scholarly journals Investigation of the radiopacity and cytotoxicity of albodent - novel strontium carbonate incorporated calcium silicate based dental cement

2021 ◽  
Vol 68 (2) ◽  
pp. 68-78
Author(s):  
Ana Despotovic ◽  
Djordje Antonijevic ◽  
Dragan Ilic ◽  
Nevena Zogovic ◽  
Vukoman Jokanovic
Keyword(s):  
Cell Viability ◽  
Calcium Silicate ◽  
Phase Contrast ◽  
L929 Cell ◽  
Dental Cement ◽  
Pulp Capping ◽  
Contrast Microscopy ◽  
Diphenyl Tetrazolium Bromide ◽  
Mtt Assays

Introduction. Calcium silicate (CS) dental cements have numerous clinical indications in dentistry including pulp capping, root end surgery, perforation repair and apexification/apexogenesis treatment. Materials and methods. Novel CS based dental cement with incorporation of SrCO3 radiopacifier named ALBO-DENT was used as an experimental cement material while Portland cement (Aalborg, Denmark) and ProRoot MTA (Tulsa Dental, USA) were used as controls. The radiopacity evaluation was performed using digital Trophy Radiographic system with an intention to precisely determine the minimum of radiopaque agent needed to confer to ISO radiopacity requirement. Thereafter, biocompatibility of material was tested in in vitro conditions in mouse fibrosarcoma L929 cell culture treated with materials? extracts. Cell morphology was observed using phase-contrast microscopy, while cell viability was measured using crystal violet (CV) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assays. Results. Radiopacity evaluation revealed that 30%wt addition of SrCO3 was necessary to achieve satisfactory radiopacity (3.45 mm Al). Cytotoxicity analysis using CV and MTT assays revealed that pure extracts of ALBO-DENT presented superior biocompatibility when compared to PC and MTA controls while serial dilutions of experimental cements? extracts as well as that of PC and MTA did not influence L929 cell viability. Conclusions. Novel formulation of CS cement - ALBO-DENT presented satisfactory radiopacity and adequate biocompatibility.

scholarly journals Comparison of Cytotoxic Effects of Calcium Silicate-based Materials on Human Pulp Fibroblasts

2019 ◽  
Vol 13 (4) ◽  
pp. 241-246 ◽  
Author(s):  
Mehmet Adıgüzel ◽  
Fuat Ahmetoğlu ◽  
Ayçe Ünverdi Eldeniz ◽  
Mehmet Gökhan Tekin ◽  
Bülent Göğebakan
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Mtt Assay ◽  
Calcium Silicate ◽  
Culture Medium ◽  
In Vitro Cytotoxicity ◽  
The Other ◽  
Cytotoxic Effects ◽  
Pulp Capping

Background. This study aimed to compare the in vitro cytotoxicity of Theracal LC, BiodentineTM, iRoot BP Plus, and MTA Angelus on human pulp fibroblasts (HPF). Methods. Fifteen discs from each calcium silicate-based material were prepared in sterile Teflon molds. After setting, the fabricated discs were eluated with a culture medium for 24 h. HPF cells were plated onto 24-well plates at 5×103 cells/well, and the cells were exposed to the material eluates. The cell viability was evaluated with MTT assay at three different times (24, 48, and 72 h). Data were statistically analyzed. The apoptotic/necrotic status of HPF cells exposed to material eluates was determined by flow cytometry. Results. The differences between the effects of Theracal LC, BiodentineTM, MTA Angelus, and iRoot BP Plus on HPF cells were found to be statistically significant (P<0.05). Theracal LC was found to be more cytotoxic considering other vital pulp capping materials at 24- (28.3%), 48- (44.9%), and 72-hour (49.2%) intervals. On the other hand, BiodentineTM showed the least cytotoxic effects (97.1%, 130.0%, and 103.7%, respectively) According to flow cytometry results, Theracal LC material increased apoptosis/necrosis ratios compared to the other materials. Conclusion. Based on the results of the present study, BiodentineTM, MTA Angelus, and iRoot BP Plus can be classified as biocompatible materials in vital endodontic treatments. However, the Theracal LC materials should be used carefully due to their cytotoxic effects.

