OBJECTIVES

To evaluate the cytotoxicity and genotoxicity induced

by root canal sealers-
1. Endoflas (Control group)
2. MTA fillapex,
3. Rokeoseal,
4. Diapex and Experimental groups
5. R-C seal
on Human Embryonic Kidney-293 cell cultures by MTT
assay and MNT assay.
To compare the biocompatibility of the above groups
EXPERIMENTAL DESIGNING
IN VITRO
HEK-293 cells(n=40)


Endoflas MTA filapex Rokeoseal Diapex RC seal
(n=8) (n=8) (n=8) (n=8) (n=8)


Cytotoxicity Genotoxicity cytotoxicity genotoxicity cytotoxicity genotoxicity cytotoxicity genotoxicity cytotoxicity genoyoxi
.

MTT MNT MTT MNT MTT MNT MTT MNT MTT MNT
STATISTICAL ANALYSIS
APPLICATION
Biocompatibility of dental materials is of great importance
as cytotoxic materials may cause inflammatory reactions
when in direct contact with adjacent tissues, compromising
the treatment outcome.
Endodontic therapy aims at elimination of residual pulp,
tissue breakdown products, microorganisms present inside
the root canal system, followed by hermetic filling as
possible, perfect apical seal, tissue mineralization
induction, immunological compatibility, antimicrobial
activity.
In addition Grossman recommended endodontic sealers
not to provoke an immune response in periradicular tissue
and neither be mutagenic nor carcinogenic
STATEMENT OF PROBLEM

Root canal sealers are routinely used in
conventional root canal therapy.
Some endodontic materials have been shown
to have a mutagenic potential to the
surrounding periradicular tissues.
SIGNIFICANCE OF THE PROBLEM

Adverse material effects may play an important role in
the failure of endodontic treatment, even if no major
fault can be identified in treatment.
Therefore, only those root canal sealers that are
biocompatible and display no mutagenic potential
should be used in conventional root canal therapy.
This demands that each endodontic material should be
evaluated for its mutagenic potential before clinical
application.
For example,sealers with inferior biocompatibility, such
should no longer be applied during treatment
METHODOLOGY
CELL LINE MAINTENANCE
(Cells will be grown &maintained in DMEM medium supplement with 10% FBS & 1% antibiotic
solution at 37
0
C with 5% CO
2
in a humidified atmosphere)


CELL LINE HARVESTING/ SUB-CULTURING OF CELL
(Cells will be trypsinized by using 0.25% trypsin-EDTA solution )


CRYOPRESERVATION OF CELL LINE

REVIVING OF FROZEN CELL
Extract/Sample preparations
(All the five endodontic sealers included in this study will be mixed according to
manufacturers instructions under aseptic environment. The materials will be then
placed in non-reactive 12 well plates and allowed to set according to the
manufacturers instructions at 37C)
FRESH EXTRACT
24 HOUR EXTRACT
7 DAYS EXTRACT
Will be collected.
All extracts will be stored at -20C till further use.


Cytotoxicity assay of root canal sealer extracts


MTT ASSAY FOR DETERMINING CELL GROWTH INHIBITION


Data Interpretation of MTT assay



Genotoxicity assay of root canal sealer extracts

MNT ASSAY

Data Interpretation of MNT assay

STATISTICAL ANALYSIS