Tissue-non-specific alkaline phosphatase mRNA expression and alkaline phosphatase activity following application of retinoic acid in cultured human dental pulp cells

Regenerative endodontic procedures using autologous platelet-rich plasma (PRP) can improve the biologic outcome of treatment. However, its mechanism of action on improving pulp regeneration is not fully elucidated. Autophagy was recently shown to be related to tissue repair and osteogenesis. Therefore, the objective of this study was to investigate the effect of PRP in dental pulp regeneration and to elucidate the role of autophagy involved in this process. Human dental pulp cells (hDPCs) were isolated from healthy dental pulp and co-cultured with an increasing concentration of PRP. Cellular migration and proliferation were determined by scratch assay, transwell assay, and cell-counting kit 8 assay. Osteogenic differentiation was clarified by using alkaline phosphatase staining, alizarin red staining, and real-time polymerase chain reaction (RT-PCR) to measure the gene expression levels of alkaline phosphatase, collagen-1, osteocalcin, dentin matrix protein 1, and dentin sialophosphoprotein. Autophagic bodies were observed by transmission electron microscopy and the expression of autophagy marker light chain 3B (LC3B) was determined by immunofluorescence staining. The mRNA and protein expression level of LC3B and Beclin-1 were quantified by qRT-PCR and western blotting. The effect of PRP on cellular migration, proliferation, and osteogenic differentiation was further investigated in the milieu of autophagy activator, rapamycin, and inhibitor, 3-methyladenine. Results showed that PRP promoted cell migration, proliferation, and osteogenic differentiation. Autophagic bodies were strongly activated and the expression level of LC3B and Beclin-1 was significantly promoted by PRP. Autophagy inhibition suppressed PRP-induced hDPCs migration, proliferation, and osteogenic differentiation, whereas autophagy activator substantially augmented PRP-stimulated migration, proliferation, and differentiation. Taken together, these findings suggested that PRP could effectively promote regenerative potentials associated with autophagy.

Download Full-text

Induction of mRNA expression of osteogenesis-related genes by guaiacol in human dental pulp cells

Odontology ◽

10.1007/s10266-010-0129-0 ◽

2010 ◽

Vol 98 (2) ◽

pp. 165-169 ◽

Cited By ~ 3

Author(s):

Takashi Kato ◽

Kumiko Shirayama ◽

Takeo W. Tsutsui ◽

Takeki Tsutsui

Keyword(s):

Mrna Expression ◽

Dental Pulp ◽

Dental Pulp Cells ◽

Human Dental Pulp Cells ◽

Human Dental Pulp ◽

Pulp Cells

Download Full-text

Investigation of PPARβ/δ within Human Dental Pulp Cells: A Preliminary In Vitro Study

PPAR Research ◽

10.1155/2021/8854921 ◽

2021 ◽

Vol 2021 ◽

pp. 1-10

Author(s):

Caroline L. de Lima ◽

Bruna R. Amorim ◽

Carine Royer ◽

Augusto P. Resende ◽

Maria F. Borin ◽

...

Keyword(s):

Mrna Expression ◽

Dental Pulp ◽

Raw264.7 Cells ◽

Dental Pulp Cells ◽

Human Dental Pulp Cells ◽

Pulp Inflammation ◽

Human Dental Pulp ◽

Pulp Cells ◽

Anti Inflammatory

Controlling the inflammatory response to restore tissue homeostasis is a crucial step to maintain tooth vitality after pathogen removal from caries-affected dental tissues. The nuclear peroxisome proliferator-activated receptor beta/delta (PPARβ/δ) is a ligand-activated transcription factor with emerging anti-inflammatory roles in many cells and tissues. However, its expression and functions are poorly understood in human dental pulp cells (hDPCs). Thus, this study evaluated PPARβ/δ expression and assessed the anti-inflammatory effects evoked by activation of PPARβ/δ in lipopolysaccharide- (LPS-) induced hDPCs. Our results showed that hDPCs constitutively expressed PPARβ/δ mRNA/protein, and treatment with LPS increased PPARβ/δ mRNA expression. The selective PPARβ/δ agonist GW0742 significantly decreased inflammation-related mRNA expression in hDPCs (IL6, IL1β, TNFα, MMP1, and MMP2) and RAW264.7 cells (Il6 and Tnfα). Further, PPARβ/δ agonist attenuated MMP2/9 gelatinolytic activity in hDPCs. Previously LPS-conditioned hDPCs increased the migration of RAW264.7 cells through the membrane of a Transwell coculture system. Conversely, pretreatment with GW0742 markedly decreased macrophage recruitment. These findings provide among the first evidence that hDPCs express PPARβ/δ. In addition, they suggest that activation of PPARβ/δ by GW0742 can attenuate some cellular and molecular in vitro aspects related to the inflammatory process, pointing out to investigate its potential target role in dental pulp inflammation.

Download Full-text

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

Materials ◽

10.3390/ma14092449 ◽

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.

Download Full-text