An in vitro evaluation of ice apple water, Aloe vera, and propolis as a storage medium to preserve viability of human periodontal ligament fibroblasts

Background: A number of media that create the best possible conditions to maintain periodontal ligament (PDL) cell viability after dental avulsion have been reported. Aim: The aim of this study is to evaluate ice apple water (IAW), Aloe vera, and propolis as a storage medium to preserve the viability of human PDL fibroblasts. Methods: An in vitro comparative type of study was performed on a PDL cell culture model. PDL fibroblasts obtained from the roots of healthy premolars were cultured in Dulbecco's Modified Eagle's Medium (DMEM) and treated with ice apple water (IAW), 7% propolis extract (PE), 30% Aloe vera extract (AVE), positive control DMEM supplemented with fetal bovine serum, negative control (NC) without any agent, and incubated at 37°C for 1 h, 3 h, and 24 h. Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay after every test period. Optical density was measured at a wavelength of 490 nm. Statistical Analysis Used: The effects of the test storage media were evaluated by one-way analysis of variance test, followed by post hoc Tukey's multiple comparison test (P < 0.05). Results: Seven percent PE demonstrated the highest capacity of maintaining PDL cell viability at 1 h and 24 h. IAW showed a statistically significantly lower percentage of viable cells at all three test periods as compared to 7% PE. After 3 h, 30% AVE demonstrated maximum viable cells. Conclusions: Within the limitations of this study, propolis at a concentration of 7% was the most effective medium for maintaining PDL cell viability.

Medios de almacenamiento para dientes avulsionados. Una revisión / Media storage for avulsed teeth. A review

Resumen La avulsión dental es el desplazamiento completo del diente de su alvéolo que puede ser causado por un traumatismo, en el que se produce la ruptura de las fibras del ligamento periodontal; además puede estar acompañado de lesiones que comprometan el cemento, el hueso alveolar y los tejidos periodontales. Las células del ligamento periodontal, luego de la avulsión, crecen óptimamente en un pH neutro; los medios de almacenamiento deben tener características ideales para que haya crecimiento celular, como: ser líquidos estériles, poseer componentes que nutran las células del ligamento periodontal, estar disponibles en el lugar del accidente, ser de larga duración y vida útil. El índice de trauma dentoalveolar ha aumentado en nuestro país y la principal causa son los accidentes en motocicleta y deportes. Se pierden muchos dientes avulsionados por desconocimiento de colocar este diente en un medio favorable antes de que sea reimplantado por el odontólogo; existen muchos medios y cada día se experimentan más, pero ¿cuáles de estos son realmente benéficos para mejorar el pronóstico de los dientes avulsionados? El propósito de esta revisión es describir los diferentes medios de almacenamiento para dientes avulsionados investigados hasta la fecha, con el fin de determinar cuál es el medio de elección para la conservación de las células del ligamento periodontal en los dientes que serán reimplantados. Se realizó una revisión de la literatura de estudios publicados en idioma inglés y español en la bibliografía médica, utilizando el buscador Pubmed/Medline, Lilacs, BBO y Scielo. La leche fresca pasteurizada sigue siendo el medio de elección, pero teniendo en cuenta que el tiempo extraoral es importante, si no se tiene la leche, los nuevos medios, como la clara de huevo, propóleo, aloe vera y agua de coco, son una buena alternativa en casos de ser los disponibles en el lugar del accidente. Las cajas de rescate son el medio ideal y deben estar disponibles en todos los países.

