Publications by authors named "Elisandra Márcia Rodrigues"

10 Publications

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Physicochemical properties and effect of bioceramic root canal filling for primary teeth on osteoblast biology.

J Appl Oral Sci 2021 3;29:e20200870. Epub 2021 May 3.

Universidade Estadual Paulista "Julio de Mesquita Fillho" (UNESP) - Faculdade de Odontologia de Araraquara - Departamento de Odontologia Restauradora - Araraquara - SP - Brasil.

Objective: Bio-C Pulpecto (Bio-CP) was recently developed as the first bioceramic root filling material for primary teeth. To evaluate the physicochemical properties of radiopacity, setting time, pH, cytocompatibility and potential of Bio-CP to induce mineralisation, compared with (1) Calen thickened with zinc oxide (Calen-ZO), and (2) zinc oxide and eugenol (ZOE).

Methodology: Physicochemical properties were evaluated according to ISO 6876. Saos-2 (human osteoblast-like cell line) exposed to extracts of the materials were subjected to assays of methyl thiazolyl tetrazolium, neutral red, alkaline phosphatase (ALP) activity and mineralised nodule production. The results were analysed using one-way or two-way ANOVA and Tukey's or Bonferroni's post-tests (α=0.05).

Results: All the materials showed radiopacity higher than 3 mm Al. Bio-CP had lower pH than Calen-ZO, but higher pH than ZOE. Calen-ZO and Bio-CP did not set. The setting time for ZOE was 110 min. The cytocompatibility order was Calen-ZO > Bio-CP > ZOE (1:2, 1:4 dilutions) and Calen-ZO > Bio-CP = ZOE (1:12, 1:24 dilutions) and Calen-ZO = Bio-CP > ZOE (1:32 dilution). Bio-CP induced greater ALP activity at 7 days, and greater mineralised nodule production, compared to Calen-ZO (p<0.05). Conclusions Bio-CP showed adequate physicochemical properties, cytocompatibility and potential to induce mineralisation.
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http://dx.doi.org/10.1590/1678-7757-2020-0870DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8092806PMC
May 2021

Physicochemical, biological, and antibacterial evaluation of tricalcium silicate-based reparative cements with different radiopacifiers.

Dent Mater 2021 02 13;37(2):311-320. Epub 2020 Dec 13.

Department of Restorative Dentistry, São Paulo State University (UNESP), School of Dentistry, Araraquara, SP, Brazil. Electronic address:

Objective: To evaluate tricalcium silicate-based (TCS) experimental materials, associated with zirconium oxide (ZrO), calcium tungstate (CaWO) or niobium oxide (NbO) radiopacifiers, in comparison with MTA Repair HP (Angelus).

Methods: Physicochemical tests: setting time, radiopacity, pH and solubility. In vitro assays: cytotoxicity: MTT and Neutral Red - NR; cell bioactivity: alkaline phosphatase activity (ALP), Alzarin red staining (ARS) and real time PCR (qPCR). Antibacterial activity: direct contact on Enterococcus faecalis in the planktonic form. Physicochemical and ARS data were submitted to ANOVA/Tukey tests; antibacterial activity, to Kruskall-Wallis and Dunn tests; MTT, NR, ALP and qPCR were analyzed by ANOVA/Bonferroni tests (α = 0.05).

Results: TCS + CaWO presented the longest setting time and MTA HP the shortest. Except for TCS, all the materials presented radiopacity above 3 mmAl. The cements had alkaline pH, antibacterial activity, low solubility and no cytotoxic effects. The highest ALP activity occurred in 14 days, especially to TCS, TCS + ZrO and TCS + CaWO. TCS + ZrO, TCS + NbO and MTAHP had higher mineralized nodule formation than those of the negative control (NC). After 7 days, there was no difference in mRNA expression for ALP, when compared to NC. However, after 14 days there was no overexpressed ALP mRNA, especially TCS + NbO, in relation to the CN. All the materials presented antimicrobial action.

