Effect of a calcium hydroxide-based intracanal medicament containing N-2-methyl pyrrolidone as a vehicle against Enterococcus faecalis biofilm

Abstract This study investigated the effect of a calcium hydroxide (CH) paste (CleaniCal®) containing N-2-methyl pyrrolidone (NMP) as a vehicle on Enterococcus faecalis (E. faecalis) biofilms compared with other products containing saline (Calasept Plus™) or propylene glycol (PG) (Calcipex II®). Methodology Standardized bovine root canal specimens were used. The antibacterial effects were measured by colony-forming unit counting. The thickness of bacterial microcolonies and exopolysaccharides was assessed using confocal laser scanning microscopy. Morphological features of the biofilms were observed using field-emission scanning electron microscopy (FE-SEM). Bovine tooth blocks covered with nail polish were immersed into the vehicles and dispelling was observed. The data were analyzed using one-way analysis of variance and Tukey tests (p<0.05). Results CleaniCal® showed the highest antibacterial activity, followed by Calcipex II® (p<0.05). Moreover, NMP showed a higher antibacterial effect compared with PG (p<0.05). The thickness of bacteria and EPS in the CleaniCal® group was significantly lower than that of other materials tested (p<0.05). FE-SEM images showed the specimens treated with Calasept Plus™ were covered with biofilms, whereas the specimens treated with other medicaments were not. Notably, the specimen treated with CleaniCal® was cleaner than the one treated with Calcipex II®. Furthermore, the nail polish on the bovine tooth block immersed in NMP was completely dispelled. Conclusions CleaniCal® performed better than Calasept Plus™ and Calcipex II® in the removal efficacy of E. faecalis biofilms. The results suggest the effect might be due to the potent dissolving effect of NMP on organic substances.

Impact of calcium aluminate cement with additives on dental pulp-derived cells

Abstract Calcium aluminate cement (CAC) has been highlighted as a promising alternative for endodontic use aiming at periapical tissue repair. However, its effects on dental pulp cells have been poorly explored. Objective: This study assessed the impact of calcium chloride (CaCl2) and bismuth oxide (Bi2O3) or zinc oxide (ZnO) additives on odontoblast cell response to CAC. Methodology: MDPC-23 cells were exposed for up to 14 d: 1) CAC with 2.8% CaCl2 and 25% ZnO (CACz); 2) CAC with 2.8% CaCl2 and 25% Bi2O3 (CACb); 3) CAC with 10% CaCl2 and 25% Bi2O3 (CACb+); or 4) mineral trioxide aggregate (MTA), placed on inserts. Non-exposed cultures served as control. Cell morphology, cell viability, gene expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), and dentin matrix protein 1 (DMP-1), ALP activity, and extracellular matrix mineralization were evaluated. Data were compared using ANOVA (α=5%). Results: Lower cell density was detected only for MTA and CACb+ compared with Control, with areas showing reduced cell spreading. Cell viability was similar among groups at days one and three (p>0.05). CACb+ and MTA showed the lowest cell viability values at day seven (p>0.05). CACb and CACb+ promoted higher ALP and BSP expression compared with CACz (p<0.05); despite that, all cements supported ALP activity. Matrix mineralization were enhanced in CACb+ and MTA. Conclusion: In conclusion, CAC with Bi2O3, but not with ZnO, supported the expression of odontoblastic phenotype, but only the composition with 10% CaCl2 promoted mineralized matrix formation, rendering it suitable for dentin-pulp complex repair.

Purification of inulinases by changing the ionic strength of the medium and precipitation with alcohols

ABSTRACT The present study evaluated the purification of inulinase by changing the ionic strength of the medium by addition of NaCl and CaCl2 followed by precipitation with n-propyl alcohol or iso-propyl alcohol. The effects of the concentration of alcohols and the rate of addition of alcohols in the crude extract on the purification yield and purification factor were evaluated. Precipitation caused an activation of enzyme and allowed purification factors up to 2.4-fold for both alcohols. The purification factor was affected positively by the modification of the ionic strength of the medium to 0.5 mol.L-1 NaCl before precipitation with the alcohol (n-propyl or iso-propyl). A purification factor of 4.8-fold and an enzyme yield of 78.1 % could be achieved by the addition of 0.5 mol.L-1 of NaCl to the crude extract, followed by the precipitation with 50 % (v/v) of n-propyl alcohol, added at a flow rate of 19.9 mL/min.

