Cell Viability Values Research Articles

Cytotoxicity of Peruvian propolis and Psidium guajava on human gingival fibroblasts, PBMCs and HeLa cells.

It is indisputable that every day it is demonstrated that natural products present diverse therapeutic benefits, which has boosted their incorporation within various products for clinical use. However, this must be accompanied by knowledge of their effect on cell lines to ensure their use is safe. The objective of this study was to evaluate the cytotoxic effect of two ethanolic extracts based on Peruvian natural products, on three human cell lines. Cervical cancer cell lines (HeLa), human gingival fibroblasts (HGF-1 - ATCC CRL-2014) (HGF-1) and peripheral blood mononuclear cells (PBMCs) were cultured and subsequently treated with preparations of ethanolic extracts of propolis (EEP) and Psidium guajava (EEG) from a concentration of 50 mg/mL to 0.024 mg/mL, by the 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazole bromide reduction assay. At a concentration of 0.24 mg/mL EEG, viability of 99.7±1.24%, 99.8±2.2% and 99.7±2.7% was observed in HeLa, HGF-1 and PBMCs, respectively; >90% cell viability values were observed with EPP at 0.024 mg/mL, with HGF-1 showing the highest viability (96.9±1.15%). A dose-dependent effect was observed for both extracts with a decrease in cell viability as concentrations increased (up to 50 mg/mL). EEP and EEG extracts at low concentrations do not show cytotoxicity in human cell lines, these findings are an advance in the preclinical evaluation on their safety and open a continuity to further studies for their potential applications in dentistry and medicine.

EFFECT OF KARAMUNTING LEAF EXTRACT (Melastoma malabathricum) ON THE PROLIFERATION OF BABY HAMSTER KIDNEY-21 (BHK-21) FIBROBLAST CELLS

Background: Pulpitis is an inflammation of dental pulp due to infection of microorganisms from dental caries. Pulpitis is clinically divided into reversible and irreversible pulpitis. Treatment of reversible pulpitis can be done by applying a pulp capping material. The use of calcium hydroxide as a pulp capping material is still not optimal, so an alternative material is needed, namely karamunting leaf (Melastoma malabathricum). The secondary metabolites in karamunting leaf extract have antibacterial activity. Antibacterial activity is needed in the regeneration of pulp tissue. Purpose: To analyze the effect of karamunting leaf extract (Melastoma malabathricum) on viability and proliferation of Baby Hamster Kidney-21 (BHK-21) fibroblast cells using the MTT Assay method. Methods: This study is a purely experimental study with post test only with control group design which were divided into 8 groups. The group was treated with karamunting leaf extract consisted of 5 groups, were 5%, 10%, 15%, 20%, and 25% concentrations also 3 control groups, were positive control (calcium hydroxide), cell control, and media control. Absorbance value was read using ELISA Reader and the percentage of cell viability was calculated. Results: All treatment groups had cell viability values higher than cell control and cause proliferation of BHK-21 fibroblast cells. The One Way Anova showed a value of sig=0,000 (p<0,05), which means that there was a minimum difference in one treatment group. Conclusion: Karamunting leaf extract and calcium hydroxide can maintain viability and cause proliferation of BHK-21 fibroblast cells. Karamunting leaf extract 25% concentration more effective in causing proliferation of BHK-21 fibroblast cells.

Hsa_circ_0017639 regulates cisplatin resistance and tumor growth via acting as a miR-1296-5p molecular sponge and modulating sine oculis homeobox 1 expression in non-small cell lung cancer

