Silver Nanoparticles Alone or in Combination with Calcium Hydroxide Modulate the Viability, Attachment, Migration, and Osteogenic Differentiation of Human Mesenchymal Stem Cells.

This study aimed to evaluate the effect of silver nanoparticles (AgNPs) alone or in combination with calcium hydroxide (Ca(OH)2) on the proliferation, viability, attachment, migration, and osteogenic differentiation of human mesenchymal stem cells (hMSCs). Different concentrations of AgNPs alone or mixed with Ca(OH)2 were prepared. Cell proliferation was measured using AlamarBlue, and hMSCs attachment to dentin disks was evaluated using scanning electron microscopy. Live-dead imaging was performed to assess apoptosis. Wound healing ability was determined using the scratch-migration assay. To evaluate osteogenic differentiation, the expression of Runt-related transcription factor (RUNX2), Transforming growth factor beta-1 (TGF-ß1), Alkaline Phosphatase (ALP), and Osteocalcin (OCN) were measured using real-time reverse transcriptase polymerase chain reaction. ALP staining and activity were also performed as indicators of osteogenic differentiation. AgNPs alone seemed to favor cell attachment. Lower concentrations of AgNPs enhanced cell proliferation. AgNP groups showed markedly less apoptosis. None of the medicaments had adverse effects on wound closure. The expression of TGF-ß1 was significantly upregulated in all groups, and OCN was highly expressed in the AgNP groups. AgNPs 0.06% showed the most enhanced ALP gene expression levels, activity, and marked cytochemical staining. In conclusion, AgNPs positively affect hMSCs, making them a potential biomaterial for various clinical applications.

Evaluation of ozonized calcium hydroxide as an effective intracanal medicament during root canal procedures: an in vitro observational study.

Achieving thorough disinfection is regarded as one of the pillars in endodontics. Although calcium hydroxide (CH) is one of the routinely used intracanal medicament in endodontics; alternative approaches are gaining popularity to mitigate endodontic pathology. However, CH has to be tested for its dissociation which is a rate-limiting attribute essential for its therapeutic action. The dissociation of CH into OH- and Ca2+ depends on the vehicle used to prepare the paste. This in-vitro study evaluated the use of ozonized olive oil in facilitating calcium ion release and change in pH when combined with CH. Fifty single rooted extracted human mandibular premolars were instrumented with NiTi rotary files (40/6). The teeth were divided into two groups (n = 25 per group) on the basis of vehicle: olive oil (CH + olive oil) and ozonized olive oil (CH + ozonized olive oil) groups. Both olive and ozonized olive oil vehicles allowed the diffusion of ions. However, pastes prepared with ozonized oil showed more ion diffusion, with marked calcium ion release after 15 days and alkalinity was maintained for complete period of 15 days, depicting better support for CH action. The change in calcium ion release and alkalinity were statistically significant in ozonized oil vehicle compared to olive oil vehicle. The present in-vitro study supports the use of ozonized olive oil as a vehicle to be used with CH as an intracanal medicament, considering its anti-microbial potential and sustainable release of calcium ions. The study was approved by the Institutional Ethical Committee of Manubhai Patel Dental College (approval No. MPDC_130/CONS-25/17) on June 4, 2018.

Combined effects of calcium hydroxide and photobiomodulation therapy on apexogenesis of immature permanent teeth in dogs.

Treatment of high-risk traumatic immature teeth due to incomplete root development is challenging. Apexogenesis is currently the ideal treatment option that allows normal root development. The purpose of this study was to evaluate the apexogenesis process of immature permanent teeth of dogs when co-administered with calcium hydroxide and photobiomodulation therapy (PBMT). A total of 36 immature permanent anterior and premolar teeth were selected from three 4-6-month-old dogs of Iranian mixed generation. The teeth were categorized into two groups, calcium hydroxide with laser irradiation (CHL) and calcium hydroxide without laser irradiation (CH). All the selected teeth received calcium hydroxide pulpotomy. After restoring teeth with amalgam, the CHL group received gallium­aluminum-arsenide (GaAlAs) diode laser (810 nm, 4.2 J/cm2, 0.3 W, 9 s,CW) on apical one-third of both buccal and lingual roots. The irradiation was repeated every 48 h for fourteen days. Intravenous tetracycline was used to observe newly formed dentin in the first, third, seventh, and fourteenth days. The distance between tetracycline lines (DTL) was examined by Fluorescence microscopy. Generalized estimating equations (GEE) were used for data analysis. In all assessments, the mean DTL were greater in the CHL group. However, the two groups had no significant differences in the amount of deposited dentin between the first and third, third and seventh, and first and seventh lines. Meanwhile, there was a significant difference between the two groups in terms of the distances between lines 7 and 14, 1 and 14 and also 3 to 14 (P < .001). In other words, from the 7th day onwards, there was a significant difference between the two groups. Within the limitation of this study, the combination therapy of PBMT and pulpotomy with calcium hydroxide accelerated apexogenesis in immature permanent dogs' teeth.