Porosity of temporary denture soft liners containing antifungal agents

ABSTRACT Incorporation of antifungals in temporary denture soft liners has been recommended for denture stomatitis treatment; however, it may affect their properties. Objective: To evaluate the porosity of a tissue conditioner (Softone) and a temporary resilient liner (Trusoft) modified by minimum inhibitory concentrations (MICs) of antifungal agents for Candida albicans biofilm. Material and Methods: The porosity was measured by water absorption, based on exclusion of the plasticizer effect. Initially, it was determined by sorption isotherms that the adequate storage solution for specimens (65×10×3.3 mm) of both materials was 50% anhydrous calcium chloride (S50). Then, the porosity factor (PF) was calculated for the study groups (n=10) formed by specimens without (control) or with drug incorporation at MICs (nystatin: Ny-0.032 g, chlorhexidine diacetate: Chx-0.064 g, or ketoconazole: Ke-0.128 g each per gram of soft liner powder) after storage in distilled water or S50 for 24 h, seven and 14 d. Data were statistically analyzed by 4-way repeated measures ANOVA and Tukey's test (α=.05). Results: Ke resulted in no significant changes in PF for both liners in water over 14 days (p>0.05). Compared with the controls, Softone and Trusoft PFs were increased at 14-day water immersion only after addition of Ny and Chx, and Chx, respectively (p<0.05). Both materials showed no significant changes in PF in up to 14 days of S50 immersion, compared with the controls (p>0.05). In all experimental conditions, Softone and Trusoft PFs were significantly lower when immersed in S50 compared with distilled water (p<0.05). Conclusions: The addition of antifungals at MICs resulted in no harmful effects for the porosity of both temporary soft liners in different periods of water immersion, except for Chx and Ny in Softone and Chx in Trusoft at 14 days. No deleterious effect was observed for the porosity of both soft liners modified by the drugs at MICs over 14 days of S50 immersion.

Candida albicans biofilms and MMA surface treatment influence the adhesion of soft denture liners to PMMA resin

The effect of Candida albicans biofilms and methyl methacrylate (MMA) pretreatment on the bond strength between soft denture liners and polymethyl methacrylate (PMMA) resin was analyzed. Specimens were prepared and randomly divided with respect to PMMA pretreatment, soft liner type (silicone-based or PMMA-based), and presence or absence of a C. albicans biofilm. Samples were composed of a soft denture liner bonded between two PMMA bars. Specimens (n = 10) were incubated to produce a C. albicans biofilm or stored in sterile PBS for 12 days. The tensile bond strength test was performed and failure type was determined using a stereomicroscope. Surface roughness (SR) and scanning electron microscopy (SEM) analysis were performed on denture liners (n = 8). Highest bond strength was observed in samples containing a silicone-based soft liner and stored in PBS, regardless of pretreatment (p < 0.01). Silicone-based specimens mostly underwent adhesive failures, while samples containing PMMA-based liners predominantly underwent cohesive failures. The silicone-based specimens SR decreased after 12 days of biofilm accumulation or PBS storage, while the SR of PMMA-based soft liners increased (p < 0.01). The PMMA-based soft liners surfaces presented sharp valleys and depressions, while silicone-based specimens surfaces exhibited more gentle features. In vitro exposure to C. albicans biofilms reduced the adhesion of denture liners to PMMA resin, and MMA pretreatment is recommended during relining procedures.

The sealing ability of MTA apical plugs exposed to a phosphate-buffered saline

OBJECTIVE: The aim of this study was to evaluate the influence of exposure of the mineral trioxide aggregate (MTA) - with and without calcium chloride (CaCl2) - to phosphate-buffered saline (PBS) on the apical microleakage using a glucose leakage system. MATERIAL AND METHODS: Sixty root segments were randomly divided into 4 experimental groups (n=15). After resecting the apical segments and enlarging the canals with Gates-Glidden drills, the apical cavities were filled with MTA with or without CaCl2 and the root canals were dressed with a moistened cotton pellet or PBS, as follows: 1) MTA/cotton pellet; 2) MTA/PBS; 3) MTA+10%CaCl2/cotton pellet; 4) MTA+10%CaCl2/PBS. All root segments were introduced in floral foams moistened with PBS. After 2 months, all root segments were prepared to evaluate the glucose leakage along the apical plugs. The amount of glucose leakage was measured following an enzymatic reaction and quantified by a spectrophotometer. Four roots were used as controls. The data were analyzed using Kruskal-Wallis and Mann-Whitney tests (p<0.05). RESULTS: There were no differences between groups 1 and 2 (p>0.05), and 3 and 4 (p>0.05). The addition of CaCl2 to the MTA significantly decreased its sealing ability (p<0.05). CONCLUSION: The interaction with PBS did not improve the MTA sealing ability. The addition of CaCl2 to the MTA negatively influenced the apical seal. .

