Hank's Balanced Salt Research Articles

Enhanced bioactivity and interfacial marginal adaptation of mineral trioxide aggregate and Biodentine materials modified by eggshell powder at root dentine

Objectives: Mineral trioxide aggregate (MTA) as silicate-based cement material may provide better bioactivity than Biodentine material which used as a replacement to MTA in dental practice in different endodontic uses. Also the modifying MTA and Biodentine by adding eggshell powder (ESP) may improve their bioactivity and promote the formation of a reliable sealing through formation a thick and continuous interfacial layer at root dentine/material interface. Aim: This study was aimed to investigate marginal adaptability, the presence or absence of interfacial layer, and enhanced bioactivity's of MTA and Biodentine modified by addition of ESP, at the root dentine/material interface. Materials and methods: Thirteen root dentine segments of about 2 mm thickness were obtained from extracted human teeth and mechanically prepared by endodontic files to enlarge their canals. The specimens were then divided into six groups (n=5), each group were filled with unmodified MTA and Biodentine and modified with 0.5 and 1% ESP. The specimens were immersed in Hank's Balanced Salt Solution simulating body fluid for 30 days. The specimens were then, examined by scanning electron microscopy and energy disperse x-ray spectroscopy (EDX) to examine the presence or absence of interfacial layer and Ca/P ratio. Results: The interfacial layer can be seen in MTA and Biodentine groups that modified with 1% ESP. Also, MTA showed better marginal adaptability than Biodentine groups. There was a statistically significant difference in the Ca/P ratio among the all investigated groups. Conclusion: Addition of ESP to MTA and Biodentine can improve the mineral deposition at root dentine/material interface and also improve their sealing ability through the formation of interfacial layer.

Correction: Efficacy of Hank's balanced salt solution compared to other solutions in the preservation of the periodontal ligament. A systematic review and meta-analysis.

[This corrects the article DOI: 10.1371/journal.pone.0200467.].

Co-sputtered Antibacterial and Biocompatible Nanocomposite Titania-Zinc Oxide thin films on Si substrates for Dental Implant applications

ABSTRACTThe present work explores the possible applications of co-sputtered titania(Ti)-zinc(Zn)-oxide nanocomposite-(nC) thin films to improve the bonding of metallic fixture with bone. As a proof of principle study, the nC-titania-zinc-oxide thin films have been deposited on Si-(100) substrates by reactive magnetron co-sputtering at various substrate temperatures. In vitro, static immersion test in Hanks Balanced Salt Solution reveals the ability of the composite films to encourage the formation of bone-like apatite. Results of the cytocompatibility tests with MG-63 human osteoblast and L929 mouse fibroblast cell lines demonstrate superior adhesion and proliferation ability on the nC-titania-zinc-oxide coated substrates as compared to uncoated ones. The developed films material was observed to strongly inhibit the growth of both Staphylococcus aureus and Escherichia coli. The microstructural, morphological and surface properties of the films were studied, using various characterization techniques. Overall, our results favorably support the use of nanocomposite titania-zinc-oxide films material for the dental applications.

Effects of myogenic precursor cells (C2C12) transplantation and low-level laser therapy on muscle repair.

The aim of the present study was to evaluate the effects of myoblast inoculation in combination with photobiomodulation therapy (PBMT) on skeletal muscle tissue following injury. Sixty-five Wistar rats were divided into five groups: Control-animals not submitted to any procedure; Injury-cryoinjury of the tibialis anterior muscle; HBSS-animals submitted to cryoinjury and intramuscular Hank's Balanced Salt Solution; Injury + Cells-animals submitted to cryoinjury, followed by myogenic precursor cells (C2C12) transplantation; Injury + Cells + LLLT-animals submitted to cryoinjury, followed by myogenic precursor cells (C2C12) transplantation and PBMT (780 nm, 40 mW, 3.2 J in 8 points). The periods analyzed were 1, 3, and 7 days. The tibialis anterior muscle was harvest for histological analysis, collagen analysis, and immunolabeling of macrophages. No differences were found between the HBSS group and injury group. The Injury + Cells group exhibited an increase of inflammatory cells and immature fibers as well as a decrease in the number of macrophages on Day 1. The Injury + Cells + LLLT group exhibited a decrease in myonecrosis and inflammatory infiltrate at 7 days, but an increase in inflammatory infiltrate at 1 and 3 days as well as an increase in blood vessels at 3 and 7 days, an increase in macrophages at 3 days and better collagen organization at 7 days. Cell transplantation combined with PBMT led to an increase in the number of blood vessels, a reduction in myonecrosis and total inflammatory cells as well as better organization of collagen fibers during the skeletal muscle repair process. Lasers Surg. Med. © 2018 Wiley Periodicals, Inc.

