scholarly journals Combined Fluorescence-Based in Vitro Assay for the Simultaneous Detection of Cell Viability and Alkaline Phosphatase Activity during Osteogenic Differentiation of Osteoblast Precursor Cells

2020 ◽  
Vol 3 (2) ◽  
pp. 30 ◽  
Author(s):  
Sebastian Wilkesmann ◽  
Fabian Westhauser ◽  
Joerg Fellenberg
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Osteogenic Differentiation ◽  
Simultaneous Detection ◽  
Cell Number ◽  
Precursor Cells ◽  
Vitro Assay ◽  
Alp Activity ◽  
Bone Substitute Materials

Novel bone substitute materials need to be evaluated in terms of their osteogenic differentiation capacity and possible unwanted cytotoxic effects in order to identify promising candidates for the therapy of bone defects. The activity of alkaline phosphatase (ALP) is frequently quantified as an osteogenic marker, while various colorimetric assays, like MTT assay, are used to monitor cell viability. In addition, the DNA or protein content of the samples needs to be quantified for normalization purposes. As this approach is time consuming and often requires the analysis of multiple samples, we aimed to simplify this process and established a protocol for the combined fluorescence-based quantification of ALP activity and cell viability within one single measurement. We demonstrate that the fluorogenic substrate 4-methylumbelliferone-phosphate (4-MUP) and the commonly used para-nitrophenylphosphate (p-NPP) produce comparable and highly correlating results. We further show that fluorescein–diacetate (FDA) can be used to quantify both cell viability and cell number without interfering with the quantification of ALP activity. The measurement of additional normalization parameters is, therefore, unnecessary. Therefore, the presented assay allows for a time-efficient, simple and reliable analysis of both ALP activity and cell viability from one sample and might facilitate experiments evaluating the osteogenic differentiation of osteoblast precursor cells.

Osteogenic differentiation of equine cord blood multipotent mesenchymal stromal cells within coralline hydroxyapatite scaffolds in vitro

2011 ◽  
Vol 24 (05) ◽  
pp. 354-362 ◽  
Author(s):  
R. J. Figueroa ◽  
T. G. Koch ◽  
D. H. Betts
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Osteogenic Differentiation ◽  
Alkaline Phosphatase Activity ◽  
Stromal Cells ◽  
Culture Medium ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Coralline Hydroxyapatite ◽  
Osteogenic Induction

SummaryObjective: To investigate the osteogenic differentiation potential of equine umbilical cord blood-derived multipotent mesenchymal stromal cells (CB-MSC) within coralline hydro-xyapatite scaffolds cultured in osteogenic induction culture medium.Methods: Scaffolds seeded with equine CBMSC were cultured in cell expansion culture medium (control) or osteogenic induction medium (treatment). Cell viability and distribution were confirmed by the MTT cell viability assay and DAPI nuclear fluorescence staining, respectively. Osteogenic differentiation was evaluated after 10 days using reverse transcription polymerase chain reaction, alkaline phosphatase activity, and secreted osteocalcin concentration. Cell morphology and matrix deposition were assessed by scanning electron microscopy (SEM) after 14 days in culture.Results: Cells showed viability and adequate distribution within the scaffold. Successful osteogenic differentiation within the scaffolds was demonstrated by the increased expression of osteogenic markers such as Runx2, osteopontin, osteonectin, collagen IA increased levels of alkaline phosphatase activity increased osteocalcin protein secretion and bone-like matrix presence in the scaffold pores upon SEM evaluation.Clinical significance: These results demonstrate that equine CB-MSC maintain viability and exhibit osteogenic potential in coralline hydroxyapatite scaffolds when induced in vitro. Equine CB-MSC scaffold constructs deserve further investigation for their potential role as biologically active fillers to enhance bone-gap repair in the horse.

