Aim
To evaluate in a laboratory setting the effects of Endosequence BC Sealer HiFlow (Brasseler USA, Savannah, GA, USA), a novel calcium silicate‐based sealer developed for use in warm canal filling ...techniques, on human periodontal ligament stem cells (hPDLSCs).
Methodology
Eluates of EndoSequence BC Sealer HiFlow (BCHiF) (Brasseler USA), EndoSequence BC Sealer (BCS) (Brasseler USA) and AH Plus (AHP) (Dentsply DeTrey GmbH, Konstanz, Germany) were placed in contact with hPDLSCs. The characterization of the chemical elements of the root canal sealers was assessed using scanning electron microscopy and energy‐dispersive X‐ray analysis (SEM‐EDX). Inductively coupled plasma‐mass spectrometry (ICP‐MS) was used to determine the ion release of the sealers. MTT assay and wound healing techniques were used to determine cell viability and migration, respectively. Cell morphology and cell attachment were assessed using a direct contact technique of hPDLSCs onto the surface of the sealers and analysed by SEM. The bioactivity potential was carried out with the Alizarin Red and qPCR testing methods. The statistical differences were evaluated using one‐way anova and Tukey’s test (P < 0.05).
Results
ICP‐MS and EDX revealed significantly more zirconium in BCHiF than BCS (P < 0.05), whereas BCS had slightly higher levels of Ca2+ than BCHiF (P < 0.05). The cell viability assay revealed no relevant differences between BCS and BCHiF when compared with the control group (P > 0.05). Both BCS and BCHiF had similar rates of cell migration to the control group at 24 and 48 h. Cell morphology and adhesion capacity were also similar for BCS and BCHiF groups, whilst the AHP group was associated with reduced adhesion capacity. The Alizarin Red assay revealed a significant difference between the BCS and the control group (P < 0.001), as well as for the BCHiF group (P < 0.001). Finally, BCS and BCHiF promoted overexpression of osteo/cementogenic genes.
Conclusions
In general, EndoSequence BC Sealer HiFlow possesses suitable biological properties to be safely used as a root canal filling material and promote increased expression of oste/cementogenic genes by hPDLSCs.
Aims
To investigate the cytotoxicity and bioactivity of several pulpotomy materials: Biodentine (Septodont, Saint‐Maur‐des‐Fosses, France) MTA (Angelus, Londrina, PR, Brazil), Theracal LC (Bisco ...Inc., Schamburg, IL, USA) and IRM (Dentsply DeTrey GmbH, Konstanz, Germany), after contact with stem cells isolated from human exfoliated primary teeth (SHEDs).
Methodology
SHEDs were cultured in the presence of the eluates of various pulpotomy materials for 24, 48 and 72 h. Cell viability was determined by mitochondrial dehydrogenase enzymatic (MTT) assay. Apoptosis and changes in cell phenotype were evaluated by flow cytometry. Also, an in vitro scratch wound‐healing assay was used to determine their effects on cell migration. To assess cell morphology and attachment to the different pulpotomy materials, SHEDs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). Finally, the deposition of a calcified matrix in presence of these materials was verified by Alizarin Red staining. Statistical analysis was performed with analysis of variance and Bonferroni or Tukey post‐test (α = 0.05).
Results
Cell viability in the presence of Biodentine eluates was significantly higher to that obtained using complete medium alone (control; P < 0.01) and was also significantly higher than using MTA Angelus from 48 h of incubation (P < 0.01). However, Theracal LC and IRM were associated with low rates of cell viability (P < 0.001). Similar results were obtained in an apoptosis assay. In addition, SHEDs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of Biodentine. SEM studies revealed a suitable proliferation rate, cell spreading and attachment, especially when using Biodentine and MTA Angelus discs. Finally, Biodentine eluates significantly induced calcified matrix deposition from 7 days of culture (P < 0.01).
Conclusions
Biodentine exhibited better cytocompatibility and bioactivity than MTA Angelus, Theracal LC and IRM.
Aim
To evaluate the biological effects in vitro of MTA‐Angelus (MTA‐Ang; Angelus, Londrina, PR, Brazil), MTA Repair HP (MTA‐HP; Angelus) and NeoMTA Plus (NeoMTA‐P; Avalon Biomed Inc, Bradenton, FL, ...USA) on human dental pulp stem cells (hDPSCs).
Methodology
Cell viability and cell migration assays were performed using eluates of each material. To evaluate cell morphology and cell attachment to the different materials, hDPSCs were directly seeded onto the material surfaces and analysed by immunocytofluorescence and scanning electron microscopy, respectively. The chemical composition of the materials was determined by energy‐dispersive X‐ray (EDX), and eluates were analysed by inductively coupled plasma–mass spectrometry (ICP‐MS). Statistical analysis was performed with the analysis of variance and Bonferroni or Tukey post‐test (α < 0.05).