Biophysical Characteristics of Human Neuroblastoma Cell in Oligomeric $\beta $ -Amyloid (1-40) Cytotoxicity.

Beta amyloid ( ) peptide, which is a common neuropathological hallmark deposit in the brain of patients with Alzheimer's disease, typically comprises 39-43 amino acid residues. peptides exist as isoforms of and with various lengths. In this research, atomic force microscopy (AFM) was applied to investigate aggregations in Hank's Balanced Salt Solution. Toxic effect of oligomer was investigated in live SH-SY5Y neuroblastoma cells by characterizing cell morphology and cell mechanics using high-resolution AFM scanning. oligomer-induced cytoskeleton reorganization was also observed under confocal microscopy, and it can account for reduction in Young's modulus of cells. Meanwhile, phosphorylation of tau increased after oligomer treatment, possibly resulting in microtubule disassembly. This paper demonstrates the linkage between cellular mechanical changes and neurodegeneration mediated by . The method used implies promising applications of real-time monitoring of cellular mechanical properties given the toxic effects of on living neuronal cells.

Significantly enhanced mechanical properties of ZrAlCo bulk amorphous alloy by microalloying with Ta

The alloy composition of Zr54Al17Co29 with high glass forming ability (GFA) in the Zr-Al-Co system was predicted by using the multiple binary eutectic clusters from the phase diagram. Then the microalloying effects of Ta on the microstructure, thermal properties, mechanical properties, and corrosion resistance of (Zr54Al17Co29)100-xTax (x = 0, 1, 1.5, 2, 2.5, 3) bulk amorphous alloy (BAA) were systematically investigated. The alloy composition with 1 at.% Ta alloy can be cast into BAA rods with diameters of 4 mm. The (Zr54Al17Co29)99Ta1 exhibited high yield stress of 2553 MPa, high fracture strength of 3170 MPa, plastic strain of 8.6%, and low Young's modulus of 111 GPa with better corrosion resistance in comparison with the ZrCuNiAl BAA, ZrCuAlAg BAA, and 316L stainless steel in the Hank's Balanced Salt solution. The Ni-, Cu-, Be-free (Zr54Al17Co29)99Ta1 bulk amorphous alloys with a critical diameter up to 4 mm, excellent mechanical properties, superior corrosion resistance are believed to be useful in orthopedic applications.

Properties of NeoMTA Plus and MTA Plus cements for endodontics.

To test a novel calcium silicate cement mixed with a water-based gel (NeoMTA Plus) with regard to chemical-physical properties and apatite-forming ability. NeoMTA Plus (Avalon Biomed Inc. Bradenton, FL, USA; lot. 2014090301) and a commercial MTA-based material with similar properties (MTA Plus, Prevest Denpro Limited, Jammu, India; lot. 41001) were tested for ion-releasing ability, initial and final setting times, radiopacity, open and impervious porosity and apparent porosity, water sorption, weight loss, solubility, ability to nucleate calcium phosphates (CaP) after immersion in HBSS (Hank's Balanced Salt Solution) by ESEM-EDX and micro-Raman spectroscopy. The results were analysed statistically with the anova test (P < 0.05). NeoMTA Plus had a prolonged setting time (315min) and a satisfactory radiopacity (3.76mm Al). Calcium and hydroxyl ion release was significantly greater and more prolonged in comparison with MTA Plus (P<0.05). Both NeoMTA Plus and MTA Plus had high values of open porosity and solubility. ESEM-EDX and micro-Raman confirmed the ability to nucleate calcium phosphates on their surface after immersion in HBSS. NeoMTA Plus is a new calcium silicate-based cement for root filling with an adequate radiopacity and prolonged setting time. The ion release and CaP-forming ability could increase stability of the root filling and promote endodontic and periodontal tissue regeneration, enhancing the bioactivity and biocompatibility of the material.