ABSTRACT Cisplatin (DDP)-induced chemoresistance is an important reason for the failure of non-small cell lung cancer (NSCLC) treatment. Circular RNAs (circRNAs) participate in the chemoresistance of diverse cancers. However, the function of hsa_circ_0017639 (circ_0017639) in the DDP resistance of NSCLC is unclear. Forty-one NSCLC samples (21 DDP-resistant samples and 20 DDP-sensitive samples) were utilized in the research. The relative expression levels of some genes were determined by real-time quantitative polymerase chain reaction (RT-qPCR). 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay for half-maximal inhibitory concentration (IC50) value of DDP and cell viability, colony formation and 5-ethynyl-2’-deoxyuridine (EDU) assays for cell proliferation, flow cytometry assay for cell apoptosis, transwell assay for cell invasion and wound-healing assay for cell migration were performed. The regulation mechanism of circ_0017639 was demonstrated by a dual-luciferase reporter assay. We observed higher levels of circ_0017639 in DDP-resistant NSCLC samples and cells. Functionally, circ_0017639 silencing decreased tumor growth and elevated DDP sensitivity in vivo and induced apoptosis, repressed proliferation, invasion, and migration of DDP-resistant NSCLC cells in vitro. Mechanically, circ_0017639 modulated sine oculis homeobox 1 (SIX1) expression via sponging microRNA (miR)-1296-5p. Also, miR-1296-5p inhibitor restored circ_0017639 knockdown-mediated impacts on cell DDP resistance in DDP-resistant NSCLCs. Furthermore, SIX1 overexpression counteracted the inhibiting impact of miR-1296-5p upregulation on DDP resistance and malignant phenotypes of DDP-resistant NSCLC cells. In conclusion, circ_0017639 conferred DDP resistance and promoted tumor growth via elevating SIX1 expression through sequestering miR-1296-5p in NSCLC, providing a new mechanism for understanding the chemoresistance and progression of NSCLC.

Influence of Copper on the Microstructural, Mechanical, and Biological Properties of Commercially Pure Zn-Based Alloys for a Potential Biodegradable Implant.

Three Zn-based alloys (Zn1Cu, Zn2Cu, and Zn3Cu) were developed by the addition of Cu (1, 2, and 3 wt %) into commercially pure Zn. This report systematically investigates the potential for these newly developed Zn-based alloys as biodegradable materials. Microstructural studies reveal the presence of spherical-shaped nanosized precipitates of ε-CuZn4 in the Zn1Cu alloy, whereas Zn2Cu and Zn3Cu alloys exhibit the presence of both micron- and nanosized precipitates of ε-CuZn4. The mechanical properties such as hardness, tensile and compressive strengths improve significantly with an increase in the amount of Cu in the alloy. The Zn3Cu alloy exhibits the highest yield strength (225 ± 9 MPa) and ultimate tensile strength (330 ± 12 MPa) among all of the alloys, which are ∼2.7 and 2 times higher than those of pure Zn. In vitro degradation behavior is evaluated by the potentiodynamic polarization study and immersion testing in Hank's solution for 20 and 75 days. The corrosion rate after both polarization and immersion testing follows the order of pure Zn < Zn1Cu < Zn3Cu < Zn2Cu. An electrochemical impedance spectroscopy (EIS) study also concludes that Zn2Cu shows the lowest corrosion resistance. The % cell viability values of 3T3 fibroblasts cells after 5 days of culture in a 50% diluted extract of pure Zn, Zn2Cu, and Zn3Cu alloys are 76 ± 0.024, 86.18 ± 0.033, and 92.9 ± 0.026%, respectively, establishing the improved cytocompatibility of the alloys as compared to pure Zn. Furthermore, an antibacterial study also reveals that the Zn3Cu alloy exhibits 80, 67, and 100% increases in the zone of inhibition (ZOI) for Escherichia coli, Bacillus subtilis, and Pseudomonas aeruginosa bacteria, respectively, as compared to that of pure Zn.

