Calcium Chloride Solution Research Articles

Contrasting effects of polysaccharide components on the cooking properties of roots, tubers and bananas.

Consumer preferences for boiled or fried pieces of roots, tubers and bananas (RTBs) are mainly related to their texture. Different raw and cooked RTBs were physiochemically characterized to determine the effect of biochemical components on their cooking properties. Firmness in boiled sweetpotato increases with sugar and amylose contents but no significant correlation was observed between other physicochemical characteristics and cooking behaviour. Hardness of boiled yam can be predicted by dry matter (DM) and galacturonic acid (GalA) levels. For cassava, no significant correlation was found between textural properties of boiled roots and DM, but amylose and Ca2+ content were correlated with firmness, negatively and positively, respectively. Water absorption of cassava root pieces boiled in calcium chloride solutions was much lower, providing indirect evidence that pectins are involved in determining cooking quality. A highly positive correlation between textural attributes and DM was observed for fried plantain, but no significant correlation was found with GalA, although frying slightly reduced GalA. The effect of main components on texture after cooking differs for the various RTBs. The effect of global DM and major components (i.e. starch, amylose) is prominent for yam, plantain and sweetpotato. Pectins also play an important role on the texture of boiled yam and play a prominent role for cassava through interaction with Ca2+. © 2023 Bill and Melinda Gates Foundation. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Triaxial bioprinting large-size vascularized constructs with nutrient channels

Bioprinting has demonstrated great advantages in tissue and organ regeneration. However, constructing large-scale tissue and organs in vitro is still a huge challenge due to the lack of some strategies for loading multiple types of cells precisely while maintaining nutrient channels. Here, a new 3D bioprinting strategy was proposed to construct large-scale vascularized tissue. A mixture of gelatin methacrylate (GelMA) and sodium alginate (Alg) was used as a bioink, serving as the outer and middle layers of a single filament in the triaxial printing process, and loaded with human bone marrow mesenchymal stem cells and human umbilical vein endothelial cells, respectively, while a calcium chloride (CaCl2) solution was used as the inner layer. The CaCl2 solution crosslinked with the middle layer bioink during the printing process to form and maintain hollow nutrient channels, then a stable large-scale construct was obtained through photopolymerization and ion crosslinking after printing. The feasibility of this strategy was verified by investigating the properties of the bioink and construct, and the biological performance of the vascularized construct. The results showed that a mixture of 5% (w/v) GelMA and 1% (w/v) Alg bioink could be printed at room temperature with good printability and perfusion capacity. Then, the construct with and without channels was fabricated and characterized, and the results revealed that the construct with channels had a similar degradation profile to that without channels, but lower compressive modulus and higher swelling rate. Biological investigation showed that the construct with channels was more favorable for cell survival, proliferation, diffusion, migration, and vascular network formation. In summary, it was demonstrated that constructing large-scale vascularized tissue by triaxial printing that can precisely encapsulate multiple types of cells and form nutrient channels simultaneously was feasible, and this technology could be used to prepare large-scale vascularized constructs.

Development of alginate-based film incorporated with anthocyanins of red cabbage and zinc oxide nanoparticles as freshness indicator for prawns

The objective of this study was to develop pH-sensitive film indicators for intelligent food packaging by incorporating red cabbage anthocyanins (RCA) and zinc oxide nanoparticles (ZnO NPs) into an alginate (Alg) film, aiming to mitigate the risk of foodborne illnesses. The films were fabricated using a solvent-casting method and crosslinked with a calcium chloride (CaCl2) solution. Thorough evaluations of the films' physical, mechanical, and structural properties demonstrated significant improvements in elastic modulus and UV/vis light barrier characteristics, reduced water vapor permeability (WVP), and moisture content attributed to integrating RCA and ZnO NPs. The resulting film displayed discernible color changes when exposed to various pH buffer solutions and ammonia vapor, indicating heightened sensitivity to pH fluctuations due to the presence of ZnO NPs. Visual assessment using prawns as test specimens revealed a color shift from violet (indicating satisfactory condition) to blue-greenish (indicating spoilage), corroborated by colorimetric analysis. Moreover, the Alg/ZnO/RCA film exhibited antioxidant and antibacterial properties, demonstrated biodegradation activity, and showed no toxic effects on RSC96 cells, further underscoring its potential as an effective freshness indicator for food products.

