Biological Effects of Provisional Resin Materials on Human Dental Pulp Stem Cells

2017 ◽  
Vol 42 (2) ◽  
pp. E81-E92 ◽  
Author(s):  
S-K Jun ◽  
C Mahapatra ◽  
H-H Lee ◽  
H-W Kim ◽  
J-H Lee

SUMMARY Objectives: This study investigated the in vitro cytotoxicity as well as the proinflammatory cytokine expression of provisional resin materials on primary cultured human dental pulp stem cells (hDPSCs). Methods: Five commercially available provisional resin materials were chosen (SNAP [SN], Luxatemp [LT], Jet [JE], Revotek LC [RL], and Vipi block [VB]). Eluates that were either polymerizing or already set were added to hDPSCs under serially diluted conditions divided into three different setting times (25% set, 50% set, and 100% set) and incubated for 24 hours with 2× concentrated culture media. Cell cytotoxicity tests were performed by LDH assay and live and dead confocal microscope images. The expression of proinflammatory cytokines in SN and VB was measured using cytokine antibody arrays. Data were analyzed using repeated measures analysis of variance (ANOVA) or ANOVA followed by the Tukey post hoc test at a significance level of p<0.05. Results: Cytotoxicity greater than 30% was observed in the 50% diluted culture in SN, LT, and JE in the already set stage (p<0.05), while it was detected in SN and LT in early or intermediate stage samples. The cytotoxicity of SN, JE, and LT was greater with eluates from the polymerizing phase compared to that from already set samples (p<0.05), as observed by live and dead images. On the other hand, RL and VB did not exhibit cytotoxicity greater than 30%. Proinflammatory cytokines were not detected in 12.5% diluted culture with eluates from VB and early set stage SN. Conclusions: The eluates from chemical-activated provisional resin materials during polymerization (SN, LT, and JE) were cytotoxic to hDPSCs and may adversely affect pulp tissue.

2021 ◽  
Vol 30 ◽  
pp. 096368972110344
Author(s):  
Xiaohui Fu ◽  
Yimiao Feng ◽  
Bingyi Shao ◽  
Yanzhen Zhang

Background: Dental pulp stem cells (DPSCs) are a unique source for future clinical application in dentistry such as periodontology or endodontics. However, DPSCs are prone to apoptosis under abnormal conditions. Taxifolin is a natural flavonoid and possesses many pharmacological activities including anti-hypoxic and anti-inflammatory. We aimed to elucidate the mechanisms of taxifolin protects DPSC under hypoxia and inflammatory conditions. Methods: DPSCs from human dental pulp tissue was purchased from Lonza (cat. no. PT-5025. Basel, Switzerland)) and identified by DPSC’s biomarkers. DPSC differentiation in vitro following the manufacturers’ instructions. ARS staining and Oil red staining verify the efficiency of differentiation in vitro after 2 weeks. The changes of various genes and proteins were identified by Q-PCR and western-blot, respectively. Cell viability was determined by the CCK-8 method, while apoptosis was determined by Annexin V/PI staining. Results: DPSC differentiation in vitro shows that hypoxia and TNF-α synergistically inhibit the survival and osteogenesis of DPSCs. A final concentration of 10 μM Taxifolin can significantly reduce the apoptosis of DPSCs under inflammation and hypoxia conditions. Taxifolin substantially increases carbonic anhydrase IX (CA9) expression but not HIF1a, and inhibitions of CA9 expression nullify the protective role of taxifolin under hypoxia and inflammatory condition. Conclusion: Taxifolin significantly increased the expression of CA9 when it inhibits DPSC apoptosis and taxifolin synergistically to protect DPSCs against apoptosis with CA9 under hypoxia and inflammatory conditions. Taxifolin can be used as a potential drug for clinical treatment of DPSC-related diseases.


Biomedicines ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 483
Author(s):  
Jon Luzuriaga ◽  
Jon Irurzun ◽  
Igor Irastorza ◽  
Fernando Unda ◽  
Gaskon Ibarretxe ◽  
...  

The generation of vasculature is one of the most important challenges in tissue engineering and regeneration. Human dental pulp stem cells (hDPSCs) are some of the most promising stem cell types to induce vasculogenesis and angiogenesis as they not only secrete vascular endothelial growth factor (VEGF) but can also differentiate in vitro into both endotheliocytes and pericytes in serum-free culture media. Moreover, hDPSCs can generate complete blood vessels containing both endothelial and mural layers in vivo, upon transplantation into the adult brain. However, many of the serum free media employed for the growth of hDPSCs contain supplements of an undisclosed composition. This generates uncertainty as to which of its precise components are necessary and which are dispensable for the vascular differentiation of hDPSCs, and also hinders the transfer of basic research findings to clinical cell therapy. In this work, we designed and tested new endothelial differentiation media with a fully defined composition using standard basal culture media supplemented with a mixture of B27, heparin and growth factors, including VEGF-A165 at different concentrations. We also optimized an in vitro Matrigel assay to characterize both the ability of hDPSCs to differentiate to vascular cells and their capacity to generate vascular tubules in 3D cultures. The description of a fully defined serum-free culture medium for the induction of vasculogenesis using human adult stem cells highlights its potential as a relevant innovation for tissue engineering applications. In conclusion, we achieved efficient vasculogenesis starting from hDPSCs using serum-free culture media with a fully defined composition, which is applicable for human cell therapy purposes.