scholarly journals Uptake and intracellular accumulation of diamond nanoparticles – a metabolic and cytotoxic study

2017 ◽  
Vol 8 ◽  
pp. 1649-1657 ◽  
Author(s):  
Antonín Brož ◽  
Lucie Bačáková ◽  
Pavla Štenclová ◽  
Alexander Kromka ◽  
Štěpán Potocký
Keyword(s):  
Cell Viability ◽  
Phase Contrast ◽  
Cell Number ◽  
Rapid Reduction ◽  
Diamond Nanoparticles ◽  
Cultivation Period ◽  
High Pressure High Temperature ◽  
Accessible Surface ◽  
In Vitro Interactions

Diamond nanoparticles, known as nanodiamonds (NDs), possess several medically significant properties. Having a tailorable and easily accessible surface gives them great potential for use in sensing and imaging applications and as a component of cell growth scaffolds. In this work we investigate in vitro interactions of human osteoblast-like SAOS-2 cells with four different groups of NDs, namely high-pressure high-temperature (HPHT) NDs (diameter 18–210 nm, oxygen-terminated), photoluminescent HPHT NDs (diameter 40 nm, oxygen-terminated), detonation NDs (diameter 5 nm, H-terminated), and the same detonation NDs further oxidized by annealing at 450 °C. The influence of the NDs on cell viability and cell count was measured by the mitochondrial metabolic activity test and by counting cells with stained nuclei. The interaction of NDs with cells was monitored by phase contrast live-cell imaging in real time. For both types of oxygen-terminated HPHT NDs, the cell viability and the cell number remained almost the same for concentrations up to 100 µg/mL within the whole range of ND diameters tested. The uptake of hydrogen-terminated detonation NDs caused the viability and the cell number to decrease by 80–85%. The oxidation of the NDs hindered the decrease, but on day 7, a further decrease was observed. While the O-terminated NDs showed mechanical obstruction of cells by agglomerates preventing cell adhesion, migration and division, the H-terminated detonation NDs exhibited rapid penetration into the cells from the beginning of the cultivation period, and also rapid cell congestion and a rapid reduction in viability. These findings are discussed with reference to relevant properties of NDs such as surface chemical bonds, zeta potential and nanoparticle types.

In vitro Maturation of the Exoerythrocytic Stage of Plasmodium knowlesi Observed under Phase Contrast Microscopy

1990 ◽  
Vol 76 (6) ◽  
pp. 923 ◽  
Author(s):  
Pascal Millet ◽  
William E. Collins ◽  
Claude E. Monken ◽  
Bobby G. Brown
Keyword(s):  
Phase Contrast ◽  
In Vitro Maturation ◽  
Plasmodium Knowlesi ◽  
Phase Contrast Microscopy ◽  
Contrast Microscopy

scholarly journals 209FUNCTIONAL ASSESSMENT OF WHITE RHINOCEROS CERATHOTERIUM SIMUM EPIDIDYMAL SPERMATOZOA BEFORE AND AFTER CRYOPRESERVATION

2004 ◽  
Vol 16 (2) ◽  
pp. 226 ◽  
Author(s):  
F. Martinez-Pastor ◽  
F. Olivier ◽  
T. Spies ◽  
L. Anel ◽  
P. Bartels
Keyword(s):  
Plasma Membrane ◽  
Assisted Reproduction ◽  
Phase Contrast ◽  
Egg Yolk ◽  
Phase Contrast Microscopy ◽  
White Rhinoceros ◽  
Contrast Microscopy ◽  
Before And After ◽  
Epididymal Spermatozoa

