EVALUATING THE POTENTIAL OF ELECTROSPUN MEMBRANES DECLARATION I, Dexy Joseph, hereby declare that I

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Transcript of EVALUATING THE POTENTIAL OF ELECTROSPUN MEMBRANES DECLARATION I, Dexy Joseph, hereby declare that I

  • EVALUATING THE POTENTIAL OF ELECTROSPUN MEMBRANES FOR TISSUE SPECIFIC SIGNAL DELIVERY

    IN CARTILAGE AND BONE TISSUE ENGINEERING

    A DISSERTATION SUBMITTED

    By

    DEXY JOSEPH

    IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF

    MASTER OF PHILOSOPHY

    SREE CHITRA TIRUNAL INSTITUTE FOR MEDICAL SCIENCES AND TECHNOLOGY

    THIRUVANANTHAPURAM– 695 012

    July 2013

  • DECLARATION

    I, Dexy Joseph, hereby declare that I had personally carried out the work depicted

    in the thesis entitled “Evaluating the potential of electrospun membranes for

    tissue specific signal delivery in cartilage and bone tissue engineering” under

    the direct supervision of Dr. Neethu Mohan, CHVF-D, Division of Tissue

    Engineering and Regenerative Technologies, Biomedical Technology Wing, Sree

    Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram,

    Kerala, India. External help sought are acknowledged.

    Signature

    Dexy Joseph

  • SREE CHITRA TIRUNAL INSTITUTE FOR MEDICAL SCIENCES & TECHNOLOGY

    THIRUVANANTHAPURAM – 695011, INDIA (An Institute of National Importance under Govt.of India)

    CERTIFICATE

    This is to certify that the dissertation entitled “Evaluating the potential of

    electrospun membranes for tissue specific signal delivery in cartilage and

    bone tissue engineering” submitted by Dexy Joseph in partial fulfilment for the

    degree of Master of Philosophy in Biomedical Technology to be awarded by this

    Institute. The entire work was done by her under my supervision and guidance at,

    Division of Tissue Engineering and Regenerative Technologies, Biomedical

    Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology

    (SCTIMST), Thiruvananthapuram, 695012.

    Thiruvananthapuram Signature Date Dr. Neethu Mohan CHVF- D DTERT, SCTIMST

  • The Dissertation

    Entitled

    EVALUATING THE POTENTIAL OF ELECTROSPUN MEMBRANES FOR TISSUE

    SPECIFIC SIGNAL DELIVERY IN CARTILAGE AND BONE TISSUE

    ENGINEERING

    Submitted

    By

    Dexy Joseph

    For

    Master of Philosophy in Biomedical Technology

    Of

    SREE CHITRA TIRUNAL INSTITUTE FOR MEDICAL SCIENCES AND TECHNOLOGY

    THIRUVANANTHAPURAM

    Evaluated and approved

    By

    Supervisor Examiner’s Name and Designation Dr. Neethu Mohan CHVF- D DTERT

  • ACKNOWLEDGEMENTS

    I would like to express my heartfelt gratitude to my guide Dr. Neethu Mohan and

    HOD, DTERT, Dr. Prabha D Nair, for their guidance, constant encouragement,

    inspiring discussions and valuable suggestions to do this work with confidence. I

    would forever be grateful to you especially for giving me this incredible opportunity to

    be a part of DTERT.

    I express my sincere gratitude to Dr. K. Radhakrishnan (Former Director), Dr.

    Jagan Mohan Tharakan (Present Director), Dr. A.V George (Registrar), Dr. C P

    Sharma (Head, BMT wing) and Dr. S. Sundar Jaya Singh (Deputy Registrar),

    SCTIMST for granting me permission to work in this institute and to make use of the

    facilities needed during the course.

    I am indebted, to Dr. Jayabalan, Dr. Sunitha, our course coordinators, for

    providing me this opportunity. I thank Dr. Kallyana Krishnan, Dr. Anoop Kumar T, Dr.

    K.Sreenivasan, Dr. Harikrishna Varma P. R., Dr. Lissy K Krishnan, Dr. Maya

    Nandakumar and Dr. Shabareeswaran A. for allowing me to use various equipments

    for my studies. I thank all other Scientists and staff at BMT campus. I would like to

    thank all MPhil faculties who had conducted various course works. I thank Mrs.

    Radha, Mr. Renjith, Mr. Joseph and Mr. Nishad for helping me to do my works.

    On personal note, I would like to thank every member of DTERT for their support

    and encouragement. I would like to thank Dr. Babitha, Dr. Dhanya, Dr. Bindu, Dr.

    Lynda, Mrs. Neena, Mr. Merlin, Mr. Dhanesh, Ms. Rakhi, Mrs. Geetha, Ms. Anupama

    and Ms. Neelima, Division of Tissue Engineering and Regenerative Technology for

    their friendly support and helping hands.

    I thank my beloved parents and all those who have directly or indirectly

    contributed to the success of my work.

    Above all, I thank God almighty for His blessings

    Dexy Joseph

  • LIST OF ABBREVIATIONS

    OA Osteoarthritis

    ACT Autologous cell transplantation

    ECM Extracellular matrix

    PCL Poly(caprolactone)

    PEG Poly(ethylene glycol)

    PLA Poly(lactic) acid

    PLGA Poly(lactic-co-glycolic)acid

    FDA Food and drug administration

    HCA Hydroxyl carbonate apatite

    CS Chondroitin sulfate

    FTIR Fourier transform infrared spectroscopy

    ESEM Environmental scanning electron microscope

    ADMSC Adipose derived mesenchymal stem cell

    BG Bioactive glass

    SEM Scanning electron microscope

    GAG Glycosaminoglycans

    AC Articular cartilage

    MACT Matrix assisted autologous cartilage transplantation

    HA Hydroxy apatite

    TGF Transforming growth factor

    BMP Bone morphogenic protein

    IGF Insulin like growth factor

    FGF Fibroblastic growth factor

  • ASC Adipose stem cell

    MSC Mesenchymal stem cell

    CSL Chondroitin sulfate low concentration

    CSH Chondroitin sulfate high concentration

    BGL Bioactive glass low concentration

    BGH Bioactive glass high concentration

    FACS Fluorescent activated cell sorting

    FBS Fetal bovine serum

    CD Cluster of differentiation

  • LIST OF CONTENTS SYNOPSIS..................................................................................................................1 1. INTRODUCTION ....................................................................................................5

    1.1 Background............................................................................................................5

    1.2 Literature review....................................................................................................6

    1.2.1 Composition and anatomy of articular cartilage [AC] and subchondral bone.................6

    1.2.2 Articular cartilage injury...................................................................................................9

    1.2.3 Treatments strategies.....................................................................................................9

    1.2.4 Tissue engineering.......................................................................................................10

    1.2.5 Osteochondral tissue engineering................................................................................11

    1.2.6 Signals used for osteochondral tissue engineering......................................................12

    1.2. 7 Cells sources for osteochondral tissue engineering....................................................13

    1.2.8 Chondroitin sulfate and its role in cartilage tissue engineerin ………………………....13

    1.2.9 Bioactive glass in bone tissue engineering...................................................................14

    1.2.10 Electrospinning...........................................................................................................14 .

    1.2 Hypothesis...........................................................................................................16

    1.3 Objectives............................................................................................................16

    2. MATERIALS AND METHODS...............................................................................17

    2.1Fabrication of biosignal incorporated membranes................................................17

    2.2 Characterizations of membranes.......................................................................