Effectiveness of Free and Liposome-Entrapped Antitumoral Drugs against Hepatocellular Carcinoma: A Comparative In vitro Study

Hepatocellular carcinoma (HCC) is a highly prevalent form of liver cancer. Sorafenib is the first-line treatment for patients who are not eligible for surgical procedures. Still, it causes important adverse effects, does not provide symptoms relief, and is marketed at expensive costs. Here we investigated the effectiveness of imatinib, sunitinib, and bortezomib against an in vitro model of HCC. These drugs are less expensive than Sorafenib and are approved for other malignant neoplasia. The drugs were tested as free and liposome-entrapped forms against HepG2 cells. Liposomes were prepared without using organic solvents and were tested for physicochemical stability. Entrapment efficiency was determined by spectrophotometric methods. Liposomes' average size was 238±33 nm, with an average entrapment efficiency of 34.3%. All drugs significantly decreased the viability of HepG2 cells in their free form and liposome-entrapped forms compared to the control. In spite of the low entrapment efficiency, no organic solvent was used in liposomes preparation, and cell viability values after 18h treatments with liposome-entrapped drugs were only surpassed by the highest concentration of free drugs. Our data open doors for more studies to assess the safety of the liposomes in vivo.

Potential Translational Thioflavin T Methodology as a Complement of Cell-Based Assays and after Drug Exposition

Protein aggregation is a common characteristic of several human diseases such as Alzheimer’s disease. Recent evidence has indicated that the aggregation of peptides such as p53 is also marked in cancer cells. The aim of this study was to correlate Thioflavin T (ThT) data with different cellular viability assays (Neutral Red and MTT) in SH-SY5Y neuroblastoma cells and HT-29 colon cancer cells treated with doxorubicin, a classical antineoplastic agent. We also studied the effects of the well-known peptide Aβ42 on the aggregation process in these cells. Our data suggest that both cancer cell lines are responsive to doxorubicin and formed aggregates, highlighting a relationship between ThT and cellular viability methodologies. We observed that lower values of cell viability corresponded with pronounced aggregation. Thus, these results indicated that the ThT methodology used in cells may complement the cell viability assays. In addition, this methodology may be of interest to evaluate the role of protein aggregation in other cancer cells.

3D Bioprinting of Novel κ-Carrageenan Bioinks: An Algae-Derived Polysaccharide.

Novel green materials not sourced from animals and with low environmental impact are becoming increasingly appealing for biomedical and cellular agriculture applications. Marine biomaterials are a rich source of structurally diverse compounds with various biological activities. Kappa-carrageenan (κ-c) is a potential candidate for tissue engineering applications due to its gelation properties, mechanical strength, and similar structural composition of glycosaminoglycans (GAGs), possessing several advantages when compared to other algae-based materials typically used in bioprinting such as alginate. For those reasons, this material was selected as the main polysaccharide component of the bioinks developed herein. In this work, pristine κ-carrageenan bioinks were successfully formulated for the first time and used to fabricate 3D scaffolds by bioprinting. Ink formulation and printing parameters were optimized, allowing for the manufacturing of complex 3D structures. Mechanical compression tests and dry weight determination revealed young’s modulus between 24.26 and 99.90 kPa and water contents above 97%. Biocompatibility assays, using a mouse fibroblast cell line, showed high cell viability and attachment. The bioprinted cells were spread throughout the scaffolds with cells exhibiting a typical fibroblast-like morphology similar to controls. The 3D bio-/printed structures remained stable under cell culture conditions for up to 11 days, preserving high cell viability values. Overall, we established a strategy to manufacture 3D bio-/printed scaffolds through the formulation of novel bioinks with potential applications in tissue engineering and cellular agriculture.

Cytotoxic Effect of Nano Fast Cement and ProRoot Mineral Trioxide Aggregate on L-929 Fibroblast Cells: an in vitro Study.

