Evaluation of the Circulatory Levels of Heat Shock Protein 60 Levels in Periodontitis and Cardiovascular Disease Patients

ABSTRACT

Introduction:

HSP is arguably the most thoroughly studied self-antigens connected to Cardio Vascular Diseases (CVD) and periodontal disease. Hence, the major goal of this analysis was to determine the amount of HSP60 in patients’ Chronic Periodontitis (CP) patients’ serum.

Materials and Methods:

The current investigation involved 100 patients in all. Based on the patients’ periodontal and cardiovascular health, the patients were divided. The patients were made aware that this research had no direct bearing on disease treatment or cure.

Results:

In contrast to periodontal disease, which had a mean serum HSP60 of 59.94 ng/dl, CVD had a mean serum HSP60 of 85.98 ng/dl. When compared to periodontal disease, the CVD increased significantly (P < 0.05, 0.03).

Discussion and Conclusion:

We emphasize the function of HSP60 in the pathophysiology of individuals with chronic periodontitis based on the findings of the current investigation. Serum HSP60 concentrations can serve as a biomarker for periodontal inflammation. More longitudinal and interventional research with a larger sample size is required to validate the present findings. In periodontal therapies, targeting HSP60 may enhance results.

INTRODUCTION

Specific gram-negative bacteria colonizing the gingival crevice are the main cause of periodontitis, a persistent tissue-destructive inflammatory disease.[1,2] It has long been believed that exogenous antigens from red-complex bacteria like Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola play a significant role in the etiology of illness. Endogenous antigens have attracted a lot of study in recent years.[3,4]

It is yet unclear how precisely periodontal tissue is harmed. Heat shock proteins (HSPs) are positively expressed in the basal layer of the periodontal pockets, according to immunohistochemical investigations. Under the base layer of periodontal pockets, mononuclear inflammatory cells are infiltrating the tissues more frequently. Such activation of macrophages and other inflammatory cells results in the production of pro-inflammatory cytokines by these cells, which is a result of periodontal bacteria prompting the periodontal cells to create HSPs. The tissue deterioration brought on by periodontal disease is facilitated by this mechanism.[5,6]

Perhaps the most thoroughly studied self-antigen linked to periodontal disease is HSPs. As a result, they are known as molecular chaperones. A chaperonin found in mitochondria called HSP60 helps fold linear amino acid chains into the appropriate three-dimensional structures. More than 50% of the amino acids in human HSP60 and their bacterial counterparts are homologous. Due to this structural similarity, immunological responses may unintentionally be mediated. HSPs are arguably the most thoroughly studied self-antigens connected to Cardio Vascular Diseases (CVD) and periodontal disease.[1,2] Hence, the major goal of this research was to determine the amount of HSP60 in patients with periodontitis and cardiovascular diseases.

MATERIALS AND METHODS

A total of 100 patients were accounted for in the current investigation. All patients were chosen from the institute’s outpatient clinic. The hospital’s department of cardiovascular disorders was involved in the analysis. Before being enrolled in the trial, included patients were given information about it and asked for their written informed consent.

Two groups of included patients were created. The patients were separated based on their cardiovascular and periodontal health. The patients were made aware that this research had no direct bearing on disease treatment or cure.

Patients in group A have periodontal disease if at least six of their teeth have radiographic evidence of bone loss, a probing pocket depth of at least 5 mm, and clinical attachment loss of at least 3 mm.

Group B: Patients with CVD who have active chronic periodontitis and positive coronary angiography results were included in the group.

In the study, patients of all ages were involved.

Exclusion criteria: Patients who underwent scaling or extraction 3 months before to sample collection were not included in the study. The experiment was closed to patients with weak immune systems.

The antecubital fossa was venipunctured to get two milliliters of blood. The blood was centrifuged for 10 minutes at 3000 rpm after being allowed to clot for 30 minutes at room temperature. After that, it was used for more analysis after being kept at -70°C.

A suitable kit for an enzyme-linked immunosorbent test was bought. The ELISA system was used to analyze serum samples in accordance with the manufacturer’s suggested methodology, and a 96-well plate that was pre-coated with the necessary antibodies was employed. Serum samples and standards were added first, and then they were incubated for 3 hours. Following the addition of the conjugate antibody, it was allowed to sit at room temperature for a full hour. After a second wash, the plates received the substrate and were incubated for 30 minutes at room temperature and in the dark. The optical densities were measured at 450 nm, and the findings for HSP60 were expressed by comparing the samples and standards.

The circulatory HSP60 values in health and sickness, which were presented as mean and standard deviation, were statistically examined using the Student’s t-test.

RESULTS

In the current investigation, serum levels of HSP60 were assessed using the sandwich enzyme-linked immune sorbent assay (ELISA) method in patients with periodontal disease and cardiovascular disease.

Patients in group A were in good overall health, while those in group B had cardiovascular disease. The serum HSP60 levels were evaluated using the sandwich ELISA method. 100 samples of saliva were taken from each of the two groups.

While group B had 52 individuals with periodontitis and cardiovascular disease, group A had 48 patients with periodontitis and general health. The serum samples were centrifuged using the ELISA technique to compare the amounts of HSP60 in the two groups [Table 1].

Table 1.

Serum HSP60 levels comparison between the two groups

Groups	No. of cases	Mean value (ng/dl)	P
Group A	48	59.94	0.03
Group B	52	85.98

In contrast to periodontal disease, which had a mean serum HSP60 of 59.94 ng/dl, CVD had a mean serum HSP60 of 85.98 ng/dl. When compared to periodontal disease, the CVD increased significantly (P < 0.05, 0.03).

DISCUSSION

When bacterial antigens in the subgingival biofilm trigger an inflammatory reaction in the gingiva, periodontal disease results. Through immunological or inflammatory interactions, the pathogenic species found in subgingival biofilm harm host tissue. These immune or inflammatory interactions frequently involve neutrophils, monocytes/macrophages, T cells, and B cells.[3,7]

Since ELISA is more adaptable, accurate, repeatable, and affordable than immunoblotting, it was chosen to evaluate HSP60. There is evidence to support the adoption of this strategy from earlier research by Tabeta et al. (2001),[8] Watanabe S et al. (2003),[9] and Handley HH et al. (1995).[10]

The link between periodontal disease and CVD has received a great deal of attention recently. Numerous researchers have discovered a clear correlation between the two disorders, and the American Association of Cardiologists and the American Association of Periodontologists have written a joint consensus report to address this issue. Numerous writers have proposed various pathogenic mechanisms by which periodontal disease may affect CVD. Our research suggests that HSP60 may be another avenue by which periodontal disease affects the state of systemic inflammation and, ultimately, CVD. It is now well-known how inflammation contributes to the formation of atheromatous plaque.

Due to the study’s numerous limitations, which include the following, the results must also be interrupted with caution: A role for developing pathophysiology could not be determined due to the small sample size and the fact that HSP60 levels were not evaluated in relation to the severity of CVD.

CONCLUSION

We emphasize the function of HSP60 in the pathophysiology of individuals with chronic periodontitis based on the findings of the current investigation. Serum HSP60 concentrations can serve as a biomarker for periodontal inflammation. More longitudinal and interventional research with a bigger sample size is required to validate our findings. In periodontal therapies, targeting HSP60 may enhance results.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.