Efecto antimicrobiano de la pasta 3-ATB y el hidróxido de calcio con distintos solventes / Antimicrobial effect of triple antibiotic paste and calcium hydroxide with different solvents

Objetivo: El objetivo de este estudio ha sido valorar cómo el hidróxido de calcio y la pasta triantibiótica (3-ATB) con agua destilada o propilenglicol actúan frente a una de las principales bacterias anaerobias facultativas presentes en dientes permanentes jóvenes con pulpa necrótica. Material y Método: Se llevó a cabo la siembra de la cepa bacteriana E.Coli (ATCC(R) 25922) en placas Petri para probar la susceptibilidad de los materiales mediante el método de difusión en agar. Se realizó la lectura de los resultados a las 48 horas mediante la medición de los halos de inhibición (mm) presentes en las placas. Resultados: La pasta tri-antibiótica mostró mayor inhibición que el hidróxido de calcio. Respecto a los solventes, la pasta 3-ATB ofreció mejor resultado combinada con propilenglicol, mientras que la respuesta del hidróxido de calcio fue más favorable al mezclar con agua destilada. Conclusiones: La pasta 3-ATB es más efectiva contra microorganismos anaerobios facultativos que el hidróxido de calcio, independientemente del solvente utilizado. En cuanto al solvente, el propilenglicol es más efectivo con pasta tri-antibiótica y el hidróxido de calcio lo es con agua destilada

Objective: The objective of this study was to assess how calcium hydroxide and triple antibiotic paste (3-ATB) with distilled water or propylene glycol act against one of the main facultative anaerobic bacteria present in young permanent teeth with necoritc pulp. Material and Methods: The seeding of the E.Coli (ATCC(R) 25922) was carried out in Petri dishes to test the susceptibility of the materials by the agar diffusion method. The results were read at 48 hours by measuring the inhibition zones (mm) present in the plates. Results: 3-ATB paste showed greater inhibition than calcium hydroxide. Regarding the solvents, 3-ATB paste offered better re-sults with propylene glycol, while the calcium hydroxide response was more favorable mixed with distilled water. Conclusions: 3-ATB paste is more effective against facultative anaerobic microorganisms than calcium hydroxide, regardless of the solvent used. As for the solvent, propylene glycol is more effective with 3-ATB paste while calcium hydroxide with distilled water

Benefit of solid-liquid separation on volatile fatty acid production from grass clipping with ultrasound-calcium hydroxide pretreatment.

Ultrasound-calcium hydroxide (US-Ca(OH)2) pretreatment effectively enhanced volatile fatty acid (VFA) production from lignocellulosic biomass. In this paper, solid and liquid fraction of pretreated grass clipping was for the first time separately fermented in order to improve organic recovery from liquid fraction and reduce inhibition due to alkaline pretreatment. The total VFA yield and VS removal reached 515 mg/g TS and 59.7% after solid-liquid separation, exhibiting an increase of 116.7% and 91.9% comparing to that of mixture sample. The dominate components of VFAs are acetic and propionate acid, accounting for 80-90% of total VFAs. Kinetic analysis showed that the highest maximum VFA production rate of 690 mg/L·d and the highest cumulative VFA production potential of 3299 mg/L were achieved in the fermentation of solid fraction. Microbial analysis showed that the dominate genera for VFA production were Halocella and Ruminiclostridium, both with a relative abundance of 20.1% in fermentation of solid fraction.

The role of Foxq1 in proliferation of human dental pulp stem cell.