Biological Resource Banks represent a potentially valuable tool for species conservation. It is, however, necessary to understand the species-specific cryopreservation process and its consequences for spermatozoa to aid in the development of assisted reproduction as a future conservation tool. The aim of this study was to assess the in vitro functionality of white rhinoceros Cerathoterium simum epididymal spermatozoa both before and after cryopreservation. Testes from a harvested white rhino bull were removed and transported at 5°C to the laboratory within 4h. The cauda epididymis was dissected out and flushed with 2mL of Tris-citrate egg yolk extender (fraction A, Biladyl, Minitüb, Germany). A 0.1mL aliquot was removed for analysis and the balance (9mL; 2mL fraction A+7mL sperm sample) mixed with an additional 27.2mL of Tris-citrate egg yolk with glycerol (fraction B, Bidadyl). The extended sample was allowed to cool to 4°C over a 6-h period before an additional 29.2mL of cooled fraction B were added (final sperm concentration=150×106mL−1). Sperm samples were loaded into 0.25-mL straws and frozen over LN2 vapor (4cm for 20min) for later assessment. Sperm straws were thawed by placing the straws in water at 37°C for 30s. Pre-freeze and post-thaw evaluations were carried out in the same manner. Media used included: HEPES for washing (20mM HEPES, 355mM sucrose, 10mM glucose, 2.5mM KOH) and HEPES saline (197mM NaCl, instead of sucrose). An aliquot was diluted with HEPES (washing) and centrifuged for 5min at 600×g; the pellet was resuspended in HEPES saline. Sperm motility (total motility %, TM;; and progressive motility %, PM) was assessed using phase contrast microscopy (×200; 37°C). Sperm plasma membrane status was assessed using the fluorescent dye, propidium iodide (50ngmL−1 in HEPES saline;; 10min, RT). Percentage of cells with plasma membranes intact (unstained;; PMI) was recorded. Mitochondrial status was assessed with the fluorescent dye, JC-1 (7.5μM in HEPES saline;; 30min, 37°C). The % of cells with an orange-stained midpiece was recorded (active mitochondria;; MIT). Resilience to hypoosmotic shock (HOS test) was assessed by diluting a sample in 100mOsm/kg HEPES saline (1:20; 15min, RT). An aliquot was stained with PI to assess plasma membrane status (HOSPMI), and the rest was fixed with formaldehyde, and % coiled tails (positive endosmosis;; HOST) was estimated using phase contrast microscopy (×400). Evaluations of PMI, MIT and HOSPMI were performed using fluorescence microscopy (×400, 450–490nm excitation filter). The results indicated that quality was good pre-freezing (TM: 60%; PMI: 86%; MIT: 100%), except for a PM value of 15%. After thawing, although there was a drop in TM (30%), there was no decrease in PM (20%). Our in vitro functional assessment indicated a loss of quality between the pre-freeze and post-thaw evaluations, but PMI and MIT maintained their pre-thaw levels (60% and 72%, respectively). The HOS test, which indicates plasma membrane integrity, decreased from the pre-freeze level (91%) to a post-thaw value of 70%. HOSTPMI was 72% pre-freeze, and decreased to 54% post-thaw. In conclusion, epididymal spermatozoa from the white rhino may retain its functionality after cryopreservation in a commerically available cryo-extender (Bidadyl). The use of assisted reproduction techniques could someday play a role in the management and conservation of the white rhinoceros and related species.

scholarly journals Biological Characteristics and Odontogenic Differentiation Effects of Calcium Silicate-Based Pulp Capping Materials

Materials ◽  
2021 ◽  
Vol 14 (16) ◽  
pp. 4661
Author(s):  
Yemi Kim ◽  
Donghee Lee ◽  
Hye-Min Kim ◽  
Minjoo Kye ◽  
Sin-Young Kim
Keyword(s):  
Cell Viability ◽  
Calcium Silicate ◽  
Cell Attachment ◽  
Biological Properties ◽  
Cell Counting ◽  
Control Group ◽  
Runx2 Expression ◽  
Pulp Capping ◽  
Alp Activity ◽  
Odontogenic Differentiation

