Dental caries vaccine : induction of protective immunity against streptococcus mutans after immunization with cell free and cell associated with glucosyltransferase

Dissertant

al-Hajj, Yaqub Hamadt Allah

Thesis advisor

al-Sanusi, Sulayman Muhammad
Uthman, Zakiyah Abd al-Samad

University

Omdurman Islamic University

Faculty

Faculty of Medical Laboratory

Department

Department of Medical Microbiology

University Country

Sudan

Degree

Ph.D.

Degree Date

2011

English Abstract

This study was done to evaluate the effect of glucosyltransferase (GTF) enzyme and killed Streptococcus mutans to use as a dental caries vaccine in albino rats.

To isolate S.

mutans saliva was collected from persons suffering from dental caries.

All specimens were inoculated on mitis salivarius bacitracin agar, which is a selective medium for isolation of S.

mutans.

All biochemical tests required to identify S.

mutans were done.

For preparing of GTF enzyme, S.

mutans is grown on BHI broth medium containing 1% glucose at 370c for 18-20 hours.

The supernatant from broth culture media was incubated with 10% sucrose at 370c for 18 hours, to provide water insoluble polysaccharide, and then incubated at 40c for 1 hour.

Water insoluble polysaccharide was removed by centrifugation and urea solution to obtain pure and ready to use GTF enzyme.

Whilst to obtain the killed S.

mutans cells, the organism was cultured on mitis salivarius agar at 370c for 72hours.

Some colonies were then transferred to a test tube containing 5ml of BHI broth and incubated at 370c for 16 hours.

Twenty five microliter transferred to another test tube containing 10 ml of BHI broth with 2% sucrose.

These tubes were incubated at 370c for 22hours and then the bacterial cells were collected by centrifugation.

The bacterial cells were washed by normal saline and killed with 10% formalin.

Thirty albino rats were bred in a suitable place, specially prepared for this target.

The age range of these rats was between 50-60 days and of almost similar weights.

The rats were then divided into 5 groups ; each group containing 6 rats.

Four groups of these were vaccinated using 2types of vaccines, which have been previously prepared (GTF and killed S.

mutans cells).

Immunization has been achieved via oral and intranasal route.

Meanwhile the other group was left not immunized as a control group.

Blood and saliva specimens were collected from all groups before immunization and also one week, 2 weeks, and three weeks after immunization.

The blood samples were used for a complete blood count by using autohematolyzer (sysmex), whereby saliva specimens were used for S.

mutans cells count and the measurement immunoglobulin-A level.

This study has shown a significant increase in white blood cells’ count, which composes the front defense line in the immune system, accompanied by an increase in the immunoglobulin- A level.

This increase has led to a substantial decrease in Streptococcus mutans cell count.

This decrease appeared obviously in three weeks ; the thing that proves that the use of GTF enzyme and the killed Streptococcus mutans have a great role in protecting teeth against caries, which is mainly caused by Streptococcus mutans.

It is noticeable, through this study, that there has been a stability in the several components of blood (as for instance the hemoglobin concentration, red blood cells count and the platelets count), which indicates that there are no adverse side effects from the employment of these vaccines.

So, it is possible to state that the use of both these vaccines can provide sufficient protection against dental cavity.

Main Subjects

Dental

Topics

• Microbiology
• Diseases

No. of Pages

180

Table of Contents

• APA
• MLA
• AMA

Language

English

Data Type

Arab Theses

Record ID

BIM-365039