This study aimed to investigate the role for Foxq1 in proliferation activity regulation of dental pulp stem cells (DPSCs). Proliferation of DPSC was induced by calcium hydroxide, then expression alteration of Foxq1 was evaluated. Lentivirus was employed to manipulate Foxq1 level in DPSC, and proliferation activities were evaluated. To look into mechanism regulating Foxq1 level after calcium hydroxide stimulation, expressions of various microRNAs were evaluated, then bioinformatics study and dual-luciferase study were carried out to confirm targeting relationship between microRNA and Foxq1. The result of our study indicated that proliferation activities of DPSCs were enhanced after calcium hydroxide stimulation, during which expression of Foxq1 was also up-regulated. Cell viability and progression from G1 to S phase were both improved with overexpression of Foxq1, and microRNAs profiling study and dual-luciferase result suggested miR-320b contributed to the up-regulation of Foxq1 after calcium hydroxide stimulation. These results suggested that miR-320b mediated Foxq1 up-regulation promote proliferation of dental pulp stem cells.

Inclusion of calcium hydroxide-treated corn stover as a partial forage replacement in diets for lactating dairy cows.

Chemical treatment may improve the nutritional value of corn crop residues, commonly referred to as corn stover, and the potential use of this feed resource for ruminants, including lactating dairy cows. The objective of this study was to determine the effect of prestorage chopping, hydration, and treatment of corn stover with Ca(OH)2 on the feeding value for milk production, milk composition, and dry matter intake (DMI). Multiparous mid-lactation Holstein cows (n = 30) were stratified by parity and milk production and randomly assigned to 1 of 3 diets. Corn stover was chopped, hydrated, and treated with 6% Ca(OH)2 (as-fed basis) and stored in horizontal silo bags. Cows received a control (CON) total mixed ration (TMR) or a TMR in which a mixture of treated corn stover and distillers grains replaced either alfalfa haylage (AHsub) or alfalfa haylage and an additional portion of corn silage (AH+CSsub). Treated corn stover was fed in a TMR at 0, 15, and 30% of the diet DM for the CON, AHsub, and AH+CSsub diets, respectively. Cows were individually fed in tiestalls for 10 wk. Milk production was not altered by treatment. Compared with the CON diet, DMI was reduced when the AHsub diet was fed and tended to be reduced when cows were fed the AH+CSsub diet (25.9, 22.7, and 23.1 ± 0.88 kg/d for CON, AHsub, and AH+CSsub diets, respectively). Energy-corrected milk production per unit of DMI (kg/kg) tended to increase with treated corn stover feeding. Milk composition, energy-corrected milk production, and energy-corrected milk per unit of DMI (kg/kg) were not different among treatments for the 10-wk feeding period. Cows fed the AHsub and AH+CSsub diets had consistent DMI over the 10-wk treatment period, whereas DMI for cows fed the CON diet increased slightly over time. Milk production was not affected by the duration of feeding. These data indicate that corn stover processing, prestorage hydration, and treatment with calcium hydroxide can serve as an alternative to traditional haycrop and corn silage in diets fed to mid-lactation dairy cows.

Effect of treating sugarcane bagasse with urea and calcium hydroxide on feed intake, digestibility, and rumen fermentation in beef cattle.

Four beef cattle with initial body weight of 283 ± 14 kg were randomly allocated according to a 4 × 4 Latin square design to study on the effect of feeding sugarcane bagasse (SB) treated with urea and/or calcium hydroxide (Ca(OH)2) on feed intake, digestibility, and rumen fermentation. The treatments were as follows: rice straw (RS), untreated SB (SB), 4 % urea-treated SB (SBU), and 2 % urea + 2 % Ca(OH)2-treated SB (SBUC), respectively. The results revealed that cattle fed with SBU and SBUC had higher feed intake and apparent digestibility. Ammonia nitrogen and blood urea nitrogen were increased in cattle fed with SB as roughage source (P < 0.05). Feeding SBU and SBUC to cattle resulted in higher propionic acid and lower acetic acid, acetic to propionic ratio, and methane production (P < 0.05). Moreover, the number of fungi was increased in SBU- and SBUC-fed groups while protozoa population was unchanged. This study concluded that the nutritive value of SB was improved by urea and/or Ca(OH)2 treatment, and feeding treated SB could increase feed intake, digestibility, and rumen fermentation. This study suggested that SB treated with 2 % urea + 2 % Ca(OH)2 could be used as an alternative roughage source for ruminant feeding.

