Periodontitis is an inflammatory disease of the periodontium. Any imbalance between the matrix metalloproteinases (MMPs) secreted by neutrophils and tissue inhibitors initiates the destruction of collagen in gum tissue, leading to chronic periodontitis. This study aimed to correlate salivary levels of MMP-8 and periodontal parameters of chronic periodontitis to establish MMP-8 as a noninvasive marker for the early diagnosis of chronic periodontitis. The study involved 40 subjects visiting the periodontic OPD of Dr. Ziauddin Ahmad Dental College and Hospital, located in Aligarh, U.P., India, from 2011 to 2012. The subjects were divided into two groups: group I consisted of 20 periodontally healthy subjects (controls) while group II consisted of 20 patients with chronic periodontitis. Chronic periodontitis was assessed on the basis of several periodontal parameters, including pocket probing depth (PPD), clinical attachment level (CAL), gingival index (GI), and plaque index (PI). Around 3ml of unstimulated and whole expectorated saliva was collected for MMP-8 estimation by ELISA using Quantikine human total MMP-8 immunoassay kits. Data were analyzed using STATISTICA (Windows version 6) software. Salivary MMP-8 levels of groups I and II were 190.91 ± 143.89 ng/ml and 348.26 ± 202.1 ng/ml, respectively. The MMP-8 levels and periodontal status (PPD, CAL, GI, and PI) of groups I and II showed positive and significant correlations (for PPD, r = 0.63, P < 0.001; for CAL, r = 0.54, P < 0.001; for GI, r = 0.49, P < 0.001; and for PI, r = 0.63, P < 0.001). The results of this study demonstrate elevated concentrations of MMP-8 in individuals with chronic periodontitis.

Aims and Objectives:(1) To evaluate the effect of type 2 diabetes mellitus on salivary TNF-α level in chronic periodontitis. (2) To evaluate the effect of smoking on salivary TNF-α level in chronic periodontitis. (3) To compare and correlate TNF-α level with the healthy individuals.Materials and Methods:Subjects aged 30-35 years were included for the study and divided into four groups as a group of 20 systemically and periodontally healthy individuals (group I), a group of 20 subjects with pocket probing depth (PPD) ≥5 mm and clinical attachment loss (CAL) of ≥2 mm (group II), a group of 20 diabetic subjects (of more than 5 years) with periodontal parameters as of group II as (group III) and a group of 20 subjects smoking (≥10 cigarettes a day) with periodontal parameters of group II as (group IV). Periodontal parameters of PPD, CAL, gingival index (GI), and plaque index (PI) were measured using standard indices and criteria. Three milliliter of unstimulated saliva was taken and salivary TNF-α determined by using ELISA technique (Quantikine Human total TNF-A immunoassay kit).Results:Data revealed highest mean TNF-α in group III followed by group IV, group II, and group I. Mean TNF-α of both group III (76.1%) and group IV (48.8%) was significantly higher as compared to group I (P < 0.001). Mean TNF-α of group III was also found to be significantly different and higher (68.1%) as compared to group II (P < 0.001). Although higher mean TNF-α (31.5%) was found in group IV in comparison to group II, the difference was not statistically significant. Besides above, TNF-α also showed a direct positive correlation with PPD in group II (r = 0.30, P > 0.05) and a significant negative correlation was observed between CAL and TNF-α in group IV.Conclusion:Our study clearly underlines a profound impact of diabetes and smoking on salivary TNF-α in chronic periodontitis subjects in comparison to healthy subjects. Moreover, diabetes status increased TNF-α significantly in comparison to smoking in chronic periodontitis patients.

Saliva represents an ideal matrix for diagnostic biomarker development as it is readily available and requires no invasive collection procedures. However, salivary RNA is labile and rapidly degrades. Previous attempts to isolate RNA from saliva have yielded poor quality and low concentrations. Here we compare collection and processing methods and propose an approach for future studies. The effects of RNA stabilisers, storage temperatures, length of storage and fasting windows were investigated on pooled saliva samples from healthy volunteers. Isolated RNA was assessed for concentration and quality. Bacterial growth was investigated through RT-PCR using bacterial and human primers. Optimal conditions were implemented and quality controlled in a clinical setting. The addition of RNAlater increased mean RNA yield from 4912 ng/μl to 15,473 ng and RNA Integrity Number (RIN) from 4.5 to 7.0. No significant changes to RNA yield were observed for storage at room temperature beyond 1 day or at -80˚C. Bacterial growth did not occur in samples stored at ambient temperature for up to a week. There was a trend towards higher RNA concentration when saliva was collected after overnight fasting but no effect on RIN. In the clinic, RNA yields of 6307 ng and RINs of 3.9 were achieved, improving on previous reports. The method we describe here is a robust, clinically feasible saliva collection method using preservative that gives high concentrations and improved RINs compared to saliva collected without preservative.

This study suggests that MMP-8 is involved in periodontal destruction associated with smoking. Additionally, smoking exerts disastrous effects on immune response and can affect the pathogenesis of disease; hence, smoking results in increased severity of periodontal destruction.

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