Biochemical characterization of a calcium-sensitive protein kinase LeCPK2 from tomato.

LeCPK2 (GenBank GQ205414), a versatile calcium-dependent protein kinase (CDPK or CPK) gene was isolated from tomato in our previous study. In this study, the biochemical properties of LeCPK2 were further investigated. To examine the role of the C-terminal calmodulin-like domain (CLD) of LeCPK2 with respect to Ca2+ activation, the kinase activities of recombinant full-length and truncated LeCPK2 were measured by Kinase-Glo® Luminescent kinase assay (Promega). The results showed that LeCPK2 activity was Ca2+-dependent and the C-terminal CLD of 161 residues was essential for the activation of LeCPK2. The activity of LeCPK2 was sharply stimulated by Ca2+ with K0.5 (concentration of Ca2+ for half-maximal activity) of 48.8 and 45.5 nM with substrate histone IIIs and syntide 2, respectively. The optimal concentration of Mg2+ for LeCPK2 activity was 20 and 10 mM for substrate histone IIIs and syntide 2, respectively. The Km value of LeCPK2 towards histone IIIs and syntide 2 was 44.9 μg/ml and 89.52 μM, respectively. The determination of biochemical properties of LeCPK2 would provide some clues on how its activity was regulated in vivo.

Effect of additives on the compressive strength and setting time of a Portland cement

Improvements in strength and setting time of Portland cements (PC) are needed to enhance their performance as endodontic and load bearing materials. This study sought to enhance the compressive strength and setting time of a PC by adding one of the following additives: 20 percent and 30 percent poly-methylmethacrylate (PMMA), 20 percent and 30 percent irregular and spherical amalgam alloys, and 10 percent CaCl2. The control consisted of unreinforced PC specimens. Setting time was determined using a Gillmore apparatus according to standardized methods while compressive strength was measured using a universal testing machine after 21 hours or 60 days of water storage. Data were analyzed by ANOVA, Tukey and Games-Howell tests (á = 5 percent). All additives significantly decreased both initial and final setting times as compared with the PC-control (p < .05). 30 percent PMMA and 30 percent irregular alloy had the lowest values of initial setting time. 30 percent irregular alloy also produced the lowest values of final setting time while 30 percent spherical alloy yielded the highest (p < .05). No differences were detected between the compressive strength values of 21 hours and 60 days. While 10 percent CaCl2, 20 percent and 30 percent PMMA produced values significantly lower than the PC-control, 30 percent spherical alloy significantly improved the compressive strength of the reinforced PC (p < .05). In summary, all additives significantly reduced the setting time and 30 percent spherical amalgam alloy yielded a significant increase in compressive strength for the tested PC, which might represent an improved composition for PCs to expand their use as endodontic and potentially load bearing materials.

Novel multi-layer APPPA microcapsules for oral delivery: preparation condition, stability and permeability.

Oral therapy utilizing cell microencapsulation has shown promise in the treatment of many diseases. Current obtainable microcapsule membranes, however, show inadequate stability in the gastrointestinal (GI) environment, thus restricting the general application of live cells for oral therapy. To overcome this limitation, we have previously developed a novel multi-layer alginate/poly-L-lysine/pectin/poly-L-lysine/alginate microcapsule (APPPA) with demonstrated improvement on membrane stability over the frequently reported alginate/poly-L-lysine/alginate (APA) microcapsules. In this study, we further examined the effects of preparation conditions on microcapsule formation, and assessed the membrane strength and GI stability. Results showed that increased membrane strength of the APPPA microcapsules was attained by using pectin with low degree of esterification as the mid-layer material, saline as the solvent for the preparation solutions and washing medium, and 0.1 M CaCl2 as the gelling solution for alginate cores. Resistance of this membrane to the simulated GI fluids was also investigated. Permeability of and release profiles from the APPPA microcapsules were found comparable to the APA microcapsules. These findings suggested that the multi-layer APPPA microcapsule formulation may have potential in oral delivery of proteins, live bacterial cells and other biomedical applications.

