Salivary Soluble CD44 Level in Chronic Periodontitis – A Comparative Assessment between Preoperative and Postoperative Scaling and Root Planing

ABSTRACT

Background:

Salivary biomarkers can be used to diagnose illnesses, track their course, and gauge a patient’s response to therapy. Inflammatory circumstances result in higher sCD44 levels. The most typical inflammatory disorder connected to periodontal tissue is called periodontitis. Therefore, the purpose “of the current study was to compare the levels of sCD44 in patients with chronic periodontitis before and after scaling and root planing.

Objectives:

1) Determining CD44 concentrations in healthy individuals’ spit. 2) To quantify CD44 levels in the saliva of individuals with persistent periodontitis. The third objective is to compare the levels of CD44 that can be dissolved in saliva between those with healthy gums and those with active periodontitis. 4) Assessing CD44 levels in saliva after “scaling and root planing in patients with chronic periodontitis.”

Materials and Methods:

Half of the study participants were classified as having chronic periodontitis, while the other half had healthy gums. Participants with healthy gums and those with chronic periodontitis had their plaque levels, gingival status, and testing depths measured. All the same, data were calculated in persons with chronic periodontitis one month after they had “scaling and root planing done. Clinical attachment levels were evaluated before scaling and root planing was performed on patients with chronic periodontitis and again one month afterwards. Patients with chronic periodontitis,” those whose condition had not improved following scaling and root planing (SRP), and healthy individuals all had spit tests conducted after a month. The concentration of sCD44 in the saliva was determined with a chemically coupled immunosorbent assay.

Results:

Salivary sCD44 levels were substantially greater in people with chronic periodontitis than in people without the condition. After one month of scaling and root planing in patients with chronic periodontitis, sCD44 levels drastically dropped (p < 0.0001).

Conclusion:

Chronic periodontitis is linked to elevated levels of salivary sCD44. sCD44 can be employed as a possible salivary biomarker for chronic Periodontitis.

INTRODUCTION

Periodontitis refers to an inflamed, destructive state of the supporting structures that worsens with time. The lack of consistency in periodontology’s established clinical guidelines makes it difficult to detect areas of actively deteriorating health, evaluate the efficacy of treatment, and gauge patients’ susceptibility to the spread of disease. Progress in periodontal diagnostics has led to the development of tools like the “Biomarker” that enable the detection and categorization of periodontal risk.[1] CD44, a biomarker with a flaming reputation, has been linked to the severity of the disease. It is a glycoprotein found on the surface of many different cell types. CD44 is also involved in inflammation, the immune response, and blood cell production. Its presence is crucial during the early stages of B and Lymphocyte development, and it is also upregulated during antigen receptor activation and takes part in cell dealing during inflammation. Ultimately, proteolytic cleavages release CD44 from the cell surface and dissolve it (sCD44). Previous research has discovered that elevated CD44 levels are present in persons with underlying inflammatory diseases.[2] As a possible biomarker for periodontal disorders, sCD44 has recently come under consideration.[3]

Biomarkers have been looked for in serum, GCF, and saliva. An array of biomarkers can be detected in spit and used as a roadmap to pinpoint the source of a periodontal infection. Examining its true potential as a symptomatic marker for periodontitis necessitates looking at salivary sCD44 levels in persistent periodontitis during scaling and root planing and evaluating salivary sCD44 levels in periodontal health and disease.

MATERIAL AND METHODS

The organization’s ethical advisory board approved this assessment. Review subjects were people who sought treatment at the Periodontology Department randomized on the basis of computer-generated list. One hundred people, ranging in age from 30 to 60, participated in the study. The participants were separated into two groups. Group B (chronic periodontitis) included fifty individuals treated with scaling and root planing before and again one month later, while Group A (periodontal disease) included fifty subjects treated with scaling and root planing before and again one month afterward (periodontic health) Group A, which consisted of participants with healthy gums, and Group B consisted of subjects with persistent periodontitis, and were evaluated both before and after scaling and root planing. Exclusion criteria: Subjects who had anti-infection and periodontal therapy within the last three months, those who were current smokers, and those with medical histories indicating any primary infections were excluded. Nursing mothers, expectant mothers, and anyone with oral premalignant disorders were also excluded.

Besides doing a periodontal evaluation and meticulous documentation the characteristics chosen for the research were executed. Overall, the accounting was done by a single inspector. Certain locations with maximum inflammation were chosen, and the following criteria were recorded: Silness and Loe index were used for the plaque scores,[4] the gingival status, the Periodontal pocket depth, and the clinical attachment loss.

