This study aimed to evaluate carbonated hydroxyapatite (CHA)’s ability for mineral induction and its
Precursors for the study include di-ammonium hydrogen phosphate and calcium nitrate tetrahydrate, with sodium hydrogen carbonate added to achieve different levels of carbonate substitution. The synthesized CHA samples are characterized using X-ray diffraction, Fourier transform infrared spectroscopy, and Raman spectroscopy. Scanning electron microscopy (SEM) was used to observe morphology. For 14 days at 37°C, samples were submerged in simulated body fluid to assess their mineral induction capabilities. SEM was used to confirm apatite formation on sample surfaces. The cytotoxicity assay was used to assess the vitality of the cells following their exposure to various concentrations of CHA.
The Joint Committee on Powder Diffraction Standards data for HA aligned well with the results from X-ray diffraction analysis of CHA across 3 different concentrations, indicating strong agreement. Fourier transform infrared spectra indicated the presence of phosphate, hydroxyl, and carbonate groups within the samples. SEM and Energy-dispersive X-ray analysis show agglomerated and flaky nanoparticles. All the samples are bioactive, but the formation of apatite differs from one another.
The results of this study may provide insight into the potential use of carbonated HA as a dental pulp-capping material for vital pulp therapy.
The aim of this study was to investigate the expression of 7 different sirtuin genes (SIRT1-SIRT7) in human dental pulp cells (HDPCs), and to determine the role of SIRTs in the odontoblastic differentiation potential of HDPCs.
HDPCs were isolated from freshly extracted third molar teeth of healthy patients and cultulred in odontoblastic differentiation inducing media. Osteocalcin (OCN) and dentin sialophosphoprotein (DSPP) expression was analyzed to evaluate the odontoblastic differentiation of HDPCs by reverse transcription-polymerase chain reaction (RT-PCR), while alizarin red staining was used for the mineralization assay. To investigate the expression of SIRTs during odontoblastic differentiation of HDPCs, real time PCR was also performed with RT-PCR.
During the culture of HDPCs in the differentiation inducing media, OCN, and DSPP mRNA expressions were increased. Mineralized nodule formation was also increased in the 14 days culture. All seven SIRT genes were expressed during the odontogenic induction period. SIRT4 expression was increased in a time-dependent manner.
Our study identified the expression of seven different SIRT genes in HDPCs, and revealed that SIRT4 could exert an influence on the odontoblast differentiation process. Further studies are needed to determine the effects of other SIRTs on the odontogenic potential of HDPCs.
Genetic information such as DNA sequences has been limited to fully explain mechanisms of gene regulation and disease process. Epigenetic mechanisms, which include DNA methylation, histone modification and non-coding RNAs, can regulate gene expression and affect progression of disease. Although studies focused on epigenetics are being actively investigated in the field of medicine and biology, epigenetics in dental research is at the early stages. However, studies on epigenetics in dentistry deserve attention because epigenetic mechanisms play important roles in gene expression during tooth development and may affect oral diseases. In addition, understanding of epigenetic alteration is important for developing new therapeutic methods. This review article aims to outline the general features of epigenetic mechanisms and describe its future implications in the field of dentistry.
We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization.
Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data.
Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated.
Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.
This study investigated changes in gene expressions concerning of differentiation, proliferation, mineralization and inflammation using Human-8 expression bead arrays when white Mineral Trioxide Aggregate and calcium hydroxide-containing cement were applied
wMTA (white ProRoot MTA, Dentsply) and Dycal (Dentsply Caulk) in a Teflon tube (inner diameter 10 mm, height 1 mm) were applied to HDPCs. Empty tube-applied HDPCs were used as negative control. Total RNA was extracted at 3, 6, 9 and 24 hr after wMTA and Dycal application. The results of microarray were confirmed by reverse transcriptase polymerase chain reaction.
Out of the 24,546 genes, 43 genes (e.g., BMP2, FOSB, THBS1, EDN1, IL11, COL10A1, TUFT1, HMOX1) were up-regulated greater than two-fold and 25 genes (e.g., SMAD6, TIMP2, DCN, SOCS2, CEBPD, KIAA1199) were down-regulated below 50% by wMTA. Two hundred thirty nine genes (e.g., BMP2, BMP6, SMAD6, IL11, FOS, VEGFA, PlGF, HMOX1, SOCS2, CEBPD, KIAA1199) were up-regulated greater than two-fold and 358 genes (e.g., EDN1, FGF) were down-regulated below 50% by Dycal.
Both wMTA and Dycal induced changes in gene expressions related with differentiation and proliferation of pulp cells. wMTA induced changes in gene expressions related with mineralization, and Dycal induced those related with angiogenesis. The genes related with inflammation were more expressed by Dycal than by wMTA. It was confirmed that both wMTA and Dycal were able to induce gene expression changes concerned with the pulp repair in different ways.
This study was performed to investigate the biocompatibility of newly introduced Bioaggregate on human pulp and PDL cells.
Cells were collected from human pulp and PDL tissue of extracted premolars. Cell culture plate was coated either with Bioaggregate or white MTA, then the same number of cells were poured to cell culture dishes. Cell attachment and growth was examined under a phase microscope after 1,3 and 7 days of seeding. Cell viability was measured and the data was analyzed using Student
Both types of cells used in this study were well attached and grew healthy on Bioaggregate and MTA coated culture dishes. No cell inhibition zone was observed in Bioaggregate group. There was no statistical difference of viable cells between bioaggreagte and MTA groups.