Increased proliferation of osteoblast precursor cells in estrogen-deficient rats

1993 ◽  
Vol 264 (2) ◽  
pp. E190-E196 ◽  
Author(s):  
D. Modrowski ◽  
L. Miravet ◽  
M. Feuga ◽  
P. J. Marie
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Number ◽  
Estrogen Deficiency ◽  
Precursor Cells ◽  
Proliferation Rate ◽  
Bone Surface ◽  
Post Surgery ◽  
Ovx Rats

We have evaluated the in vivo and in vitro changes in osteoblast characteristics induced by estrogen deficiency and 17 beta-estradiol (E2) treatment in ovariectomized (OVX) rats. Estrogen deficiency induced osteopenia and increased bone turnover, as evidenced by bone histomorphometry at 1, 3, and 6 mo postovariectomy. Bone surface osteoblastic cells (OB) isolated from tibias of OVX rats, OVX rats treated with E2 (10 micrograms/kg body wt), and sham rats showed no difference in alkaline phosphatase activity and osteocalcin production in vitro. In contrast the proliferation rate of OB cells was higher in OVX rats compared with sham rats at all time points post-surgery, as shown by [3H]thymidine incorporation and cell number. The proliferation rate of alkaline phosphatase-positive marrow cells was also higher in OVX rats compared with sham rats. E2 treatment of OVX rats corrected histologic indexes of bone resorption and formation and normalized OB cell proliferation. induced by estrogen deficiency in OVX rats is related to an increased proliferation of osteoblast precursor cells present in the marrow stroma and along the endosteal bone surface.

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.

scholarly journals Biological characterization of implant surfaces - in vitro study

2015 ◽  
Vol 44 (4) ◽  
pp. 195-199 ◽  
Author(s):  
Priscilla Barbosa Ferreira Soares ◽  
Camilla Christian Gomes Moura ◽  
Huberth Alexandre da Rocha Júnior ◽  
Paula Dechichi ◽  
Darceny Zanetta-Barbosa
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Viability ◽  
Total Protein ◽  
Cell Attachment ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Serum Total Protein ◽  
Trypan Blue Exclusion ◽  
Experimental Surface

<title>Abstract</title><sec><title>Objective</title><p>Evaluate the biological performance of titanium alloys grade IV under different surface treatments: sandblasting and double etching (Experimental surface 1; Exp1, NEODENT); surface with wettability increase (Experimental surface 2; Exp2, NEODENT) on response of preliminary differentiation and cell maturation.</p></sec><sec><title>Material and method</title><p>Immortalized osteoblast cells were plated on Exp1 and Exp2 titanium discs. The polystyrene plate surface without disc was used as control group (C). Cell viability was assessed by measuring mitochondrial activity (MTT) at 4 and 24 h (n = 5), cell attachment was performed using trypan blue exclusion within 4 hours (n = 5), serum total protein and alkaline phosphatase normalization was performed at 4, 7 and 14 days (n = 5). Data were analyzed using one-way ANOVA and Tukey test.</p></sec><sec><title>Result</title><p>The values of cell viability were: 4h: C– 0.32±0.01<sup>A</sup>; Exp1– 0.34±0.08<sup>A</sup>; Exp2– 0.29±0.03<sup>A</sup>. 24h: C– 0.43±0.02<sup>A</sup>; Exp1– 0.39±0.01<sup>A</sup>; Exp2– 0.37±0.03<sup>A</sup>. The cell adhesion counting was: C– 85±10<sup>A</sup>; Exp1- 35±5<sup>B</sup>; Exp2– 20±2<sup>B</sup>. The amounts of serum total protein were 4d: C– 40±2<sup>B</sup>; Exp1– 120±10<sup>A</sup>; Exp2– 130±20<sup>A</sup>. 7d: C– 38±2<sup>B</sup>; Exp1– 75±4<sup>A</sup>; Exp2– 70±6<sup>A</sup>. 14 d: C– 100±3<sup>A</sup>; Exp1– 130±5<sup>A</sup>; Exp2– 137±9<sup>A</sup>. The values of alkaline phosphatase normalization were: 4d: C– 2.0±0.1<sup>C</sup>; Exp1– 5.1±0.8<sup>B</sup>; Exp2– 9.8±2.0<sup>A</sup>. 7d: C– 1.0±0.01<sup>C</sup>; Exp1– 5.3±0.5<sup>A</sup>; Exp2– 3.0±0.3<sup>B</sup>. 14 d: C– 4.1±0.3<sup>A</sup>; Exp1– 4.4±0.8<sup>A</sup>; Exp2– 2.2±0.2<sup>B</sup>. Different letters related to statistical differences.</p></sec><sec><title>Conclusion</title><p>The surfaces tested exhibit different behavior at dosage of alkaline phosphatase normalization showing that the Exp2 is more associated with induction of cell differentiation process and that Exp1 is more related to the mineralization process.</p></sec>

