Eagle's Medium Research Articles

Establishment and characterization of a liver cell line, ALL, derived from yellowfin sea bream, Acanthopagrus latus, and its application to fish virology.

Yellowfin sea bream (Acanthopagrus latus) is an important economic fish, which is seriously threatened by various fish viruses. In this study, a cell line designated as ALL derived from the liver of yellowfin sea bream was developed and characterized. The cell line grew well in Dulbecco's modified Eagle's medium containing 10%-20% foetal bovine serum at 28°C. Amplification of the cytochrome B gene indicated that ALL cells originated from yellowfin sea bream. The modal chromosome number of ALL cells was 48. ALL cells were efficiently transfected with pEGFP-N3 plasmids, indicating the potential application of ALL cells in exogenous gene manipulation studies. ALL cells were susceptive to three main fish viruses, including viral haemorrhagic septicaemia virus (VHSV), red-spotted grouper nervous necrosis virus (RGNNV) and largemouth bass virus (LMBV). The replication of VHSV, RGNNV and LMBV in ALL cells was confirmed by quantitative real-time polymerase chain reaction, virus titre and transmission electron microscopy assays. Moreover, ALL cells could respond to VHSV, RGNNV and LMBV infections, as indicated by the differential expression of antiviral genes involving in the innate immune response. In conclusion, the newly established ALL cell line will be an excellent in vitro platform for the study of the virus-yellowfin sea bream interaction.

Glucose-responsive oral insulin delivery platform for one treatment a day in diabetes

Glucose-responsive oral insulin delivery platform for one treatment a day in diabetes

Retinoic acid and taurine enhance differentiation of the human bone marrow stem cells into cone photoreceptor cells and retinal ganglion cells

Degeneration and apoptotic death of the photoreceptor cell-layer of retina are a major cause of irreversible blindness in the development era. The stem cell replacement therapy is one of the strategies for the retinal repairing. In addition, exogenous signals critically contribute to the direction of lineage decisions that causes the fate-restricted photoreceptor progenitors from stem cell progeny in culture. It has been found that epidermal growth factor (EGF), taurine, and retinoic acid (RA) initially act in the instructive as well as lineage-restricted way in the progenitor lineage for producing neuroretinal cells or photoreceptor like cells from stem cell. The study aims to investigate the effect of RA and taurine in differentiation of the human bone marrow stem cell into cone photoreceptors cells and retinal ganglion cells. Mesenchymal stem cell was derived from human bone marrow of the term delivery. Therefore, the cultured cells have been treated with Dulbecco's modified Eagle's medium (DMEM)/high glucose (H+ ). After the four-cell passage, basal medium was replaced with DMEM/F12 complemented with 50 μmol/L taurine, RA (1 µM) and EGF (1 µg/ml). Subsequently cellular change morphology was detected following 7 and 14 days. Then, gene expression of neuroretinal and photoreceptor cell biomarkers (CRX, OTX2, PKC-α, recoverin, and Rho) were examined by quantitative polymerase chain reaction (Q-PCR). Also, cells were cultured, fixed, and then immunocytochemical analyzed. Primary antibodies included CRX and Rho. Cellular morphology demonstrated spindle elongated morphology. Taurine alone and combination of RA upregulate neuroretinal and photoreceptor cell biomarkers in messenger RNA and protein levels but along with EGF have not significant effect. Our data showed that taurine combination with RA can differentiate bone marrow mesenchymal stem cells into neuroretinal or photoreceptor like cells in vitro that can offer an attractive treatment ground for transplantation in the cell-replacement therapy for some forms of the retinal degeneration.

Implications of the Coffee-Ring Effect on Virus Infectivity.