We compared calcium silicate-based pulp capping materials to conventional calcium hydroxide in terms of their biological properties and potential effects on odontogenic differentiation in human dental pulp stem cells (hDPSCs). We cultured hDPSCs on disks (7-mm diameter, 4-mm high) of ProRoot MTA (Dentsply Tulsa Dental Specialties), Biodentine (Septodont), TheraCal LC (Bisco), or Dycal (Dentsply Tulsa Dental Specialties). Cell viability was assessed with cell counting (CCK) and scanning electron microscopy (SEM). Odontogenic activity was assessed by measuring alkaline phosphatase (ALP) activity and gene expression (quantitative real-time PCR). CCK assays showed that Dycal reduced cell viability compared to the other materials (p < 0.05). SEM showed low and absent cell attachment on TheraCal LC and Dycal disks, respectively. hDPSCs exposed to TheraCal LC and Dycal showed higher ALP activity on day 6 than the control group (p < 0.05). The day-9 Runx2 expression was higher in the ProRoot MTA and TheraCal LC groups than in the control group (p < 0.05). On day 14, the ProRoot MTA group showed the highest dentin sialophosphoprotein levels (not significant; p > 0.05). In conclusion, various pulp capping materials, except Dycal, exhibited biological properties favorable to hDPSC viability. ProRoot MTA and TheraCal LC promoted higher Runx2 expression than Biodentine. Future studies should explore the odontogenic potential of pulp capping materials.

In vitro induction of the acrosome reaction in ovine spermatozoa by calcium ionophore A23187

2003 ◽  
Vol 51 (1) ◽  
pp. 103-109 ◽  
Author(s):  
Ö. Uçar ◽  
T. J. Parkinson
Keyword(s):  
Phase Contrast ◽  
Acrosome Reaction ◽  
Calcium Ionophore ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
Contrast Microscopy ◽  
The Relationship ◽  
In Vitro Induction ◽  
Artificial Vagina

The relationship between concentration of calcium ionophore A23187 and incubation time upon the proportion of spermatozoa undergoing acrosome reaction (AR) in vitro was investigated in rams from a commercial artificial insemination (AI) program. Two ejaculates were collected by artificial vagina from each of nine rams of three breeds (Finn Dorset, Charolais and Suffolk) aged 8-36months. Each ejaculate was diluted in a skimmed milk extender. Spermatozoa were thereafter incubated for 45 or 60min in modified Tyrode's medium (TALP) which contained either zero, 0.1 or 1.0µM/l A23187. After fixing in 10% formaldehyde, the number of spermatozoa that had undergone AR was determined by phase contrast microscopy. In pre-incubation samples, 21.3± 3.3% of spermatozoa had undergone AR. Percentages of acrosome reacted spermatozoa were significantly (P<0.001) increased after incubation with A23187. After incubation with 0.1µM/l A23187 for 45 and 60min there were 22.4±3.0% and 31.7±4.3% acrosome reacted spermatozoa, respectively. After incubation with 1.0µM/l A23187 for 45 and 60min there were 46.2±6.5% and 53.8±5.9% acrosome reacted spermatozoa, whilst corresponding numbers in control samples were 17.0±2.7% and 22.3±4.2%. There was also a significant (P<0.001) effect of individual animals upon the responses to different concentrations of A23187. These findings indicate that (i) A23187 can be used to assess the AR of ovine spermatozoa in vitro and (ii) there are effects of individual animals upon the proportion of spermatozoa undergoing AR.

scholarly journals Cytotoxicity assessment of Bio-C Repair Íon+: A new calcium silicate-based cement

2021 ◽  
Vol 15 (3) ◽  
pp. 152-156
Author(s):  
Celso Afonso Klein-Junior ◽  
Roberto Zimmer ◽  
Tãnyre Dobler ◽  
Vanessa Oliveira ◽  
Daniel Rodrigo Marinowic ◽  
...  
Keyword(s):  
Cell Viability ◽  
Calcium Silicate ◽  
Metallic Matrix ◽  
Physical And Mechanical Properties ◽  
Mean Values ◽  
Significance Level ◽  
Pulp Capping ◽  
Cytotoxicity Assessment ◽  
And Control ◽  
Nih 3T3

