AimTo explore a set of inflammatory biomarkers obtained from dentinal fluid (DF) from patients with symptomatic irreversible pulpitis (IP), reversible pulpitis (RP) and normal pulp (NP).MethodologyA cross‐sectional exploratory study was performed, recruiting 64 patients on the basis of their respective pulp condition. DF samples were obtained from all patients (23, from IP patients; 20, from RP patients; and 21, from NP patients). Quantification of biomarkers was performed using a Luminex® MAGPIX platform system and multiplex assay kits. The Kruskal–Wallis test was used for comparisons with regard to pulp state. A simple logistic regression model and the odds ratio (OR) with a 95% level of confidence (P = 0.05) were used to evaluate associations between biomarker levels and pulpal diagnosis. The performance discrimination of the biomarkers was evaluated through the construction of a receiver operating characteristic (ROC) curve by calculating the area under the curve (AUC) for IP versus RP after logistic regression modelling. Youden criteria were used to establish cut‐off points for biomarkers alone with AUC > 70 and P‐value < 0.05, or estimated probabilities from the multivariable logistic model.ResultsThe biomarkers that had significantly higher values in participants with IP versus RP were IL‐1α, VEGF‐α and FGF acid (P < 0.05). FGF acid (OR: 12.62; P = 0.0085; CI 95% 1.91–83.29) and VEGF‐α (OR: 2.61; P = 0.0252; CI 95% 1.13–6.03) were associated with pulp diagnoses of IP versus RP. The AUC‐ROC curve for FGF acid was 0.79. The model containing FGF acid, IL‐1α, IL‐6 and TIMP‐1 had an AUC‐ROC of 0.92 for IP versus RP with a significant difference from the FGF acid ROC curve (P = 0.0231).ConclusionsDentinal fluid could be used to assay pulpal mediators in the molecular diagnosis of pulpitis. Despite the limitation of the clinical diagnostics used in the present study, it was possible to detect a difference between irreversible symptomatic pulpitis and reversible pulpitis associated with the following combined biomarkers: FGF acid + IL‐6 + IL‐1α, +TIMP‐1.

Na+-K+-ATPase gene expression and activity were studied in aortas from adrenalectomized (ADX) rats and ADX rats with deoxycorticosterone supplement (ADX-DOCA). Northern analysis of RNA from ADX rats revealed a significant decrease in α2-mRNA levels (38.5 ± 8.3% of control, P < 0.01) that was prevented by DOCA ( P < 0.05). A decrease to 55.8 ± 7.7% in α2-isoform protein was observed 8 days after adrenal removal ( P < 0.05); DOCA reversed this effect (90.8 ± 10.5%). Adrenalectomy induced a decrease of 68.5 ± 4.5% in β1-mRNA ( P < 0.01) and 52.7 ± 8.3% in ADX-DOCA rats ( P < 0.01). Also, a reduction in β1-isoform protein that was not prevented by DOCA was detected after adrenalectomy (47.1 ± 11%, P < 0.01). In contrast, no differences in α1-mRNA or -protein levels were observed. Vascular sodium pump activity was reduced to 59.8 ± 4.6% of control values after adrenalectomy ( P < 0.01); this reduction was reversed by DOCA. Our data indicate that corticosteroids regulate Na+-K+-ATPase isoform expression and activity in vascular tissue in vivo, suggesting a mineralocorticoid-dependent modulation of α2-Na+-K+-ATPase gene expression in aorta, with β1-isoform expression dependent on the presence of glucocorticoids.

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High NaCl increases mRNA and protein abundance of numerous gene products, including the osmoprotective transcription factor TonEBP/OREBP. High NaCl increases TonEBP/OREBP abundance by stabilizing its mRNA. miRNAs regulate gene expression post‐transcriptionally by destabilizing target mRNAs and inhibiting translation. The purposes of the present studies were 1) to find whether miRNAs regulate TonEBP/OREBP abundance and 2) to identify miRNAs that are regulated by the level of NaCl. DICER and Ago2 are necessary for maturation and activity of miRNAs. We knocked down DICER and Ago2 in HEK293 cells by conditional expression of specific sh_miRNAs. Knocking down either DICER or Ago2 increases TonEBP/OREBP protein abundance, suggesting that miRNAs are involved in its regulation. In order to begin investigating the role of miRNAs in high NaCl‐induced changes of expression of proteins, including TonEBP/OREBP, we conducted a microarray screen of miRNAs affected by high NaCl. We adapted HEK293 cells to 500 mosmol/kg (NaCl added) through many passages or kept them at 300 mosmol/kg. We isolated miRNAs from them with a mirVana Kit, then screened with microarrays containing 470 probes (LC Sciences). Levels of 11 miRNAs are significantly greater at 500 mosmol/kg and 9 are less. Several of these miRNAs have likely target sites in the 3′‐UTRs of TonEBP/OREBP and other tonicity regulated genes. We conclude that the level of NaCl regulates several miRNAs and that these miRNAs may be involved in post‐transcriptional regulation of TonEBP/OREBP and other proteins.