Background
Though the use of salivary miRNAs as potential biomarkers has been reported in few diseases/conditions such as rheumatoid arthritis and oral cancer, there are no reported studies on their utility in periodontal diagnostics. Thus, the aim of the present study was to profile salivary miRNAs and identify the most suitable salivary miRNA biomarker in chronic periodontitis.
Methods
In this study, we have explored the potential application of next generation sequencing (NGS) technology for profiling miRNAs in two unstimulated saliva samples collected by passive drool method from a patient diagnosed with generalized chronic periodontitis and a healthy control. Subsequently, the validation of most highly expressed known miRNA in periodontitis was performed in saliva samples collected from an independent set of 16 chronic periodontitis patients and 16 periodontally healthy controls using quantitative real‐time PCR (qRT‐PCR). Target gene prediction and pathway mapping were performed using bioinformatic tools.
Results
NGS analysis identified 40 upregulated and 40 downregulated known miRNAs in chronic periodontitis compared to healthy controls, of which miR‐143‐3p was the most highly expressed miRNA in periodontitis (Read count – 227630; fold change – 5.82). Validation using qRT‐PCR showed significant upregulation of miR‐143‐3p expression in the test group compared with controls (P < 0.05). K‐RAS (V‐Ki‐ras2 Kirsten rat sarcoma viral oncogene) gene was predicted as the target gene for miR‐143‐3p in humans. KEGG (Kyoto Encyclopedia of genes and genomes) pathway mapping revealed the involvement of K‐RAS in mitogen‐activated protein kinases (MAPK) pathway.
Conclusions
The application of NGS for miRNA expression profiling can be considered a valuable tool in detection of novel biomarkers in periodontal diagnostics. Also, the results of the study points to the potential utility of miR143‐3p as a novel salivary biomarker for chronic periodontitis.
There is a substantial increase in the concentration of both MIP-1α and MCP-1 with increasing severity of periodontal disease. Both the analytes showed promising results as biomarkers for discriminating periodontal disease from health.
Background
Fetuin‐A has garnered recognition in the etiopathogenesis of several systemic disorders. It has been recently acknowledged as an anti‐inflammatory marker for periodontal disease. This study aimed to compare and correlate salivary and serum fetuin‐A levels in health and patients with stages II‐III periodontitis along with evaluating the effect of non‐surgical periodontal therapy (NSPT) on the same.
Methods
Group 1 comprised of 30 healthy subjects. Group 2 embodied 30 patients with stages II‐III periodontitis. Clinical periodontal parameters were recorded. Saliva and serum samples were assembled. Periodontitis patients received non‐surgical periodontal treatment. They were recalled after 6 months, and collection of samples and recording of clinical parameters were reiterated. Fetuin‐A levels were analyzed using ELISA.
Results
Salivary and serum fetuin‐A levels were significantly lower in periodontitis patients when compared with the healthy subjects (P < 0.001) at baseline. Their concentrations significantly upregulated 6 months after active periodontal therapy (P < 0.001). Salivary fetuin‐A levels revealed a significant positive correlation with their serum levels in Group 1 at baseline (P < 0.001). They also displayed a positive correlation in Group 2 at baseline and 6 months post periodontal therapy, nevertheless failed to establish a statistically significant association.
Conclusion(s)
Our study concluded that salivary and serum fetuin‐A levels diminished with increasing severity of periodontal inflammation, and NSPT remarkably improved their levels. They also displayed a significant positive correlation in health, and a non‐significant, yet positive correlation in patients with periodontitis.
The diagnosis and management of periodontal diseases can be enhanced by the identification of biomolecules which can predict disease susceptibility, indicate current disease activity, and monitor response to therapy. Salivary proteomics is a major avenue in the ongoing search for a biomarker in periodontal research. Saliva is a valuable diagnostic vehicle which “mirrors” oral and systemic health and disease. Standardized methods of saliva sampling and processing will increase diagnostic test accuracy and decrease bias in measurements. Prominent databases such as PubMed/Medline, PMC, Scopus, and Google Scholar were searched and literature evidence from January 2000 till September 2022 were studied to identify the methodological considerations employed in salivary proteomics for periodontal research. Evidence and recommendations in this regard were collated into this narrative review. The methods of saliva collection and processing presented in this review will help researchers conduct salivary proteomic studies with standardized protocols.
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