Bioaggregate appeared to be biocompatible compared with white MTA on human pulp and PDL cells.
This study investigated the changes in gene expression when mineral trioxide aggregate (MTA) was applied
The purpose of this study was to characterize functional distinction between human dental pulp cells(PC) and periodontal ligament cells(PDLC) using cDNA microarray assay and to confirm the results of the microarray assay using RT-PCR. 3 genes out of 51 genes which were found to be more expressed(>2 fold) in PC were selected, and 3 genes out of 19 genes which were found to be more expressed(>2 fold) in PDLC were selected for RT-PCR as well.
According to this study, the results were as follows:
1. From the microarray assay, 51 genes were more expressed (2 fold) from PC than PDLC.
2. RT-PCR confirmed that ITGA4 and TGF β2 were more expressed in PC than in PDLC.
3. From the microarray assay, 19 genes were more expressed (2 fold) from PDLC than PC.
4. RT-PCR confirmed that LUM, WISP1, and MMP1 were more expressed in PDLC than in PC.
From the present study, different expression of the genes between the PC and PDLC were characterized to show the genes which play an important role in dentinogenesis were more expressed from PC than PDLC, while the genes which were related with collagen synthesis were more expressed from PDLC than PC.
The purpose of this study is to investigate the response of human pulp cell on Portland cement mixed with β-glycerophosphate. To investigate the effect of β-glycerophosphate and/or dexamethasone on human pulp cell, ALP activity on various concentration of β-glycerophosphate and dexamethasone was measured and mineral nodule of human pulp cell was stained with Alizarin red S. MTS assay and ALP activity of human pulp cell on Portland cement mixed with various concentration of β-glycerophosphate (10 mM, 100mM, 1M) was measured and the specimens were examined under SEM.
Addition of β-glycerophosphate or dexamethasone alone had no effect however, the addition of 5 mM β-glycerophosphate and 100 nM dexamethasone had the largest increasement in ALP activity. There was no toxicity in all samples and the data showed that Portland cement mixed with 10 mM β-glycerophosphate had more increase in ALP activity compared with control.
In conclusion, Portland cement mixed with β-glycerophosphate has no toxicity and promotes differentiation and mineralization of pulp cell compared with additive-free Portland cement. This implicated that application of Portland cement mixed with β-glycerophosphate might form more reparative dentin and in turn it would bring direct pulp capping to success.
Dental pulp is a loose, mesenchymal tissue almost entirely enclosed in the dentin. It consists of cells, ground substance, and neural and vascular supplies. Damage to the dental pulp by mechanical, chemical, thermal, and microbial irritants can provoke various types of inflammatory response. Pulpal inflammation leads to the tissue degradation, which is mediated in part by Matrix metalloproteinase leads to accelerate extracellular matrix degradation with pathological pathway. We have now investigated the induction of MMPs and inflammatory cytokines by Lipopolysaccharide (LPS) control of inflammatory mediators by peroxisome proliferator-activated receptors (PPARs).
Human dental pulp cells exposed to various concentrations of LPS (1-10 µg/ml) revealed elevated levels of MMP-2 and MMP-9 at 24 hrs of culture. LPS also stimulated the production of ICAM-1, VCAM-1, IL-1β, and TNF-α. Adenovirus PPARγ (Ad/PPARγ) and PPARγ agonist rosiglitazone reduced the synthesis of MMPs, adhesion molecules and pro-inflammatory cytokines. The inhibitory effect of Ad/PPARγ was higher than that of PPARγ agonist.
These result offer new insights in regard to the anti-inflammatory potential of PPARγ in human dental pulp cell.
The purpose of this study was to regenerate human dental pulp tissues similar to native pulp tissues. Using the mixture of type I collagen solution, primary cells collected from the different tissues (pulp, gingiva, and skin) and NIH 3T3 (1 × 105 cells/ml/well) were cultured at 12-well plate at 37℃ for 14 days. Standardized photographs were taken with digital camera during 14 days and the diameter of the contracted collagen gel matrix was measured and statistically analyzed with student t-test. As one of the pulp tissue engineering, normal human dental pulp tissue and collagen gel matrix cultured with dental pulp cells for 14 days were fixed and stained with Hematoxyline & Eosin.
According to this study, the results were as follows:
1. The contraction of collagen gel matrix cultured with pulp cells for 14 days was significantly higher than other fibroblasts (gingiva, skin) (p < 0.05).
2. The diameter of collagen gel matrix cultured with pulp cells was reduced to 70.4% after 7 days, and 57.1% after 14 days.
3. The collagen gel without any cells did not contract, whereas the collagen gel cultured with gingiva and skin showed mild contraction after 14 days (88.1% and 87.6% respectively).
4. The contraction of the collagen gel cultured with NIH 3T3 cells after 14 days was higher than those cultured with gingival and skin fibroblasts, but it was not statistically significant (72.1%, p > 0.05).
5. The collagen gel matrix cultured with pulp cells for 14 days showed similar shape with native pulp tissue without blood vessels.
This approach may provide a means of engineering a variety of other oral tissue as well and these cell behaviors may provide information needed to establish pulp tissue engineering protocols.