Statement of the Problem: Endodontic materials that are placed in direct contact with living tissues should be biocompatible. The cytotoxicity of Nano Fast Cement (NFC) compared to ProRoot Mineral Trioxide Aggregate (ProRoot MTA) must be evaluated.Purpose: This In vitro study aimed to assess the cytotoxic effects of NFC in comparison to ProRoot MTA on L-929 mouse fibroblast cells.Materials and Method: In this animal study, L-929 mouse fibroblast cells were grown in Dulbecco's Modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS) in an atmosphere of 5% co2/95% air at 37 C̊. A total of 104 cells from the fourth collection were plated in each well of a 96-well micro-titer plate. Materials were mixed according to the manufacturer’s instruction and placed into the related plastic molds with 5 mm diameter and 3 mm height. After 24 hours and a complete setting, the extracts of the tested materials were produced at six different concentrations and placed in the related wells. Cells in DMEM served as the negative control group. DMEM alone was used as the positive control group. Methyl-thiazoltetrazolium (MTT) colorimetric assay was conducted after 24, 48, and 72 hours. The absorbance values were measured by ELISA plate reader at 540 nm wavelength. Three-way analysis of variance, post-hoc Tukey, LSD, and independent t-test were used for the statistical analyses using SPSS software, version 16.0.Results: There was no statically significant difference between MTA and NFC in cell viability values at different concentrations and different time intervals (p= 0.649). Viability values were significantly decreased after 72 hours, but there was no significant difference between the first and second MTT assays (p= 0.987). Cytotoxicity significantly increased at concentrations higher than 6.25 µɡ/ml.Conclusion: Cytotoxicity depends on time, concentration, and cement composition. There was no statistically significant difference between NFC and MTA concerning their cytotoxic effects on L-929 mouse fibroblast cells.

Antiproliferative Effects of Curcumin Different Types of Breast Cancer

Objective:Breast cancer is one of the most frequently diagnosed malignancy among women. Turmeric is isolated from Curcuma longa. Curcumin is main curcuminoid of the turmeric which is a member of Zingiberaceae. In this current study antiproliferative effects of curcumin were investigated in luminal A breast cancer cell line MCF-7 and triple negative breast cancer cell line MDA-MB-231. Methods:For this purpose cell viability, cell index values by xCELLigence Real-Time Cell Analysis DP instrument, mitotic index and apoptotic index analysis were used. Results:Cell viability and cell index values showed that 75 µM concentration of curcumin was IC50 concentration. When IC50 concentration was applied to both cell lines, a significant decrease was observed in the mitotic index values, while a significant increase was observed in the apoptotic index values (p<0.05). Conclusion:Curcumin, which has antiproliferative effects on breast cancer cells, is thought to be effective in cancer treatment.

The cell viability assay analysis and physicochemical characterization of porous hydroxyapatite scaffold using honeycomb and paraffin wax as polymeric porogen for bone tissue engineering

To fabricate and characterize the porous hydroxyapatite-based scaffold, honeycomb as a natural polymer (HA/HCB) and paraffin wax (HA/Wax) were used. The fabrication of scaffold using the porogen leaching method was varied temperatures between 700, 900, and 1100 °C. Theoretically, the temperature of calcination influenced the morphology of the scaffold, crystallite size, functional group, and porosity. According to the previous study, the crystallite size of the polymer scaffold is less than 100 nm. The HA-based scaffold was analyzed by the Energy Dispersive X-Ray Spectroscopy (EDS), Scanning Electron Microscopy (SEM), X-Ray Diffractometer (XRD), Fourier Transform Infrared Spectroscopy (FTIR), and tested by the cell viability. According to the XRD results, the crystallite size of HA/HCB scaffold decreases, while scaffold HA/Wax crystallite size tends to decrease when calcination temperature increases. As calcination temperature increases, porosity tends to be small for both HA/HCB and HA/wax scaffolds. The scaffold HA/HCB 900 °C has interconnected pores, uniform, and small porosity. In contrast, the scaffold HA/Wax 900 °C has fewer interconnected pores and non-uniform particles. The FTIR result of the HA/HCB 900 °C has C-H functional group, affecting cell viability. Through MTT (3-(4,5-Dimethylthiazol-2-yl)−2,5 diphenyltetrazolium bromide) assays, the cell viability value of the HA/Wax 900 °C was greater than the HA/HCB 900 °C for 48 h incubated time. It is caused by the alkane chains on HA/HCB, causing the death of cells. Considering cell viability assay studies for the nanocomposite scaffold, the obtained results confirm the non-toxicity of the material.