Significance: The pure tricalcium silicate associated with ZrO, CaWO or NbO had appropriate physicochemical properties, antibacterial activity, cytocompatibility and induced mineralization in Saos-2, indicating their use as reparative materials.
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http://dx.doi.org/10.1016/j.dental.2020.11.014DOI Listing
February 2021

In vivo and in vitro anti-inflammatory and pro-osteogenic effects of citrus cystatin CsinCPI-2.

Cytokine 2019 11 18;123:154760. Epub 2019 Jun 18.

Department of Diagnosis and Surgery, School of Dentistry at Araraquara, Sao Paulo State University - UNESP, Araraquara, São Paulo, Brazil. Electronic address:

Cystatins are natural inhibitors of cysteine peptidases. Recently, cystatins derived from plants, named phytocystatins, have been extensively studied. Among them, CsinCPI-2 proteins from Citrus sinensis were identified and recombinantly produced by our group. Thus, this study described the recombinant expression, purification, and inhibitory activity of this new phytocystatin against human cathepsins K and B and assessed the anti-inflammatory effect of CsinCPI-2 in vitro in mouse and in vivo in rats. In addition, the pro-osteogenic effect of CsinCPI-2 was investigated in vitro. The inflammatory response of mouse macrophage cells stimulated with P. gingivalis was modulated by CsinCPI-2. The in vitro results showed an inhibitory effect (p < 0.05) on cathepsin K, cathepsin B, IL-1β, and TNF-α gene expression. In addition, CsinCPI-2 significantly inhibited in vivo the activity of TNF-α (p < 0.05) in the blood of rats, previously stimulated by E. coli lipopolysaccharide (LPS). CsinCPI-2 had a pro-osteogenic effect in human dental pulp cells, demonstrated by the increase in alkaline phosphatase (ALP) activity, deposition of mineralized nodules, and the gene expression of the osteogenic markers as bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (Runx-2), ALP, osteocalcin, and bone sialoprotein (BSP). These preliminary studies suggested that CsinCPI-2 has a potential anti-inflammatory, and at the same time, a pro-osteogenic effect. This may lead to new therapies for the control of diseases where inflammation plays a key role, such as periodontal disease and apical periodontitis.
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http://dx.doi.org/10.1016/j.cyto.2019.154760DOI Listing
November 2019

Effects of Calcium Hypochlorite and Octenidine Hydrochloride on L929 And Human Periodontal Ligament Cells.

Braz Dent J 2019 Jun 3;30(3):213-219. Epub 2019 Jun 3.

Department of Restorative Dentistry, Araraquara School of Dentistry, Universidade Estadual Paulista, Araraquara, SP, Brazil.

The aim of this study was to assess cytotoxicity and cell migration of calcium hypochlorite [Ca(OCl)2] and octenidine hydrochloride - OCT (Octenisept®, Schülke & Mayr, Norderstedt, Germany) in L929 and human periodontal ligament (hPDL) cells. The cells were exposed to different doses of different solutions: 2.5% and 5% Ca(OCl)2, 0.1% OCT, 2.5% NaOCl and 2% CHX for 10 min. Cell viability was assessed by methyl-thiazol-tetrazolium (MTT) and neutral red (NR) assays, and cell migration was determined by wound-healing assay. Statistical analysis was performed by two-way ANOVA and Bonferroni tests (α=0.05). The MTT and NR assays revealed that 0.1% OCT was less cytotoxic in hPDL cells (p<0.05), followed by 2% CHX and 2.5% Ca(OCl)2 (p<0.05). There was no significant difference between 2.5% NaOCl and 5% Ca(OCl)2 (p>0.05), but these solutions showed greater cytotoxicity than the others. The result was the same for L929 cells, except that there was no significant difference between 2% CHX and 2.5% Ca(OCl)2 (p>0.05). Wound-healing assay in L929 and hPDL cells showed that cell migration of 0.1% OCT, 2% CHX and 2.5% Ca(OCl)2 groups was higher than 5% Ca(OCl)2 and 2.5% NaOCl groups at 24 h (p<0.05). In conclusion, 0.1% OCT had lower cytotoxicity in tested cell lines than CHX, Ca(OCl)2 and NaOCl. Cell migration was higher for 0.1% OCT, 2% CHX and 2.5% Ca(OCl)2. Therefore, in terms of cytotoxicity, OCT and Ca(OCl)2 have the potential to be used as root canal irrigants.
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http://dx.doi.org/10.1590/0103-6440201902280DOI Listing
June 2019

Addition of zirconium oxide to Biodentine increases radiopacity and does not alter its physicochemical and biological properties.

