresentar relato de caso clínico, de paciente de gênero feminino, envolvendo a colagem de fragmento após fratura coronária do dente 11. O fragmento foi reposicionado e colado adesivamente com resina composta ao remanescente dental, resgatando a estética, função e autoestima da paciente. Portanto, a colagem de fragmentos dentários representa uma técnica eficaz, pois é rápida, fácil, de excelente resultado estético e funcional (AU).

Dental trauma often affects children and adolescents, compromising mainly permanent teeth. Among traumatic injuries, uncomplicated coronary fractures are the most common. In such cases, the ideal treatment is the bonding of the dental fragment to reestablish function and esthetics, restoring patients' quality of life. The aim of this study was to present a clinical case report of bonding of a fragment after coronary fracture of incisor central tooth in a female patient. The fragment was repositioned and adhesively bonded using composite resin to the dental structure, restoring aesthetics, function and self-esteem. Therefore, the bonding of dental fragments represents an effective technique, since it is fast and simple, with excellent esthetic and functional result (AU).

Effect of lectin (ScLL) on fibroblasts stimulated with LPS - an in vitro study.

The lectin (ScLL) extracted from the Synadenium carinatum plant has been evaluated as an immunomodulator in diseases such as asthma, neosporosis and leishmaniasis. However, it has not yet been evaluated in the oral cavity. This study evaluated the effect of ScLL on viability, proliferation and release of IL-10 in human gingival fibroblasts (HGF) stimulated with lipopolysaccharide (LPS). HGF were stimulated with LPS 1 µg/ml and treated with ScLL in concentrations of 10, 5 and 2 µg/ml for 1 and 5 h, and evaluated by flow cytometry for viability, apoptosis (initial/advanced) and necrosis. The supernatant was collected to detect release of IL-10 by ELISA. The proliferation was assessed with the BrdU assay. Positive control consisted of cells maintained in Dulbecco's Modified Eagles Medium (DMEM), and the negative control, of those kept in tap water. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found for ScLL concentrations regarding viability or initial and advanced apoptosis (p=0.455). All the groups, including the positive control, had a significantly lower necrosis parameter than negative control at 5 h (p < 0.001). No difference was found for proliferation among the experimental groups (p = 0.832). ScLL at 5 and 2 µg/ml resulted in a lower release of IL-10 than positive and negative controls at 5 h (p = 0.047). The results indicated that ScLL concentrations tested were not cytotoxic, and had no effect on proliferation and release of IL-10 parameters. A thorough understanding of ScLL, regarding its immunomodulatory potential, may open the door to new perspectives for dentistry.

Evaluation of coconut water neutralized by different agents on the viability of human fibroblasts: an in vitro study / Avaliação da água de coco neutralizada por diferentes agentes na viabilidade de fibroblastos humanos: estudo in vitro

Objective: This study evaluated four types of pH adjustment of the coconut water (CW) on viability of human fibroblasts (HFF). Material and method: Natural and industrialized CW were adjusted to pH 7.0 using: (1) Sodium Hidroxide (NaOH), (2) Sodium bicarbonate (NaHCO3), (3) Triethanolamine (C6H15NO3), (4) 2-Amino-2-Methil-1-Propanol (C4H11NO). Fibroblasts were plated at 2×104/ well in 96 well plates and maintained in the CW solutions for 2 h and 4 h. Positive control was represented by HFF maintained in DMEM and the negative control by tap water. Cell viability was analyzed by MTT formazan method. Data were analyzed by 3-way ANOVA followed by Tukey's and Dunnet's test. Result: There are no significant effect on the cell viability regarding type of CW, period of evaluation, and the interactions between CW and period of evaluation, CW and pH adjustment method, pH adjustment method and period of evaluation (p>0.05). Conclusion The product used for CW pH adjustment did not influenced HFF viability, thought there are a tendency of better performance in natural CW.

Objetivo: Avaliar a eficácia de quatro tipos de substâncias usadas para ajuste do pH da água de coco (AC) sobre a viabilidade de fibroblastos humanos (HFF). Material e método: O pH da AC natural e industrializada foi ajustado para pH 7,0 utilizando: (1) Hidróxido de Sódio (NaOH), (2) bicarbonato de sódio (NaHCO3), (3) Trietanolamina (C6H15NO3), (4) 2-Amino-2- Methil-1-propanol (C4H11NO). Células HFF foram plaqueadas em 2×104 células/poço em placas de 96 poços e mantidas nas diferentes soluções de AC acima durante 2 h e 4 h. Controle positivo foi representado por HFF mantidas em DMEM e o controle negativo por água da torneira. A viabilidade celular foi avaliada pelo método de MTT Formazan. Os dados foram analisados por 3-way ANOVA seguido pelo teste de Tukey e Dunnett. Resultado: A viabilidade celular não é influenciada pelo período de avaliação, e as interações entre AC e período de avaliação, AC e método de ajuste de pH, método de ajuste de pH e período de avaliação (p> 0,05). Conclusão: O produto utilizado para ajuste do pH não interfere na viabilidade de FH, embora, haja uma tendência de melhor desempenho em AC natural.