Synergistic anticancer effects of everolimus (RAD001) and Rhein on gastric cancer cells via phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway

ABSTRACT Everolimus (RAD001) is a mTOR inhibitor and is widely used for the treatment of gastric cancer (GC). Evidence suggests that Rhein has anticancer effect on GC. But the synergistic effect and mechanism of RAD001 and Rhein combination on GC is not clear. The current study aims to clarify the combination of RAD001 and Rhein in GC treatment. We found Rhein dose-dependently repressed MGC-803 cell viability (50% inhibition concentration (IC50) value = 94.26 μM). Rhein (80 μM) significantly suppressed GC cell proliferation and invasion. RAD001 dose-dependently repressed MGC-803 cells viability (IC50 value = 45.41 nM). The combination of Rhein and RAD001 repressed MGC-803 cells viability, invasion, and proliferation compared to the administration of Rhein or RAD001 alone. Protein levels of epithelial-mesenchymal transition (EMT)-related molecules E-cadherin, N-cadherin and Vimentin expressions were significantly affected by the combination of Rhein and RAD001. The combination of Rhein and RAD001 significantly facilitated cell apoptosis and up-regulated expressions of cell apoptosis and cycle-related protein p53, cyclin-dependent kinase 4 (CDK4) and cyclin D1 compared to the administration of Rhein or RAD001 alone. Moreover, the combination of Rhein and RAD001 repressed the expressions of phosphorylation-phosphoinositide-3-kinase (p-PI3K), p-protein kinase B (p-AKT) and p-mammalian target of rapamycin (p-mTOR). Finally, the combination of RAD001 and Rhein significantly decreased tumor weight and volume, suppressed the expressions of p-PI3K, p-Akt and p-mTOR, and repressed cell proliferation marker Ki-67 expression, which exerted synergistic cancer prevention in GC in vivo. Overall, the combination of Rhein and RAD001 exert synergistic cancer prevention in GC via PI3K/Akt/mTOR pathway.

Effect of Three Chlorhexidine-Based Mouthwashes on Human Gingival Fibroblasts: An In Vitro Study

Mouthwashes containing chlorhexidine (CHX) are deemed to be associated with dose-dependent side effects, including burning sensation and taste alteration. To overcome these drawbacks, mouthwashes with CHX at lower concentrations with or without adjunctive agents are proposed. The aim of this in vitro study was to investigate the effects of three CHX-based mouthwashes on human gingival fibroblasts (HGFs). After 3 days of cell culture, groups were randomly treated for 30 s, 60 s or 120 s with (a) CHX 0.05% in combination with cetylpyridnium chloride (CPC) 0.05%; (b) CHX 0.1%; (c) CHX 0.2%; or (d) NaCl as control. Cell viability, cytotoxicity and apoptosis were evaluated at 2 h, 3 days and 6 days after the exposure to the different solutions. Similar cell viability values were found among the test groups at all time points. At day 0, higher cytotoxicity was measured in the group treated with CHX 0.2%, in particular after long application time (120 s), while no significant difference was found between CHX + CPC and the control group. All the investigated mouthwashes were well tolerated by HGF cells for the tested application times. The highest cytotoxic effect was observed for CHX 0.2%; therefore, clinicians should consider limiting its usage to carefully selected clinical situations.

Evaluation of the antimicrobial and cytotoxic potential of endophytic fungi extracts from mangrove plants Rhizophora stylosa and R. mucronata