The effects of calcium hydroxide-treated whole-plant and fractionated corn silage on intake, digestion, and lactation performance in dairy cows.

The objective of this trial was to evaluate, in dairy cattle, the effects of calcium hydroxide treatment of whole-plant corn and a treatment applied to the bottom stalk fraction of the corn plant, achieved by harvesting corn in 2 crop streams. The treatments were calcium hydroxide-treated corn silage (TRTCS), toplage supplemented with calcium hydroxide-treated stalklage (TPL), a positive control of brown midrib corn silage (BMR), and a negative control of conventional whole-plant corn silage (WPCS). The toplage was harvested at a height of 82 cm with 2 of the 6 rows set as ear-snapping to incorporate higher tissues into the stalklage. Stalklage was harvested at 12 cm, and other corn silages were harvested at 27 cm. Sixteen pens, each with 8 Holstein cows averaging 70±25 d in milk and 46±11 kg of milk d(-1), were assigned 4 per treatment in a completely randomized design. The diet was approximately 40% corn silage, 20% alfalfa silage, and 40% concentrate on a dry matter basis. A 2-wk covariate period with conventional corn silage was followed by an 8-wk treatment period in which the 4 corn silage treatments were the only effective difference in diets. Cows fed TPL and TRTCS consumed more (1.9 and 1.4 kg of organic matter d(-1), respectively) than did cows fed WPCS. Milk yield was greater for cows fed BMR, TPL, and TRTCS. Cows fed BMR and TPL produced 2.9 and 2.7 kg d(-1), respectively, more energy-corrected milk (ECM) than cows fed WPCS, and cows fed TRTCS had the greatest ECM production (4.8 kg of ECM d(-1) greater than cows fed WPCS). No differences in body weight or body condition scored were observed. Milk fat concentration was similar among treatments and milk protein concentration was reduced for TRTCS. Starch and neutral detergent fiber digestibility were greater for cows fed TRTCS.

Bioaggregate of photo-fermentative bacteria for enhancing continuous hydrogen production in a sequencing batch photobioreactor.

Hydrogen recovery through solar-driven biomass conversion by photo-fermentative bacteria (PFB) has been regarded as a promising way for sustainable energy production. However, a considerable fraction of organic substrate was consumed for the growth of PFB as biocatalysts, furthermore, these PFB were continuously washed out from the photobioreactor in continuous operation because of their poor flocculation. In this work, PFB bioaggregate induced by L-cysteine was applied in a sequencing batch photobioreactor to enhance continuous hydrogen production and reduce biomass washout. The effects of the hydraulic retention time (HRT), influent concentration and light intensity on hydrogen production of the photobioreactor were investigated. The maximum hydrogen yield (3.35 mol H2/mol acetate) and production rate (1044 ml/l/d) were obtained at the HRT of 96 h, influent concentration of 3.84 g COD/l, and light intensity of 200 W/m(2). With excellent settling ability, biomass accumulated in the photobioreactor and reached 2.15 g/l under the optimum conditions. Structural analysis of bioaggregate showed that bacterial cells were covered and tightly linked together by extracellular polymeric substances, and formed a stable structure. Therefore, PFB bioaggregate induced by L-cysteine is an efficient strategy to improve biomass retention capacity of the photobioreactor and enhance hydrogen recovery efficiency from organic wastes.

Hydrothermal treatment of naturally contaminated maize in the presence of sodium metabisulfite, methylamine and calcium hydroxide; effects on the concentration of zearalenone and deoxynivalenol.