Removal of fluoride from water with powdered corn cobs.

The adsorption of fluoride on corn cobs powder was investigated in the present study. Neat powdered corn cobs did not show remarkable adsorption but aluminium treated corn cobs had good adsorption capacity. The parameters studied include the contact time, concentration, temperature and pH. Near neutral pH was identified as the optimum condition of the medium, and 90 to 120 minutes was the best contact time for maximum fluoride adsorption. The adsorption process was found to follow Freundlich isotherm. The adsorption process was found to be exothermic as adsorption decreased with increasing temperature.

Como evitar a formação de substâncias tóxicas durante a absorção de dióxido de carbono pela cal sodada com uso de anestésicos halogenados / Preventing toxic substances production during carbon dioxide absorption by soda lime with halogenate anesthetics

JUSTIFICATIVA E OBJETIVOS: A cal sodada desde o início do seu uso sempre apresentou algumas complicações que resultaram em dificuldade na sua aplicabilidade. No entanto, devido as grandes vantagens que oferecia em relação a redução do fluxo de gases frescos, despoluição da sala de cirurgia e umidificação do sistema de inalação e via aérea, fizeram com que continuassem as pesquisas para que pudesse ser melhorada e corrigida de forma que a continuidade da sua utilização fosse assegurada. Atualmente existe o problema da desidratação com elevação da temperatura e da degradação metabólica dos anestésicos halogenados que necessitam de cuidados especiais para evitar a formação de produtos tóxicos. CONTEUDO: Existe uma reação em cadeia a partir da cal sodada desidratada ou ressecada com baixos volumes percentuais de água. Há aumento da temperatura, maior absorção de anestésico halogenado para o interior do granulo de cal em seguida maior degradação metabólica das moléculas destes agentes e conseqüentemente a produção de substâncias tóxicas como o Composto A pela reação dos hidróxidos com o sevoflurano. Há também formação de monóxido de carbono produzido da mesma forma pela reação entre os halogenados e as bases fortes da cal. O composto A é nefrotóxico e o monóxido de carbono leva a hipóxia e alterações graves da coagulação do sangue. Além dos cuidados para a hidratação da cal sodada é possível usá-la sem conter as bases fortes como os hidróxidos de potássio e de sódio, contendo apenas hidróxido de cálcio para evitar excessivo aumento da temperatura e grande degradação metabólica dos halogenados sem prejudicar a absorção do dióxido de carbono. CONCLUSÕES: Deve-se ter o cuidado em usar a cal sodada mais recente possível e quando ela fica exposta ao meio ambiente (ar seco) por muitas horas como por exemplo em um final de semana (mais de 48 horas) é recomendável colocar água, de preferência destilada, na relação de 25 ml para cada 500 g de cal. Atualmente a indústria está bem informada sobre o problema da composição da cal, então, deve-se preferir a cal sodada que tenha somente o hidróxido de cálcio e seja totalmente desprovida de hidróxido de potássio e hidróxido de sódio.

Leaching of As(III) and As(V) from exhausted calsico--a case study.

Batch leaching experiments were conducted to estimate the leaching of As(III) and As(V) from exhausted CalSiCo.The leaching of As(III) and As(V) was found to be function of time and concentrations of anions such as Cl-, NO3-, SO4(2-) in extraction fluid. Extraction tests were carried out to determine the maximum leachable concentration under the chosen conditions of leaching medium and leaching time. It has been observed that the leaching of As(III) is more than As(v) in all the cases. Leaching of As(III) and As(v) from exhausted CalSiCo is also carried out in rainwater and in tap water. It is observed that leaching of As is more in rainwater than tap water. Further higher leaching is observed in case of fluids containing chloride compared to those containing nitrate and sulphate.

A carbon-limited medium for growth and sporulation of Bacillus thuringiensis var. kurstaki.

A culture medium for batch production of d-endotoxin by Bacillus thuringiensis (B., t.) has been modified. Through batch and continuous cultivation studies, the original medium was diagnosed to be limited in organic nitrogen. Corn steep liquor was found to be an excellent source for the organic nitrogen and its addition resulted in a carbon limited medium and in a significant increase in the amount of spore-toxin complex formed in shake flasks. Results of bioassay, conducted on Trichoplusia ni, suggest enhancement of larvicidal efficacy under carbon-limited growth conditions.