Mangrove endophytic fungi are tolerant to numerous stresses and are inevitably capable of exhibiting excellent biological activity by producing impressive numbers of metabolites with special biological functions, based on previous work on the biological potential of mangrove-derived endophytic fungi. To obtain marked antimicrobial and cytotoxic fermentation products of culturable endophytic fungi from mangrove forests, our research evaluated the antimicrobial and cytotoxic activities of crude extracts of endophytic fungi from Rhizophora stylosa and Rhizophora mucronata. Forty-six fungal isolates were cultured on four different media, namely, dextrose agar (PDA), Czapek’s agar (CZA), rice medium (RM) and grain medium (GM) and harvested by ethyl acetate solvent at 40 days. The extracts were tested for antimicrobial activity by the microdilution method against the gram-negative bacteria Pseudomonas adaceae (PA), gram-positive bacteria Enterococcus faecalis (EF), methicillin-resistant Staphylococcus aureus (MRSA) and pathogenic fungus Monilia albicans (MA). The cytotoxic activity of the extracts was evaluated by MTT assay using A549 human lung cancer cells, HeLa human cervical carcinoma cells, and HepG2 human hepatocellular cells. The results showed that rice medium could promote the secretion of antimicrobial and antitumour secondary metabolites of endophytic fungi in comparison with other cultivation media. Seventeen strains (68%) from R. stylosa exhibited inhibitory effects on indicators, especially N. protearum HHL46, which could inhibit the growth of four microbes with MIC values reaching 0.0625 mg/mL. Fifteen strains (71.4%) from R. mucronata displayed activities against human pathogenic microbes; in particular, Pestalotiopsis sp. HQD6 and N. protearum HQD5 could resist the growth of four microbes with MIC values ranging from 0.015 to 1 mg/mL. In the cytotoxicity assay, the extracts of 10 strains (40%), 9 strains (40%) and 13 strains (52%) of R. stylosa and 13 strains (61.9%), 10 strains (47.6%) and 10 strains (47.6%) of R. mucronata displayed cytotoxicity against A549, HeLa and HepG2 cancer cells with cell viability values ≤ 50%. Neopestalotiopsis protearum HHL46, Phomopsis longicolla HHL50, Botryosphaeria fusispora HQD83, Fusarium verticillioides HQD48 and Pestalotiopsis sp. HQD6 displayed significant antitumour activity with IC50 values below 20 μg/mL. These results highlighted the antimicrobial and antitumour potential of endophytic fungi from R. stylosa and R. mucronata and the possibility of exploiting their antimicrobial and cytotoxic agents.

Water-Soluble Silicon Quantum Dots toward Fluorescence-Guided Photothermal Nanotherapy

We report carboxy-terminated silicon quantum dots (SiQDs) that exhibit high solubility in water due to the high molecular coverage of surface monolayers, bright light emission with high photoluminescence quantum yields (PLQYs), long-term stability in the PL property for monitoring cells, less toxicity to the cells, and a high photothermal response. We prepared water-soluble SiQDs by the thermal hydrosilylation of 10-undecenoic acid on their hydrogen-terminated surfaces, provided by the thermal disproportionation of triethoxysilane hydrolyzed at pH 3 and subsequent hydrofluoric etching. The 10-undecanoic acid-functionalized SiQDs (UA:SiQDs) showed long-term stability in hydrophilic solvents including ethanol and water (pH 7). We assess their interaction with live cells by means of cellular uptake, short-term toxicity, and, for the first time, long-term cytotoxicity. Results show that UA:SiQDs are potential candidates for theranostics, with their good optical properties enabling imaging for more than 18 days and a photothermal response having a 25.1% photothermal conversion efficiency together with the direct evidence of cell death by laser irradiation. UA:SiQDs have low cytotoxicity with full viability of up to 400 μg/mL for the short term and a 50% cell viability value after 14 days of incubation at a 50 μg/mL concentration.

Development of a hydroxyapatite nanoparticle-based gel for enamel remineralization -A physicochemical properties and cell viability assay analysis.

Nano-hydroxyapatite (nHA) was synthesized from abalone mussel shells (Haliotis asinina) using a precipitation method, and gel HA-Abalone was developed using the carbomer materials with concentrations of 0, 10, 20, 30, and 40 wt%. The specimens used were 25 freshly extracted caries-free premolar teeth, and the treatment was done twice a day for 14 days. Gel HA-Abalone 20 wt%, with a crystallite size of 14.70±1.21 nm, was the best concentration to achieve the best remineralization (~863 VHN) of the superficial layer. Based on the results of cell viability assay on gel HA-Abalone 20 wt%, the growth of NIH/3T3 cells was inhibited beginning at a gel concentration of 1,000 µg/mL, and the half maximal inhibitory concentration (IC50) value was 1,497 µg/mL. Based on to the one-way analysis of variance (ANOVA), the result reflected statistically significant differences in the average of the cell viability and enamel surface microhardness values (p<0.05).