J Appl Oral Sci 2019 Apr 1;27:e20180429. Epub 2019 Apr 1.

Universidade Estadual Paulista - UNESP, Faculdade de Odontologia de Araraquara, Departamento de Odontologia Restauradora, Araraquara, São Paulo, Brasil.

Objectives: To evaluate the radiopacity of Biodentine (BD) and BD associated with 15% calcium tungstate (BDCaWO4) or zirconium oxide (BDZrO2), by using conventional and digital radiography systems, and their physicochemical and biological properties.

Materials And Methods: Radiopacity was evaluated by taking radiographs of cement specimens (n=8) using occlusal film, photostimulable phosphor plates or digital sensors. Solubility, setting time, pH, cytocompatibility and osteogenic potential were also evaluated. Data were analyzed using one-way ANOVA and Tukey post-test or two-way ANOVA and Bonferroni post-test (α=0.05).

Results: BD radiopacity was lower than 3 mm Al, while BD ZrO2 and BD CaWO4 radiopacity was higher than 3 mm Al in all radiography systems. The cements showed low solubility, except for BDCaWO4. All cements showed alkaline pH and setting time lower than 34 minutes. MTT and NR assays revealed that cements had greater or similar cytocompatibility in comparison with control. The ALP activity in all groups was similar or greater than the control. All cements induced greater production of mineralized nodules than control.

Conclusions: Addition of 15% ZrO2 or CaWO4 was sufficient to increase the radiopacity of BD to values higher than 3 mm Al. BD associated with radiopacifiers showed suitable properties of setting time, pH and solubility, except for BDCaWO4, which showed the highest solubility. All cements had cytocompatibility and potential to induce mineralization in Saos-2 cells. The results showed that adding 15% ZrO2 increases the radiopacity of BD, allowing its radiography detection without altering its physicochemical and biological properties.
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http://dx.doi.org/10.1590/1678-7757-2018-0429DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6442842PMC
April 2019

Ytterbium Oxide as Radiopacifier of Calcium Silicate-Based Cements. Physicochemical and Biological Properties.

Braz Dent J 2018 Sep-Oct;29(5):452-458

Department of Restorative Dentistry, Araraquara Dental School, UNESP - Univ Estadual Paulista, Araraquara, SP, Brazil.

This study evaluated physicochemical properties, cytotoxicity and bioactivity of MTA Angelus (MTA), calcium silicate-based cement (CSC) and CSC with 30% Ytterbium oxide (CSC/Yb2O3). Setting time was evaluated using Gilmore needles. Compressive strength was evaluated in a mechanical machine. Radiopacity was evaluated using radiographs of materials and an aluminum scale. Solubility was evaluated after immersion in water. Cell viability was evaluated by means of MTT assay and neutral red staining, and the mineralization activity by using alkaline phosphatase activity and Alizarin Red staining. The data were submitted to ANOVA, Tukey and Bonferroni tests (5% significance). The bioactive potential was evaluated by scanning electron microscopy. The materials presented similar setting time. MTA showed the lowest compressive strength. MTA and CSC/Yb2O3 presented similar radiopacity. CSC/Yb2O3 showed low solubility. Saos-2 cell viability tests showed no cytotoxic effect, except to 1:1 dilution in NR assay which had lower cell viability when compared to the control. ALP at 1 and 7 days was similar to the control. MTA and CSC had greater ALP activity at 3 days when compared to control. All the materials present higher mineralized nodules when compared with the control. SEM analysis showed structures suggesting the presence of calcium phosphate on the surface of materials demonstrating bioactivity. Ytterbium oxide proved to be a properly radiopacifying agent for calcium silicate-based cement since it did not affected the physicochemical and biological properties besides preserving the bioactive potential of this material.
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http://dx.doi.org/10.1590/0103-6440201802033DOI Listing
April 2019

Influence of the Vehicle and Antibiotic Formulation on Cytotoxicity of Triple Antibiotic Paste.

