Salivary macrophage chemokines as potential biomarkers of gingivitis.

OBJECTIVE: The present study aimed to analyze the salivary levels of macrophage-activating factor (MAF), macrophage-chemotactic factor (MCF), and macrophage migration inhibitory factor (MIF) in healthy and gingivitis patients, and to correlate between the concentrations of these chemo attractants with the intensity of gingival inflammation clinically. METHODS: Sixty saliva specimens were collected from periodontally healthy (n = 30), and gingivitis patients (n = 30). Bleeding on probing (BOP), Visible Plaque Index (VPI), and Simplified Modified Gingival Index (SMGI) were recorded through clinical examination. Salivary MAF, MCF, and MIF concentrations were assayed using enzyme-linked immunosorbent assays (ELISA). Statistical analysis was performed using SPSS (version 28). Total mean score for each biomarker was determined, and descriptive bivariate statistics were conducted to characterize the levels of biomarkers among the study groups. The difference in the biomarker levels among the study groups were analyzed by independent sample t test and one-way ANOVA. The diagnostic ability of the biomarkers was further tested by ROC curve analysis. RESULTS: Salivary levels of MAF was not significantly different between periodontally healthy individuals and gingivitis patients. The difference in MCF and MIF levels between patients with gingivitis and those with healthy periodontium was statistically significant (p 0.05 and p 0.001, respectively). When examined across the various stages of disease progression, MIF showed statistically significant difference among the three biomarkers (p 0.05). ROC curve analysis further revealed that area under the curve (AUC) for MIF has a better diagnostic capacity than MCF (AUC 0.981 vs. 0.673). CONCLUSIONS: Our results suggest that MIF could be considered as a potential salivary biomarker for gingivitis.

Differential Expression Profile of Salivary oncomiRNAs among Smokeless Tobacco Users.

OBJECTIVE: The aim of this study was to evaluate the expression of selected salivary oncomiRNAs among smokeless tobacco users and nonsmokers. MATERIALS AND METHODS: Twenty-five subjects with chronic smokeless tobacco habit (> 1 year) and 25 nonsmokers were selected for this study. MicroRNA was extracted from saliva samples using the miRNeasy Kit (Qiagen, Hilden, Germany). The forward primers used in the reactions include hsa-miR-21-5p, hsa-miR-146a-3p, hsa-miR-155-3p, and hsa-miR-199a-3p. Relative expression of miRNAs was calculated using the 2-ΔΔCt method. Fold change is calculated by raising 2 to the power of the negative ΔΔCT value. STATISTICAL ANALYSIS: Statistical analysis was carried out using GraphPad Prism 5 software. A p-value less than 0.05 was considered statistically significant. RESULTS: The four tested miRNAs were found overexpressed in saliva of subjects with smokeless tobacco habit when compared with saliva from nontobacco users. miR-21 expression was 3.74 ± 2.26 folds higher among subjects with smokeless tobacco habit compared to nontobacco users (p < 0.01). The expression for miR-146a (5.56 ± 8.3 folds; p < 0.05), miR-155 (8.06 ± 23.4 folds; p < 0.0001) and miR-199a (14.39 ± 30.3 folds; p < 0.05) was significantly higher among subjects with smokeless tobacco habit. CONCLUSION: Smokeless tobacco leads to salivary overexpression of the miRs 21, 146a, 155, and 199a. Monitoring the levels of these four oncomiRs may provide insight about the future development of oral squamous cell carcinoma, especially in patients with smokeless tobacco habits.

Mesenchymal stem cells: a promising tool for targeted gene therapy of endometriosis.

Endometriosis is a leading, benign gynecological disorder around the world. Last few years have witnessed tremendous growth in the field of endometriosis and endometrial stem-cell research. Despite advancements in the biology and pathology of endometriosis, disease recurrence is still an enigma. Gene therapy holds promise in treating many pathologic conditions including endometriosis. Mesenchymal stem cells (MSCs) serve as ideal candidates for regenerative medicine and cell-based therapies. Owing to their specificity to the endometrium, residing endometrial MSC populations could be utilized as ideal candidates for targeting endometrial disorders. Recently, we demonstrated their flexibility for gene transduction using adenoviral vectors. The review highlights the potential of endometrial MSCs in devising targeted gene therapies for endometriosis.

Expression of surfactant proteins SP-A and SP-D in murine decidua and immunomodulatory effects on decidual macrophages.

Surfactant proteins SP-A and SP-D are pattern recognition innate immune molecules that belong to the C-type lectin family. In lungs, they play an important role in the clearance of pathogens and control of inflammation. SP-A and SP-D are also expressed in the female reproductive tract where they play an important role in pregnancy and parturition. However, the role of SP-A and SP-D expressed at the feto-maternal interface (decidua) remains unclear. Here, we have examined the expression of SP-A and SP-D in the murine decidua at 17.5 (pre-parturition) and 19.5dpc (near parturition) and their effect on lipopolysaccharide (LPS)-treated decidual macrophages. SP-A and SP-D were localized to stromal cells in the murine decidua at 17.5 and 19.5dpc in addition to cells lining the maternal spiral artery. Purified pre-parturition decidual cells were challenged with LPS with and without SP-A or SP-D, and expression of F4/80 and TNF-α were measured by flow cytometry. On their own, SP-A or SP-D did not affect the percentage of F4/80 positive cells while they suppressed the percentage of TNF-α positive cells. However, simultaneous addition of SP-A or SP-D, together with LPS, reduced TNF-α secreting F4/80 positive cells. It is likely that exogenous administration of SP-A and SP-D in decidua can potentially control infection and inflammation mediators during spontaneous term labor and infection-induced preterm labor. Thus, the presence of SP-A and SP-D in the murine decidua is likely to play a protective role against intrauterine infection during pregnancy.

Surfactant protein D induces immune quiescence and apoptosis of mitogen-activated peripheral blood mononuclear cells.

Surfactant protein D (SP-D) is an integral molecule of the innate immunity secreted by epithelial cells lining the mucosal surfaces. The C-type lectin domain of SP-D performs pattern recognition functions while it binds to putative receptors on immune cells to modify cellular functions. Activation of immune cells and increased serum SP-D is observed in a range of patho-physiological conditions including infections. We speculated if SP-D can modulate systemic immune response via direct interaction with activated PBMCs. In this study, we examined interaction of a recombinant fragment of human SP-D (rhSP-D) on PHA-activated PBMCs. We report a significant downregulation of activation receptors such as TLR2, TLR4, CD11c and CD69 upon rhSP-D treatment. rhSP-D inhibited production of Th1 (TNF-α and IFN-γ) and Th17 (IL-17A) cytokines along with IL-6. Interestingly, levels of IL-2, Th2 (IL-4) and regulatory (IL-10 and TGF-ß) cytokines remained unaltered. Analysis of co-stimulatory CD28 and co-inhibitory CTLA4 receptors along with their ligands CD80 and CD86 revealed a selective up-regulation of CTLA4 in the lymphocyte subset. rhSP-D induced apoptosis in the activated but not in non-activated lymphocytes. Blockade of CTLA4 inhibited rhSP-D mediated apoptosis of activated lymphocytes, confirming involvement of CTLA4. We conclude that SP-D restores immune homeostasis. It regulates expression of immunomodulatory receptors and cytokines, which is followed by induction of apoptosis in activated lymphocytes. These findings suggest a critical role of SP-D in immune surveillance against activated immune cells.