Abstract Dental avulsion is the complete displacement of a tooth from its alveolus that may be caused by trauma, producing the rupture of fibers from the periodontal ligament and other lesions involving cement, alveolar bone and periodontal tissues. After the avulsion, cells from the periodontal ligament grow optimally in a neutral pH, therefore the storage media must have ideal characteristics for cellular growth, such as: Sterile liquids, periodontal ligament cell nutrients, availability at the place of the accident and long lasting life. In our country, dento-alveolar trauma index has increased and the main cause are accidents in motorcycles and sports. Many avulsed teeth are lost because of the ignorance of placing the tooth in a favorable storage media before the dentist replaces it. There are many storage media and every day other, more are experimented, so then, which of these are beneficial for the outcome of avulsed teeth? The purpose of this review of literature is to describe the different storage media for avulsed teeth to determine which is the best storage media for the conservation of periodontal ligament cells in teeth that will be re-implanted. A literature review was done of articles published in English and Spanish using Pubmed/ Medline, Lilacs, BBO and Scielo. Conclusion: Fresh pasteurized milk is still the storage media of choice, taking into consideration the importance of extraoral time. Other storage media such as egg yolk, propolis, aloe vera and coconut water should be considered as alternatives in case there's no milk available in the place of the accident. Rescue boxes should be available in every country.

Effect of Milk Renewal on Cell Viability In Vitro at Different Time Frames

Abstract The purpose of this study was to evaluate if the renewal of milk as a storage medium, every 12, 24 and 48 h, is able to increase its ability to maintain human periodontal ligament fibroblasts (PDLF) viability over time. PDLF were soaked in Minimum Essential Medium at 37 °C (MEM-37) (positive control), tap water (Water) (negative control) and in skimmed milk (44 wells) at 5 °C and 20 °C. The skimmed milk was renewed every 12 h (Milk-12), 24 h (Milk-24) and 48 h (Milk-48) in 11 wells of each plate, and the milk in the remaining 11 wells of each plate was maintained in situ (not renewed milk) (NRM). After 24, 48, 72, 96 and 120 h, cell viability was determined by the tetrazolium salt-based colorimetric (MTT) assay. Data were statistically analyzed by Kruskal-Wallis, Scheffé and Mann-Whitney tests (a=5%). At 5 °C, only Milk-48 was significantly better than NRM. At 20 °C, NRM was more effective than Milk-12 and Milk-24 in all time periods. In relation to the temperature (5 °C or 20 °C), renewal of milk at 5 °C was better in maintaining cell viability than the renewal at 20 °C. In conclusion, the renewal of milk was able to increase its ability to maintain cell viability only when performed every 48 h in milk maintained at 5 °C.

Resumo O objetivo deste estudo foi avaliar se a renovação do leite, a cada 12, 24 e 48 h, é capaz de aumentar sua capacidade de manter a viabilidade de fibroblastos do ligamento periodontal humano (FLPH) ao longo do tempo. FLPH foram conservados em Meio Essencial Mínimo a 37 °C (MEM-37) (controle positivo), água da torneira (água) (controle negativo) e em leite desnatado (44 poços) a 5 °C e 20 °C. O leite desnatado foi renovado a cada 12 h (leite-12), 24 h (leite-24) e 48 h (leite-48) em 11 poços de cada placa, e em outros 11 poços de cada placa o leite foi deixado in situ (leite não renovado) (LNR). Depois de 24, 48, 72, 96 e 120 h, a viabilidade celular foi determinada pelo ensaio colorimétrico à base de sal tetrazólio (MTT). Os dados foram analisados estatisticamente pelos testes de Kruskal-Wallis, Scheffé e Mann-Whitney (α=5%). A 5 °C, somente o leite-48 foi significantemente melhor do que o LNR. A 20 °C, LNR foi mais efetivo do que o leite-12 e leite-24 em todos os períodos de tempo. Em relação à temperatura (5 °C ou 20 °C), a renovação do leite a 5 °C foi melhor na manutenção da viabilidade celular do que a renovação a 20 °C. Concluindo, a renovação do leite foi capaz de aumentar sua habilidade em manter a viabilidade celular apenas quando realizada a cada 48 h no leite mantido a 5 °C.

