Human pulp tissue dissolution ability of different extracts of Sapindus mukorossi: An in vitro study / 中草药·英文版

Objective: Due to the many negative properties of sodium hypochlorite used in current root canal treatment, interest in biocompatible natural agents is increasing day by day. The aim of this study was to evaluate whether various extract solutions of Sapindus mukorossi have dissolution effects on human pulp tissues. Methods: Primarily powder extracts were obtained by extracting fruit shells of S. mukorossi in different solvents (ethanol, methanol, buthanol and distilled water). The test solutions were prepared and randomly separated into six groups with 10 samples in each group: ethanol extract, methanol extract, butanol extract, distilled water extract of S. mukorossi, sodium hypochlorite (NaOCl) and the control group. Among these, S. mukorossi extracts were separated into two subgroups, depending on their concentration level (50 µg/mL and 100 µg/mL). The pulp tissues of freshly extracted human molars were used for dissolution test. The weights of the pulpal tissues were measured and recorded for two times after the samples were placed in the solutions. Statistical analysis for all descriptive statistics was performed using SPSS 22 (P < 0.05). Results: Our results showed that maximum percent yield of preparation was obtained in methanol extract of S. mukorossi. Among all of the groups, the best dissolution capacity was seen in the NaOCl group (positive control group). Among S. mukorossi groups, the best tissue solvent solution was found in SMM group at 50 µg/mL and SMB group at 100 µg/mL. Conclusion: The different extracts of S. mukorossi had a capacity to dissolve pulp tissue but this capacity was less than NaOCl. Therefore, further studies will enable the creation of a commercial solution for clinical use by increasing the effectiveness of S. mukorossi while combining it with other endodontic irrigation solutions.

Contenido orgánico de extractos parcialmente purificados de pulpa dental humana y bovina / Organic content of human and bovine dental pulp partially purified extracts / Conteúdo orgânico de extratos parcialmente purificado de polpa dentária humana e bovina

La pulpa dental contiene tejido conectivo rico en proteínas, mayormente colágeno. El objetivo de este trabajo fue describir un método de purificación de componentes orgánicos pulpares humanos y bovinos, vital y necrótico para validar el modelo experimental bovino. Se prepararon extractos con 50 mg de fracción media de pulpas unirradiculares, 20 humanas o 1 bovina, en 500 µL de Tris-HCl 50 mM pH 7,4, floruro de fenil metil sulfonilo 1mM e hidrocloruro de benzamidina 5 mM frío. Se homogeneizó y centrifugó a 10000 rpm. Los sobrenadantes fueron parcialmente purificados con sulfato de protamina 1%, los sedimentos resuspendidos y dializados 2 horas con recambio en acetato de sodio 500 mM pH 6. Se determinaron proteínas, hidroxiprolina e hidratos de carbono, y se calculó el rendimiento según el contenido de hidroxiprolina. Se aplicó electroforesis en geles de poliacrilamida. Se encontró mayor contenido orgánico en tejido humano que en bovino, y en vital que en necrótico. Por hidroxiprolina/proteínas y rendimiento, el extracto bovino contendría mayor proporción de colágeno. En pulpas humanas se obtuvieron bandas a 78 y 80 kDa, y en bovinas a 74 y 76 kDa. No hubo diferencias entre tejido vital y necrótico. Las evidencias cuali y cuantitativas expresadas validarían al modelo bovino en investigaciones endodónticas.

The dental pulp contains connective tissue and is rich in proteins, mainly collagen. The aim of this study was to describe a purification method for organic compounds of human and bovine pulp tissues, either vital as necrotic, in order to validate the experimental bovine model. Extracts of 50 mg of the middle fraction of one root pulp teeth, 20 humans or 1 bovine, in 500 µL of cold 50 mM Tris - HCl pH 7.4, 1 mM phenyl methyl sulfonyl fluoride and 5 mM benzamidine hydrochloride were prepared. Extracts were homogenized and centrifuged at 10000 rpm. Supernatants were partially purified with 1% protamine sulphate, sediments re-suspended and dialyzed during 2 hours with a spare in 500 mM sodium acetate pH 6. Proteins, hydroxiproline and carbohydrates were determined, and the yield of the process was calculated according to the hydroxiproline content. Poliacrylamide gel electrophoresis was applied. A greater organic content was found in human than in bovine tissue, and in vital rather than in the necrotic one. Hydroxyproline/ proteins and yield evidenced that bovine extract would contain a larger proportion of collagen. Bands at 78 and 80 kDa in human pulp and at 74 and 76 kDa in the bovine tissue were observed. There was no difference between vital and necrotic tissues. The qualitative and quantitative evidences expressed in this work would validate the bovine research endodontic model.