Examining the Hydration Behavior of Aqueous Calcium Chloride (CaCl2) Solution via Atomistic Simulations

Equilibrium molecular dynamics simulations are performed to examine the hydration behavior of Ca2+ and Cl− across a wide range of salt concentrations (from 1 wt.% to 60 wt.% CaCl2) in an aqueous solution. The predicted radial distribution functions (RDFs) and coordination numbers (CNs) of Ca2+–water, Cl−–water, and Ca2+–Cl− agree with the previous studies conducted at concentrations below the solubility limit at room temperature. The hydration limit of aqueous calcium chloride solution is identified at 10 wt.% CaCl2 as the CNs remain constant below it. Beyond the bulk solubility limit ~44.7 wt.% CaCl2, a noticeable decrease in the CN of Cl− and water is recorded, implying the saturation of the solution. The solvating water molecules decrease with increasing salt concentration, which can be attributed to the growth in the number of ion pairs.

Development and Evaluation of Cross-Linked Alginate-Chitosan-Abscisic Acid Blend Gel.

Abscisic acid (ABA) has been proposed to play a significant role in the ripening of nonclimacteric fruit, stomatal opening, and response to abiotic stresses in plants, which can adversely affect crop growth and productivity. The biological effects of ABA are dependent on its concentration and signal transduction pathways. However, due to its susceptibility to the environment, it is essential to find a suitable biotechnological approach to coat ABA for its application. One promising approach is to utilize alginate and chitosan, two natural polysaccharides known for their strong affinity for water and their ability to act as coating agents. In this study, an alginate-chitosan blend was employed to develop an ABA cover. To achieve this, an alginate-chitosan-abscisic acid (ALG-CS-ABA) blend was prepared by forming ionic bonds or complexes with calcium ions, or through dual cross-linking. This was done by dripping a homogeneous solution of alginate-chitosan and ABA into a calcium chloride solution, resulting in the formation of the blend. By combining the unique properties of alginate, chitosan, and ABA, the resulting ALG-CS-ABA blend can potentially offer enhanced stability, controlled release, and improved protection of ABA. These characteristics make it a promising biotechnological approach for various applications, including the targeted delivery of ABA in agricultural practices or in the development of innovative plant-based products. Further evaluation and characterization of the ALG-CS-ABA blend will provide valuable insights into its potential applications in the fields of biomedicine, agriculture, and tissue engineering.

Emergence of Power-Law Particle Size Distribution in Microfluidic Calcium Carbonate Precipitation: An Extended Yule Process with a Ripening Effect.

Precipitation of calcium carbonate in bulk solutions is well known to result in a bell-shaped or bimodal particle size distribution. However, it is unclear how the distribution behaves if precipitation occurs in a small, confined volume. In this Letter, we conduct microfluidic experiments where sodium carbonate and calcium chloride solutions are continuously injected into a microchannel to precipitate calcium carbonate particles. Results show that, regardless of the variations in reagent concentrations, mixing schemes, flow rates, and precipitation time, sizes of precipitated particles in the channel are power law distributed, with an exponent of 1.4. The data are described by an extended Yule process with the introduction of a ripening term. Since the Yule process is a general mechanism for power law generation, the extended Yule process proposed here provides a general model for systems where growth and ripening simultaneously present.

Mimicking animal adipose tissue using a hybrid network-based solid-emulsion gel with soy protein isolate, agar, and alginate

The growth of the plant-based meat analogue market has prompted the need for new approaches to replace animal adipose tissue. In this study, we developed a novel solid emulsion gel system using a hybrid gel network to simulate the thermal behavior and texture of adipose tissue. At various concentrations, emulsion gels were prepared using soybean oil, soybean protein isolate, agar, and alginate. After mixing all solutions at appropriate concentrations, cold-set gelation of agar was followed by ion-induced alginate gelation by immersion in calcium chloride solution. The resulting emulsion gel contained a dense network that trapped oil droplets and exhibited elastic behavior at all temperatures, similar to porcine adipose tissue. Furthermore, the optimized hybrid emulsion gel showed increased hardness compared with the single-agar emulsion gel. Overall, our findings suggested that the developed emulsion gel system could enhance the quality of plant-based meat analogues by mimicking the physical and thermal properties of animal adipose tissue.