Effect of lectin (ScLL) on fibroblasts stimulated with LPS - an in vitro study

Abstract: The lectin (ScLL) extracted from the Synadenium carinatum plant has been evaluated as an immunomodulator in diseases such as asthma, neosporosis and leishmaniasis. However, it has not yet been evaluated in the oral cavity. This study evaluated the effect of ScLL on viability, proliferation and release of IL-10 in human gingival fibroblasts (HGF) stimulated with lipopolysaccharide (LPS). HGF were stimulated with LPS 1 µg/ml and treated with ScLL in concentrations of 10, 5 and 2 µg/ml for 1 and 5 h, and evaluated by flow cytometry for viability, apoptosis (initial/advanced) and necrosis. The supernatant was collected to detect release of IL-10 by ELISA. The proliferation was assessed with the BrdU assay. Positive control consisted of cells maintained in Dulbecco's Modified Eagles Medium (DMEM), and the negative control, of those kept in tap water. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found for ScLL concentrations regarding viability or initial and advanced apoptosis (p=0.455). All the groups, including the positive control, had a significantly lower necrosis parameter than negative control at 5 h (p < 0.001). No difference was found for proliferation among the experimental groups (p = 0.832). ScLL at 5 and 2 µg/ml resulted in a lower release of IL-10 than positive and negative controls at 5 h (p = 0.047). The results indicated that ScLL concentrations tested were not cytotoxic, and had no effect on proliferation and release of IL-10 parameters. A thorough understanding of ScLL, regarding its immunomodulatory potential, may open the door to new perspectives for dentistry.

Soy milk as a storage medium to preserve human fibroblast cell viability: an in vitro study

Soy milk (SM) is widely consumed worldwide as a substitute for cow milk. It is a source of vitamins, carbohydrates and sugars, but its capacity to preserve cell viability has not been evaluated. The purpose of the present study was to investigate the efficacy of SM to maintain the viability of human fibroblasts at short periods compared with different cow milks. Human mouth fibroblasts were cultured and stored in the following media at room temperature: 10% Dulbecco's Modified Eagle Medium (DMEM) (positive control group); long shelf-life ultra-high temperature whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM); powdered cow milk (PM); and soy milk (SM). After 5, 15, 30 and 45 min, cell viability was analyzed using the MTT assay. Data were analyzed statistically by the Kruskal-Wallis test with post-analysis using the Dunn's method (α=0.05). SKM showed the lowest capacity to maintain cell viability in all analyzed times (p<0.05). At 30 and 45 min, the absorbance levels in control group (DMEM) and SM were significantly higher than in SKM (p<0.05). Cell viability decreased along the time (5-45 min). The results indicate that SM can be used as a more adequate storage medium for avulsed teeth. SKM was not as effective in preserving cell viability as the cell culture medium and SM.

O leite de soja (LS) é largamente consumido em todo o mundo como substituto para o leite bovino. Este é uma fonte de vitaminas, carboidratos e açúcares, mas a sua capacidade para preservar a viabilidade celular não foi avaliada. A finalidade do estudo foi investigar a eficácia do LS em manter a viabilidade de fibroblastos humanos em períodos curtos em comparação com diferentes leites bovinos. Fibroblastos de boca humanos foram cultivados e armazenados nos seguintes meios à temperatura ambiente: 10% de meio Dulbecco's Modified Eagle (DMEM) (grupo controle positivo); leite bovino integral longa vida (LI); leite bovino desnatado longa vida - LD; leite em pó - LP; leite de soja - LS. Depois de 5, 15, 30 e 45 min, a viabilidade celular foi analisada usando o método de MTT. Os dados foram analisados estatisticamente pelo teste de Kruskal-Wallis e posteriormente usando o método de Dunn (α=0,05). O grupo LD apresentou a menor capacidade para manter a viabilidade celular em todos os tempos analisados (p<0,05). Aos 30 e aos 45 min, os níveis de absorbância no grupo controle (DMEM) e LS foram significativamente maiores que no grupo LD (p<0,05). A viabilidade celular diminuiu ao longo do tempo (5-45 min). Os resultados indicaram que LS pode ser usado como meio de armazenamento mais adequado para dentes avulsionados. LD não foi eficaz na preservação da viabilidade das células como o meio de cultura de células e o LS.

Soy milk as a storage medium to preserve human fibroblast cell viability: an in vitro study.

Soy milk (SM) is widely consumed worldwide as a substitute for cow milk. It is a source of vitamins, carbohydrates and sugars, but its capacity to preserve cell viability has not been evaluated. The purpose of the present study was to investigate the efficacy of SM to maintain the viability of human fibroblasts at short periods compared with different cow milks. Human mouth fibroblasts were cultured and stored in the following media at room temperature: 10% Dulbecco's Modified Eagle Medium (DMEM) (positive control group); long shelf-life ultra-high temperature whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM); powdered cow milk (PM); and soy milk (SM). After 5, 15, 30 and 45 min, cell viability was analyzed using the MTT assay. Data were analyzed statistically by the Kruskal-Wallis test with post-analysis using the Dunn's method (α=0.05). SKM showed the lowest capacity to maintain cell viability in all analyzed times (p<0.05). At 30 and 45 min, the absorbance levels in control group (DMEM) and SM were significantly higher than in SKM (p<0.05). Cell viability decreased along the time (5-45 min). The results indicate that SM can be used as a more adequate storage medium for avulsed teeth. SKM was not as effective in preserving cell viability as the cell culture medium and SM.