The inhibitory effects of vancomycin on rat bone marrow–derived mesenchymal stem cell differentiation

2021 ◽  
pp. 1-5
Author(s):  
Kari Hanson ◽  
Carly Isder ◽  
Kristen Shogren ◽  
Anthony L. Mikula ◽  
Lichun Lu ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
High Dose ◽  
Inhibitory Effects ◽  
Alizarin Red ◽  
Osteogenic Factors

OBJECTIVE The use of intrawound vancomycin powder in spine surgery has been shown to decrease the rate of surgical site infections; however, the optimal dose is unknown. High-dose vancomycin inhibits osteoblast proliferation in vitro and may decrease the rate of solid arthrodesis. Bone marrow–derived mesenchymal stem cells (BMSCs) are multipotent cells that are a source of osteogenesis in spine fusions. The purpose of this study was to determine the effects of vancomycin on rat BMSC viability and differentiation in vitro. METHODS BMSCs were isolated from the femurs of immature female rats, cultured, and then split into two equal groups; half were treated to stimulate osteoblastic differentiation and half were not. Osteogenesis was stimulated by the addition of 50 µg/mL l-ascorbic acid, 10 mM β-glycerol phosphate, and 0.1 µM dexamethasone. Vancomycin was added to cell culture medium at concentrations of 0, 0.04, 0.4, or 4 mg/mL. Early differentiation was determined by alkaline phosphatase activity (4 days posttreatment) and late differentiation by alizarin red staining for mineralization (9 days posttreatment). Cell viability was determined at both the early and late time points by measurement of formazan colorimetric product. RESULTS Viability within the first 4 days decreased with high-dose vancomycin treatment, with cells receiving 4 mg/mL vancomycin having 40%–60% viability compared to the control. A gradual decrease in alizarin red staining and nodule formation was observed with increasing vancomycin doses. In the presence of the osteogenic factors, vancomycin did not have deleterious effects on alkaline phosphatase activity, whereas a trend toward reduced activity was seen in the absence of osteogenic factors when compared to osteogenically treated cells. CONCLUSIONS Vancomycin reduced BMSC viability and impaired late osteogenic differentiation with high-dose treatment. Therefore, the inhibitory effects of high-dose vancomycin on spinal fusion may result from both reduced BMSC viability and some impairment of osteogenic differentiation.

scholarly journals A novel in vitro assay to study chondrocyte-to-osteoblast transdifferentiation

Endocrine ◽  
2021 ◽  
Author(s):  
Miriam E. A. Tschaffon ◽  
Stefan O. Reber ◽  
Astrid Schoppa ◽  
Sayantan Nandi ◽  
Ion C. Cirstea ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Molecular Mechanisms ◽  
In Vitro Assay ◽  
Immunofluorescent Staining ◽  
Marker Genes ◽  
Osteogenic Differentiation Medium ◽  
Vitro Assay ◽  
Osteogenic Cells ◽  
Osteogenic Lineage