The factors contributing to the survival of enveloped viruses (e.g., influenza and SARS-CoV-2) on fomite surfaces are of societal interest. The bacteriophage Phi6 is an enveloped viral surrogate commonly used to study viability. To investigate how viability changes during the evaporation of droplets on polypropylene, we conducted experiments using a fixed initial Phi6 concentration while systematically varying the culture concentration and composition (by amendment with 2% fetal bovine serum (FBS), 0.08 wt % BSA, or 0.5 wt % SDS). The results were consistent with the well-founded relative humidity (RH) effect on virus viability; however, the measured viability change was greater than that previously reported for droplets containing either inorganic salts or proteins alone, and the protein effects diverged in 1× Dulbecco's modified Eagle's medium (DMEM). We attribute this discrepancy to changes in virus distribution during droplet evaporation that arise due to the variable solute drying patterns (i.e., the "coffee-ring" effect) that are a function of the droplet biochemical composition. To test this hypothesis, we used surface-enhanced Raman spectroscopy (SERS) imaging and three types of gold nanoparticles (pH nanoprobe, positively charged (AuNPs(+)), and negatively charged (AuNPs(-))) as physical surrogates for Phi6 and determined that lower DMEM concentrations, as well as lower protein concentrations, suppressed the coffee-ring effect. This result was observed irrespective of particle surface charge. The trends in the coffee-ring effect correlate well with the measured changes in virus infectivity. The correlation suggests that conditions resulting in more concentrated coffee rings provide protective effects against inactivation when viruses and proteins aggregate.

Neutralization antibody titers against SARS-CoV-2 in an infant born to a mother with COVID-19

Neutralization antibody titers against SARS-CoV-2 in an infant born to a mother with COVID-19

Abstract P171: Glucose Consumption Of Vascular Cell Types In Culture; Toward Optimization Of Experimental Conditions

Any experimental outcomes are potentially influenced by extracellular glucose availability and its cellular metabolism. However, there is a lack of attention paid to the supply and utilization of glucose in many cultured experiments. Surveillance of vascular related journals for the past 5 years demonstrated that less than 20% of published articles declared the medium glucose concentration. The present studies were designed to seek ideal medium glucose concentration(s) in various cell types with particular attention paid to changes in glucose consumption. By using distinct glucose concentrations in Dulbecco's Modified Eagle's medium (DMEM), we identified ideal glucose media for stimulation experiments. We have compared glucose consumption in three vascular cell types, endothelial cells (EC), vascular smooth muscle cells (VSMC) and adventitial fibroblasts (AF) with or without angiotensin II (AII) stimulation. In all cell types after a 48h incubation in relatively low glucose media (1 g/L), medium glucose concentration was reduced to almost 0 (EC 0.01±0.01 g/L, VSMC 0.13±0.05 g/L, AF 0.02±0.01 g/L). Whereas medium glucose concentration remained significantly higher (EC 2.77±0.05 g/L, VSMC 3.87±0.05 g/L, AF 3.32±0.01 g/L) when cells were incubated for 48h in high glucose (4.5 g/L) media. In middle glucose (2.75 g/L) media, medium glucose concentration remained in physiological ranges (EC 0.62±0.18 g/L, VSMC 1.98±0.07 g/L, AF 1.17±0.17 g/L). AII treatment enhanced glucose consumption in AF and low passage VSMC but not in EC. In AF, AII induction of target proteins varied depending on the glucose concentration used. In low glucose media induction of Grp78 or hexokinase II was highest, whereas induction of VCAM1 was lowest. Utilization of specific inhibitors further suggest essential roles of AT1 receptor and glycolysis in AII-induced fibroblast activation. Overall, the present study demonstrates a high risk of hypo- or hyperglycemic conditions when standard low or high glucose media is used with vascular cells. Moreover, these conditions may significantly alter experimental outcomes. Medium glucose concentration should be monitored during any culture experiments and utilization of middle glucose media is recommended for all vascular cell types.

The intermediate-conductance calcium-activated potassium channel KCa3.1 contributes to alkalinization-induced vascular calcification in vitro.

