Evaluation of Commercial Latex as a Positive Control for &lt;i&gt;In Vitro&lt;/i&gt; Testing of Bioceramics

Lourenço, Emanuelle Stellet; Côrtes, Juliana; Costa, Joyce; Linhares, Adriana; Alves, Gutemberg Gomes

doi:10.4028/www.scientific.net/kem.631.357

Cited by 10 publications

(9 citation statements)

References 9 publications

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“…Negative control groups were composed of cells plus culture medium (Alpha-MEM) and polystyrene plus culture medium. Extracts of latex fragments on culture medium were the positive control group, since its consistent cytotoxicity in ISO-based assays was previously demonstrated (10). Each condition was tested using three replicates in each of the three different assays.…”

Section: Sample Preparationmentioning

confidence: 99%

Cytotoxicity Evaluation of Two Bis-Acryl Composite Resins Using Human Gingival Fibroblasts

et al. 2016

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Bis-acryl resins are used for temporary dental restorations and have shown advantages over other materials. The aim of this work was to evaluate the in vitro cytotoxicity of two bis-acryl composite resins (Protemp 4 and Luxatemp Star), obtained at 1, 7 and 40 days after mixing the resin components, using a standardized assay employing human primary cells closely related to oral tissues. Human gingival fibroblast cell cultures were exposed for 24 h to either bis-acryl composite resins, polystyrene beads (negative control) and latex (positive control) extracts obtained after incubation by the different periods, at 37 °C under 5% CO2. Cell viability was evaluated using a multiparametric procedure involving sequential assessment (using the same cells) of mitochondrial activity (XTT assay), membrane integrity (neutral red test) and total cell density (crystal violet dye exclusion test). The cells exposed to the resin extracts showed cell viability indexes exceeding 75% after 24 h. Even when cells were exposed to extracts prepared with longer conditioning times, the bis-acryl composite resins showed no significant cytotoxic effects (p>0.05), compared to the control group or in relation to the first 24 h of contact with the products. There were no differences among the results obtained for the bis-acryl composite resins evaluated 24 h, 7 days and 40 days after mixing. It may be concluded that the bis-acryl resins Protemp 4 and Luxatemp Star were cytocompatible with human gingival fibroblasts, suggesting that both materials are suitable for use in contact with human tissues.

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Section: Sample Preparationmentioning

confidence: 99%

Cytotoxicity Evaluation of Two Bis-Acryl Composite Resins Using Human Gingival Fibroblasts

et al. 2016

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“…Figure 5(a) confirms, as expected, that surgical grade titanium and polystyrene are detected as being not cytotoxic, while the latex showed high cytotoxicity, in a dose dependent manner, as expected. However, we can see that the toxicity levels observed for the positive control were much lower than those usually observed in two-dimensional culture, which attain, in the same conditions, cytotoxicity approximately 3 times higher for MC3T3-E1 cells [17]. This lower toxicity may reflect results closer to the clinic, where latex gloves, tourniquets, probes, and tubes are used constantly in direct contact with the body of patients without causing necrosis or actual levels of relevant chronic toxicity or allergy, when not associated with powder [28,29].…”

Section: Resultsmentioning

confidence: 52%

“…As representatives of negative, biocompatible controls, we employed smooth surface machined titanium (SIN Implants, Brazil) and dense beads of polystyrene (INT, Brazil). Amber latex tubes obtained commercially were used as positive controls, since they are known for their increased cytotoxicity [17].…”

Section: Methodsmentioning

confidence: 99%

“…International standards for evaluation of medical and dental use materials (ISO 10993-1:2009; ASTM) recommend the use of control materials of recognized biocompatibility (negative controls) or well-known cellular toxicity (positive controls of cytotoxicity) [17]. In addition, it is recommended by ISO 10993-5:2009 [18] that testing should be performed during the late log phase of cell growth, attaining approximately 80% confluence.…”

Section: Introductionmentioning

confidence: 99%

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A 3D OsteoblastIn VitroModel for the Evaluation of Biomedical Materials

Restle

Costa-Silva

Lourenço

et al. 2015

Advances in Materials Science and Engineering

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Biomedical materials for bone therapy are usually assessed for their biocompatibility and safety employing animal models orin vitromonolayer cell culture assays. However, alternativein vitromodels may offer controlled conditions closer to physiological responses and reduce animal testing. In this work, we developed a 3D spheroidal cell culture with potential to evaluate simultaneously material-cell and cell-cell interactions. Different cell densities of murine MC3T3-E1 preosteoblasts or human primary osteoblasts (HOb) were used to determine the ideal procedure of spheroidal cultures and their adequacy to material testing. Cells were seeded on 96-well plates coated with agar and incubated in agitation from 1 to 7 days. Aggregate morphology was qualitatively evaluated considering the shape, size, repeatability, handling, and stability of spheroids. Higher cell densities induced more stable spheroids, and handling was considered appropriate starting from 2 × 104cells. Confocal microscopy and Scanning Electron Microscopy indicate that most cells within the aggregate core are viable. Exposure to positive controls has shown a dose dependent cell death as measured by XTT assay. Aggregates were stable and presented good viability when employed on standardized testing of metallic and polymer-based biomaterials. Therefore, osteoblast spheroids may provide a promising tool for material screening and biocompatibility testing.

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“…As previously described for latex, mitochondrial activity is more impacted than cell density and membrane integrity, even though all parameters remained under 25% of cell survival when compared to the control. 23 This multiparametric methodology confirmed that Biodentine is cytocompatible with human bone cells, even when challenged with long-time extractions of possible toxicants. It is not easy to compare the results of different cell culture experiments due to the numerous variations in experimental conditions, such as cell type, cell material contact method, and exposure time.…”

Section: Discussionmentioning

confidence: 59%

BiodentineTM is cytocompatible with human primary osteoblasts

et al. 2017

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Calcium silicate-based materials have been widely studied due to their resemblance to, and similar applicability of, mineral trioxide aggregate (MTA). Among these, Biodentine™ (BD) was specifically designed as a "dentin replacement" material for applications such as root perforations, apexification, treatment of resorptive lesions, and as a retrograde filling material. The present study aimed to assess the in vitro response of human primary osteoblasts to BD using MTA Angelus TM as a reference material, by simultaneously analyzing three different cell viability parameters, namely mitochondrial activity, membrane integrity, and cell density. BD and MTA extracts were prepared by incubation on culture media for 24 h or 42 days after mixing. Primary human osteoblasts were exposed to extracts for 24 h, at 37 o C with 5% CO 2 , and cell viability was evaluated by the XTT, NRU, and CVDE assays. Both materials induced cell viability levels higher than 70% when extracted for 24 h. However, when cells were exposed to extracts with increased conditioning times, MTA presented significant cytotoxic effects (p < 0.05) in comparison to the control and MTA at 24 h. After 42 days, the XTT assay identified a significant reduction in cell viability by BD when compared to the control (p<0.05), despite the fact that levels above the 70% viability cutoff were attained for biocompatible materials. It can be concluded that BD is cytocompatible with human primary osteoblasts, indicating its adequacy in direct contact with bone tissues.

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Evaluation of Commercial Latex as a Positive Control for <i>In Vitro</i> Testing of Bioceramics

Cited by 10 publications

References 9 publications

Cytotoxicity Evaluation of Two Bis-Acryl Composite Resins Using Human Gingival Fibroblasts

Cytotoxicity Evaluation of Two Bis-Acryl Composite Resins Using Human Gingival Fibroblasts

A 3D OsteoblastIn VitroModel for the Evaluation of Biomedical Materials

BiodentineTM is cytocompatible with human primary osteoblasts

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