Abstract Purpose Endochondral ossification, which involves transdifferentiation of chondrocytes into osteoblasts, is an important process involved in the development and postnatal growth of most vertebrate bones as well as in bone fracture healing. To study the basic molecular mechanisms of this process, a robust and easy-to-use in vitro model is desirable. Therefore, we aimed to develop a standardized in vitro assay for the transdifferentiation of chondrogenic cells towards the osteogenic lineage. Methods Murine chondrogenic ATDC5 cells were differentiated into the chondrogenic lineage for seven days and subsequently differentiated towards the osteogenic direction. Gene expression analysis of pluripotency, as well as chondrogenic and osteogenic markers, cell–matrix staining, and immunofluorescent staining, were performed to assess the differentiation. In addition, the effects of Wnt3a and lipopolysaccharides (LPS) on the transdifferentiation were tested by their addition to the osteogenic differentiation medium. Results Following osteogenic differentiation, chondrogenically pe-differentiated cells displayed the expression of pluripotency and osteogenic marker genes as well as alkaline phosphatase activity and a mineralized matrix. Co-expression of Col2a1 and Col1a1 after one day of osteogenic differentiation indicated that osteogenic cells had differentiated from chondrogenic cells. Wnt3a increased and LPS decreased transdifferentiation towards the osteogenic lineage. Conclusion We successfully established a rapid, standardized in vitro assay for the transdifferentiation of chondrogenic cells into osteogenic cells, which is suitable for testing the effects of different compounds on this cellular process.

scholarly journals MTT assay to evaluate the cytotoxic potential of a drug

2017 ◽  
Vol 12 (2) ◽  
pp. 8 ◽  
Author(s):  
Ashutosh Bahuguna ◽  
Imran Khan ◽  
Vivek K. Bajpai ◽  
Sun Chul Kang
Keyword(s):  
Cell Viability ◽  
Mtt Assay ◽  
Colorimetric Assay ◽  
Video Clip ◽  
Cell Number ◽  
Cytotoxic Potential ◽  
Lung Adenocarcinoma Cell Line ◽  
Full Screen ◽  
Purple Color

<p>Quantification of cell viability and proliferation form the fundamental for numerous <em>in vitro</em> assays in response to external factors. An MTT assay is a colorimetric assay based on assessing the cell metabolic activity. A549 Lung adenocarcinoma cell line was used to see the cytotoxic potential of a new drug for initial screening of apoptosis or necrosis. The biochemical mechanism behind the MTT assay involves NAD(P)H-dependent cellular oxidoreductase enzyme that converts the yellow tetrazolium MTT [3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] into insoluble (E,Z)-5-(4,5-dimethylthiazol-2-yl)-1,3-diphenylformazan (formazan). The formed formazan can be dissolved with dimethyl sulfoxide (DMSO) to give a purple color with characteristic absorption at 540 nm. Intensity of purple color is directly proportional to the cell number and thus indicating the cell viability.</p><p><strong>Video Clip of Methodology:</strong> 3 min 56 sec  <a href="https://www.youtube.com//v/eqFxzDVunt8">Full screen</a>   <a href="https://www.youtube.com/watch?v=eqFxzDVunt8">If Failed</a></p>

332 GRANULOCYTE-MACROPHAGE COLONY STIMULATING FACTOR (GM-CSF) ENHANCES CUMULUS CELLS EXPANSION OF IN VITRO-MATURED BOVINE CUMULUS - OOCYTE COMPLEXES IN A CHEMICALLY DEFINED MEDIUM

2010 ◽  
Vol 22 (1) ◽  
pp. 322
Author(s):  
D. D. Bücher ◽  
M. A. Castro ◽  
M. E. Silva ◽  
M. A. Berland ◽  
I. I. Concha ◽  
...  
Keyword(s):  
Cell Viability ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Cell Number ◽  
Control Group ◽  
Cumulus Expansion ◽  
Gm Csf ◽  
Nuclear Stage