J Endod 2018 Dec;44(12):1812-1816

Department of Restorative Dentistry, Araraquara School of Dentistry, São Paulo State University-UNESP, Araraquara, São Paulo, Brazil.

Introduction: The aim of this study was to assess the influence of antibiotic formulations (tablet/capsule [TC] or United States Pharmacopeia [USP]-grade antibiotics) and vehicles (water [HO] or macrogol + propylene glycol [MP]) on the cytotoxicity and pH of triple antibiotic pastes (TAPs).

Methods: L929 fibroblasts were exposed to TAPs prepared with TC or USP-grade antibiotics mixed with HO or MP for 72 hours. Each isolated antibiotic with each vehicle, each isolated vehicle, and the culture medium were used as controls. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and neutral red assays. The pH was measured after 3 and 8 hours of immersion of the pastes in water. Data were analyzed using analysis of variance, the Bonferroni or Tukey posttests (α = 0.05), and the Pearson correlation test (α = 0.05).

Results: The pastes prepared with TC were less cytotoxic than pastes prepared with USP-grade antibiotics (P < .05), and pastes with the MP vehicle were less cytotoxic than pastes with HO (P < .05). TC TAP + MP showed the lowest cytotoxicity, whereas USP-grade TAP + HO showed the highest cytotoxicity (P < .05). All TAPs showed a pH ranging from 4.64-5.20. Irrespective of the vehicle, USP-grade TAP showed a lower pH than TC TAP (P < .05). TAPs with HO had a lower pH than TAPs with MP (P < .05).

Conclusions: The vehicle and the antibiotic formulations influenced the cytotoxicity and pH of TAP. The pastes prepared with TC and MP were less acidic and less cytotoxic than the type prepared with USP-grade antibiotics and HO.
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http://dx.doi.org/10.1016/j.joen.2018.09.009DOI Listing
December 2018

Cytotoxicity and Bioactivity of Calcium Silicate Cements Combined with Niobium Oxide in Different Cell Lines.

Braz Dent J 2017 Jan-Feb;28(1):65-71

Department of Restorative Dentistry, Araraquara Dental School, UNESP - Universidade Estadual Paulista, Araraquara, SP, Brazil.

The aim of this study was to evaluate the cytotoxicity and bioactivity of calcium silicate-based cements combined with niobium oxide (Nb2O5) micro and nanoparticles, comparing the response in different cell lines. This evaluation used four cell lines: two primary cultures (human dental pulp cells - hDPCs and human dental follicle cells - hDFCs) and two immortalized cultures (human osteoblast-like cells - Saos-2 and mouse periodontal ligament cells - mPDL). The tested materials were: White Portland Cement (PC), mineral trioxide aggregate (MTA), white Portland cement combined with microparticles (PC/Nb2O5µ) or nanoparticles (PC/Nb2O5n) of niobium oxide (Nb2O5). Cytotoxicity was evaluated by the methylthiazolyldiphenyl-tetrazolium bromide (MTT) and trypan blue exclusion assays and bioactivity by alkaline phosphatase (ALP) enzyme activity. Results were analyzed by ANOVA and Tukey test (a=0.05). PC/Nb2O5n presented similar or higher cell viability than PC/Nb2O5µ in all cell lines. Moreover, the materials presented similar or higher cell viability than MTA. Saos-2 exhibited high ALP activity, highlighting PC/Nb2O5µ material at 7 days of exposure. In conclusion, calcium silicate cements combined with micro and nanoparticles of Nb2O5 presented cytocompatibility and bioactivity, demonstrating the potential of Nb2O5 as an alternative radiopacifier agent for these cements. The different cell lines had similar response to cytotoxicity evaluation of calcium silicate cements. However, bioactivity was more accurately detected in human osteoblast-like cell line, Saos-2.
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http://dx.doi.org/10.1590/0103-6440201700525DOI Listing
January 2018

Cytotoxicity and genotoxicity of calcium silicate-based cements on an osteoblast lineage.