Results
Undiluted MTA‐Ang, MTA‐HP and NeoMTA‐P displayed a significant increase in cell viability greater than that obtained using complete medium alone (control) (*P < 0.05; **P < 0.01; ***P < 0.001). Moreover, a cell migration assay revealed cell migration rates after incubation with extracts of MTA‐Ang, MTA‐HP and NeoMTA‐P that were similar to levels obtained in the control group. In addition, stretched cytoskeletal F‐actin fibres were detected in the cells treated with the three material extracts. SEM studies revealed a high degree of cell proliferation and attachment on all three materials. EDX analysis demonstrated similar weight percentages of C, O and Ca in all three materials, whilst other elements such as Al, Si and S were also found.
Conclusions
MTA‐Ang, MTA‐HP and NeoMTA‐P were associated with biological effects on hDPSCs in terms of cell proliferation, morphology, migration and attachment.
Aim
To evaluate the biocompatibility of three calcium silicate‐based endodontic sealers, Bioroot BC Sealer (Septodont, Saint‐Maur‐des‐Fosses, France), Endoseal MTA (EndoSeal, Maruchi, Seoul, Korea) ...and Nano‐ceramic Sealer (B&L Biotech, Fairfax, VA, USA) (NCS), on human periodontal ligament stem cells (hPDLSCs).
Methodology
Human periodontal ligament stem cells were cultured in the presence of various endodontic sealer eluates for 24 h. Cell viability was determined using the MTT assay. Cell death and changes in phenotype induced by the set endodontic sealer eluates were evaluated through flow cytometry. Also, an in vitro scratch wound‐healing model was used to determine their effects in cell migration. Finally, to assess cell morphology and attachment to the different sealers, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). One‐way analysis of variance (anova) followed by a Bonferroni post‐test was performed (P < 0.05).
Results
At 24 h, cell spreading was evident in the presence of Bioroot BC Sealer (BR) and Nano‐ceramic Sealer (NCS), but not Endoseal MTA (ES). At 72 h, BR and NCS exhibited high and moderate cell proliferation, respectively, whereas ES revealed low rates of cell proliferation (P < 0.05). Similar results were obtained in a cell death assay. In addition, hPDLSCs maintained their mesenchymal phenotype in all conditions although their capacity to migrate was higher in the presence of BR. Finally, SEM studies revealed a good degree of proliferation, cell spreading and attachment, especially when using BR and NCS discs.
Conclusions
BR and NCS were associated with better cytocompatibility than ES. Further in vitro and in vivo investigations are required to confirm the suitability of these calcium silicate‐based endodontic sealers for clinical application.
Aim
To investigate in vitro the cytocompatibility of the calcium silicate‐containing endodontic sealers MTA Fillapex and TotalFill BC Sealer on human periodontal ligament stem cells (hPDLSCs) by ...assaying their biological responses and compare them with that observed when using an epoxy resin‐based sealer (AH Plus).
Methodology
Specimens from the three different endodontic sealers were eluated with culture medium for 24 h. The cytotoxicity of these eluates was evaluated using the MTT assay. In addition, an in vitro scratch wound healing model was used to determine their effects on cell migration. Cell adhesion to collagen type I after treatment with the different sealer eluates was also measured, whereas cytotoxicity was determined using the DNA‐specific fluorochrome Hoechst 33342. Finally, to assess cell morphology and attachment to the different sealers, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy (SEM). One‐way analysis of variance (anova) followed by a Bonferroni post‐test were performed (P < 0.05).
Results
hPDLSCs exposed to different dilutions of TotalFill BC Sealer eluates had significantly higher cell proliferation compared with that observed when cells were treated with AH Plus and MTA Fillapex eluates (P < 0.001). In addition, TotalFill eluates were associated with significantly increased cell adhesion to collagen type I and migration of hPDLSCs in a concentration‐dependent manner than displayed after treatment with MTA Fillapex or AH Plus eluates (P < 0.001). Moreover, TotalFill BC Sealer‐induced cytotoxicity was significantly lower than observed using AH Plus and MTA Fillapex eluates (P < 0.001). Finally, SEM studies revealed suitable proliferation, cell spreading and attachment, especially when using TotalFill BC Sealer discs.
Conclusion
TotalFill BC Sealer exhibited a higher cytocompatibility than AH Plus and MTA Fillapex. Further investigations using in vivo animal models are required to validate the potential biological responses of TotalFill BC Sealer on hPDLSCs.