Granulocyte-macrophage colony stimulating factor (GM-CSF) is a pleiotropic cytokine that stimulates proliferation, differentiation and function in different cells types. We have previously demonstrated (Bücher DD et al. 2008 Reprod. Dom. Anim. 43 (Suppl. 3), 146 abst.) that both subunits of GM-CSF receptor are expressed in granulosa cells from antral follicles in bovine ovaries. Also, we determined that the cytokine enhances glucose uptake through facilitative hexose transporters in granulosa cells in primary culture. The goals of the present study were to characterize the expression of GM-CSF receptor in cumulus cells and oocytes from bovine antral follicles and to determine its effects on in vitro-matured bovine COCs in a chemically defined medium. To determine the presence of a and |5 subunits of GM-CSF receptor, COCs were aspirated from follicles <8 mm in diameter, fixed, and submitted to immunocytochemistry. To study the effect of GM-CSF on in vitro maturation of oocytes, COCs (n =481) were cultured using serum-free medium (SOF) containing 0, 1, 10, and 100 ng mL-1 of human recombinant GM-CSF (R&D Systems, Inc., Minneapolis, MN, USA) for 22 h at 39°C, 5% CO2 in humidified air. Nuclear stage, cumulus expansion, cumulus cell number, and viability were analyzed after in vitro maturation. Cumulus expansion was assessed using the cumulus expansion index (CEI) (Fagbohun C and Down S 1990 Biol. Reprod. 42, 413-423). Nuclear stage was evaluated using aceto-orcein stain. To determine cumulus cell viability and number, COCs (n = 10-12 per group) were transferred into an Eppendorf tube and cumulus cells were removed by vortexing for 3 min, stained with trypan blue and counted with a hemocytometer. The study was conducted in 6 replicates. Data from cumulus expansion and cell number were analyzed by Kruskal-Wallis analysis. Data for nuclear stage and cell viability were analyzed by chi-square analysis and one way ANOVA, respectively. Both receptor subunits were present in cumulus cells and oocytes from COCs. COCs cultured in 10 and 100 ng mL-1 GM-CSF had CEI scores (0.8 and 1.22, respectively) greater (P < 0.01) than controls (0.2), but the proportion of COCs displaying second metaphase did not differ (P = 0.5) among treatment groups. GM-CSF at a concentration of 100 ng mL-1 increased (P < 0.01) cumulus cell viability by more than 20% compared to the control group. Similarly, GM-CSF at concentrations of 10 and 100 ng mL-1 increased (P < 0.05) cumulus cell number by more than 20% and 45%, respectively, from the control group. The use of a specific inhibitor of PI3 kinase (Ly294002; 10 and 100 μM) blocked the stimulatory effect of GM-CSF on cumulus expansion, cell viability, and cell number. In conclusion, the results of the study suggest a plausible modulator role of GM-CSF in the metabolism and function of cumulus cells and oocytes during in vitro maturation. Funding from Faculty of Veterinary Sciences, Universidad Austral de Chile, MECESUP AUS-0005, AUS-0601, and DID D-2006-24 and from Universidad Católica de Temuco, research grant 2007 DGI-CDA-04.

RNA Synthesis by DNA Methyltransferase 1 siRNA Transfection with Cationic Liposomes into Osteoblastic Progenitor Cells Based on SiO2 Nanomagnetic Beads for Proliferation and Differentiation

2020 ◽  
Vol 20 (10) ◽  
pp. 6077-6086
Author(s):  
Qingzhen Chen ◽  
Tao Jiang ◽  
Qinshen Wang ◽  
Yongqing Huang ◽  
Min Shao
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Osteogenic Differentiation ◽  
Dna Methyltransferase ◽  
Cationic Liposomes ◽  
Precursor Cells ◽  
Dna Methyltransferase 1 ◽  
Alizarin Red ◽  
Alp Activity ◽  
Proliferation And Differentiation