How does calcium and sodium in saline water affect the potential of different rates and size of sugarcane biochar to soil water repellency remediation?

Soil water repellency (SWR) is a growing problem in arid and semiarid regions with non-saline water limitations. The objective of this study was to investigate the potential of different rates and sizes of sugarcane biochar on reducing soil water hydrophobicity when using saline and non-saline water. Eleven sugarcane biochar application rates from 0 to 10% with two different sizes including <0.25 and 0.25–1 mm were studied. The experiment was conducted in two strongly and extremely water-repellent soil. In addition, to investigate the effect of electrolyte concentration on biochar potential for SWR reduction, calcium chloride and sodium chloride electrolyte solutions with 5 concentrations including 0, 0.15, 0.3, 0.45 and 0.6 mol L−1 were considered. The results showed that both sizes of biochar reduced the soil water repellency. While in strongly repellent soil the 4% biochar was enough to change the soil from strongly repellent to hydrophilic, in extremely water-repellent soil, the use of 8% of fine biochar and 6% of coarse biochar changed the extremely water-repellent soil to slightly hydrophobic and strongly hydrophobic respectively. Increasing the electrolyte concentration caused the expansion of soil hydrophobicity and reduced the positive effect of biochar to water repellency management. Increasing the electrolyte concentration in sodium chloride solution has a greater effect on increasing hydrophobicity than in calcium chloride solution. In conclusion, biochar could suggest as a soil-wetting agent in these two hydrophobic soils. However, the salinity of water and the its dominant ion could increase the amount of biochar for soil repellency reduction.

Formation and stability of fibers obtained by cold gelation of pea protein isolate aggregates in a hydrodynamic spinning process

Using plant protein sources in the formulation of food products is an important option to reduce the overall carbon footprint of the human diet. However, the ability to convert plant proteins into functional ingredients can hamper their use by the food industry. Our objective was to assemble pea protein isolate (PPI) aggregates into edible fibers. They were obtained by co-injection of a suspension of PPI aggregates with a solution of calcium chloride by means of physico-chemical bonds, i.e. in the absence of thermal treatment or chemical reaction. As soon as specific hydrodynamic conditions were met, homogeneous fibers of few hundreds μm diameter and few cm length were obtained. Small angle X-ray scattering showed that the building blocks of these fibers were dense aggregates of 400 nm radius. Calcium and PPI concentrations required for the processing of fibers were roughly given by the sol-gel state diagram of PPI aggregates. Compared to their dairy protein based equivalent, PPI fibers were obtained with a low mass fraction of protein (3% w/w) and were stable regardless of the concentration of calcium chloride used. We concluded that the robustness of the hydrodynamic spinning process was attributed to the strong reactivity of PPI aggregates with calcium ions and the low tendency of PPI gels to syneresis.

Methyl orange dye sequestration using polyaniline nanotube-filled sodium alginate bio-composite microbeads

Dye pollution is a significant problem in today's environmental protection, and bio-adsorbents have been getting a lot of attention in wastewater treatment because of their affordability, sustainability, and eco-friendly. Bio-based sodium alginate-polyaniline nanotube (SA-PANINT) composite microbeads were fabricated by a facile cross-linking with calcium chloride solution for the adsorptive removal of methyl orange (MO) dye. Depending on the amount of PANINT in the composite composition, SA-PANINT-10, SA-PANINT-20, SA-PANINT-30, and SA-PANINT-40 composite microbeads were prepared. Among these microbeads, SA-PANINT-30 composite microbeads gave the best results in experimental tests. The obtained SA-PANINT-30 composite microbeads were characterized by FTIR-ATR, SEM, XRD, and BET analyses, and the influence of dose (0.01 g/50 mL–0.15 g/50 mL), pH (2–12), time (0–210 min), and concentration (10–50 mg/L) on the adsorption of MO was examined. From the Langmuir isotherm model, under optimum conditions, the qm value of SA-PANINT-30 microbeads was determined as 370.4 mg/g at 25 °C. It was also obtained that the adsorption process followed the PSO model and the adsorption process took place chemically. From the thermodynamic parameters, the adsorption process for SA-PANINT-30 microbeads was found to be endothermic (ΔHo=24.09 kJ/mol) and spontaneous (ΔGo=−25.56 kJ/mol at 25 °C). Considering all these results, it was seen that the prepared composite microbeads can be used as a highly efficient, low-cost, sustainable, and potential adsorbent in the removal of synthetic anionic dyes from wastewater.

