PROGRESS REPORT
CENTRE OF EXCELLENCE IN
ORTHOPEDIC TISSUE ENGINEERING
AND REHABILITATION
FUNDED BY TEQIP-II
National Institute of Technology, Rourkela
Sanction order No: F.No. 16-16/2013-TS. VII (General), (SC), (ST) Date: 13th June 2013
Name of the
Investigator
Department Designation
1 Dr. K. Pramanik Dept. of Biotechnology & Medical Engg. Professor
2 Dr. Subrat Panda Dept. of Mechanical Engg. Assistant Professor
3 Dr. A. Thirunanam Dept. of Biotechnology & Medical Engg Assistant Professor
4 Dr. S.K Sarangi Dept.of Mechanical Engineering Professor
5 Dr. Mukesh K. Gupta Dept. of Biotechnology & Medical Engg Associate Professor
6 Dr. Amit Biswas Dept. of Biotechnology & Medical Engg Assistant Professor
7 Dr S. Dasgupta Dept. of Ceramic Engineering Associate Professor
8 Dr. I. Banerjee Dept. of Biotechnology & Medical Engg Assistant Professor
9 Dr. D.P Mohapatra Dept. of Computer Engineering Associate Professor
10 Dr. Kunal Pal Dept. of Biotechnology & Medical Engg Assistant Professor
11 Dr. B.C. Roy Dept. of Metallurgical &Material Engg. Professor
Name of Investigators
Procurement
Sl no. Name of Equipment Present Status
1 RT-PCR
Equipment received
2 Force Plate 3D Motion Analysis
Equipment received
3 Hypermesh Equipment received
4 Environmental SEM
Process is initiated
5 Mimics
Process is initiated
•Research Scholar Recruitment-
Name of the student PhD/M.Tech by Reseach Darte of joining
1 Sudhanshu S. Behera PhD Dec.2013
2 Shreesan Jena PhD
Dec.2013
3 Gourishankar Saw M.Tech (R) 6th Dec.2013
4 G Gurimruti PhD July 2014
5 Tanushree Sahu PhD July 2014
Activities under CoE
Aim of the research
Development of load bearing orthopedic implants including knee & hip joints
Development of neovascularised Bone Tissue Construct
Osteochondral Tissue engineering
Design of rapid prototyping device for fabrication of 3D scaffold
Development of orthotic solution for patient having abnormal Gait pattern
cellular &
molecular
level
tissue and
organ level
Physiological
Functional
level
ORTHOPEDIC TISSUE ENGINEERING AND REHABILITATION
Our Approach…
WORK DONE SO FAR
Surface modification of load bearing titanium implants for
orthopedic application
Objective
Surface modification of Titanium alloy for improving the wear resistance femur
Surface modification of titanium stem material to reduce the stress shielding
effect and improving the osteoconductive
Development of porous titanium scaffold for load bearing bone defects
Fig 1: SEM micrographs of Thermally oxidized Ti6Al4V at 700°C for (a) 12 hrs. (b) 24
hrs and (c) 36 hrs
•The thermal oxidation of Ti6Al4V oxidized at 700°C suggests the
formation crack free oxide film
•The amount of anatase and rutile phase depends on the temperature &
duration of thermal oxidation
Fig 2: SEM micrographs showing the thickness of oxide formed at
the surface of thermally oxidized at 750°C for (a) 24 hrs (b) 36 hrs.
• The oxide formation of titanium has been studied earlier on the surface
that prevents it from further oxidation or oxygen diffusion at lower
temperature. The occurrence of mainly rutile phase on Ti6Al4V oxidized
at 700°C for 24hrs and 36 hrs suggests the formation of a thick oxide film.
Development of Neovascularised bone tissue construct
Objectives
Preparation & characterization of cobalt (Co+2) doped hydroxyapatite
in-vitro evaluation of its pro angiogenic and osteogenic properties
Generation of bone tissue construct by in vitro growth of cell-seeded scaffold
in vivo biocompatility of tissue construct by animal model test
-
S.No. Sample Notation Color Yield
(g)
Theoretical
%of doping
Experimental
% of doping
1 STD HAp HA1 White - 0
0
2 Pure HAp HA2 White 0.79 0
0
3 0.5% CoCl2-HAp HAC1 White 0.94 0.5 0.175
4 0.5% Co(NO3)2-HA HAN1 White 0.8 0.5 0.175
5 1% CoCl2-HAp HAC2 Light ash 0.79 1 0.33
6 1% Co(NO3)2-HAp HAN2 Light ash 0.83 1 0.37
7 5% CoCl2-HAp HAC3 Dark green 0.85 5 1.19
8 5% Co(NO3)2-HAp HAN3 Dark ask 0.8 5 1.2
Work done so far…
Synthesis of doped hydroxy apatite and characterization of doping
XRD Analysis FTIR Analysis
Physico-chemical Characterization….
