scholarly journals PiggyBac transposon-mediated gene delivery efficiently generates stable transfectants derived from cultured primary human deciduous tooth dental pulp cells (HDDPCs) and HDDPC-derived iPS cells

2015 ◽  
Vol 7 (3) ◽  
pp. 144-154 ◽  
Author(s):  
Emi Inada ◽  
Issei Saitoh ◽  
Satoshi Watanabe ◽  
Reiji Aoki ◽  
Hiromi Miura ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Dental Pulp ◽  
Ips Cells ◽  
Deciduous Tooth ◽  
Dental Pulp Cells ◽  
Piggybac Transposon ◽  
Stable Transfectants ◽  
Pulp Cells

scholarly journals Choice of Feeders is Important When First Establishing iPSCs Derived from Primarily Cultured Human Deciduous Tooth Dental Pulp Cells

Cell Medicine ◽  
2015 ◽  
Vol 8 (1-2) ◽  
pp. 9-23 ◽  
Author(s):  
Issei Saitoh ◽  
Emi Inada ◽  
Yoko Iwase ◽  
Hirofumi Noguchi ◽  
Tomoya Murakami ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Deciduous Tooth ◽  
Dental Pulp Cells ◽  
Pulp Cells

scholarly journals Time-Dependent Response of Human Deciduous Tooth-Derived Dental Pulp Cells Treated with TheraCal LC: Functional Analysis of Gene Interactions Compared to MTA

2020 ◽  
Vol 9 (2) ◽  
pp. 531 ◽  
Author(s):  
Ok Hyung Nam ◽  
Jae-Hwan Kim ◽  
Sung Chul Choi ◽  
Young Kim
Keyword(s):  
Functional Analysis ◽  
Dental Pulp ◽  
Enrichment Analysis ◽  
Deciduous Tooth ◽  
Functional Enrichment ◽  
Receptor Interaction ◽  
Dental Pulp Cells ◽  
Pulp Tissue ◽  
Pulp Capping ◽  
Pulp Cells

Pulp capping material should facilitate hard tissue regeneration on the injured pulp tissue. TheraCal LC (TC) was recently developed. Although TC has shown reliable clinical outcomes after direct pulp capping, there are still remaining concerns regarding its detrimental effect on pulp cells. Therefore, this study aimed to identify the gene expression of human deciduous tooth-derived dental pulp cells exposed to TC compared to mineral trioxide aggregate (MTA). The cells were cultured and exposed to TC and MTA for 24 and 72 h. Next, total RNA was isolated. QuantSeq 3′ mRNA-sequencing was used to examine differentially expressed genes (DEGs) in exposed to TC and MTA. Functional analysis of DEGs was performed using bioinformatics analysis. In gene ontology (GO) functional enrichment analysis, cells in TC for 24 h presented significantly enriched immune response (p < 0.001) and inflammatory response (p < 0.01) compared to MTA. TC showed enriched positive regulation of cell migration at 72 h (p < 0.001). In Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, neuroactive ligand–receptor interaction (p = 1.19 × 10−7) and calcium signaling pathway (p = 2.96 × 10−5) were confirmed in the shared DEGs in TC. In conclusion, DEGs in TC may be involved in pathways associated with osteoclastogenesis and osteoclastic differentiation.

scholarly journals piggyBac Transposon-Based Immortalization of Human Deciduous Tooth Dental Pulp Cells with Multipotency and Non-Tumorigenic Potential

2019 ◽  
Vol 20 (19) ◽  
pp. 4904 ◽  
Author(s):  
Emi Inada ◽  
Issei Saitoh ◽  
Naoko Kubota ◽  
Yoko Iwase ◽  
Yuki Kiyokawa ◽  
...  
Keyword(s):  
Cell Lines ◽  
Dental Pulp ◽  
Fluorescent Protein ◽  
Deciduous Tooth ◽  
Specific Factor ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Normal Medium ◽  
Cdna Expression ◽  
Pulp Cells

We aimed to immortalize primarily isolated human deciduous tooth-derived dental pulp cells (HDDPCs) by transfection with piggyBac (PB)-based transposon vectors carrying E7 from human papilloma virus 16 or complementary DNA (cDNA) encoding human telomerase reverse transcriptase (hTERT). HDDPCs were co-transfected with pTrans (conferring PB transposase expression) + pT-pac (conferring puromycin acetyltransferase expression) + pT-tdTomato (conferring tdTomato cDNA expression) and pT-E7 (conferring E7 expression) or pTrans + pT-pac + pT-EGFP (conferring enhanced green fluorescent protein cDNA expression) + pT-hTERT (conferring hTERT expression). After six days, these cells were selected in medium containing 5 μg/mL puromycin for one day, and then cultured in normal medium allowing cell survival. All resultant colonies were harvested and propagated as a pool. Stemness and tumorigenic properties of the established cell lines (“MT_E7” for E7 and “MT_hTERT” for hTERT) with untransfected parental cells (MT) were examined. Both lines exhibited proliferation similar to that of MT, with alkaline phosphatase activity and stemness-specific factor expression. They displayed differentiation potential into multi-lineage cells with no tumorigenic property. Overall, we successfully obtained HDDPC-derived immortalized cell lines using a PB-based transfection system. The resultant and parental cells were indistinguishable. Thus, E7 and hTERT could immortalize HDDPCs without causing cancer-associated changes or altering phenotypic properties.

scholarly journals RNA analysis based on a small number of manually isolated fixed cells (RNA-snMIFxC) to profile stem cells from human deciduous tooth-derived dental pulp cells