Effect of Colchicum ritchii derived nanoparticles on apple ring rot disease without changing the taste through the prevented metabolites changes and increased sialic acid synthesis

ObjectiveIn this current study, we were planned to produce Colchicum ritchii flower extract associated calcium chloride nanoparticle (CRNPs) in a simple organic method and evaluate their apple fruit pathogenic resistance as well as their shelf-life extension properties during storage. MethodsThis CRNPs was confirmed by analysis of various established physical and chemical methods. Moreover, the apple fruit pathogenic resistance as well as their shelf-life extension properties during storage were evaluated by standardizing methods. ResultsOur results showed that the CRNPs were contrived and proven by physical and chemical evaluations. Moreover, the biological observation of CRNPs indicates that they have good pathogenic resistance properties against the Botryosphaeria dothidea without changing the taste through the prevented metabolites changes and increased sialic acid synthesis. The effective dose of pathogenic resistance in CRNPs was 40 µg/mL. The levels of defensive enzymes such as chitinase (CHI), polyphenol oxidase (PPO), phenylalanine ammonia lyase (PAL), and β-1,3-glucanase (GLU) significantly increased in CRNPs treated fruits. The increasing antioxidant enzymes activities including superoxide dismutase (SOD) and catalase (CAT) were shown in CRNPs treated apple fruit. The CRNPs-treated apple fruit was found to increase in salicylic acid (SA) and SA glucoside (SAG) levels, owing to increased SA synthesis. Moreover, shelf-life extension studies of fresh cut apples were conducted by immersing them in 20 and 40 µg/mL CRNPs as well as a standard solution of 2% calcium chloride for 5 mins. The standard solution of calcium chloride as well the CRNPs were proved as a good preservative and they were preserved the apple fruits shelf-life up to 25 days. ConclusionIn this study, we have concluded that the CRNPs were produced by fusion with Colchicum ritchii flower extract and calcium chloride in a superlative natural method. Our results recommended that the CRNPs is acted as a best source for apple ring rot resistance without changing the taste through the prevented metabolites changes and increased sialic acid synthesis as well as the antioxidant properties. In addition, the CRNPs is superlative source for apple shelf-life extension up to 25 days without any toxicology nature.

The Influence of Calcium Chloride on Soil Animals, Microorganisms, and Phytoplankton

Calcium chloride (CaCl2) is an inorganic chemical from the chloride group, and it is widely used as one of snow removal agents on the road in winter. Laboratory experiments were conducted to examine the effects of the road salt (NaCl) on soil organisms (a soil animal, microorganisms, and plankton). The soil dwelling Armadillidium vulgare died from high concentration of calcium chloride treatment. The longer the processing time of A. vulgare at the high concentration of calcium chloride, the lower the survival rate. A. vulgare died 16% at a calcium chloride concentration of 1 mM. In the disc diffusion test, the diameter of the inhibition zone proportionally increased with the concentration of calcium chloride. Micrococcus sp. was slightly higher in calcium chloride inhibitory activity than those of three soil microorganisms (Bacillus subtillus, Pseudomonas flurescens, and Xanthomonas maltophilia). About 90% of microorganisms (phytoplanktons) died when the 15.0 mM calcium chloride solution was used. In this study, high concentrations of calcium chloride affected the survival of soil animals, soil microorganisms, and water microorganisms. If chloride dissolves in water and flows into rivers or lakes, it can cause destruction of soil or aquatic ecosystems and threat to the survival of small organisms.

Encapsulation of activated carbon into calcium alginate microspheres toward granular-activated carbon adsorbents for elemental mercury capture from natural gas.