Background. Direct pulp capping is a method designed to preserve the exposed dental pulp. Due to good biological, physical, and mechanical properties, new versions of calcium silicate-based materials have been developed as pulp capping materials. The present study aimed to evaluate the cytotoxic effects of four calcium silicate-based pulp capping materials, of which the Bio-C Repair Íon+ is still in an experimental phase. Methods. Biodentine, MTA Repair HP, Bio-C Repair, and Bio-C Repair Íon+ cements were dispensed in a metallic matrix to produce 125-mm3 specimens, which were immersed in Dulbecco’s Modified Eagle Medium (DMEM) to obtain extracts. NIH 3T3 cells were cultured and exposed to the extracts for 24 hours and seven days. Cell viability was assessed by the methyl tetrazolium test (MTT). The mean values for the experimental and control groups (without treatment) were compared by analysis of variance (ANOVA) and post hoc Tukey tests, considering a significance level of 5%. Results. All the tested materials demonstrated a reduction in cell viability (P<0.05). According to ISO 10993-5: 2009 (E), Bio-C Repair Íon+ exhibited mild and moderate cytotoxicity in the 24- hour and 7-day analyses, respectively. Bio-C Repair and Biodentine showed mild cytotoxicity, and MTA Repair HP exhibited moderate cytotoxicity at both intervals. Conclusion. The highest cell viability was demonstrated by Biodentine, MTA, and Repair HP, in descending order. Bio-C Repair and Bio-C Repair Íon+ showed moderate cytotoxicity, similar to MTA Repair HP in the 7-day analysis.

scholarly journals Cytotoxicity and Bioactivity of Dental Pulp-Capping Agents towards Human Tooth-Pulp Cells: A Systematic Review of In-Vitro Studies and Meta-Analysis of Randomized and Controlled Clinical Trials

Materials ◽  
2020 ◽  
Vol 13 (12) ◽  
pp. 2670 ◽  
Author(s):  
Mariano S. Pedano ◽  
Xin Li ◽  
Kumiko Yoshihara ◽  
Kirsten Van Landuyt ◽  
Bart Van Meerbeek
Keyword(s):  
Calcium Hydroxide ◽  
Calcium Silicate ◽  
Dental Pulp ◽  
Meta Analysis ◽  
Capping Agents ◽  
Human Dental Pulp Cells ◽  
Pulp Capping ◽  
Human Dental Pulp ◽  
Pulp Cells

Background. In the era of biology-driven endodontics, vital pulp therapies are regaining popularity as a valid clinical option to postpone root-canal treatment. In this sense, many different materials are available in the market for pulp-capping purposes. Objectives. The main aim of this systematic review and meta-analysis was to examine literature regarding cytotoxicity and bioactivity of pulp-capping agents by exposure of human dental pulp cells of primary origin to these materials. A secondary objective was to evaluate the inflammatory reaction and reparative dentin-bridge formation induced by the different pulp-capping agents on human pulp tissue. Data sources. A literature search strategy was carried out on PubMed, EMBASE and the Web of Science databases. The last search was done on 1 May 2020. No filters or language restrictions were initially applied. Two researchers independently selected the studies and extracted the data. Study selection included eligibility criteria, participants and interventions, study appraisal and synthesis methods. In vitro studies were included when human dental pulp cells of primary origin were (in) directly exposed to pulp-capping agents. Parallel or split-mouth randomized or controlled clinical trials (RCT or CCT) were selected to investigate the effects of different pulp-capping agents on the inflammation and reparative bridge-formation capacity of human pulp tissue. Data were synthesized via odds ratios (95% confidence interval) with fixed or random effects models, depending on the homogeneity of the studies. The relative risks (95% confidence interval) were presented for the sake of interpretation. Results. In total, 26 in vitro and 30 in vivo studies were included in the systematic review and meta-analysis, respectively. The qualitative analysis of in vitro data suggested that resin-free hydraulic calcium-silicate cements promote cell viability and bioactivity towards human dental pulp cells better than resin-based calcium-silicate cements, glass ionomers and calcium-hydroxide cements. The meta-analysis of the in vivo studies indicated that calcium-hydroxide powder/saline promotes reparative bridge formation better than the popular commercial resin-free calcium-silicate cement Pro-Root MTA (Dentsply-Sirona), although the difference was borderline non-significant (p = 0.06), and better than calcium-hydroxide cements (p < 0.0001). Moreover, resin-free pulp-capping agents fostered the formation of a complete reparative bridge better than resin-based materials (p < 0.001). On the other hand, no difference was found among the different materials tested regarding the inflammatory effect provoked at human pulp tissue. Conclusions. Calcium-hydroxide (CH) powder and Pro-Root MTA (Dentsply-Sirona) have shown excellent biocompatibility in vitro and in vivo when tested on human cells and teeth. Their use after many years of research and clinical experience seems safe and proven for vital pulp therapy in healthy individuals, given that an aseptic environment (rubber dam isolation) is provided. Although in vitro evidence suggests that most modern hydraulic calcium-silicate cements promote bioactivity when exposed to human dental pulp cells, care should be taken when these new materials are clinically applied in patients, as small changes in their composition might have big consequences on their clinical efficacy. Key findings (clinical significance). Pure calcium-hydroxide powder/saline and the commercial resin-free hydraulic calcium-silicate cement Pro-Root MTA (Dentsply-Sirona) are the best options to provide a complete reparative bridge upon vital pulp therapy. Systematic review registration number. PROSPERO registration number: CRD42020164374.