Braz Oral Res 2016 May;30(1)

Universidade Estadual Paulista - UNESP, Araraquara School of Dentistry, Department of Restorative Dentistry, Araraquara, SP, Brazil.

Several calcium silicate-based biomaterials have been developed in recent years, in addition to Mineral Trioxide Aggregate (MTA). The aim of this study was to evaluate the cytotoxicity, genotoxicity and apoptosis/necrosis in human osteoblast cells (SAOS-2) of pure calcium silicate-based cements (CSC) and modified formulations: modified calcium silicate-based cements (CSCM) and three resin-based calcium silicate cements (CSCR1) (CSCR 2) (CSCR3). The following tests were performed after 24 hours of cement extract exposure: methyl-thiazolyl tetrazolium (MTT), apoptosis/necrosis assay and comet assay. The negative control (CT-) was performed with untreated cells, and the positive control (CT+) used hydrogen peroxide. The data for MTT and apoptosis were submitted to analysis of variance and Bonferroni's posttest (p < 0.05), and the data for the comet assay analysis, to the Kruskal-Wallis and Dunn tests (p < 0.05). The MTT test showed no significant difference among the materials in 2 mg/mL and 10 mg/mL concentrations. CSCR3 showed lower cell viability at 10 mg/mL. Only CSC showed lower cell viability at 50 mg/mL. CSCR1, CSCR2 and CSCR3 showed a higher percentage of initial apoptosis than the control in the apoptosis test, after 24 hours exposure. The same cements showed no genotoxicity in the concentration of 2 mg/mL, with the comet assay. CSC and CSCR2 were also not genotoxic at 10 mg/mL. All experimental materials showed viability with MTT. CSC and CSCR2 presented a better response to apoptosis and genotoxicity evaluation in the 10 mg/mL concentration, and demonstrated a considerable potential for use as reparative materials.
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http://dx.doi.org/10.1590/1807-3107BOR-2016.vol30.0048DOI Listing
May 2016

Biocompatibility and bioactivity of calcium silicate-based endodontic sealers in human dental pulp cells.

J Appl Oral Sci 2015 Oct;23(5):467-71

Departamento de Odontologia Restauradora, Escola de Odontologia, Universidade Estadual Paulista, Araraquara, SP, Brazil.

Unlabelled: Mineral Trioxide Aggregate (MTA) is a calcium silicate-based material. New sealers have been developed based on calcium silicate as MTA Fillapex and MTA Plus.

Objective: The aim of this study was to evaluate biocompatibility and bioactivity of these two calcium silicate-based sealers in culture of human dental pulp cells (hDPCs).

Material And Methods: The cells were isolated from third molars extracted from a 16-year-old patient. Pulp tissue was sectioned into fragments with approximately 1 mm3 and kept in supplemented medium to obtain hDPCs adherent cultures. Cell characterization assays were performed to prove the osteogenic potential. The evaluated materials were: MTA Plus (MTAP); MTA Fillapex (MTAF) and FillCanal (FC). Biocompatibility was evaluated with MTT and Neutral Red (NR) assays, after hDPCs exposure for 24 h to different dilutions of each sealer extract (1:2, 1:3 and 1:4). Unexposed cells were the positive control (CT). Bioactivity was assessed by alkaline phosphatase (ALP) enzymatic assay in cells exposed for one and three days to sealer extracts (1:4 dilution). All data were analyzed by ANOVA and Tukey post-test (p≤0.05%).

Results: MTT and NR results showed suitable cell viability rates for MTAP at all dilutions (90-135%). Cells exposed to MTAF and FC (1:2 and 1:4 dilutions) showed significant low viability rate when compared to CT in MTT. The NR results demonstrated cell viability for all materials tested. In MTAP group, the cells ALP activity was similar to CT in one and three days of exposure to the material. MTAF and FC groups demonstrated a decrease in ALP activity when compared to CT at both periods of cell exposure.

Conclusions: The hDPCs were suitable for the evaluation of new endodontic materialsin vitro. MTAP may be considered a promising material for endodontic treatments.
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http://dx.doi.org/10.1590/1678-775720150170DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4621938PMC
October 2015