Subjects were instructed to fast for 60 minutes prior to collecting unstimulated spit samples (water was an exemption). The tests were conducted in sealed, germ-free Uricol containers. Spit method unstimulated saliva was used to collect. Dissolvable CD44 in spit tests was differentiated using a sandwich-type ELISA. An explicit monoclonal neutralizer for CD44 had been applied to the microtiter strips provided. Guidelines with established CD44 fixations and queries were pipetted into these wells. Underlying hatching binds CD44 antigen, standards, and zeros; subsequent brooding presents a biotinylated monoclonal immunizer explicit for CD44. Once the sample had been cleaned, the chemical (streptavidin-peroxidase) was shown. After broiling and washing to remove any free catalyst, a substrate arrangement was known to provide a brilliant reaction. The sample CD44 concentration is directly related to the intensity of this color change. A total of 100 ml of each standard, test, control, and standard diluent was poured into a well for this experiment. The meat was then broiled at room temperature for a further hour. At that moment, 100 ml of Streptavidin-HRP was added and incubated, and then 50 ml of diluted biotinylated unfriendly to CD44 was applied to all wells and incubated for 30 minutes at room temperature. After 10-20 minutes of incubation, 100 ml of a ready-to-use Tetramethylbenzidine (TMB) substrate arrangement was added. Following the placement of the microtiter plate in the LISA, the microplate peruser was used to examine the resulting diversity.

Statistical analysis

The sociological quantitative software package SPSS 20 was used to analyze the data. Plaque and gingival scores, testing pocket depth, clinical partnership failure, and sCD44 levels were analyzed across all audit groups to identify their respective means and standard deviations. We used the Mann-Whitney U test to compare the groups without disease and the group with persistent Periodontitis, and we used the Wilcoxon matched matches test to compare the pre-and post-employment data from the last group.

RESULTS

There were a total of 100 participants in this study; 50 were healthy, while the other 50 suffered from persistent Periodontitis. Twenty-six men and twenty-four women made up the healthy group, whereas twenty-five men and thirty women were among the people with chronic periodontitis. The average age of the review’s participants was 42.75 ± 8.01. The average ages of participants who suffered from chronic Periodontitis and those who were otherwise in excellent health were 37.05 ± 3.10 and 49.55 ± 4.50, respectively.

The Mann-Whitney U test was used to analyze the plaque document score, and the result was a statistically significant P-value of 0.0001. Ongoing cases of Periodontitis were associated with greater mean plaque record scores compared to healthy people. The Wilcoxon matched-matches test used to compare results from before and after surgery, showed a statistically significant difference (p < 0.001). One-month scaling and root arranging were shown to be significantly diminished by an increase in plaque score [Table 1].

Table 1.

Comparison of Healthy and Chronic periodontitis and pre and post -operative with respect to Plaque index scores

Group	Mean	SD	P
Healthy group	0.70	0.12	0.0001*
Chronic periodontitis group	2.02	0.25
Pre operative	2.02	0.25
Post operative	1.14	0.21

*P<0.05

To compare the gingiva index score between patients with healthy gums and those with chronic periodontitis, a statistically significant P-value of 0.0001 was discovered. In comparison to participants without chronic periodontitis, the gingival index score was considerably greater. The “data were compared using the Wilcoxon matched pairs test, and statistical significance was observed at a P-value of 0.0001. One month after scaling and root planing, the” gingival index score considerably decreased [Table 2].

Table 2.

Comparison of Healthy and Chronic periodontitis groups and pre and post operative with respect to gingival index scores

Group	Mean	SD	P
Healthy group	0.69	0.12	0.0001*
Chronic periodontitis group	2.21	0.20
Pre operative	2.21	0.20
Post operative	1.46	0.39

*P<0.05

Mann-Whitney U test was used to compare these participants, and the P-value was discovered to be 0.0001. When compared to participants without chronic Periodontitis, the probing depth of the pocket was considerably greater. Using the Wilcoxon matched pairs test, a P-value of 0.0001 was discovered for the comparison within chronic Periodontitis individuals. One month post-operatively, probing pocket depth considerably decreased [Table 3].

Table 3.

Comparison of Healthy and Chronic periodontitis groups and pre and post operative with respect to Probing pocket depth (in mm) scores

Group	Mean	SD	P
Healthy group	2.30	0.31	0.0001*
Chronic periodontitis group	4.51	0.44
Pre operative	4.51	0.44
Post operative	3.82	0.41

*P<0.05

The data within the chronic periodontitis group were compared using the Wilcoxon matched pairs test, and a P-value of 0.0001 was discovered. One month after surgery, clinical attachment loss greatly improves [Table 4].

Table 4.

Comparison of pre and postoperative clinical attachment loss (in mm) scores in chronic periodontitis group by Wilcoxon matched pairs test (Table 4)

Time	Mean	Std. Dev.	P
Pre operative	4.93	0.57	0.0001*
Post operative	4.53	0.58

*P<0.05

“Mann- Whitney” A P value of 0.0407 was discovered using the U test to compare the sCD44 levels between the groups. It was more pronounced in participants with chronic periodontitis. The gap between the two events was enormous by any standard. Using the Wilcoxon matched pairs test, we discovered a truly enormous difference (p < 0.0001) between the sCD44 levels of healthy members and those with chronic Periodontitis. The levels of sCD44 dropped significantly one month after root arrangement and scaling [Table 5].