Biological Characterization……
Cell proliferation study Cell Cycle analysis
Biological Characterization……
Study of Osteoblast Differentiation
Ru
nx 2
Ostr
ex
SE
M s
tud
y
Biological Characterization……
Study of Angiogenesis
VEGF expression
OBJECTIVES
To develop natural gum modified bio-polymeric hydrogel & tissue engineered scaffold
To study physico-chemical and mechanical properties of the hydrogel and scaffold
To study invivo and invitro biocompatibility of the hydrogel and scaffold
DEVELOPMENT OF CARBOXY-METHYL TAMARIND & TAMARIND
GUM MODIFIED HYDROGELS AND TISSUE ENGINEERED SCAFFOLD
FOR BONE TISSUE REGENERATION
Sample Gelatin(20% soln)
(ml added)
Carboxymethyl
Tamarind gum (20%
soln) (ml added)
Tamarind gum(20%
soln)(ml added)
Glutaraldehyde
(25%) Reagent
(ml)
C1 16 4 - 1
C2 12 8 - 1
C3 8 12 - 1
T1 16 - 4 1
T2 12 - 8 1
T3 8 - 12 1
C1 C2 C3 T1 T2 T3
0.5h 1h 2h 3h 4h 5h 6h 7h 8h
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
% s
we
llin
g r
atio
time
CMT-1
CMT-2
CMT-3
0.5h 1h 2h 3h 4h 5h 6h 7h 8h
0.0
0.1
0.2
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0.7
0.8
0.9
1.0
% s
we
llin
g r
atio
time
CMT-1
CMT-2
CMT-3
Sw
elli
ng
ra
tio
Time
0 200 400
0 200 400
A (##Temp./ّ C)
T1
T2
T3
PG
0.5h 1h 2h 3h 4h 5h 6h 7h 8h
0.0
0.1
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0.9
1.0
% s
we
llin
g r
atio
time
TG-1
TG-2
TG-3
Sw
elli
ng
ra
tio
Time
0.5h 1h 2h 3h 4h 5h 6h 7h 8h
0.0
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1.0
% s
we
llin
g r
atio
time
TG-1
TG-2
TG-3
0 200 400
0 200 400
A (##Temp./ّ C)
C1
C2
C3
PG
DSC Thermogram DSC Thermogram
Swelling study Swelling study
Physico-chemical characterization of the hydrogel
0 10 20 30 40 50 60 70 80
0
10
20
30
40
50
60
70
80
90
Fo
rce
Time(sec)
Force(T1)
Force(T2)
Force(T3)
0 10 20 30 40 50 60 70 80
0
10
20
30
40
50
60
70
80
90
100
110
120
130
140
Fo
rce
(gm
)
Time(sec)
c(1)
c(2)
c(3)
pg(4)
0 2 4 6 8 10 12
0
50
100
150
200
250
300
350
400
450
500
550
600
650
Fo
rce
Time
c(1)
c(2)
c(3)
pg(4)
0 2 4 6 8 10 12
0
50
100
150
200
250
300
350
400
450
500
550
600
650
Fo
rce
Time
c(1)
c(2)
c(3)
pg(4)
0 2 4 6 8 10 12
0.00
0.05
0.10
0.15
0.20
0.25
0.30
Fo
rce
Time(sec)
T(1)
T(2)
T(3)
0 2 4 6 8 10 12
0.00
0.05
0.10
0.15
0.20
0.25
0.30
Fo
rce
Time(sec)
T(1)
T(2)
T(3)
Stress Relaxation Stress Relaxation
Compression Compression
Physico-chemical characterization of the hydrogel (continued…)
Hemocompatibility
Cytocompatibility
60 min 210 min 360 min 510 min 660 min
-5
0
5
10
15
20
25
30
35
40
CP
DR
time(min)
c1
c2
c3
60 min 210 min 360 min 510 min 660 min
-5
0
5
10
15
20
25
30
35
40
CP
DR
time(min)
c1
c2
c3
60 min 210 min 360 min 510 min 660 min
-5
0
5
10
15
20
25
30
35
40
CP
DR
TIME(Min)
T(1)
T(2)
T(3)
60 min 210 min 360 min 510 min 660 min
-5
0
5
10
15
20
25
30
35
40
CP
DR
TIME(Min)
T(1)
T(2)
T(3)
Cell viability of MG-63
Biological characterization of the hydrogel
In vitro drug release study
In vitro drug release study
OBJECTIVES
Development of orthotic solution for people having
abnormal Gait pattern
Studying gait parameters of control group
Designing computational model of orthotic solution
Fabrication of orthotic solution
Gait Analysis of Human Locomotion
Gait Analysis refers to the study of various motions executed by the human
body during day-to-day movement.