A polpa dentária contém tecido conectivo rico em proteínas, na sua maioria colágeno. O objetivo deste trabalho foi descrever um método de puriIcação de componentes orgânicos da polpa dos humanos e bovinos, vitales e necróticos para validar o modelo experimental bovino. Foram preparados extratos com 50 mg de fração média de polpas unirradiculares, 20 humanas ou 1 bovina em 500 µL de Tris-HCl 50 mM pH 7,4, fenil-metil-sulfonil Juoreto 1 mM e hidrocloreto de benzamidina 5 mM frio. Foi homogeneizado e centrifugado a 10.000 rpm. Os sobrenadantes foram parcialmente puriIcados com sulfatos de protamina 1%, os sedimentos ressuspensos e dialisados 2 horas com recâmbio em acetato de sódio 500 mM pH 6. Foram determinadas proteínas, hidroxiprolina e hidratos de carbono e se calculou o rendimento conforme o conteúdo de hidroxiprolina. Foi aplicada eletroforese em géis de poliacrilamida. Maior conteúdo orgânico foi encontrado em tecido humano que em bovino e em vital que em necrótico. Por hidroxiprolina/proteínas e rendimento, o extrato bovino conteria maior proporção de colágeno. Em polpas humanas foram obtidas faixas a 78 e 80 kDa e em bovinas a 74 e 76 kDa. Não houve diferenças entre tecido vital e necrótico. As evidências qualitativas e quantitativas expressas validariam o modelo bovino em pesquisas endodônticas.

Demyelination of neurofilament protein 200 immune positive never fibers in human pulp

INTRODUCTION: Mammalian tooth pulp is densely innervated by sensory nerves that are mostly C fibers and A delta fibers. However, there is evidence suggesting that many unmyelinated axons in the pulp are in fact parent meylinated axons. MATERIALS AND METHODS: The pulp was removed from healthy premolars and 3rd molars extracted from juveniles and adults undergoing orthodontic treatment, and immunohistochemical staining were applied with NPF200 antibodies, which specifically dye myelinated axons. The specimens underwent an electron microscopy examination with diaminobenzidine (DAB) immunostaining after observation and analysis by fluorescence and confocal laser scanning microscopy. RESULTS: The NPF200 immuno-positive axons in the radicular pulp areas were observed as bundles of many nerve fibers. Many small bundles were formed with fewer axons when firing to the coronal pulp areas and then reachrd a different direction. CONCLUSION: Myelinated fibers innervated to the dental pulp maintain their myelins in the radicular portion, but these fibers lost myelins in the coronal portion. Afterthe loss of myelin, the size of the axoplasm also decreased.

A bioactivity study of Portland cement mixed with beta-glycerophosphosphate on human pulp cell / 대한치과보존학회지

The purpose of this study is to investigate the response of human pulp cell on Portland cement mixed with beta-glycerophosphate. To investigate the effect of beta-glycerophosphate and/or dexamethasone on human pulp cell, ALP activity on various concentration of beta-glycerophosphate and dexamethasone was measured and mineral nodule of human pulp cell was stained with Alizarin red S. MTS assay and ALP activity of human pulp cell on Portland cement mixed with various concentration of beta-glycerophosphate (10 mM, 100mM, 1M) was measured and the specimens were examined under SEM. Addition of beta-glycerophosphate or dexamethasone alone had no effect however, the addition of 5 mM beta-glycerophosphate and 100 nM dexamethasone had the largest increasement in ALP activity. There was no toxicity in all samples and the data showed that Portland cement mixed with 10 mM beta-glycerophosphate had more increase in ALP activity compared with control. In conclusion, Portland cement mixed with beta-glycerophosphate has no toxicity and promotes differentiation and mineralization of pulp cell compared with additive-free Portland cement. This implicated that application of Portland cement mixed with beta-glycerophosphate might form more reparative dentin and in turn it would bring direct pulp capping to success.