Objective
Compositional modifications may alter the biological and physicochemical characteristics of calcium silicate-based sealers (CSBS) and, ultimately, their bioactivity. The main objective of ...this study was to evaluate the biological properties of three CSBS: EndoSequence BC Sealer, Ceraseal, and Endoseal mineral trioxide aggregate.
Materials and methods
Human periodontal ligament stem cells (hPDLSCs) were exposed to several eluates of CSBS. The ion release profile and pH were determined, and metabolic activity and cell migration were assessed using the MTT and wound healing assays. hPDLSCs were cultured in direct contact with the surface of each material, and cell morphology and attachment were analyzed by scanning electron microscopy (SEM). Bioactivity potential was assessed by RT-qPCR and mineralization assays. Statistical differences between biomaterials were assessed using one- or two-way ANOVA (
α
< 0.05).
Results
All materials showed an alkaline pH, although Endoseal exhibited a significantly higher pH compared with the other CSBS (
p
< 0.05). Ceraseal released significantly more Ca
2+
(
p
< 0.05) than EndoSequence BC Sealer and Endoseal. Interestingly, Endoseal induced a significant reduction in cell viability and cell migration compared with the control (
p
< 0.001). Moreover, SEM showed abundant cells adhering to EndoSequence BC Sealer and Ceraseal surfaces, whereas very few round cells were detected on the surface of Endoseal. Finally, Ceraseal and EndoSequence induced ALP, CAP, and CEMP-1 expression and a significantly higher mineralization capacity than Endoseal (***
p
< 0.001).
Conclusions
The eluates from EndoSequence BC Sealer and Ceraseal displayed higher cell viability, cell attachment, cell migration rates, and ion release rates than Endoseal. Ceraseal and EndoSequence BC Sealer exhibited significantly more gene expression and mineralization capacity than Endoseal.
Clinical relevance
The results obtained in the present work suggest that EndoSequence BC Sealer and Ceraseal possess biological properties that make them suitable materials for root canal treatment.
Aim
To analyse in vitro changes in ion release and biological properties of Endocem‐MTA (Maruchi, Wonju, Korea) and NeoMTA‐Plus (Avalon Biomed Inc, Bradenton, FL, USA) exposed to acidic or neutral ...environment on human dental periodontal ligament stem cells (hPDLSCs).
Methodology
Cell viability and wound healing assays were performed using eluates of each material. Cell death and changes in phenotype induced by the set endodontic sealer eluates were evaluated through flow cytometry. To evaluate cell attachment to the different materials, hPDLSCs were directly seeded onto the material surfaces and analysed by scanning electron microscopy. The chemical composition of the materials was determined by energy‐dispersive X‐ray (EDX), and ion release was evaluated by inductively coupled plasma‐mass spectrometry. Statistical analysis was performed with analysis of variance and a Bonferroni or Tukey post‐test (α < 0.05).
Results
The MTT assay revealed non‐cytotoxic effects of NeoMTA‐Plus and Endocem‐MTA at pH 5.2 and 7.4. However, there were minor differences compared with the control, especially at pH 5.2, where both materials were associated with significantly greater cell viability (P < 0.05). In both environments, the materials stimulated hPDLSCs to migrate. hPDLSCs were attached to the bioactive cements, with multiple prolongations proliferated on the surface of the samples. Moreover, there were no changes to cell phenotype or apoptosis/necrosis rates, indicating that the acidic environment did not induce cell death. Prismatic crystalline structures were seen on the surface of the cements exposed to butyric acid and EDX analysis identified a marked peak of Ca2+ from NeoMTA‐Plus and Endocem‐MTA in acidic and physiological environments.
Conclusions
An acidic environment favoured the release of Ca2+ ions from both bioactive cements, and the cytotoxicity of these bioactive cements was low in both environments studied.
The term sepsis refers to a complex and heterogeneous syndrome. Although great progress has been made in improving the diagnosis and treatment of this condition, it continues to have a huge impact on ...morbidity and mortality worldwide. Mesenchymal stem cells are a population of multipotent cells that have immunomodulatory properties, anti-apoptotic effects, and antimicrobial activity. We studied these capacities in a porcine model of peritoneal sepsis.
We infused human adipose-derived mesenchymal stem cells (ADSCs) into a porcine model of peritoneal sepsis. Twenty piglets were treated with antibiotics alone (control group) or antibiotics plus peritoneal infusion of ADSCs at a concentration of 2 × 10
cells/kg or 4 × 10
cells/kg (low- and high-dose experimental groups, respectively). The animals were evaluated at different time points to determine their clinical status, biochemical and hematologic parameters, presence of inflammatory cytokines and chemokines in blood and peritoneal fluid, and finally by histologic analysis of the organs of the peritoneal cavity.
