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  • NANOFIBER INCORPORATED INTRACANAL MEDICAMENTS AND ITS ANTIBACTERIAL

    EFFECT AGAINST ENTEROCOCCUS FAECALIS BIOFILM - AN INVITRO STUDY

    Dissertation submitted to

    THE TAMIL NADU DR M.G.R. MEDICAL UNIVERSITY

    In partial fulfillment for the degree of

    MASTER OF DENTAL SURGERY

    BRANCH — IV

    CONSERVATIVE DENTISTRY AND ENDODONTICS

    2016- 2019

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  • ACKNOWLEDGEMENT

    First of all, I thank GOD, THE ALMIGHTY, for blessing me abundantly and for giving

    me the confidence and inclination to complete this Dissertation.

    I express my sincere thanks to Chairman Thiru. Lion. Dr.K.S.Rangasamy, MJF.,

    Principal Dr.G.S.Kumar, M.D.S., K.S.R Institute of Dental Science and Research,

    Thiruchengode, for allowing me to pursue this course and avail the facilities of this college.

    I express my deepest gratitude to my Professor, Head of the Department and my guide

    Dr. Sebeena Mathew, M.D.S., for her expert and never-failing guidance, valuable suggestions,

    constant encouragement and support with kindness in all aspects of my career.

    I am extremely thankful to Dr.K.Karthick,M.D.S., Dr.T.Boopathi.,M.D.S., and

    Dr.Deepa.N.T for their caring advices which has driven me to work with confidence in both

    academic and clinical studies.

    I would like to thank my biggest source of strength, my parents ,

    Mr.Prakash.P.K and Mrs .Sudha whose unwavering, unselfish love, their expeditious

    encouragement and prayers have always been a pillar of support fo r m e.

    I would like to thank my seniors Dr.Kumar, Dr.Jayakumar,

    Dr.Haribaskar,Dr.Sreedev,Dr.Iswarya,Dr.Nishan,Dr.AbithaBanu,Dr.Loganathan

    and Dr.Poojitha for their constant support and motivation, friends for my life time.

    I extend my heartfelt thanks to my co PG’s Dr.Elangovan and Dr.Kamesh for

    supporting me throughout the course.

  • I would like to express my sincere gratitude to my Dr.Mythili,

    Dr.Mayilanandham, Dr.Kanimozhi, Dr.Aarthi, Dr.Janani and Dr.Nivedha for

    their support.

    I thank my colleague Dr.Suhail for being there with me all the time. I

    thank the non-teaching faculty from the Department of Conservative

    Dentistry and Endodontics for their prompt and patient help throughout the

    course.

  • CONTENTS

    TABLE OF CONTENTS

    SL NO

    TITLE

    PAGE NO.

    1.

    INTRODUCTION

    1

    2.

    AIM AND OBJECTIVES

    4

    3.

    REVIEW OF LITERATURE

    5

    4.

    MATERIALS AND METHODS

    24

    5,

    RESULTS

    40

    6.

    DISCUSSION

    50

    7.

    SUMMARY

    56

    8.

    CONCLUSION

    57

    9.

    BIBLIOGRAPHY

    58

    10.

    ANNEXURE

    67

  • CONTENTS

    LIST OF FIGURES

    S NO TITLE PAGE NO

    1.

    70 samples of coronal third of radicular

    Dentin

    28

    2. Polyvinylpyrrolidone 29

    3. After dispensing of polymer and monomer 29

    4.

    Magnetic stirring for triple antibiotic

    nanofiber for 24 hours

    30

    5.

    After completion of magnetic stirring for

    triple antibiotic nanofiber for 24 hours

    31

    6.

    Magnetic stirring for calcium hydroxide

    nanofiber

    31

    7.

    After 24 hours, stirred solution loaded into

    the syringe

    32

    8. Nano Fiber Electrospinning Unit 32

    9. Placed in the electrospinning 33

  • CONTENTS

    10.

    After electrospinning, nanofibers are

    obtained

    33

    11. After removal of nanofibers 34

    12.

    Field Emission Scanning Electron

    Microscope

    34

    13.