Table 5.

Comparison of Healthy and Chronic periodontitis groups with respect to sCD44 count (in ng/ml) scores

Group	Mean	SD	P
Healthy group	2.43	2.23	0.0407*
Chronic periodontitis group	3.97	2.23
Pre operative	3.97	2.23	0.0001*
Post operative	0.25	0.59

*P<0.05

DISCUSSION

Clinical agents and specialists alike may find it challenging to identify persons at risk for periodontal disease and to diagnose the disease in its active phases while symptoms are still vague. The use of oral fluids, such as spit, in the diagnosis of periodontal disease, is currently being researched by scientists. Biomarkers are large particles that may be used to track health, the onset of illness, the effectiveness of treatment, and other relevant outcomes. Anyone, from generally sound people to those with particular underlying goof-ups, can make them.[3,5]

In addition to serving as the main hyaluronan receptor, the cell-bound particle CD44 also serves as a receptor for the bulk of the other extracellular grid components, including laminin, fibronectin, and type I and IV collagens. As a result, it interacts with flagging particles such as oncogene items with protein tyrosine kinase activity, ties solvent development elements, and cytokines, operating as a phone grip atom, a sign transmitter, and a development introducing particle.[6,7] As a transmembrane receptor, a crucial part of the matrix, and a soluble protein in bodily fluids, CD44 exists in three different forms.[4] Inflammation, immune activation, and malignancy all have higher levels of sCD44 than healthy conditions, according to studies, whereas immunodeficiency conditions have lower levels of sCD44.[8] There are few studies on soluble CD44 in connection to periodontitis, the most frequent type of periodontal disorder in the oral cavity that is also related to elevated levels of salivary sCD44.

Dental plaque is the primary etiological factor for the onset and progression of periodontal diseases. We found that the plaque list score for the group with chronic Periodontitis was substantially higher than that of the healthy group. Results showed that the gingival index score was considerably higher in the group with chronic Periodontitis compared to the healthy group. After one month, both the plaque file score and the gingival list score had decreased considerably after scaling and root planing. Similar findings were observed by Badersten et al.,[9] who investigated the effects of supra- and subgingival debridement on plaque scores and found that the files were significantly expanded after the procedures.

The average testing pocket depth of the active Periodontitis group was significantly greater than that of the healthy group. Patients with chronic Periodontitis were studied to get a mean value for clinical attachment loss (CAL). This might be linked to the initial bacterial infection and its byproducts, which set off a resistant provocative reaction in the host. This leads to the destruction of soft tissue and bone. Previous studies[10] have demonstrated that the clinical parameters will drop after periodontal non-surgical treatment, and the tissue conditions will improve. In our current investigation, the probing pocket depth and CAL in the chronic Periodontitis group significantly decreased one month after SRP which was consistent with the past findings by Badersten et al., and Ishikawa et al.[9,10]

The primary ligand for CD44 is hyaluronan. Through a connection with CD44 and TLR4 receptors, HA activates “immune cells and increases regulator cytokines of the inflammatory process during inflammation.[11] According to their findings, Khan et al.[12] conclude that CD44 contributed to the severe inflammatory response by facilitating Neutrophil grip and migration.

Zhuo et al.[13] supported the capacity of CD44 in neutrophil-endothelial” connections by showing that it could roll and initiate bonding when subjected to physiological shearing powers. Salivary sCD44 is theorized to be released due to neutrophil cell damage evidently caused by cytokine excitation, given that neutrophils are likely mediators of tissue passage in periodontal disease. Steady periodontitis patients had elevated sCD44 levels, indicating a higher risk of cell damage and tissue degeneration. In this ongoing research, we compared the sCD44 subsets of people with chronic periodontitis to those of healthy controls. Research done previously by EJ Franzmann et al.,[14] and Kaur et al.,[15] the average values of sCD44 in healthy individuals were 1.09 and 1.66 ng/ml, respectively. In our research, we noticed a clear disparity between those with healthy gums and those with chronic periodontitis. Consistent with the findings of other studies, the current analysis showed that one month after scaling and root planing, sCD44 levels in people with chronic periodontitis significantly decrease. This strengthens the case that the spit sCD44 profile might be useful in diagnosing periodontal disease.

Badersten et al.[9] recommended focusing post-stage one therapy assessments within the first 90 days and intermittently afterward for the next nine months. Limiting factors in this regard may include the short follow-up time of the current review (one month). but in the present study follow-up was done after one month.

CONCLUSION

According to the results of our investigation, sCD44 levels can be utilized as a biomarker to identify periodontal disorders. But further research is needed to determine and implement sCD44’s function as a diagnostic biomarker.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.