Human Norovirus and Its Surrogates Induce Plant Immune Response in Arabidopsis thaliana and Lactuca sativa.

Human norovirus is the leading cause of foodborne illness worldwide with the majority of outbreaks linked to fresh produce and leafy greens. It is essential that we thoroughly understand the type of relationship and interactions that take place between plants and human norovirus to better utilize control strategies to reduce transmission of norovirus in the field onto plants harvested for human consumption. In this study the expression of gene markers for the salicylic acid (SA) and jasmonic acid (JA) plant defense pathways was measured and compared in romaine lettuce (Lactuca sativa) and Arabidopsis thaliana Col-0 plants that were inoculated with Murine Norovirus-1, Tulane Virus, human norovirus GII.4, or Hank's Balanced Salt Solution (control). Genes involving both the SA and JA pathways were expressed in both romaine lettuce and A. thaliana for all three viruses, as well as controls. Studies, including gene expression of SA- and JA-deficient A. thaliana mutant lines, suggest that the JA pathway is more likely involved in the plant immune response to human norovirus. This research provides the first pieces of information regarding how foodborne viruses interact with plants in the preharvest environment.

Characterization on melt-derived bioactive glass powder from SiO2-CaO-Na2O-P2O5 system

In the present work melt-derived route was employed to fabricate new compositions of bioactive glasses (BG) from SiO2-CaO-Na2O-P2O5 system. Thermal analyses indicated all compositions can be melted at lower temperature than 1400°C. X-ray fluorescence (XRF) revealed that the amount of SiO2 and Na2O detected within the fabricated glass was significantly different compared to the initial weight measured during glass compositions preparation. X-ray diffraction (XRD) confirmed the amorphous glass structure for all glass compositions. Formation of silica network is established with the existence of functional group Si-O-Si (tetrahedral), confirmed with Fourier transform infrared spectroscope (FTIR) analyses. The bioactivity of all BG is evaluated by incubating the BGs in Hank's Balanced Salt Solution (HBSS) for 1h, 24h, 7 and 30days. The finding showed that all glass compositions remained in amorphous structure even after 30days of immersion with weak characteristic of carbonate group (C-O) and P-O band detected in FTIR analysis on carbonated hydroxyapatite (CHA) formation.

Nature's Benefaction as a Life Saver for an Avulsed Tooth: An In vitro Study.

Dentoalveolar trauma is a very common and avulsion is the most frequent injury. Treatment of choice following avulsion is immediate reimplantation. However, in cases where this might not be possible the prognosis of reimplanted teeth can be improved by the selection of an appropriate storage media. Prevention of ankylosis and replacement resorption depends more on the potential of storage media to maintain cell viability rather than the extraalveolar time period. To compare and evaluate the efficacy of different storage media in maintaining the viability of Periodontal Ligament (PDL) cells in an avulsed tooth. A total of 58 freshly extracted human premolar teeth were divided into four experimental groups and two control groups. The positive and negative controls corresponded to 0 minute and an eight hours dry time, respectively. The experimental teeth were stored dry for 30 minutes and then immersed in one of the four media: Hank's Balanced Salt Solution (HBSS), coconut water, aloe vera and saline for 45 minutes. The teeth were then treated with Collagenase Type II and Dispase for 30 minutes. The number of viable PDL cells were counted with a haemocytometer and analysed. The statistical analysis was done using SPSS version 15.0. The Analysis of Variance (ANOVA) and Post-hoc tests (Tukey-HSD) were performed to reveal the statistical significance. The values were represented in number (%) and mean±SD. Statistical analysis demonstrated that among the experimental groups maximum percentage of viable cells were seen in HBSS (87.33%) followed by coconut water (79.87%), aloe vera (70.59%) and saline (50.56%). Within the parameters of this study, it can be concluded that the HBSS is the most effective storage media in maintaining the viability of PDL cells. However, in Indian scenario where availability of HBSS is questionable, coconut water can be considered as the best natural storage media for an avulsed tooth followed by aloe vera.