ObjectiveIn order to find new strategies for the prevention of vascular calcification in uremic individuals especially treated by dialysis and develop novel therapeutic targets in vascular calcification, we explore the role of KCa3.1 in alkalinization‐induced VSMCs calcification in vitro.MethodRat VSMCs calcification model was established by beta‐glycerophosphate (β‐GP, 10 mM) induction. The pH of Dulbecco's modified Eagle's medium (DMEM) was adjusted every 24 h with 10 mM HCl or 10 mM NaHCO3. The mineralization was measured by Alizarin Red staining and O‐cresolphthalein complex one method. mRNA and protein expression were detected by RT‐PCR and Western blot or immunofluorescence. Ca2+ influx was measured by Elisa.ResultThe results indicated that alkalization induced an increase in Ca2+ influx to enhance VSMCs calcification. Furthermore, the increase of calcification was associated with the expression of KCa3.1 via advanced expression of osteoblastic differentiation markers alkaline phosphatase (ALP) and Runt‐related transcription factor 2 (Runx2). Blocking KCa3.1 with TRAM‐34 or shRNA vector can significantly lowered the effects of calcification in the activity of ALP and Runx2 expression.ConclusionTogether all, our studies suggested that alkalinization can promote vascular calcification by upregulating KCa3.1 channel and enhancing osteogenic/chondrogenic differentiation by upregulating Runx2. The specific inhibitor TRAM‐34 and KCa3.1‐shRNA ameliorated VSMCs calcification by downregulating KCa3.1.

Hypaconitine confers protection on ketamine-induced neuronal injury in neonatal rat brain via a mechanism involving PI3K/Akt/Bcl-2 pathway

Purpose: To investigate the neuroprotective effect of hypaconitine against ketamine-induced neuronal injury in the brains of neonatal rats, and the underlying mechanism of action.
 Methods: Seven day-old Sprague-Dawley pups weighing 15.0 to 20.0 g (mean weight = 17.5 ± 2.5 g), and aged 7 days were used for this study. The pups were sacrificed, and their forebrains isolated and used to prepare cell suspensions. The isolated cells were treated with ketamine (100 µM) or varied concentrations of hypaconitine (0.1 – 2 µM) or LY294002 (10 µM). The cells were trypsinized and cultured at 37 °C in 10 % fetal bovine serum (FBS) supplemented Dulbecco's modified Eagle's medium (DMEM) in a humidified incubator containing 5 % CO2. Cell viability was determined using MTT assay, while TUNEL assay was used to determine the extent of apoptosis in the cells. The expressions of pAkt, Bcl-2 and caspase-3 were determined using Western blotting.
 Results: There were only few viable cells in the ketamine-treated group, and cell viability was significantly and dose-dependently increased in hypaconitine-treated groups (p < 0.05). The extent of apoptosis was significantly higher in ketamine-treated cells than in control cells, but treatment with hypaconitine significantly reduced the number of apoptotic cells (p < 0.05). However, in the presence of LY294002 (a PI3K-specific inhibitor), the effect of hypaconitine on neuronal cell apoptosis was significantly reversed (p < 0.05). The expressions of p-Akt and Bcl-2 were significantly down-regulated while the expression of caspase-3 was significantly upregulated in ketamine-treated neuronal cells, when compared with control group (p < 0.05). However, in cells treated with hypaconitine, the expressions of p-Akt and Bcl-2 were significantly upregulated, while the expression of caspase-3 was significantly down-regulated (p < 0.05). Treatment of neuronal cells with hypaconitine in the presence of LY294002 significantly reversed the effect of hypaconitine on the expressions of p-Akt, Bcl-2 and caspase-3 (p < 0.05).
 Conclusion: These results suggest that hypaconitine ameliorates ketamine-induced neuronal injury in neonatal rats via a mechanism involving the PI3K/Akt/Bcl-2 pathway.