Activated carbon (AC) is an effective adsorbent for removing environmental pollutants. However, the traditional powder form of AC shows difficulty in handling during application which widely limits its utilization on the industrial scale. Herein, to avoid such limitation, traditional AC powder was encapsulated into calcium alginate (CA) microspheres. Calcium alginate/activated carbon (CAA) composite microspheres were prepared via cross-linking of sodium alginate/activated carbon composite solutions in a calcium chloride solution. Furthermore, in order to boost adsorption affinity of CAA composite microspheres toward elemental mercury (Hg°), ammonium iodide (NH4I)-treated calcium alginate/activated carbon (NCA) composite microspheres were obtained by a simple impregnation method using NH4I treatment. The morphological, structural, and textural properties of the microspheres were characterized and their Hg° adsorptive capacity was tested at different temperatures. Interestingly, the maximum adsorption capacity of NCA adsorbent composite microspheres was determined as 36,056.5μg/g at a flow rate of 250mL/min, temperature of 25°C, and 500μg/Nm3 of Hg° initial concentration. The Gibbs free energy (ΔG°) for NCA adsorbent composite microspheres varied from - 8.59 to - 10.54kJ/mol indicating a spontaneous adsorption process with an exothermic nature. The experimental Hg° breakthrough curve correlated well with Yoon‒Nelson and Thomas models. The breakthrough time (tb) and equilibrium time (te) were found to be 7.5days and 23days, respectively. Collectively, the findings of this work indicate a good feasibility of using NCA composite microspheres as potential adsorbents for removing Hg° from natural gas.

Evaluation of Different Extractants to Estimate Bioavailable Arsenic in Soil

ABSTRACT Owing to the similar chemistry of phosphorus (P) and arsenic (As), sodium bicarbonate (0.5 N NaHCO3) is commonly used to extract plant accessible As in soil. This extractant has neither been tested widely in relation to plant As, nor is this extractant compatible with inductively coupled plasma mass spectrometry (ICP-MS) due to the high concentration of dissolved solid. Subsequently, it is of utmost important to design a suitable chemical extraction method in order to estimate plant available As compatibility with ICP-MS. For this purpose, paired soil and plant samples were collected from paddy fields located in Nadia, West Bengal, India. Soil was extracted with 0.5 M NaHCO3, 0.1 N and 0.5 N phosphoric acid (H3PO4), 0.1 N and 0.5 N sulfuric acid (H2SO4), 0.1 N, 0.5 N, 1.0 N, 1.5 N HNO3 and 0.01 M calcium chloride (CaCl2) solution. Arsenic extracted with NaHCO3, H3PO4 and H2SO4 was determined in hydride generation-atomic absorption spectrophotometer (HG-AAS), while ICP-MS served to determine As extracted from soil with HNO3. Olsen-extractable As in soils ranged from 0.48 to 3.57 mg kg−1 with a mean value of 1.45 mg kg−1. The extractable As content in soil varied from 0.01 to 10.1 mg kg−1 across the extractants. In the case of grain As, 0.1 N H3PO4, 0.5 N NaHCO3 and 1.5 N HNO3 extractable As had distinctly higher correlation coefficients (r = 0.49**, r = 0.47**, r = 0.45**) when compared to other extractants. More or less similar relationships of extractable As were obtained with straw As content like that of rice grain. In view of rapidity of the soil test method for As, 1.5 N HNO3 can be recommended for assessing available As in soil.

Hydroxyapatite stabilized silver nanoparticles and their catalytic activity

Abstract In the current work, hydroxyapatite (HAP) particles were synthesized by using Citrus Limon. Pectin was extracted from lemon and was used as stabilizing agent during particles synthesis. HAP particles were synthesized by mixing calcium chloride solution and ammonium dihydrogen phosphate solution and pectin in a definite ratio. Fabrication of silver nanoparticles was carried out in the presence of HAP particles to obtain Ag-HAP hybrid particles. HAP and Ag-HAP systems were characterized using Fourier transform infrared spectroscopy (FTIR), UV–visible spectroscopy (UV–vis), scanning electron microscopy (SEM), X-ray diffraction (XRD) and Energy dispersive X-ray spectroscopy (EDX). The catalytic activity of Ag-HAP system was evaluated by reducing 4-nitrophenol and by degrading organic dyes (Rhodamine b, Brilliant blue and Methyl orange). The experimental results showed that Ag-HAP hybrid particles have good catalytic activity and may be used for catalysis of different organic reactions.