Fusarium toxin-contaminated ground maize was hydrothermally treated in the presence of different combinations of chemicals in order to simultaneously reduce zearalenone (ZEA) and deoxynivalenol (DON) concentrations. Treatments were carried out in a laboratory conditioner at 80 °C and 17 % moisture. Six different treatments were performed, consisting of 3 doses of methylamine (MMA; 2.5, 5 and 10 g/kg maize) at a constant dose of 5 g sodium metabisulfite (SBS)/kg, either with or without the addition of 20 g calcium hydroxide (Ca(OH)2)/kg. The used maize was contaminated with approximately 45.99 mg DON/kg and 3.46 mg ZEA/kg. Without the addition of Ca(OH)2, DON reductions reached approximately 82% after 1-min treatment and the toxin disappeared nearly completely after 10 min when 2.5 or 5 g MMA were applied. ZEA concentrations were only marginally affected. In the presence of Ca(OH)2, reductions in DON concentrations were lower, but were enhanced by increasing doses of MMA. ZEA concentrations were reduced by 72, 85 and 95% within the first 5 min of the treatment at MMA dosages of 2.5, 5 and 10 g/kg maize, respectively. The application of SBS in combination with a strong alkaline during hydrothermal treatment seems to be a promising approach to simultaneously decontaminate even high amounts of DON and ZEA in ground maize and may contribute to reduce the toxin load of diets.

Effect of feeding of calcium hydroxide-treated or vitamin E-supplemented cottonseed meal on plasma gossypol levels, blood parameters, and performance of Bikaneri lambs.

To study the effect of feeding calcium hydroxide-treated or vitamin E-supplemented cottonseed meal (CSM) incorporated diets on plasma gossypol, blood parameters and animal performance, 24 male Bikaneri lambs of 6-7 months of age and of uniform body weight were divided into four groups of six animals each in a completely randomized design and respectively fed isonitrogenous and isocaloric concentrate mixtures containing 20% soybean meal (CON) or 40% raw CSM (RCSM), 40% raw CSM supplemented with 500 IU of vitamin E per head per day (ERCSM), and 40%, 1.5% calcium hydroxide-treated CSM (CaCSM) along with ad libitum wheat straw throughout 510 days of experimental feeding. The lambs on all the diets grew linearly throughout the experimental period. The total weight gain, in turn the average daily gain (ADG), was not affected by dietary variations. The daily intake of dry matter, crude protein (CP), digestible crude protein (DCP) and total digestible nutrients (TDN) were found comparable among lambs of all the groups. Though total gossypol intake was similar in RCSM, ECSM and CaCSM groups, however, free gossypol intake was significantly higher (P < 0.01) in RCSM, ECSM groups as compared to CaCSM group. Serum iron and blood hemoglobin levels were significantly (P < 0.05) lower in RCSM group as compared to CaCSM and CON groups, and ALT activity was significantly (P < 0.05) higher on RCSM group as compared to other groups. Plasma gossypol and osmotic fragility of erythrocytes were significantly (P < 0.05) increased in RCSM group as compared to CaCSM and ERCSM groups. However, there was no significant difference in the concentration of other blood/serum biochemical constituents among the lambs on different groups. Based on the results, it may be concluded that feeding of 40% CSM in the concentrate mixture of the diet in Bikaneri lambs did not have any major adverse effect in blood parameters and animal performance. Either calcium hydroxide treatment or vitamin E supplementation did not produce any major additional benefits.

Bioactivity of EndoSequence root repair material and bioaggregate.

AIM: To evaluate the bioactivity of Bioaggregate (BA), EndoSequence Root Repair Material (ERRM), and white ProRoot Mineral trioxide aggregate (MTA). METHODOLOGY: Sixty horizontal root sections with standardized canal spaces were divided randomly into 3 groups (n = 20) and filled with white ProRoot MTA (groups 1 and 2), BA (groups 3 and 4) or ERRM putty (groups 5 and 6). The specimens of groups 1, 3 and 5 (each of 10) were immersed in phosphate-buffered saline (PBS) for 1 week and those of groups 2, 4 and 6 (each of 10) for 2 months. After the experimental periods, the specimens were processed for scanning electron microscopy (SEM) observations. Precipitation of apatite crystals on the surfaces of the cements and/or at the dentine-cement interface was evaluated and analysed elementally by energy dispersive X-ray (EDX) instrument. RESULTS: Analysis of specimens revealed various surface morphologies that were dependent on the material and immersion time in PBS. The formation of precipitates was observed on the surfaces of all materials at 1 week, which increased substantially over time. After 2 months, the surface of the cements was changed dramatically and consisted of a substantially greater amount of apatite aggregates. Interfacial layers in some areas of the dentine-cement interface were found only following 2 months of immersion. Precipitates on MTA revealed high peaks of Ca, Si and O after 1 week of immersion; after 2 months, high peaks of Ca, P and O were present. Precipitates on BA and ERRM displayed high Ca, P O peaks after both 1 week and 2 months. CONCLUSION: Exposure of MTA, BA and ERRM to PBS resulted in precipitation of apatite crystalline structures that increased over time. This suggests that the tested materials are bioactive.