Trichlorophenyl-benzoxime induces apoptosis in human colon carcinoma cells via activation of mitochondria dependent pathway

Purpose: To determine the apoptotic effect of trichlorophenyl-benzoxime (TCPB) on colorectal cancer (CRC) cells, and to elucidate the mechanism of action.
 Methods: Colon carcinoma cell lines (DLD-1 and HT-29) were used in this study. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin at 37 ˚C in an atmosphere of 5 % CO2 and 95 % air. When the cells attained 60 - 70 % confluency, they were treated with serum-free medium and graded concentrations of TCPB (1.0 – 6.0 μM) for 24 h. Cell viability and apoptosis were assessed using 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) and flow cytometric assays, respectively. Western blotting and 2', 7' dichlorofluorescein diacetate (DCFH DA) assays were used for the determination of expression levels of apoptotic proteins, and levels of reactive oxygen species (ROS), respectively.
 Results: Treatment of DLD-1 and HT-29 cells with TCPB led to significant and dose-dependent reductions in their viability, as well as significant and dose-dependent increases in the number of apoptotic cells (p < 0.05). Treatment of HT-29 cells with TCPB led to significant increases in the population of cells in the G0/G1 phase, but significant reduction of cell proportion in S and G2/M phases (p < 0.05). It also significantly and dose-dependently upregulated the expressions of caspase-3 and bax, down-regulation of the expression of bcl-2 (p < 0.05). TCPB treatment upregulated the expressions of p53, cytochrome c (cyt c), procaspase-3, and procaspase-9, but down-regulated the expression of pAkt dose-dependently (p < 0.05). The expression of Akt in HT-29 cells was not significantly affected by TCPB (p > 0.05). However, TCPB significantly enhanced the cleavage of PARP1, and significantly and dose-dependently increased the levels of ROS in HT-29 cells (p < 0.05).
 Conclusion: These results suggest that TCPB exerts apoptotic effect on CRC cells via activation of mitochondria-dependent pathway, and thus can be suitably developed for the management of colon cancer.

Inhibitory effects of tamoxifen and tanshinone, alone or in combination, on the proliferation of breast cancer cells via activation of p38 MAPK signalling pathway

Purpose: To investigate the effects of tamoxifen and tanshinone administered individually or in combination, on the proliferation of breast cancer (BC) cells, and the underlying mechanism(s) of action.
 Methods: Human breast cancer cell lines (SNU-306, SNU-334 and SNU-1528), and normal primary mammary epithelial cell line (HMEC) were cultured at 37 °C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 5 % fetal bovine serum (FBS), l glutamine (2 mM), penicillin (100 U/ml) and streptomycin (100 μg/ml) in a humidified incubator containing 5 % CO2. Cell proliferation was determined using MTT assay, while real-time quantitative polymerase chain reaction (qRT-PCR) was used to determine the expressions of apoptosis-related genes. The expressions of p38 mitogenactivated protein kinases (p38 MAPK) were determined by Western blotting.
 Results: There were only few viable cells in tamoxifen- and tanshinone-treated wells, and cell viability was concentration-dependently reduced. Treatment of SNU-306 cells with tamoxifen (30 µM) or tanshinone (20 µM) alone significantly reduced the expression of Wip1 after 72 h of incubation, and the level of expression was significantly reduced in SNU-306 cells treated with combination of tamoxifen and tanshinones, relative to those treated with tamoxifen or tanshinone alone (p < 0.05). The extent of apoptosis was significantly higher in SNU-306 cells treated with tamoxifen or tanshinone alone or in combination than in control cells (p < 0.05). Expressions of Bax, caspase 3 and p53 were significantly higher in SNU-306 cells than in control cells, and were significantly higher in SNU-306 cells treated with combination of tamoxifen and tanshinone than in those treated with tamoxifen or tanshinone alone (p < 0.05). The level of expression of MAPK was significantly higher in SNU-306 cells treated with tamoxifen or tanshinone alone, and in combination treatment, than in control cells (p < 0.05).
 Conclusion: Tamoxifen and tanshinone administered alone or in combination promote apoptosis in BC cells via mechanisms involving the up-regulation and phosphorylation of MAPK.