scholarly journals THE CIRCUMFUSION SYSTEM FOR MULTIPURPOSE CULTURE CHAMBERS

1968 ◽  
Vol 39 (2) ◽  
pp. 430-450 ◽  
Author(s):  
George G. Rose ◽  
M. Kumegawa ◽  
M. Cattoni
Keyword(s):  
Phase Contrast ◽  
Cultured Cells ◽  
Morphological Characteristics ◽  
Time Lapse ◽  
Newborn Mouse ◽  
Contrast Microscopy ◽  
Free Environment ◽  
Parenchymal Cells

The circumfusion system is a complex in vitro pumping unit incorporating 12 multipurpose culture chambers through which a serum-supplemented fluid nutrient is recirculated at a rate of 4.5 ml/min per chamber. This system was used to study the differentiative responses of fetal and newborn mouse liver explants placed in the serum-free environment formed between the sheets of unperforated cellophane and cover glasses of the chambers. Hepatocytes (parenchymal cells) were discernible in 3–5 days. They retained many of their features of differentiation in the circumfusion system for more than 120 days of cultivation. The living morphological characteristics of the hepatocytes were studied by phase-contrast microscopy (direct viewing and time-lapse cinemicrography) and by special cytochemical staining. Electron micrographs were made of both fresh liver specimens and the cultured cells. Comparisons of the cultured parenchymal cells with their in vivo progenitors showed a remarkable preservation of their differentiated state.

scholarly journals Detection of Cell Aggregation and Altered Cell Viability by Automated Label-Free Video Microscopy

2014 ◽  
Vol 20 (3) ◽  
pp. 372-381 ◽  
Author(s):  
Obaid Aftab ◽  
Mårten Fryknäs ◽  
Ulf Hammerling ◽  
Rolf Larsson ◽  
Mats G. Gustafsson
Keyword(s):  
Cell Viability ◽  
Phase Contrast ◽  
Growth Factor Receptor ◽  
Cell Aggregation ◽  
Well Being ◽  
Time Lapse ◽  
Video Microscopy ◽  
Label Free ◽  
Monitoring Method

Automated phase-contrast video microscopy now makes it feasible to monitor a high-throughput (HT) screening experiment in a 384-well microtiter plate format by collecting one time-lapse video per well. Being a very cost-effective and label-free monitoring method, its potential as an alternative to cell viability assays was evaluated. Three simple morphology feature extraction and comparison algorithms were developed and implemented for analysis of differentially time-evolving morphologies (DTEMs) monitored in phase-contrast microscopy videos. The most promising layout, pixel histogram hierarchy comparison (PHHC), was able to detect several compounds that did not induce any significant change in cell viability, but made the cell population appear as spheroidal cell aggregates. According to recent reports, all these compounds seem to be involved in inhibition of platelet-derived growth factor receptor (PDGFR) signaling. Thus, automated quantification of DTEM (AQDTEM) holds strong promise as an alternative or complement to viability assays in HT in vitro screening of chemical compounds.

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