Synthesis and characterization of biodegradable microcapsules for the controlled delivery of calcium hydroxide.

This study aimed to synthesize and characterize biodegradable microcapsules based on poly(lactic acid) (PLA) and ethylcellulose (EC) for a controlled delivery of calcium hydroxide. Phase separation technique was adopted to synthesize calcium hydroxide-loaded PLA/EC microcapsules. Four PLA/EC blends (4/1, 1/1, 1/4, pure EC) were used as shell materials and the input ratio of calcium hydroxide to shell polymer was 4:1 for all microcapsules. The morphology and composition were studied using SEM-EDS and TEM. Particle size distribution, glass-transition temperature, drug loading, and encapsulation efficiency were characterized. In vitro release of the microcapsules was evaluated using a pH microelectrode and an auto-biochemistry analyzer. SEM images of microcapsules showed uniform spherical structures with smooth surfaces. Core-shell, hetero-structures were confirmed using TEM. The presence of calcium in the microcapsules was verified with EDS. Pure calcium hydroxide was 160 nm in diameter and the particle size of the microcapsules ranged between 500 nm and 4 µm. With an increase of PLA in PLA/EC blend, the size of microcapsules increased accordingly. Encapsulation efficiency of these microcapsules was higher than 57% and drug loading was higher than 80%, which were not significantly different among four microcapsules. Pure calcium hydroxide powder was used as a control and 90% was released within 48 h, while release of calcium hydroxide from microcapsules took between 168 and 456 h, depending on the PLA/EC ratio. Compared with calcium hydroxide powder, the calcium hydroxide-loaded microcapsules showed a sustained and prolonged release, which could be controlled via the regulation of the PLA/EC ratio.

Pilot-scale waste activated sludge alkaline fermentation, fermentation liquid separation, and application of fermentation liquid to improve biological nutrient removal.

The use of sludge fermentative short-chain fatty acids (SCFA) as an additional carbon source of biological nutrient removal (BNR) has drawn much attention recently as it can reuse sludge organics, reduce waste activated sludge production, and improve BNR performance. Our previous laboratory study had shown that the SCFA production was significantly enhanced by controlling sludge fermentation at pH 10 with NaOH. This paper focused on a pilot-scale study of alkaline fermentation of waste activated sludge, separation of the fermentation liquid from the alkaline fermentation system, and application of the fermentation liquid to improve municipal biological nitrogen and phosphorus removal. NaOH and Ca(OH)(2) were used respectively to adjust the alkaline fermentation pH, and their effects on sludge fermentation and fermentation liquid separation were compared. The results showed that the use of Ca(OH)(2) had almost the same effect on SCFA production improvement and sludge volatile suspended solids reduction as that of NaOH, but it exhibited better sludge dewatering, lower chemical costs, and higher fermentation liquid recovery efficiency. When the fermentation liquids, adjusted with Ca(OH)(2) and NaOH respectively, were added continuously to an anaerobic-anoxic-aerobic municipal wastewater BNR system, both the nitrogen and phosphorus removals, compared with the control, were improved to the same levels. This was attributed to the increase of not only influent COD but also denitrifying phosphorus removal capability. It seems that the use of Ca(OH)(2) to control sludge fermentation at pH 10 for efficiently producing a carbon source for BNR is feasible.

Modeling the kinetics of calcium hydroxide catalyzed methanolysis of sunflower oil.

The kinetics of Ca(OH)(2)-catalyzed methanolysis of sunflower oil was studied at a moderate temperature (60 degrees C), a methanol-to-oil molar ratio (6:1) and different catalyst amounts (from 1% to 10% based on oil weight). The methanolysis process was shown to involve the initial triglyceride (TG) mass transfer controlled region, followed by the chemical reaction controlled region in the latter period. The TG mass transfer limitation was caused by the low available active specific catalyst surface due to the high adsorbed methanol concentration. Both the TG mass transfer and chemical reaction rates increased with increasing the catalyst amount.

