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Evaluation of the adhesion of human dental pulp stem cells to different endodontic biomaterials before and after setting.

ABSTRACT: Background. Stem cell-based treatment modalities have been potential strategies for tissue regeneration in many conditions. Several studies have evaluated the biologic properties of DPSCs and their efficacy in the treatment of a variety of diseases. The present study was undertaken to evaluate the adhesion behavior of DPSCs on different endodontic materials before and after setting. Methods. The crowns of the selected teeth were removed, and the root canals were prepared and obturated with gutta-percha and AH26 sealer. A retrograde cavity was prepared at root ends. Different materials were placed in the cavities. Then the samples were attached to the wells with the use of a chemical glue. Dental pulp stem cells were allowed to proliferate to reach a count of 2 million and transferred to -12well plates in association with a culture medium. Finally, the samples attached to the wells were exposed to the stem cells immersed in the culture medium before and after setting. Then adhesion of the stem cells was evaluated using SEM. Results. The SEM results showed cellular adhesion in the samples containing CEM cement both before and after setting. The samples containing MTA Angelus and ProRoot MTA exhibited cellular adhesion before setting, with no cellular adhesion after setting. The samples containing AH26 and MTA Fillapex sealers exhibited cellular adhesion after setting, with no adhesion before setting. The samples containing simvastatin exhibited no cellular adhesion before setting; this material had dissolved in the culture medium after setting evaluation. Conclusion. The results of the present study showed that of all the materials tested, CEM cement had the highest capacity for dental pulp stem cell adhesion.

SUBMITTER: Aghazade M

PROVIDER: S-EPMC7464226 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Evaluation of the adhesion of human dental pulp stem cells to different endodontic biomaterials before and after setting.

Aghazade Marzie M Samiei Mohammad M Imani Marjan M Aghazadeh Zahra Z Alizadeh Effat E Rezaie Fereshte F

Journal of dental research, dental clinics, dental prospects 20200617 2

<b>Background.</b> Stem cell-based treatment modalities have been potential strategies for tissue regeneration in many conditions. Several studies have evaluated the biologic properties of DPSCs and their efficacy in the treatment of a variety of diseases. The present study was undertaken to evaluate the adhesion behavior of DPSCs on different endodontic materials before and after setting. <b>Methods.</b> The crowns of the selected teeth were removed, and the root canals were prepared and obtura ...[more]

PMID: 32908650

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Project description:BackgroundBiomaterials are widely used to regenerate or substitute bone tissue. In order to evaluate their potential use for clinical applications, these need to be tested and evaluated in vitro with cell culture models. Frequently, immortalized osteoblastic cell lines are used in these studies. However, their uncontrolled proliferation rate, phenotypic changes or aberrations in mitotic processes limits their use in long-term investigations. Recently, we described a new pluripotent-like subpopulation of dental pulp stem cells derived from the third molars (DPPSC) that shows genetic stability and shares some pluripotent characteristics with embryonic stem cells. In this study we aim to describe the use of DPPSC to test biomaterials, since we believe that the biomaterial cues will be more critical in order to enhance the differentiation of pluripotent stem cells.MethodsThe capacity of DPPSC to differentiate into osteogenic lineage was compared with human sarcoma osteogenic cell line (SAOS-2). Collagen and titanium were used to assess the cell behavior in commonly used biomaterials. The analyses were performed by flow cytometry, alkaline phosphatase and mineralization stains, RT-PCR, immunohistochemistry, scanning electron microscopy, Western blot and enzymatic activity. Moreover, the genetic stability was evaluated and compared before and after differentiation by short-comparative genomic hybridization (sCGH).ResultsDPPSC showed excellent differentiation into osteogenic lineages expressing bone-related markers similar to SAOS-2. When cells were cultured on biomaterials, DPPSC showed higher initial adhesion levels. Nevertheless, their osteogenic differentiation showed similar trend among both cell types. Interestingly, only DPPSC maintained a normal chromosomal dosage before and after differentiation on 2D monolayer and on biomaterials.ConclusionsTaken together, these results promote the use of DPPSC as a new pluripotent-like cell model to evaluate the biocompatibility and the differentiation capacity of biomaterials used in bone regeneration.

Project description:BackgroundDental pulp stem cells (DPSCs) have been developed as a potential source of mesenchymal stem cells (MSCs) for regeneration of dental pulp and other tissues. However, further strategies to isolate highly functional DPSCs beyond the colony-forming methods are required. We have demonstrated the safety and efficacy of DPSCs isolated by G-CSF-induced mobilization and cultured under normoxia (mobilized DPSCs, MDPSCs) for pulp regeneration. The device for isolation of MDPSCs, however, is not cost-effective and requires a prolonged cell culture period. It is well known that MSCs cultured under hypoxic-preconditions improved MSC proliferation activity and stemness. Therefore, in this investigation, we attempted to improve the clinical utility of DPSCs by hypoxia-preconditioned DPSCs (hpDPSCs) compared with MDPSCs to improve the potential clinical utility for pulp regeneration in endodontic dentistry.MethodsColony-forming DPSCs were isolated and preconditioned with hypoxia in a stable closed cultured system and compared with MDPSCs isolated from the individual dog teeth. We examined the proliferation rate, migration potential, anti-apoptotic activity, and gene expression of the stem cell markers and angiogenic/neurotrophic factors. Trophic effects of the conditioned medium (CM) were also evaluated. In addition, the expression of immunomodulatory molecules upon stimulation with IFN-γ was investigated. The pulp regenerative potential and transplantation safety of hpDPSCs were further assessed in pulpectomized teeth in dogs by histological and immunohistochemical analyses and by chemistry of the blood and urine tests.ResultshpDPSCs demonstrated higher proliferation rate and expression of a major regulator of oxygen homeostasis, HIF-1α, and a stem cell marker, CXCR-4. The direct migratory activity of hpDPSCs in response to G-CSF was significantly higher than MDPSCs. The CM of hpDPSCs stimulated neurite extension. However, there were no changes in angiogenic, migration, and anti-apoptotic activities compared with the CM of MDPSCs. The expression of immunomodulatory gene, PTGE was significantly upregulated by IFN gamma in hpDPSCs compared with MDPSCs. However, no difference in nitric oxide was observed. The regenerated pulp tissue was quantitatively and qualitatively similar in hpDPSC transplants compared with MDPSC transplants in dog teeth. There was no evidence of toxicity or adverse events of the hpDPSC transplantation.ConclusionsThese results demonstrated that the efficacy of hpDPSCs for pulp regeneration was identical, although hpDPSCs improved stem cell properties compared to MDPSCs, suggesting their potential clinical utility for pulp regeneration.

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Evaluation of the adhesion of human dental pulp stem cells to different endodontic biomaterials before and after setting.

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Evaluation of the adhesion of human dental pulp stem cells to different endodontic biomaterials before and after setting.

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