Effect of glutamate on autophagy induced by energy stress in SH-SY5Y neurons

Introduction: Autophagy is a cytoprotective mechanism by which cells adapt to stress. In neurons, the question whether enhanced autophagy promotes cell death or cell survival is a subject of considerable debate, especially in excitotoxic conditions such as cerebral ischemia. Excitotoxicity is a pathological process that involves overstimulation of the excitatory receptors, primarily the ionotropic and metabotropic glutamate receptors. Material and methods: The viability of neuroblastoma SH-SY5Y cells was investigated by MTT assay for mitochondrial dehydrogenase activity. The appearance of autophagolysosomes and autophagic flux were determined by acridine orange staining (AO) and immunoblot, and the expression of autophagic genes was assessed by quantitative RT-PCR (Real time polymerase chain reaction). Results: Viability test showed a decrease in cell numbers, while AO test demonstrated a decrease in number of acidic vesicles in HBSS (Hank's Balanced Salt Solution) medium with glutamate, compared to HBSS medium alone. Accumulation of autophagosome-asociated LC3-II protein was observed by immunoblot in HBSS medium, while LC3-II levels were lower in HBSS medium with glutamate. Quantitative RT-PCR showed reduced expression of the autophagy genes in HBSS medium with glutamate, compared to the HBSS medium. Conclusion: The research confirmed that glutamate, during energy deprivation in vitro, leads to a decrease in autophagy in neurons, possibly preventing the induction of autophagy by reducing the transcription of autophagy genes.

Distribution of particles, small molecules and polymeric formulation excipients in the suprachoroidal space after microneedle injection

The purpose of this work was to determine the effect of injection volume, formulation composition, and time on circumferential spread of particles, small molecules, and polymeric formulation excipients in the suprachoroidal space (SCS) after microneedle injection into New Zealand White rabbit eyes ex vivo and in vivo. Microneedle injections of 25–150 μL Hank's Balanced Salt Solution (HBSS) containing 0.2 μm red-fluorescent particles and a model small molecule (fluorescein) were performed in rabbit eyes ex vivo, and visualized via flat mount. Particles with diameters of 0.02–2 μm were co-injected into SCS in vivo with fluorescein or a polymeric formulation excipient: fluorescein isothiocyanate (FITC)-labeled Discovisc or FITC-labeled carboxymethyl cellulose (CMC). Fluorescent fundus images were acquired over time to determine area of particle, fluorescein, and polymeric formulation excipient spread, as well as their co-localization. We found that fluorescein covered a significantly larger area than co-injected particles when suspended in HBSS, and that this difference was present from 3 min post-injection onwards. We further showed that there was no difference in initial area covered by FITC-Discovisc and particles; the transport time (i.e., the time until the FITC-Discovisc and particle area began dissociating) was 2 d. There was also no difference in initial area covered by FITC-CMC and particles; the transport time in FITC-CMC was 4 d. We also found that particle size (20 nm–2 μm) had no effect on spreading area when delivered in HBSS or Discovisc. We conclude that (i) the area of particle spread in SCS during injection generally increased with increasing injection volume, was unaffected by particle size, and was significantly less than the area of fluorescein spread, (ii) particles suspended in low-viscosity HBSS formulation were entrapped in the SCS after injection, whereas fluorescein was not and (iii) particles co-injected with viscous polymeric formulation excipients co-localized near the site of injection in the SCS, continued to co-localize while spreading over larger areas for 2–4 days, and then no longer co-localized as the polymeric formulation excipients were cleared within 1–3 weeks and the particles remained largely in place. These data suggest that particles encounter greater barriers to flow in SCS compared to molecules and that co-localization of particles and polymeric formulation excipients allows spreading over larger areas of the SCS until the particles and excipients dissociate.