A novel ophthalmic latanoprost 0.005% nanoemulsion: a cytotoxicity study.

Benzalkonium chloride (BAK), the most commonly used preservative in anti-glaucoma eye drops, inflicts damage to the ocular surface. A novel anti-glaucoma formulation that avoids the use of BAK has been developed. The aim of this study was to evaluate the cytotoxicity of this formulation and to compare it with an ophthalmic solution containing BAK. Two different latanoprost eye drops were used: one ophthalmic solution (LSc) containing BAK 0.02% and one ophthalmic nanoemulsion (LNe) with a soft preservative (potassium sorbate 0.18%). Human epithelial conjunctival cells were incubated for 15, 30, and 60min with either LSc or LNe. The cytotoxicity was determined by MTT assay. Cell death was measured by flow cytometry using annexin V-FITC and propidium iodide. The values of cell viability and proliferation obtained from cells exposed to LNe were between 80 and 90% relative to the control group, whereas values obtained from cells exposed to LSc were around 30% at all study times (p < 0.05 at 15 and 30min; p < 0.01 at 60min). The percentage of viable cells decreased significantly when cells were incubated with LSc compared with cells incubated with LNe at all the study times, while the percentage of cells in late apoptosis/necrosis increased significantly in cells exposed to LSc compared to LNe. The new latanoprost nanoemulsion is significantly less cytotoxic on human conjunctival cells than LSc. These results suggest that the new formulation might be gentler on the eye surface than currently available BAK-preserved latanoprost solutions.

The Study of Steaming Durations and Temperatures on the Chemical Characterization, Neuroprotective, and Antioxidant Activities of Panax notoginseng.

Panax notoginseng (PN) is one of the most valuable traditional Chinese medicines and has extensive pharmacological effects. Recent studies demonstrated that PN exhibited pharmacological effect related to Alzheimer's disease (AD). However, whether steaming process can boost its anti-AD activity is still unexplored. To fill this gap, effects of steaming durations and temperatures on the chemical characterization, neuroprotective and antioxidant activities of PN were systematically investigated in this study. HPLC fingerprint coupled with quantitative analysis demonstrated striking conversion of original saponins to less polar ones with the increase in the steaming time and temperature. In the viewpoint of anti-AD activity on neuroprotective and antioxidant effects, several steamed PN samples (110°C-6/8/10 h, 120°C ‐4/6 h samples) displayed a significant increase both in cell viability and oxygen radical absorption capacity (ORAC) values compared with the no steamed one (P < 0.01 or P < 0.005). Steaming temperature had the greater impact on the change of chemical composition and anti-AD activity of PN. Moreover, the spectrum-effect relationship analysis revealed that the transformed saponins were partially responsible for the increased neuroprotective and antioxidant effects of steamed PN. Therefore, steamed PN could be used as a potential crude drug for prevention and treatment of AD.

Cytotoxicity evaluation of a novel latanoporst 0.005% nanoemulsion

AbstractPurposeTo evaluate the cytotoxicity of a new Latanoprost Benzalkonium chloride (BAK)‐free nanoemulsion and to compare it with a Latanoprost solution containing BAK.MethodsNormal Human Conjunctival epithelial cells (IOBA‐NHC) were incubated for 15, 30, and 60 minutes with 1:6 dilutions of either latanoprost solution containing BAK (LSc), (latanoprost 0.005%, BAK 0.02%, Xalatan, Pfizer) or latanoprost nanoemulsion (LNe), (latanoprost 0.005%, potassium sorbate 0.18%; Louten Emulsion, POEN). Cytotoxicity was determined by MTT assay. Cell death was measured by flow cytometry using annexin V‐FITC and propidium iodide.ResultsCytotoxicity evaluation: the values of cell viability and proliferation obtained from cells exposed to LNe were between 80 and 90% relative to control group, whereas values obtained from cells exposed to LSc were around 30% relative to control group at all study times (p &lt; 0.05 at 15 and 30 minutes; p &lt; 0.01 at 60 minutes).Cell death evaluation: the percentage of viability was significantly lower in cells exposed to LSc compared with those incubated with LNe at all study times (p &lt; 0.05 at 15 and 30 minutes; p &lt; 0.01 at 60 minutes). Viability of cells exposed to LNe was higher than 90% at all study times, whereas there was a time‐dependent decrease in cells exposed to Lsc. The percentage of viable cells in the LNe group was comparable to the viability of control group (93.1%) at all study times. No statistical differences between groups reached in early apoptotic cells. A significant increase of late apoptosis and necrosis cells were observed at all study times for those incubated with Lsc (p &lt; 0.05; p &lt; 0.01 at 30 and 60 minutes).ConclusionsThe new latanoprost nanoemulsion is significantly less cytotoxic on human conjunctival cells than LSc. It suggests that the new formulation could be gentler on the eye surface than currently available BAK preserved latanoprost solutions, with levels of cytotoxicity equivalent to control.