A normal gait cycle can be divided into the following phases, as shown
below:
Force plate system
for Gait Analysis
Ground reaction force curve
Ground Reaction Force
(GRF) is the reaction force
offered by the surface on
which the human executes
the act of locomotion.
GRF is the force which
propulses the body
forward.
Representative ground reaction force curve of a normal
Representative ground reaction force curve of a pathological gait
Representative centre of pressure curve of
a) normal and (b) pathological gait
Centre of Pressure graph
Line of action upon which the
gravitational force is acting.
The Torque Curve
Variation of the torque acting about
the ankle-foot joint with respect to
time.
Resultant force acting at a specific
point on the surface of the force
plate and a torque about the
vertical axis.
Representative torque curve of
a) normal and (b) pathological gait
Power expenditure curve of
a) normal and b) pathological gait
The power expenditure curve
Variation of power with time.
(-) shows energy stored during
swing phase. (+) value implies
release of energy (enabling the
forward motion)
The coefficient of friction curve
plots the variation of COF with
time.
Coefficient of friction curve of
a) normal and b) pathological gait
Impulse curve of a) normal and b) pathological gait
Impulse is the change in linear
momentum of the body.
May be defined as the product
of average force multiplied by
the time over which the force
is exerted.
GRF curve in slow gait
Slow-speed gait does not
produce sharp peaks at heel-
strike and toe-off phases of
gait as compared to fast and
normal walking speeds.
Slower gait patterns of
a) normal and b) physically challenged subject
Steps involved in the solid modeling process
Design of an Orthotic foot ware using Solidworks
Orthotic design for the individual
with ankle-foot joint distortion -
(a) three dimensional view (b) front
view and (c) side view.
OTHER ACTIVITIES DONE DURING THE PERIOD
MoU signed:-
•Ispat General Hospital, Rourkela
•University of South Carolina, Columbia, USA (www.usc.edu)
•Mondragon Unibertsitatea, Mondragon, Spain
(www.mondragon.edu)
•Konkuk University, Seoul, South Korea (www.konkuk.ac.kr).
Starting of PG Degree Course:-
A PG Degree course “M. Tech in Biotechnology & Medical
engineering (specialization: Tissue Engineering) has been
planned to be started from the coming academic year
•Publication-
List of journal paper (2013-2014)
•Nadeem Siddiqui, Pramanik, K. (2014), Effects of micro and nano β-TCP fillers in
freeze gelled chitosan scaffold for bone tissue engineering. Journal of applied polymer
science (in Press)-2014
•Senthilguru K, Pramanik K, Pal Kunal, Maiti T. K. & I. Banerjee, Development of
proangiogenic hydroxyapatite for bone tissue engineering communicated to Acta
Biomaterila (2014)
•Panda, N. N., Jonnalagadda, S., & Pramanik, K. (2013). Development and evaluation
of cross-linked collagen-hydroxyapatite scaffolds for tissue engineering. Journal of
Biomaterials Science, Polymer Edition, 24(18), 2031-2044.
•Panda, N. N., Pramanik, K., & Sukla, L. B. (2013). Extraction and characterization of
biocompatible hydroxyapatite from fresh water fish scales for tissue engineering
scaffold. Bioprocess and biosystems engineering, 1-8.
•Kaur, R., Pramanik, K., & Sarangi, S. K. (2013). Cryopreservation-induced stress on
long-term preserved articular cartilage. ISRN Tissue
Engineering,.http://dx.doi.org/10.1155/2013/973542
Publication in conferences
•PrajnaKabiraj, Indranil Banerjee(2014) “Alginate Bead Based Implant
For Drug Release And Tissue Engineering Application” oral paper
presentation national conference on Bio-mechanical science(NCBMS-
2014)
• Parinita Agrawal1, K. Pramanik, “Preparation Of Non-Woven Silk
Fibroin, Chitosan And Poly Ethylene Oxide Nanofibers By Free-Surface
Electrospinning For Cartilage Tissue Engineering” at second
International conference on Tissue Engineering and Regenerative
Medicine (ICTERM-13), NIT Rourkela, Nov-2013
Thank You
OBJECTIVE
• Development of EMG based wireless control system.
• Application of above control system in rehabilitation devices such as
wheel chairs.
Development of wireless EMG based control system
for rehabilitative device (wheel chair)
Basic Block Diagram of EMG based control system
CIRCUIT DESIGN IN MULTISIM
PCB DESIGNING AND DEVELOPMENT
BOTTOM VIEW TOP VIEW
PCB
Index
flexion
Middle
flexion
Thumb
flexion
All finger
abduction
EMG signal for
different finger movement
Signal classification
in Labview Glowing of LED as
per the signal
Classification of EMG signals for controlling the device
TESTING OF THE CLASSIFICATION
EFFICIENCY
PROTOTYPE WHEELCHAIR MOVEMENTS
(a) initial position (b) Forward (c) left and (d) right
COMPLETE SETUP FOR THE WIRELESS CONTROL OF
WHEELCHAIR MOVEMENT USING EMG SIGNAL
CONCLUSION
The ultimate target of the rehabilitation research work to be carried out
under CoE at National Institute of Technology Rourkela in the next three
years are as follows:
• To develop well-researched and optimized orthotic remedies for
individuals with altered gait patterns.