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

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

In-vitro depth-dependent hyperthermia of human mammary gland adenocarcinoma

Nanoparticle mediated photothermal ablation of cancerous tissue shows promising results and applicability as a highly efficacious treatment method. As a majority of the photothermal work has been conducted with minimal attenuation of the laser before reaching the nanoparticles within surface seeded tumors in-vivo or through buffered media in-vitro, it is important to understand the effects of greater laser attenuation on photothermal efficacy mediated by changes in the scattering and absorption of the laser. Photothermal efficacy using a near infrared (NIR) 785nm laser irradiating polystyrene (PS) stabilized magnetite (Fe3O4) nanoparticles (PS-Fe3O4) is examined on MDA-MB-231 human mammary gland adenocarcinoma in-vitro. Agarose gel columns of various heights were created to simulate soft tissue and subsequently used for NIR laser attenuation. Polystyrene was found to significantly improve magnetite nanoparticle stability in serum containing media and modified Hank's Balanced Salt Solution and was able to induce significant hyperthermic ablation at mass concentrations which also did not elicit significant innate toxicity. Furthermore it was found that the polystyrene coating significantly reduced innate toxicity over 48h compared to uncoated magnetite. Agar gel layers provided similar optical attenuation in the NIR region to skin and prostate.

C-Rel is Required for the Induction of pTregs in the Eye but Not in the Gut Mucosa

ABSTRACTRegulatory T (Treg) cells play an integral role in maintaining immune homeostasis and preventing autoimmune diseases. Forkhead box P3 expression marks the commitment of progenitor cells to the Treg lineage. Although the essential function of the nuclear factor (NF)-κB family transcription factor c-Rel in the regulation of natural Treg cells has been firmly established, little is known about whether c-Rel is involved in the in vivo generation of peripheral Treg cells (pTregs), which develop from mature CD4+ conventional T cells outside of the thymus. We sought to answer this question through the induction of pTregs in the eye and gut mucosa using ovalbumin-specific T cell receptor transgenic mice that do or do not express c-Rel. Our results showed that Tregs can be induced in the eye in a c-Rel-dependent manner when immune-mediated inflammation occurs. However, c-Rel is dispensable for the induction of pTregs in the gut mucosa after oral antigen administration. Thus, c-Rel may play distinct roles in regulating the development of pTregs in different organs.Abbreviations ACAID: Anterior Chamber-Associated Immune Deviation; ATF: activating transcription factor; CREB: cAMP responsive element-binding protein; DMEM: Dulbecco minimum essential medium; HBSS: Hanks Balanced Salt Solution; NFAT: Nuclear Factor of Activated T cells; PBS: Phosphate-buffered saline; PE: Phycoerythrin; WT: wild type.

Nutrient Starvation Decreases Cx43 Levels and Limits Intercellular Communication in Primary Bovine Corneal Endothelial Cells.

Connexin (Cx) proteins form large conductance channels which function as regulators of communication between neighboring cells via gap junctions and/or hemichannels. Intercellular communication is essential to coordinate cellular responses in tissues and organs, thereby fulfilling an essential role in the spreading of signaling, survival and death processes. Connexin 43 (Cx43), a major connexin isoform in brain and heart, is rapidly turned over. Recent studies implicated that autophagy, a lysosomal degradation pathway induced upon nutrient starvation, mediates connexins, including Cx43, degradation. Here, we examined the impact of nutrient starvation on endogenous Cx43-protein levels and endogenous Cx43-driven intercellular communication in primary bovine corneal endothelial cells (BCECs). Hank's Balanced Salt Solution (HBSS) was used as a starvation condition that induces autophagic flux without impacting the survival of the BCECs. Nutrient starvation of BCECs caused a rapid decline in Cx43-protein levels, both as gap junctions and as hemichannels. The time course of the decline in Cx43-protein levels coincided with the time course of the decline in intercellular communication, assessed as intercellular Ca(2+)-wave propagation in BCECs exposed to a single-cell mechanical stimulus. The decline in Cx43-protein levels, both as gap junctions and as hemichannels, could be prevented by the addition of bafilomycin A1, a lysosomal inhibitor, during the complete nutrient starvation period. Consistent with this, bafilomycin A1 significantly alleviated the decrease in intercellular Ca(2+)-wave propagation. This study further underpins the importance of autophagy as an important degradation pathway for Cx43 proteins during periods of nutrient deprivation, thereby impacting the ability of cells to perform intercellular communication.