In Vitro Periodontal Ligament Cell Viability in Different Storage Media

Abstract The aim of this study was to evaluate the viability of periodontal ligament cells of avulsed teeth in three different storage media. Forty-five mature premolars extracted for orthodontic therapeutic purposes were randomly and equally divided into three groups according to the storage medium: milk (control), rice water and egg white. After placing extracted teeth for 30 min in storage media, the scrapings of the periodontal ligament (PDL) were collected in Falcon tubes containing collagenase in 2.5 mL of phosphate buffer saline and were incubated for 30 min and centrifuged for 5 min at 800 rpm. Cell viability was analyzed by Trypan blue exclusion. Rice water had a significantly higher number of viable cells compared to egg white and milk. There was no statistically significant difference between egg white and milk. Rice water may be able to maintain PDL cell viability of avulsed teeth better than egg white or milk.

Resumo O objetivo deste estudo foi avaliar a viabilidade das células do ligamento periodontal de dentes avulsionados armazenados em três diferentes meios. Quarenta e cinco pré-molares com formação radicular completa extraídos por razões terapêuticas foram aleatoriamente distribuídos em três grupos, de acordo com o meio de armazenagem: leite (controle), água de arroz e clara de ovo). Após armazenar os dentes avulsionados por 30 min no meio, raspas do ligamento periodontal (LPD) foram coletadas em tubos Falcon contendo 2,5 mL de solução tamponada de soro fosfatado, incubadas por 30 min e a seguir centrifugadas por 5 min a 800 rpm. A viabilidade celular foi analisada pelo método de exclusão do Azul de Trypan. A água de arroz teve um número significativamente maior de células viáveis em comparação com o leite e a clara de ovo. Não houve diferença estatisticamente significativa entre o leite e a clara de ovo. A água de arroz pode ser capaz de manter a viabilidade das células do PDL de dentes avulsionados, melhor que o leite ou a clara de ovo.

Fibroblast Viability after Storage at 20 °C in Milk, Hank's Balanced Salt Solution and Coconut Water

Abstract The objective of this study was to evaluate the effectiveness of various storage media at 20 °C in maintaining the viability of human periodontal ligament fibroblasts (HPLF) over time. HPLF were maintained at 20 °C in skim milk (SM), whole milk (WM), freshly prepared Hank's balanced salt solution (HBSS), Save-A-Tooth(r), natural coconut water (NCW), coconut water industrialized (ICW) and tap water (negative control) for 3, 6, 24, 48, 72, 96 and 120 h. Cells maintained in Minimal Essential Medium (MEM-37) at 37 °C served as a positive control. Cell viability was determined by MTT assay. Statistical analysis was performed by Kruskal-Wallis test and Scheffe test (α = 5%). From 24 h, NCW was significantly better in maintaining cell viability than all other tested storage media (p<0.05). SM and WM were significantly better than HBSS for up to 72 h. Save-A-Tooth(r) and ICW were the worst conservation storage media. In conclusion, the effectiveness of the tested storage media to maintain the viability of the periodontal ligament cells was as follows, in a descending order: NCW > MEM-37> SM and IM> HBSS> ICW > Save-A-Tooth(r)> tap water.

Resumo O objetivo deste estudo foi avaliar a efetividade de vários meios de conservação a 20 °C em manter a viabilidade de fibroblastos do ligamento periodontal humano (FLPH) ao longo do tempo. FLPH foram conservados a 20 °C em leite desnatado (LD), leite integral (LI), solução salina balanceada de Hank (HBSS) recém preparada, Save-A-Tooth(r) (Save), água de coco natural (ACN), água de coco industrializada (ACI) e água de torneira (água - controle negativo) por 3, 6, 24, 48, 72, 96 e 120 h. Células conservadas em Meio Essencial Mínimo (MEM-37) a 37 °C serviram como controle-positivo. A viabilidade celular foi determinada pelo ensaio MTT. A análise estatística dos dados foi realizada pelos testes Kruskal-Wallis e Scheffé (α=5%). A partir de 24 h, ACN foi significantemente melhor em manter a viabilidade celular do que todos os outros meios testados (p<0,05). LD e LI foram significantemente melhores do que a HBSS por até 72 h. Save e ACI foram os piores meios de conservação. Concluindo, a efetividade dos meios de conservação testados em manter a viabilidade das células do ligamento periodontal foi a seguinte em ordem decrescente: ACN > MEM-37 > LD e LI > HBSS > ACI > Save > água.