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Emi Inada ◽  
Issei Saitoh ◽  
Naoko Kubota ◽  
Yoko Iwase ◽  
Yuki Kiyokawa ◽  
...  
Keyword(s):  
Cell Sorting ◽  
Dental Pulp ◽  
Housekeeping Gene ◽  
Surface Expression ◽  
Deciduous Tooth ◽  
Small Scale ◽  
Specific Gene ◽  
Dental Pulp Cells ◽  
Target Molecule ◽  
Pulp Cells

Abstract Background Expression of stemness factors, such as octamer-binding transcription factor 3/4 (OCT3/4), sex determining region Y-box 2 (SOX2), and alkaline phosphatase (ALP) in human deciduous tooth-derived dental pulp cells (HDDPCs) can be assessed through fixation and subsequent immuno- or cytochemical staining. Fluorescence-activated cell sorting (FACS), a powerful system to collect cells of interest, is limited by the instrument cost and difficulty in handling. Magnetic-activated cell sorting is inexpensive compared to FACS, but is confined to cells with surface expression of the target molecule. In this study, a simple and inexpensive method was developed for the molecular analysis of immuno- or cytochemically stained cells with intracellular expression of a target molecule, through isolation of a few cells under a dissecting microscope using a mouthpiece-controlled micropipette. Results Two or more colored cells (~ 10), after staining with a chromogen such a 3,3′-diaminobenzidine, were successfully segregated from unstained cells. Expression of glyceraldehyde 3-phosphate dehydrogenase, a housekeeping gene, was discernible in all samples, while the expression of stemness genes (such as OCT3/4, SOX2, and ALP) was confined to positively stained cells. Conclusion These findings indicate the fidelity of these approaches in profiling cells exhibiting cytoplasmic or nuclear localization of stemness-specific gene products at a small-scale.

scholarly journals The Preparation and Characterization of Chitosan-Based Hydrogels Cross-Linked by Glyoxal

Materials ◽  
2021 ◽  
Vol 14 (9) ◽  
pp. 2449
Author(s):  
Beata Kaczmarek-Szczepańska ◽  
Olha Mazur ◽  
Marta Michalska-Sionkowska ◽  
Krzysztof Łukowicz ◽  
Anna Maria Osyczka
Keyword(s):  
Thermal Stability ◽  
Tannic Acid ◽  
Dental Pulp ◽  
Blood Compatibility ◽  
Dental Pulp Cells ◽  
Preliminary Assessment ◽  
Human Dental Pulp Cells ◽  
Human Dental Pulp ◽  
Pulp Cells

In this study, hydrogels based on chitosan cross-linked by glyoxal have been investigated for potential medical applications. Hydrogels were loaded with tannic acid at different concentrations. The thermal stability and the polyphenol-releasing rate were determined. For a preliminary assessment of the clinical usefulness of the hydrogels, they were examined for blood compatibility and in the culture of human dental pulp cells (hDPC). The results showed that after immersion in a polyphenol solution, chitosan/glyoxal hydrogels remain nonhemolytic for erythrocytes, and we also did not observe the cytotoxic effect of hydrogels immersed in tannic acid (TA) solutions with different concentration. Tannic acid was successfully released from hydrogels, and its addition improved material thermal stability. Thus, the current findings open the possibility to consider such hydrogels in clinics.

scholarly journals Cytotoxicity assessment of different doses of ozonated water on dental pulp cells

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ferdiye Küçük ◽  
Sibel Yıldırım ◽  
Serap Çetiner
Keyword(s):  
Cell Viability ◽  
Repeated Measures ◽  
Dental Pulp ◽  
Control Group ◽  
Dental Pulp Cells ◽  
Time Points ◽  
Significant Difference ◽  
Pulp Cells ◽  
Time Point ◽  
Ozonated Water

Abstract Background The purpose of this study was to assess the cytotoxicity of various concentrations of ozonated water (OW) on human primary dental pulp cells. Methods Human primary dental pulp cells were isolated from exfoliated primary canine teeth of an 11-year-old patient with good systemic and oral health. Afterwards, cells were divided into 6 experimental groups; four groups of OW in concentrations of 2 mg/L, 4 mg/L, 8 mg/L, and 16 mg/L, untreated control group, and cell culture without cells. Cytotoxicity was evaluated after exposure for 5-min exposure using Mosmann’s Tetrazolium Toxicity (MTT) assay at 0 h and 48 h time points. Data were analyzed using a repeated measures analysis of variance and Post-hoc tests were performed using Bonferroni correction for multiple comparisons. Results All experimental groups showed proliferation at 0 h time point. However, all groups also experienced a decrease in overtime at 48 h time point (p < 0.05). At both time points 2 mg/L OW showed the highest cell viability as well as proliferation. At 0 h time point, the increase in cell viability for all experimental groups was found statistically significant when compared to positive control group (p < 0.05). At 48 h time point, although 8 mg/L and 16 mg/L OW showed statistically significant reduction in compare to 0 h time point, 2 mg/L and 4 mg/L OW groups didn’t experience any statistically significant difference (p < 0.05). Conclusion Considering our findings, due to ozonated water's induced a higher proliferation rate of dental pulp cells, indicating their biocompatibility and a possible adjuvant on irrigating agent in regenerative endodontic procedures.

The molecular functions of Biodentine and MTA in LPS‐induced inflamed dental pulp cells

2021 ◽  
Author(s):  
M.C. Wang ◽  
H.F. Tu ◽  
K.W. Chang ◽  
S.C. Lin ◽  
L.Y. Yeh ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Pulp Cells ◽  
Molecular Functions

HGF Enhanced Proliferation and Differentiation of Dental Pulp Cells

2006 ◽  
Vol 32 (8) ◽  
pp. 736-741 ◽  
Author(s):  
Ling Ye ◽  
Li Peng ◽  
Hong Tan ◽  
Xuedong Zhou
Keyword(s):  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Proliferation And Differentiation ◽  
Pulp Cells
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