Bioactivity and corrosion behavior of magnesium barrier membranes

AbstractIn the current research, magnesium and its alloys have been intensively studied as resorbable implant materials. Magnesium materials combine their good mechanical properties with bioactivity, which make them interesting for guided bone regeneration and for the application as barrier membranes. In this study, the in vitro degradation behavior of thin magnesium films was investigated in cell medium and simulated body fluid. Three methods were applied to evaluate corrosion rates: measurements of (i) the gaseous volume evolved during immersion, (ii) volume change after immersion, and (iii) polarization curves. In this comparison, measurements of H2 development in Dulbecco's modified Eagle's medium showed to be the most appropriate method, exhibiting a corrosion rate of 0.5 mm·year−1. Observed oxide and carbon contamination have a high impact on controlled degradation, suggesting that surface treatment of thin foils is necessary. The bioactivity test showed positive results; more detailed tests in this area are of interest.

Comparison of the in vitro corrosion behavior of biodegradable pure Zn in SBF, 0.9% NaCl, and DMEM

AbstractZn‐based alloys are considered to be the new biodegradable implant materials due to their suitable degradation rate and good biocompatibility. The biocorrosion behavior of pure Zn in 0.9% NaCl, simulated body fluid, and Dulbecco's modified Eagle's medium (DMEM) was investigated by electrochemical and immersion tests. These tests revealed that pure Zn has the lowest corrosion rate in DMEM and the highest in 0.9% NaCl. Aggressive Cl− has an important effect on the corrosion process. Buffering agents, amino acids, and glucose have a close connection with corrosion resistance. Among the three solutions, DMEM with a similar ion concentration and necessary nutriments is recommended as the more suitable choice for estimating biodegradable alloys' in vitro degradation.

Effect of CO 2 on the Expression of cyclooxigenase in Vascular Smooth Muscle Cells

Objectives Carbon dioxide(CO2) is one of the factors that affect the constriction and dilatation of smooth muscle cells. Cyclooxigenase(COX) catalyzes have important roles in inducing inflammatory responses and prostanoids, generated from arachidonic acid by COX-1 and COX-2, are involved in modulating of vascular tone. This study aimed to clarify the role of CO2 in regulating COX expression in vascular smooth muscle cells and elucidate its effects for vascular endothelial function. Methods A7r5 rat smooth muscle cells were maintained in Dulbecco's modified Eagle's medium, supplemented with 10% fetal bovine serum, at 37ºC in a 5% CO2 incubator. The cells were exposed to 10% and 20% CO2 for 24 h. The expression of COX-1 and COX-2 was subsequently evaluated by real time PCR, and cell viability was analyzed following exposure to 10% and 20% CO2. Results The expression levels of COX-2 were significantly reduced by concentration dependence in the vascular smooth muscle cells following exposure to 10% and 20% CO2, while no significant difference was found in those of COX-1 before and after exposure. Cell viability also decreased on the depending on the concentration of CO2. Conclusions These results suggested that exposure to CO2 may suppress the vasodilatation via the downregulation of COX-2 expression in vascular smooth muscle cells.

Evaluating the morphology of the degradation layer of pure magnesium via 3D imaging at resolutions below 40 nm

Magnesium is attractive for the application as a temporary bone implant due to its inherent biodegradability, non-toxicity and suitable mechanical properties. The degradation process of magnesium in physiological environments is complex and is thought to be a diffusion-limited transport problem. We use a multi-scale imaging approach using micro computed tomography and transmission X-ray microscopy (TXM) at resolutions below 40 nm. Thus, we are able to evaluate the nanoporosity of the degradation layer and infer its impact on the degradation process of pure magnesium in two physiological solutions. Magnesium samples were degraded in simulated body fluid (SBF) or Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS) for one to four weeks. TXM reveals the three-dimensional interconnected pore network within the degradation layer for both solutions. The pore network morphology and degradation layer composition are similar for all samples. By contrast, the degradation layer thickness in samples degraded in SBF was significantly higher and more inhomogeneous than in DMEM+10%FBS. Distinct features could be observed within the degradation layer of samples degraded in SBF, suggesting the formation of microgalvanic cells, which are not present in samples degraded in DMEM+10%FBS. The results suggest that the nanoporosity of the degradation layer and the resulting ion diffusion processes therein have a limited influence on the overall degradation process. This indicates that the influence of organic components on the dampening of the degradation rate by the suppression of microgalvanic degradation is much greater in the present study.