DNA methylation regulated gene expression is important for osteoblast proliferation and differentiation during bone remodeling and its deregulation leads to the development of osteoporosis. DNA methyltransferase 1 (DNMT1) is an important regulator of DNA methylation. To explore the effect and mechanism of differential expression of DNMT1 in osteoblast precursor cells, DNMT1 siRNAs were designed and synthesized to interfere with DNMT1 expression in the osteoblast precursor cells, MC3T3E1 (Clone 24; MC3T3E1-24). The expression of the target gene, DNMT1, and osteogenic differentiation indicators osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting (WB). MTT assay was used to detect the effect on cell proliferation. Alkaline phosphatase (ALP) activity and alizarin red staining were used to detect the effect of DNMT1 on osteogenic differentiation. Hematoxylin and eosin (H&E) staining was used to detect the morphological changes in MC3T3E1-24 cells. Twenty-four hours following the transfection of MC3T3E1-24 cells with DNMT1 siRNA using cationic liposomes, DNMT1 mRNA and protein levels decreased significantly (P <0.001 for both). The reduced expression of DNMT1 promoted the OPG mRNA and protein expression (P <0.05), increased the ratio of OPG to RANKL (P <0.05), inhibited the expression of RANKL (P <0.01) without affecting the RANKL gene expression (not significant, P >0.05). The reduced expression of DNMT1 also promoted the proliferation of osteoblast precursor cells. In addition, ALP activity test and alizarin red staining showed that reduced expression of DNMT1 resulted in an increase in OPG/RANKL ratio and promoted the differentiation of the precursor cells. The cultured cells were found to have fibroblast-like appearance, and calcium nodules were observed after 7 days of conventional culture. In addition, to improve the efficiency of RNA extraction and save time, a type of silica nanomagnetic beads was used in the early stage of this study to extract RNA and assist qPCR detection of the target genes. The results showed that the magnetic beads could effectively extract RNA from the cells. In conclusion, low expression of DNMT1 affects proliferation and maturation of osteoblasts by upregulating OPG and OPG/RANKL ratio.

scholarly journals Viability and Alkaline Phosphatase Activity of Human Dental Pulp Cells after Exposure to Yellowfin Tuna Bone-Derived Hydroxyapatite In Vitro

2020 ◽  
Vol 2020 ◽  
pp. 1-5
Author(s):  
Tetiana Haniastuti ◽  
Heni Susilowati ◽  
Margareta Rinastiti
Keyword(s):  
Alkaline Phosphatase ◽  
Dental Pulp ◽  
Colorimetric Assay ◽  
Third Molar ◽  
Yellowfin Tuna ◽  
Dental Pulp Cells ◽  
Alp Activity ◽  
High Calcium ◽  
Pulp Cells

The bone of yellowfin tuna (Thunnus albacares) contains high calcium and phosphor and can be synthesized into hydroxyapatite (HA). Due to its mineral content and similarity in chemical composition with human hard tissue, HA may have potency as a pulp capping material. The aim of this in vitro study was to evaluate the viability and alkaline phosphatase (ALP) activity of dental pulp cells after exposure to HA synthesized from yellowfin tuna bone (THA). Pulp cells were isolated from human-impacted third molar. To evaluate the viability of the pulp cells, the cells were cultured and exposed to various concentrations (6.25 to 200 μg/ml) of THA for 24, 48, and 72 hours. For ALP activity assay, pulp cells were cultured with odontoblastic differentiation media and exposed to THA for 7, 11, and 15 days. ALP activity was then determined using an ALP colorimetric assay kit. Results showed that the viability of the cells was more than 91% after exposure to various concentrations of THA and the cells demonstrated normal cell morphology in all observation periods. The ALP activity test revealed that groups exposed to THA for 7, 11, and 15 days showed higher ALP activity than the control groups ( p < 0.05 ). It is concluded that THA had no cytotoxic effect on pulp cells; furthermore, it enhanced proliferation as well as ALP activity of the pulp cells.

Sign in / Sign up

Export Citation Format

Share Document