In vitro biocompatibility testing of different base materials used for elevation of proximal subgingival margins using human gingival epithelial cells.

To analyze the biological effects of four base materials used for elevation of proximal subgingival margins on gingival epithelial cells. Twenty-eight specimens for each of the four base materials (total 112 specimens) were used: resin-modified glass ionomer (RMGI), glass hybrid (HV-GIC), flowable bulk fill resin composite (Bulk Flow) and bioactive ionic resin (Activa). Proximal enamel and root dentin were used as controls. Gingival epithelial cell viability was calculated after direct incubation on all four types of material for either 24 h or 72 h using both the methyl tetrazolium and trypan blue dye exclusion assays. Data were analyzed statistically using one-way analysis of variance, Tukey post hoc test and independent sample t-test (P < 0.05). Cell viability values in both assays showed significant differences among the study groups. Bulk Flow showed the highest values, followed in order by Activa and the control groups. Both HV-GIC and RMGI had the lowest values. Cell viability in all of the study groups was higher after incubation for 72 h than after 24 h. In terms of biocompatibility with epithelial tissues, bulk fill resin composite appears to be most suitable, followed by bioactive composite, for subgingival placement than glass ionomer-based materials, especially that containing 2-hydroxy-ethyl methacrylate.

Calcium-doped zinc oxide nanocrystals as an innovative intracanal medicament: a pilot study.

This study investigated the cytotoxicity, radiopacity, pH, and dentinal tubule penetration of a paste of 1.0% calcium-doped zinc oxide nanocrystals (ZnO:1.0Ca) combined with propylene glycol (PRG) or polyethylene glycol and propylene glycol (PEG-PRG). The pastes were prepared by mixing calcium hydroxide [Ca(OH)2] or ZnO:1.0Ca with PRG or a PEG-PRG mixture. The pH was evaluated after 24 and 96 hours of storage in deionized water. Digital radiographs were acquired for radiopacity analysis and bubble counting of each material. The materials were labeled with 0.1% fluorescein and applied to root canals, and images of their dentinal tubule penetration were obtained using confocal laser scanning microscopy. RAW264.7 macrophages were placed in different dilutions of culture media previously exposed to the materials for 24 and 96 hours and tested for cell viability using the MTT assay. Analysis of variance and the Tukey test (α = 0.05) were performed. ZnO:1.0Ca materials showed lower viability at 1:1 and 1:2 dilutions than Ca(OH)2 materials (p < 0.0001). Ca(OH)2 had higher pH values than ZnO:1.0Ca at 24 and 96 hours, regardless of the vehicle (p < 0.05). ZnO:1.0Ca pastes showed higher radiopacity than Ca(OH)2 pastes (p < 0.01). No between-material differences were found in bubble counting (p = 0.0902). The ZnO:1.0Ca pastes had a greater penetration depth than Ca(OH)2 in the apical third (p < 0.0001). ZnO:1.0Ca medicaments presented higher penetrability, cell viability, and radiopacity than Ca(OH)2. Higher values of cell viability and pH were present in Ca(OH)2 than in ZnO:1.0Ca.