Authenticating the Anti-cancer Properties of <i>Couroupita guianensis</i> in Western Ghats using HL60 Humanleukemia Cell Line

The target of this work is to evaluate the <em>in–vitro</em> anticancer activity and cellular toxicity of <em>Couroupita guianensis</em> (CG) towards the Human promyelocytic leukemia HL60 cells contrasted with the normal cell lines. The anti-cancer properties of<em> Couroupita guianensis</em> extracted with Chloroform and DMSO (Dimethyl sulfoxide) were analysed and studied throughout this paper. HL60 cell lines were permitted to develop in DMEM (Dulbecco's Modified Eagle's medium) and incubated with differed concentration of DMSO CG extract. MTT assay drew out the best approach to decide the cell feasibility and assessment was done with the optical absorbance at 570nm and 620nm as reference. The floral extract inhibited 50% growth (IC<sub>50</sub>) of HL60 cell lines at 10 mg/ml of extract concentration. Inhibitory adequacy of CG showed the cell – plausibility in time and dosage relied route with predictable morphological changes. Flow cytometer assessed the apoptosis insisting the cell toxicity esteem for MTT at IC<sub>50</sub> with 70 % cell suitability. Morphological investigation likewise plainly expresses that no apoptosis has been found in charge and comparably in CG treated when contrasted with destructive HL60 cell – line. Assessment of cell toxicity impact of CG demonstrated it can be an inert source of great transformation in human Acute promyelocytic leukemia indicating (revealing) that chemotherapeutic agent.

The role of NMT induction on odontogenic proliferation and differentiation of dental pulp stem cells

This study was conducted to investigate the odontogenic proliferation and differentiation of dental pulp stem cells (DPSCs) after induction by nanoparticle mineral trioxide (NMT). DPSCs were isolated from permanent teeth and placed in tubes containing Dulbecco's modified Eagle's medium, followed by immunocytochemistry analysis. The viability of DPSCs exposed to NMT was measured using MTT assay with trypan blue dye exclusion. Alkaline phosphatase (ALP) activity was evaluated using ALP colorimetric reactions by reacting NMT supernatants with fluorescent-specific ALP substrates. The concentration of osteocalcin was determined using an instant human osteocalcin enzyme-linked immunosorbent assay (ELISA) kit. A human dentin sialophosphoprotein (DSPP) ELISA kit coated with anti-human DSPP antibody was employed to measure DSPP levels. There was a significant difference between ALP activity after exposing the cells to NMT and trioxide mineral aggregate on days 3, 7, and 21. Osteocalcin activity showed a significant difference on days 3, 7, 14, and 21. There was a significant difference in DSPP levels on days 7 and 21. DPSCs exposed to NMT and to trioxide mineral aggregate showed extracellular matrix formation on day 7 and 14, respectively. Furthermore, NMT may effectively increase the proliferation and differentiation of DPSCs as well as their maturation toward odontoblasts.

Apoptotic and Anti-inflammatory Effects of Hypericum Perforatum Extract in Human Basal Cell Carcinoma TE 354.T Cell Line

Objectives: Basal cell carcinoma is the most common malignancy in in the Caucasian race and is a serious health problem with its ever-increasing incidence and high cost. Although the treatment is surgical, some elderly patients do not accept surgical intervention. In this case, non-surgical treatments come into prominence. The Hypericum perforatum (HP) / St. John's Wort plant has been used in the treatment of many diseases for centuries among local people. Here, we aimed to aimed to elucidate the effect and molecular mechanism of the HP plant on BCC cells.
 Methods: Human BCC cell lines TE 354.T were acquired from ATCC®, then were cultured in Dulbecco's Modified Eagle's Medium. BCC cells were seeded at a concentration of 2x106cells in each flask (n=12). After incubation, while six of the flasks were applied with a 0.8% HP extract and 99.2% medium for 48 hours, no application was made to the control group. The effects of HP on mitochondrial mediated intrinsic and extrinsic apoptotic pathway agents, cell cycle G2 / M checkpoint kinases and inflammatory mediators were investigated in BCC cancer cells. The expression and activities of these mediators in both groups were evaluated by ELISA test. 
 Results: It was determined that HP extract treatment increased the expression of apoptotic proteins (AIF, GADD153, GRP78, caspase-3, Bax) and cell cycle G2 / M checkpoint kinase (Wee1), though it reduced antiapoptotic protein (Bcl-2) and inflammatory mediators (iNOS, COX-2, cPLA2, NFkB) in BCC cell lines.
 Conclusion: HP extract can support routine chemotherapy on BCC cells with its apoptotic, anti-inflammatory and anti-cancer effect, and may be a beacon of hope in reducing resistance to chemotherapeutic drugs in the treatment of BCC.