2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Shion Orikasa ◽  
Nobuyuki Kawashima ◽  
Kento Tazawa ◽  
Kentaro Hashimoto ◽  
Keisuke Sunada-Nara ◽  
...  

AbstractAccelerated dental pulp mineralization is a common complication in avulsed/luxated teeth, although the mechanisms underlying this remain unclear. We hypothesized that hypoxia due to vascular severance may induce osteo/odontoblast differentiation of dental pulp stem cells (DPSCs). This study examined the role of B-cell CLL/lymphoma 9 (BCL9), which is downstream of hypoxia-inducible factor 1α (HIF1α) and a Wnt/β-catenin transcriptional cofactor, in the osteo/odontoblastic differentiation of human DPSCs (hDPSCs) under hypoxic conditions. hDPSCs were isolated from extracted healthy wisdom teeth. Hypoxic conditions and HIF1α overexpression induced significant upregulation of mRNAs for osteo/odontoblast markers (RUNX2, ALP, OC), BCL9, and Wnt/β-catenin signaling target genes (AXIN2, TCF1) in hDPSCs. Overexpression and suppression of BCL9 in hDPSCs up- and downregulated, respectively, the mRNAs for AXIN2, TCF1, and the osteo/odontoblast markers. Hypoxic-cultured mouse pulp tissue explants showed the promotion of HIF1α, BCL9, and β-catenin expression and BCL9-β-catenin co-localization. In addition, BCL9 formed a complex with β-catenin in hDPSCs in vitro. This study demonstrated that hypoxia/HIF1α-induced osteo/odontoblast differentiation of hDPSCs was partially dependent on Wnt/β-catenin signaling, where BCL9 acted as a key mediator between HIF1α and Wnt/β-catenin signaling. These findings may reveal part of the mechanisms of dental pulp mineralization after traumatic dental injury.


Author(s):  
Mansoore Saharkhiz ◽  
Fariba Emadian Razavi ◽  
Seyed Mohammad Riahi ◽  
Malaksima Ayadilord ◽  
Zeinab Rostami ◽  
...  

Biomolecules ◽  
2020 ◽  
Vol 10 (7) ◽  
pp. 972
Author(s):  
Roberta Souza D’Almeida Couto ◽  
Maria Fernanda Setubal Destro Rodrigues ◽  
Leila Soares Ferreira ◽  
Ivana Márcia Alves Diniz ◽  
Fernando de Sá Silva ◽  
...  

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.


2019 ◽  
Vol 52 (6) ◽  
Author(s):  
Alessio Zordani ◽  
Alessandra Pisciotta ◽  
Laura Bertoni ◽  
Giulia Bertani ◽  
Antonio Vallarola ◽  
...  

2019 ◽  
Vol 13 (1) ◽  
pp. 3-10 ◽  
Author(s):  
Saeed Rahimi ◽  
Sadegh Salarinasab ◽  
Negin Ghasemi ◽  
Reza Rahbarghazi ◽  
Shahriar Shahi ◽  
...  

Background. The aim of this in vitro study was to investigate the effect of zinc oxide (ZnO) and zirconium oxide (ZrO2) microparticles (MPs) and nanoparticles (NPs) in combination with white Portland cement (WPC) on odontogenic capacity of human dental pulp stem cells over a period of 21 days. Methods. Synthesized ZnO and ZrO2 particles were characterized using scanning electron microscopy and transmission electron microscopy. The viability of human dental pulp stem cells was measured by a 3-(4,5-dimethylthiazolyl-2-yl)-2,5- diphenyltetrazolium bromide assay at 7-, 14- and 21-day intervals after seeding on WPC disks enriched with ZnO and ZrO2 MPs and NPs. Odontogenic potential of ZnO and ZrO2 particles in combination with WPC was investigated by alkaline phosphatase (ALP) activity and ionized calcium level of supernatant culture media at different time intervals. Data were analyzed using one-way ANOVA and post hoc Tukey tests. Results. All the materials exhibited cell viability over a 21-day period, except for WPC with ZnO NPs on day 7, although it was not statistically significant (P>0.05). The ALP activity and ionized calcium level increased in all the groups compared to the control group (P<0.05). ZnO NPs had superior effect on odontogenic activity and calcium ion release compared to ZnO MPs (P=0.046). There was no significant difference between ZrO2 MPs and NPs in odontogenic activity (P>0.05). Conclusion. WPC enriched with ZnO and ZrO2 increased ALP activity and calcium ion release of human dental pulp stem cells over a period of 21 days in vitro.


Sign in / Sign up

Export Citation Format

Share Document