    Nanofiber confirmed in SEM at lower

    magnification

    35

    14.

    Nanofiber confirmed in SEM at higher

    magnification

    35

    15. Laminar flow chamber 36

    16. Enterococcus faecalis innoculation 36

    17. Placed in the incubator shaker for 2 days 37

    18. After 2 days, enterococcus faecalis culture 37

    19.

    After 5 days, conformation of enterococcus

    faecalis growth in SEM

    38

    20. Fluoroscent dye 38

    21. Confocal Laser Scanning Microscope 39

  • CONTENTS

    22.

    Live Enterococcus faecalis in control

    group (Group I)

    40

    23.

    Live Enterococcus faecalis in calcium

    hydroxide paste group (Group II)

    40

    24.

    Live Enterococcus faecalis in calcium

    hydroxide nanofiber group(Group III)

    41

    25.

    Live Enterococcus Faecalis in Triple

    antibiotic paste group (Group IV)

    41

    26.

    Live Enterococcus faecalis in Triple

    antibiotic nanofiber group (Group V)

    42

  • CONTENTS

    LIST OF TABLES

    S NO TITLE PAGE NO

    1. Mean standard deviation for live bacteria 43

    2.

    Tukey HSD for multiple comparisons of Group I

    versus Group II, Group III, Group IV, Group V

    44

    3.

    Tukey HSD for multiple comparisons of Group II

    versus Group I, Group III, Group IV, Group V

    45

    4.

    Tukey HSD for multiple comparisons of Group III

    versus Group I, Group II, Group IV, Group V

    46

    5.

    Tukey HSD for multiple comparisons of Group IV

    versus Group I, Group II, Group III, Group V

    47

    6.

    Tukey HSD for multiple comparisons of GroupV

    versus Group I, Group II, Group III, Group IV

    48

  • CONTENTS

    LIST OF BAR DIAGRAM

    SL NO

    TITLE

    PAGE NO.

    1.

    Live bacterial count

    49

  • INTRODUCTION

    1

    The success of endodontic therapy depends on complete disinfection of root canal system

    which can be accomplished by thorough chemomechanical preparation and three dimensional

    obturation of the root canal system.1 Infected teeth have formation of bacterial biofilm on

    canal walls, lateral canals, in the isthumus, inside dentinal tubules and on apical extra

    radicular root surface.2

    Endodontic infection consists of between 1 and 12 bacterial species. This colonization

    comprises of mostly facultative and strict anaerobic bacteria which have the ability to

    proliferate via interactions among bacterial cell proteins that establish a complex spatial

    structure known as biofilm. Several bacterial species have been identified including

    Enterococcus faecalis and Streptococcus sanguinis Meanwhile, a recent study found

    Actinomyces naeslundii to be the most prevalent organism in traumatized immature

    permanent teeth with necrotic pulps3.

    Enterococcus faecalis is a mushroom shaped microorganism seen in dentinal tubules,

    isthumus, rami, and in a lateral canals. Enterococcus faecalis is the most commonly seen

    microorganism in asymptomatic persistent infections and also poorly endodontic treated

    teeth . Enterococcus faecalis is the predominant bacteria that causes root canal failuers. It is a

    facultative anaerobic bacteria. Enterococcus faecalis is present in a small proportion of the

    flora in canals that are untreated. Enterococcus faecalis biofilms are resistant against

    antibodies, antimicrobial agents and phagocytes.4

    A Triple antibiotic mixture of metronidazole, ciprofloxacin, and minocycline is used for root

    canal disinfection. When using a mixture of metronidazole, ciprofloxacin, and minocycline

    bacterial elimination is seen in the deep root canal dentin. Metronidazole being bactericidal,

  • INTRODUCTION

    2

    Minocycline being bacteriostatic and Ciprofloxacin being a bactericidal broad spectrum

    synthetic quinolone, triple antibiotic paste is able to eradicate both gram positive aerobes and

    anaerobic bacteria. Antibiotic ineffectiveness in systemic route of administration has led to

    the intra canal application to increase its efficacy.5

    Calcium hydroxide is the most commonly used