The effect of cryoprotectant vehicle solution on cartilage cell viability following vitrification.

Osteochondral allografts are often used to repair large articular cartilage defects to prevent or delay the onset of osteoarthritis. This approach is limited by the timely acquisition and use of allograft tissue since standard hypothermic protocols allow for a maximum storage of 4weeks. Vitrification is a proven technique for the long-term preservation of cells and tissues, but requires careful determination of parameters to be successful, particularly for articular cartilage. One parameter that is infrequently considered is the choice of cryoprotectant vehicle solution. The aim of this study was to evaluate the impact of a subset of vehicle solutions on an established vitrification protocol for articular cartilage. These solutions were phosphate-buffered saline (PBS), Dulbecco's Modified Eagle's Medium: Nutrient Mixture F-12 (DMEM), X-VIVO, and Unisol-CV (UCV). Both the solution pH at various points throughout vitrification and the cell viability of porcine articular cartilage slices following vitrification were measured. Using randomized block ANOVA, it was found that the normalized cell viability of articular cartilage vitrified in UCV was significantly greater than that of PBS (p < 0.05) and may be greater than those of DMEM and X-VIVO (p < 0.1). There was no correlation between pH parameters and cell viability, although significant differences between calculated pH parameters were identified. These results provide information to guide the design of effective vitrification protocols for articular cartilage.

A dual responsive probe based on bromo substituted salicylhydrazone moiety for the colorimetric detection of Cd2+ ions and fluorometric detection of F‒ ions: Applications in live cell imaging

A dual responsive probe based on bromo substituted salicylhydrazone moiety for the colorimetric detection of Cd2+ ions and fluorometric detection of F‒ ions: Applications in live cell imaging

The Viability and Growth of HaCaT Cells After Exposure to Bioactive Glass S53P4-Containing Cell Culture Media.

Bioactive glass (BG) S53P4 reduces the viability of epidermal keratinocyte-derived immortalized cell line, HaCaT in sufficient concentrations in vitro. Although used in mastoid obliteration surgery, there is no data available on whether BG S53P4 granules have an inhibitory or excitatory effect on keratinocytes, found in normal skin and ear cholesteatoma in vivo. HaCaT cell cultures were incubated with a direct BG S53P4 granule contact. Microscopic evaluation of the cultures was performed and interleukin-6 (IL-6) and -8 (IL-8) concentrations were measured from the medium samples. In addition, BG granules were incubated in two cell culture media for 6 days and the pure media were used in confluent HaCaT cultures preceding cell viability assay. Finally, a scratch assay test was performed to reveal the possible BG effect on HaCaT cultures. Eight to ten cell thick layers of dead HaCaT cells were noticed after a 2-day BG granule contact. With a BG concentration of 2.5%, IL-6 and IL-8 concentrations were smaller compared with the control group without BG after 2 days' incubation. Overall, HaCaT cell viability decreased when BG was incubated in keratinocyte growth medium, but did not change in Dulbecco's modified Eagle's medium. In a scratch assay test, cell regrowth in the scratch area was notable in cultures without BG. BG S53P4 seems to have an inhibitory effect on HaCaT cell growth. Although further studies are needed, this observation seems advantageous for cholesteatoma treatment.