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.

Influência do método de polimerização na microdureza de compósitos microhíbridos armazenados em água destilada / Influence of curing method on the micro hardness of microhybrid composites immersed in distilled water

O objetivo do presente estudo é comparar e avaliar a microdureza Vickers de um compósito restaurador microhíbrido ativado por dois tipos de unidades polimerizadoras e armazenados em água destilada. Trinta espécimes foram feitos com resina Charisma B1, para cada um dos regimes de polimerização: fotopolimerização por luz halógena, fotopolimerização por LED e fotopolimerização mais ciclo adicional em autoclave. Foram feitas duas leituras de microdureza Vickers por corpo de prova em 1 dia, 7 dias e 14 dias de imersões. O ciclo adicional de polimerização mostrou uma tendência de aumentar os valores de microdureza dos compósitos restauradores, não mostrando diferenças estatisticamente significantes entre LED e Luz Halógena (p > 0,05).

The purpose of this study is to evaluate and compare the Vickers microhardness of one microhybrid composite polymerized with different sources and stored in distilled water for up to 14 days. Thirty samples have been prepared with Charisma composite, shade B1, for each polymerization method: halogen light photopolymerization, LED photopolymerization, photopolymerization plus post-cured cycles in autoclave. Two readings of Vichers micro hardness have been done in each sample on 24h, 7days and 14 days of storage. The post curing method tended to improve the microhardness, but it was not statistically different from halogen or LED curing methods (p > 0.05). After 7 days, the hardness values were higher than the first day, but statistically not different to 14 days (p < 0.05). Post-cured samples in autoclave had an improved mean value, however, without differing from those of the LED and halogen.

Endothelial Function of Cornea Preserved in Korean Corneal Storage Media

PURPOSE: To evaluate the endothelial function of cornea preserved in newly developing korean corneal storage media (CS002, CS003) by estimating the permeability of corneal endothelium and the change of corneal thickness. METHODS: The cornea were divided into six experimental groups - fresh group immediately after enucleation, 4degrees Cmoist chamber group preserved for 24 hours and 48 hours, Optisol & CS002 group for 1, 3, 5, and 7 days, and Likorol & CS003 group for 7, 10, and 14 days after enucleation, and then corneal endothelial permeability(Pac) was measured using carboxyfluorescein solution. Corneal thickness was measured using pachymeter(fine focus adjustment) of the specular microscope. RESULTS: Corneal endothelial Pac (x1 0(- 4) cm/min) was 3.64+/-0.33 in fresh group, 4.79+/-0.28 in 4degrees Cmoist chamber group for 24 hours. Each endothelial Pac of CS002 group at 5 and 7 days was 5.81+/-0.55 and 5.65+/-0.58, which were different with 4degrees Cmoist chamber preservation group for 24 hours(p<0.05) but not different with Optisol groups at same days. Each endothelial Pac of CS003 group at 7, 10, and 14 days was 4.34+/-0.34, 4.66+/-0.59, and 4.66+/-0.27, which were not different from those of Likorol. Each corneal thickness of CS002 and Optisol group at 7days was 417.80+/-19.37 mu m and 421.00+/-19.75mu m, which were resemble increment. Corneal thickness was 426.75+/-22.43mu m in CS003 group and 476.00+/- 40.08mu m in Likorol group at 7days. There was statistical difference between the two group(P<0.05), and this difference was sustained for 14days (P<0.05). CONCLUSION: There was no difference in the effect on corneal endothelial permeability between korean corneal storage media such as CS002 and CS003, and that of previous corneal storage media such as Optisol and Likorol. Corneal thickness of cornea preserved in korean corneal storage media was thinner than that of Likorol.