An explanation of the mineralization mechanism in osteoblasts induced by calcium hydroxide.

Calcium hydroxide (Ca(OH)(2)) has been broadly used in endodontics, including apexification to obtain apical closure by mineralization. However, the detailed mechanism of mineralization induced by Ca(OH)(2) is still unclear. This study focuses on the function of calcium and hydroxyl ions which dissociate from Ca(OH)(2) during the mineralization process. Though primary osteoblasts cultured in the medium without or with 0.025mgml(-1) Ca(OH)(2) did not show mineralization, they did exhibit mineralization when they were cultured with a higher concentration of Ca(OH)(2) (0.25mgml(-1)). Mineralization induced in the presence of 0.25mgml(-1) Ca(OH)(2) was greater at pH 7.4 than at pH 8.5. The high mineralization activity observed under neutral conditions was caused by the prolonged activation of p38 and JNK. Hydroxyl ions did not have any effect on the mineralization. The results demonstrate that calcium ions dissociated from Ca(OH)(2) are critical for inducing the mineralization of osteoblasts.

pH and base counterion affect succinate production in dual-phase Escherichia coli fermentations.

Succinate production was studied in Escherichia coli AFP111, which contains mutations in pyruvate formate lyase (pfl), lactate dehydrogenase (ldhA) and the phosphotransferase system glucosephosphotransferase enzyme II (ptsG). Two-phase fermentations using a defined medium at several controlled levels of pH were conducted in which an aerobic cell growth phase was followed by an anaerobic succinate production phase using 100% (v/v) CO(2). A pH of 6.4 yielded the highest specific succinate productivity. A metabolic flux analysis at a pH of 6.4 using (13)C-labeled glucose showed that 61% of the PEP partitioned to oxaloacetate and 39% partitioned to pyruvate, while 93% of the succinate was formed via the reductive arm of the TCA cycle. The flux distribution at a pH of 6.8 was also analyzed and was not significantly different compared to that at a pH of 6.4. Ca(OH)(2) was superior to NaOH or KOH as the base for controlling the pH. By maintaining the pH at 6.4 using 25% (w/v) Ca(OH)(2), the process achieved an average succinate productivity of 1.42 g/l h with a yield of 0.61 g/g.

Dynein cleavage and microtubule accumulation in okadaic acid-treated neurons.

Impairment of protein phosphatase 2A (PP2A) activity is implicated in tau hyperphosphorylation and microtubule (MT) instability in Alzheimer's disease (AD). Here, we report that okadaic acid, an effective PP2A inhibitor, suppresses the levels of acetylated and detyrosinated tubulins, but enhances tyrosinated tubulins in rat primary cortical neuron cultures. Immunocytochemistry experiments reveal that MTs accumulate intensely around soma and proximal neurites, implying impairment of MT transport to distal neurites which is mediated by dynein and dynactin. Here, we reveal that they can be cleaved by calpain. Notably, shortening of process length in OA-treated neurons is alleviated when calpain cleavage activity is inhibited. Based on these results, we propose that calpain-mediated dynein cleavage in OA-treated neurons is responsible for the MT transport deficit, and consequently, neurite retraction.

Human tooth culture: a study model for reparative dentinogenesis and direct pulp capping materials biocompatibility.

In a previous work, based on an in vitro entire tooth culture model of human immature third molars, we demonstrated that perivascular progenitor cells can proliferate and migrate to the injury site after pulp exposure. In this work, we investigated the differentiation of cells after direct capping with biomaterials classically used in restorative dentistry. Histological staining after direct pulp capping with Calcium Hydroxide XR(R) or MTA revealed early and progressive mineralized foci formation containing BrdU-labeled sequestered cells. The molecular characterization of the matrix and the sequestered cells by immunohistochemistry (Collagene type I, Dentin sialoprotein, and Nestin) clearly demonstrates that these areas share common characteristics of the mineralized matrix of reparative dentin formed by odontoblast-like cells. This reproduces some features of the pulp responses after applying these materials in vivo and demonstrates that the entire tooth culture model reproduces a part of the early steps of dentin regeneration in vivo. Its future development may be useful in studying the effects of biomaterials on this process.