Corrosion behavior, biocompatibility and biomechanical stability of a prototype magnesium-based biodegradable intramedullary nailing system

Implants made of degradable magnesium alloys are a potential alternative to conventional orthopaedic implant materials, e.g. stainless steel or titanium. Intramedullary nails made of the magnesium alloy LAE442 were subjected to cyclic fatigue tests in both distilled water and Hank's Balanced Salt Solution (HBSS) at 37.5°C until implant failure or a limit of 500,000cycles was reached. In distilled water, four of the five nails were still intact after the end of the biomechanical test. In HBSS, a breakage within the first 70,000 bending cycles was observed. Additionally, the degradation rate of this alloy was determined in HBSS according to the weight loss method (0.24±0.12mmyear−1) and based on gas release (0.21±0.03mmyear−1) with a standard eudiometer. A cytotoxicity test with L929 cells was carried out in accordance with EN ISO 10993-5/12. This test demonstrated sufficient cell viability of the diluted extracts (50%, 25% and 12.5%). The relative metabolic activity of the 100% extract was reduced slightly below 70%, which is classified as a threshold value for cytotoxicity. In conclusion, this in vitro study indicates that intramedullary nails made of LAE442 may not have the required fatigue resistance for load-bearing applications and the development of a corrosion-protective coating may be necessary to prevent early failure of the implant.

Pre-Treatment and Organic Coating of Al Free Mg Alloy for Controlling Degradation Rates

Biodegradable implant material for medical applications has to fulfill specific therapeutic tasks. For our investigations, synthetic polymers, polyurethanes (PUR) and polyetherimide (PEI), were used. Both systems have been used to coat an Al-free Mg alloy. Characterization of materials was performed by IR, spark spectral analysis, microscopy and EIS. Electrochemical investigations of the different treated samples in aqueous NaCl and Hank's Balanced Salt Solution indicate specific response of the polymer/substrate system to corrosion attack.

The effect of Oral Rehydration Solution on apoptosis of periodontal ligament cells.

Over the years, numerous studies have been conducted to find a storage medium with favorable properties to preserve the avulsed teeth. Oral Rehydration Solution (ORS) is a simple oral solution containing glucose and electrolytes. In addition to physiologic PH and osmolality, it has other important properties such as availability and inexpensiveness, storing potential in different conditions and places, and long lifetime. This study was carried out to more precisely investigate the storage of cells in this substance in different concentrations and in longer times, and to evaluate the programmed cell death or apoptosis as well as cell viability. The cultured PDL cells of this experimental study were exposed to three concentrations of 25%, 50%, and 100% ORS in comparison with Hanks Balanced Salt Solution (HBSS) and tap water. They were then analyzed by Tetrazolium Salt-based Colorimetric (MTT) Assay in 2-, 6-, 12-, 24-, and 48-h periods and by Apoptosis Assay in 12-, 24-, and 48-h time periods. Data were analyzed by spss software using anova and Post hoc (Duncan) tests. P ≤ 0.05 was considered significant. Overall, the maximum viable cells and minimum apoptotic cells were reported after the cells' exposure to ORS (100%). Also, the minimum level of viable cells was observed in ORS (25%) group and the highest level of apoptotic cells was observed in 25% and 50% ORS groups. ORS preserved more viable cells and induced fewer apoptotic cells in comparison with HBSS.

Hanks Balanced Salt Solution–HEPES

Hanks Balanced Salt Solution–HEPES