Optimal Preparation Protocol of Cell-Encapsulating Alginate Capsules for Long-Term Cell-Based Therapy

Cell-based therapy is an excellent therapeutic modality that involves cell transplantation into patients; however, given that most transplanted cells die immediately post-transplantation, the application of this strategy remains limited. Cell encapsulation is a promising technique for prolonging the survival of transplanted cells, although a definitive encapsulation protocol is yet to be established. Herein, we selected sodium alginate as a polymer for cell encapsulation and optimized the structure and function of cell-encapsulating alginate capsules. First, alginate capsules were prepared using various concentrations of sodium alginate and calcium chloride solution. The NanoLuc luciferase (Nluc)-expressing murine mesenchymal stem cell line C3H10T1/2 was used to prepare the alginate capsules, and cell survival was evaluated after transplantation into mice. The structural properties of the alginate capsules were dependent on the preparation conditions. Capsules with adequate hardness were obtained using 1% sodium alginate and 10% calcium chloride solutions. Alginate capsules encapsulating 5 × 103 C3H10T1/2/Nluc cells/10 μL maintained a constant cell number over time under in vitro culture conditions. After transplantation into mice, C3H10T1/2/Nluc cells encapsulated in alginate capsules exhibited a significantly longer survival (≥40 days) than suspended cells. Based on these findings, cell-encapsulating alginate capsules with optimal properties can be used for long-term cell-based therapies.

Treatment of Chitosan Modified-composite Polyamide Membranes by Sodium Hypochlorite for Enhancing Antifouling Property

To enhance the antifouling resistance, this study modified commercial thin-film composite polyamide membranes with chitosan by redox-initiated graft polymerization process before treating the modified membrane’s surface using sodium hypochlorite. The surfaces of the modified membranes were characterized using field emission scanning electron microscopy, and attenuated total reflection - Fourier transform infrared spectroscopy. The membranes’ performance and fouling resistance were shown through flux, retention, and maintained flux ratios during the filtration of calcium chloride solution. In addition, the antibacterial ability of membranes was evaluated through the growth of bacteria on their surface, and the anti-biofouling resistance of these membranes was evaluated through the maintained flux ratios, irreversible antifouling factor, and flux after 90-minute filtration of actual river water. The results showed that the modified membrane had superior antibacterial ability compared to the original membrane. Additionally, although the flux was reduced slightly, the modified membrane treated with NaClO exhibited a remarkable improvement in the maintained flux ratios (up to 99%) and the irreversible antifouling factor (~ 100%). This is similar to the filtration of actual river water, the chitosan-grafted polyamide membrane after NaClO treatment was shown to be more effective than the unmodified membrane in reducing the fouling phenomenon.

Cultivation of cells in alginate drops as a high-performance method of obtaining cell spheroids for bioprinting

Purpose of the study. Testing the protocol of obtaining cell spheroids of breast cancer cell cultures for bioprinting by growing in alginate drops.Materials and methods. Cells of breast cancer cell lines BT-20 and MDA-MB-453 were cultured in DMEM medium supplemented with 10 % FBS. Next, the cells were removed from the plastic using a trypsin-V ersene solution and resuspended in a sterile 2 % alginate solution in DPBS to the concentration of 105 cells/ml. Then the alginate solution with the cells was slowly dripped through a 30G needle into a sterile cooled solution of calcium chloride (100 mM) from a height of 10 cm. After polymerization, alginate drops were washed in DMEM and cultured for two weeks in DMEM with the addition of 10 % FBS at 37 °C and 5.0 % CO2.The spheroids formed in the alginate were photographed on the 3rd, 7th, 10th, and 14th days of cultivation, after which they were removed from the alginate by keeping in 55 mM sodium citrate solution with the addition of 20mM ethylenediaminetetraacetic acid (EDTA) and embedded in paraffin blocks according to the standard method, followed by histological examination.Results. Cellular spheroids were formed in both cell cultures already on the 3rd day of cultivation. From the 3rd to the 10th day in both cultures, a uniform growth of cell spheroids was observed with a gradual slowdown in the increase in the size of spheroids by the 14th day of cultivation. On the 10th day the proportion of cells that formed clones (more than 500 μm2 in size) was 25.2 % ± 7.1 % (n = 25) in the BT-20 culture and 38.5 % ± 9.9 % (n = 25) in MDA-MB-453 culture. On the 14th day, BT-20 culture was characterized by spheroids varying little in size and shape, with an average area of 1652 ± 175 µm2, having a dense structure with smooth edges. The spheroids in MDA-MB-453 culture turned out to be more loose and easily deformed, their size and shape varied noticeably, the average area of the spheroids was 2785 ± 345 µm2.Conclusion. The production of spheroids in alginate drops is inferior in speed to the methods of forming cell conglomerates in hanging drops or on microwells, but it surpasses these methods in productivity, which is comparable to the production of spheroids by constant medium stirring on low-adhesive substrates. In addition, the clonal nature of the obtained spheroids leads to an increase in research costs and thus limits their scalability.