• To fabricate such product(s) and make it viable enough to be made
available in the market.
• To lay the foundation of research for a world-class facility for designing
and fabrication of orthotic and prosthetic solutions.
Methodology
MSC isolation and characterization
• Isolation, culture and sub culture of mesenchymal stem cells from umbilical cord blood
• Cell Characterization using MSC-specific surface markers such as CD105, CD73, and CD90, non specific markers such as CD 44, CD 45 and HLA-DR.
Cryopreservation Experiment
• Preparation of a less toxic cryopreservation solution
• To optimize the biological freezing procedure, hCBMSCs were control-rate frozen in different concentrations of the new cryopreservation solution at different freezing rate
• Thawing at 37°C and determining cell viability by Trypan Blue exclusion test.
Methodology • To investigate the effect of control-rate freezing on
cell metabolism, MTT assay was performed and control-rate frozen hCBMSCs were thawed, cultured and differentiated into chodrocytes and osteocytes.
• Development of tissue engineered construct using
hCBMSCs seed on silk chitosan scaffold
• Development of tissue engineered cartilage
construct(TECC)
• Cryopreservation of TECC
• Verify desirable characteristics of cryopreserved
chondrocytes and cartilaginous tissue constructs-
survival and proliferation study, Type I and II
Collagen and Gag Content by
immunocytochemistry, DNA content by
Expected outcome
• A clinical grade, non toxic freezing solution is expected to be formulated using biological cryoprotectants and natural osmoprotectants.
• A cryopreservation protocol that shall serve as a standard for the preservation of tissue engineered cartilage construct.
Up-dated progress achieved vis-à-vis time schedule of objectives proposed.
Duration Progress of Work Done
Dec 2013- Feb 2014
MSCs were isolated from
umbilical cord blood and
characterized.
Less toxic cryopreservation
solution was prepared and
its potential was verified by
cryopreserving hCBMSCs.
The desirable
characteristics of
cryopreserved cells were
also verified in terms of cell
viability and cell
differentiation potential
Mar –May 2014 Development of TEC
Cell Differentiation Assessment
To construct an instrumented staircase with adjustable step height for recording ground reaction force and moment about ankle data during stair ascent or descent
Use of a 3-D motion analysis system to find out the dynamic reactions at other parts of the body (equipment to be acquired shortly).
Improvements in the existing orthotic solution.
Applying the model solution to a larger control group with similar disability or restriction in motion
Neural and myoelectric signals need to be studied in order to develop powered orthotics for amputees. This is an emerging sector in powered prosthetic limb design with the potential to revolutionize prosthetic development.
FUTURE WORK PLAN
SCHEMATICS REPRESENTATION OF
THE EMG ACQUISITION SYSTEM
SCHEMATICS REPRESENTATION OF
THE DEVELOPED WIRELESS CONTROL
SYSTEM
Conclusion
Easy implementation
Wireless control system
Easy to use
Low cost
Development of scaffold for bone tissue engineering using Curcuma longa extract
Objectives:
i. To extract turmeric essential oil from turmeric plant (Curcuma
longa L.) and their characterization for desired properties.
ii. To develop the scaffolds using essential oil for bone tissue
engineering applications.
Methodology 1. Extraction of turmeric essential oil using steam
distillation method
2. Characterization of extracted turmeric oil by Gas Chromatography Mass Spectroscopy (GC-MS)
3. Preparation of phytochemical scaffold (alginate/CS/CL-E scaffolds) using alginate, chitosan and curcumin extract
4. Characterization of prepared scaffold using XRD, FTIR, SEM, TGA-DSC methods
5. To study swelling property, porosity, cell proliferation, cell viability, cell attachment assay, protein adsorption, in-vitro degradation assay of the prepared scaffolds for bone tissue engineering applications.
Preparation of alginate/CS/CL-E scaffold
Future work plan
• Morphology, porosity and swelling properties of the scaffold will be investigated
• Protein adsorption on the scaffold will be measured and compared with that of control.
• Degradation of scaffold under physiological conditions will be determined.
• The adhesion and spreading of hMSCs on the scaffolds will be analysed by SEM (Cell attachment study).
• The viability of hMSCs on the scaffold will be tested using Alamar blue assay (cell viability assay).
• The cell proliferation assay will be carried out using Alamar blue test (Cell proliferation assay)
• Differentiation of hMSC by alkaline phosphatase (ALP) activity will be analysed.