One day after sepsis induction, all animals presented peritonitis with bacterial infection as well as elevated C-reactive protein, haptoglobin, IL-1Ra, IL-6, and IL-1b. Xenogeneic ADSC infusion did not elicit an immune response, and peritoneal administration of the treatment was safe and feasible. One day after infusion, the two experimental groups showed a superior physical condition (e.g., mobility, feeding) and a significant increase of IL-10 and TGF-β in blood and a decrease of IL-1Ra, IL-1b, and IL-6. After 7 days, all animals treated with ADSCs had better results concerning blood biomarkers, and histopathological analysis revealed a lower degree of inflammatory cell infiltration of the organs of the peritoneal cavity.
Intraperitoneal administration of ADSCs as an adjuvant therapy for sepsis improves the outcome and diminishes the effects of peritonitis and associated organ damage by regulating the immune system and reducing intra-abdominal adhesions in a clinically relevant porcine model of abdominal sepsis.
Objective
The aim of the present study was to evaluate the in vitro biocompatibility of Theracal PT, Theracal LC, and MTA Angelus, considered as bioactive materials used for vital pulp treatment, on ...human dental pulp stem cells (hDPSCs).
Materials and methods
Human dental pulp stem cells (hDPSCs) were isolated from third molars, and material eluates were prepared (undiluted, 1:2, and 1:4 ratios). The hDPSC cytotoxicity, adhesion, morphology, viability, and cell migration were assessed. The mineralization nodule formation was determined by Alizarin red S staining (ARS). The odonto/osteogenic differentiation potential was assessed by osteo/odontogenic marker expression real-time qPCR. The chemical composition and ion release of the vital pulp materials were determined by energy dispersive X-ray (EDX) and inductively coupled plasma-mass spectrometry (ICP-MS), respectively. Statistical differences were assessed by ANOVA and Tukey’s test (
p
< 0.05).
Results
The three vital pulp materials showed variable levels of calcium, tungsten, silicon, and zirconium release and in their chemical composition. Cytocompatibility assays revealed higher hDPSC viability and migration rates when treated with Theracal PT than with Theracal LC. The lowest cell adhesion and spreading were observed in all Theracal LC-treated groups, whereas the highest were observed when treated with MTA. Theracal PT and MTA promoted the upregulation of DSPP and RUNX2 gene expression (
p
< 0.05). After 21 days, both MTA Angelus and Theracal PT–treated cells exhibited a significantly higher mineralized nodule formation than the negative control (
p
< 0.05).
Conclusions
This study demonstrates the favorable in vitro cytocompatibility and bioactive properties of the recently introduced Theracal PT and the well-established MTA Angelus on hDPSCs, as opposed to Theracal LC. More studies, including in vivo animal testing are suggested before these new formulations might be used in the clinical setting.
Clinical relevance
Theracal PT is a new material that could be clinically suitable for vital pulp therapy. Further studies considering its biocompatibility and bioactivity are necessary.
In regenerative dentistry, stem cell-based therapy often requires a scaffold to deliver cells and/or growth factors to the injured site. Graphene oxide (GO) and silk fibroin (SF) are promising ...biomaterials for tissue engineering as they are both non toxic and promote cell proliferation. On the other hand, periodontal ligament stem cells (PDLSCs) are mesenchymal stem cells readily accessible with a promising use in cell therapy. The purpose of this study was to investigate the effects of composite films of GO, SF and GO combined with fibroin in the mesenchymal phenotype, viability, adhesion and proliferation rate of PDLSCs. PDLSCs obtained from healthy extracted teeth were cultured on GO, SF or combination of GO and SF films up to 10 days. Adhesion level of PDSCs on the different biomaterials were evaluated after 12 h of culture, whereas proliferation rate of cells was assessed using the MTT assay. Level of apoptosis was determined using Annexin-V and 7-AAD and mesenchymal markers expression of PDLSCs were analyzed by flow cytometry. At day 7 of culture, MTT experiments showed a high rate of proliferation of PDLSCs growing on GO films compared to the other tested biomaterials, although it was slightly lower than in plastic (control). However PDLSCs growing in fibroin or GO plus fibroin films showed a discrete proliferation. Importantly, at day 10 of culture it was observed a significant increase in PDLSCs proliferation rate in GO films compared to plastic (
P
< 0.05), as well as in GO plus fibroin compared to fibroin alone (
P
< 0.001). Flow cytometry analysis showed that culture of PDLSCs in fibroin, GO or GO plus fibroin films did not significantly alter the level of expression of the mesenchymal markers CD73, CD90 or CD105 up to 168 h, being the cell viability in GO even better than obtained in plastic. Our findings suggest that the combination of human dental stem cells/fibroin/GO based-bioengineered constructs have strong potential for their therapeutic use in regenerative dentistry.
PDF