Transformation of eggshell waste to egg white protein solution, calcium chloride dihydrate, and eggshell membrane powder

Abstract The present study investigated the possibility of complete utilization of eggshell waste (ESW) transforming it to adherent egg white protein solution, calcium chloride dihydrate, and eggshell membranes (ESM). Adherent egg white protein solution was obtained by washing ESW three times with distilled water at 25°C, followed by filtration, and analyzed for the protein content and lysozyme activity. ESM and calcium chloride were obtained simultaneously by the exposure of washed eggshells to 5% hydrochloric acid treatment at 25°C for 3 h, followed by separation by filtration. The separated ESM were washed, dried, and milled to powder and analyzed for protein and lipid content. The calcium chloride solution was exposed to the neutralization of excess hydrochloric acid by calcium hydroxide, followed by evaporation to one-tenth of volume. Calcium chloride crystals were precipitated from the concentrated solution with acetone, separated by filtration, dried at 110°C, and analyzed for chemical composition and purity. The obtained results revealed that 100 g of ESW can be transformed to 1.61 ± 0.34 g of adherent white proteins containing 485,821 U of lysozyme activity, 2.84 ± 0.16 g of ESM powder, and 108.74 ± 3.62 g of calcium chloride dihydrate of high purity.

Evaluation of osteogenic potential of demineralized dentin matrix hydrogel for bone formation

ObjectivesDentin, the bulk material of the tooth, resemble the bone’s chemical composition and is considered a valuable bone substitute. In the current study, we assessed the cytotoxicity and osteogenic potential of demineralized dentin matrix (DDM) in comparison to HA nanoparticles (n-HA) on bone marrow mesenchymal stem cells (BMMSCs) using a hydrogel formulation.Materials and methodsHuman extracted teeth were minced into particles and treated via chemical demineralization using ethylene diamine tetra-acetic acid solution (EDTA) to produce DDM particles. DDM and n-HA particles were added to the sodium alginate then, the combination was dripped into a 5% (w/v) calcium chloride solution to obtain DDM hydrogel (DDMH) or nano-hydroxyapatite hydrogel (NHH). The particles were evaluated by dynamic light scattering (DLS) and the hydrogels were evaluated via scanning electron microscope (SEM). BMMSCs were treated with different hydrogel concentrations (25%, 50%, 75% and neat/100%) and cell viability was evaluated using MTT assay after 72 h of culture. Collagen-I (COL-I) gene expression was studied with real-time quantitative polymerase chain reaction (RT-qPCR) after 3 weeks of culture and alkaline phosphatase (ALP) activity was assessed using enzyme-linked immune sorbent assay (ELISA) over 7th, 10th, 14th and 21st days of culture. BMMSCs seeded in a complete culture medium were used as controls. One-way ANOVA was utilized to measure the significant differences in the tested groups.ResultsDLS measurements revealed that DDM and n-HA particles had negative values of zeta potential. SEM micrographs showed a porous microstructure of the tested hydrogels. The viability results revealed that 100% concentrations of either DDMH or NHH were cytotoxic to BMMSCs after 72 h of culture. However, the cytotoxicity of 25% and 50% concentrations of DDMH were not statistically significant compared to the control group. RT-qPCR showed that COL-I gene expression was significantly upregulated in BMMSCs cultured with 50% DDMH compared to all other treated or control groups (P < 0.01). ELISA analysis revealed that ALP level was significantly increased in the groups treated with 50% DDMH compared to 50% NHH after 21 days in culture (P < 0.001).ConclusionThe injectable hydrogel containing demineralized dentin matrix was successfully formulated. DDMH has a porous structure and has been shown to provide a supporting matrix for the viability and differentiation of BMMSCs. A 50% concentration of DDMH was revealed to be not cytotoxic to BMMSCs and may have a great potential to promote bone formation ability.