Fabrication of biodegradable cardiovascular stent using

thermo induced poly(DL- lactide) shape memory polymer

by FDM and study its biodegradability, biocompatibility
and simulation by FEA

GUIDE -- Dr. D.SANGEETHA

PROJECT MEMBERS -- VIKAS P

LOKESHWAR R
MANOJ KUMAR S
INTRODUCTION
• The treatment of coronary arterial stenosis (vascular obstruction diseases) can be performed by the
implantation of stents during percutaneous trans-luminal coronary angioplasty.
• Since the first bare metal stent (BMS) was implanted in the human body in 1986, the cardiovascular disease
treatment entered a new stage.
• But the most feared complication related to coronary stent placement is stent thrombosis.
•  To avoid the late thrombosis and the extreme late thrombosis induced by non-degradable stents,
biodegradable stents were invented.
• The first polymer stent implanted in humans was the Igaki-Tamai stent made of PLLA in the year 1998.
•  Biodegradable vascular stents (BVSs) made from biodegradable materials are designed to disintegrate over
several months and to avoid complications from long-term mucosal irritation or extraction.
• Shape memory polymers (SMPs) have the property to store the energy of applied shapes under different
stimuli like temperature, light, electric field, magnetic field, pH, specific ions, or enzyme and quickly regain
their original shapes.
• Use of the shape memory effect in polymers is proposed for cardiovascular stent interventions to reduce the
catheter size for delivery. Stents from shape memory polymers could be triggered by suitable stimuli.
•  3D printing has emerged as an alternative system for fabricating stents . Fused deposition modeling (FDM) is
the most preferred  3D printing technology because of its low cost, high reliability, simple operation, and
personal designing.
LITERATURE SURVEY
• Research on PDLLA

[1]   Material  - PDLLA/HA

        Specimen preparation - Solvent casting and Compression molding
        Analysis - SEM, DSC, DMA, Shape recovery test

[2]  Material -PDLLA/HA,PDLLA/β-TCP, PDLLA/MCC

        Specimen preparation-Solvent casting
       Analysis - Shape recovery test

[3]   Material  - Nano-hydroxyapatite reinforced poly-D-L-lactide

        Specimen preparation -Solvent casting and Compression molding
        Analysis - SEM, DSC, DMA, Shape recovery test, invitro degradation 

[4]  Material  -poly(DL-lactide), poly(L-lactide-glycolide) and poly(e-caprolactone)

        Specimen preparation - Solvent casting and Compression molding
        Analysis - DSC, Tensile testing, invitro degradation 
LITERATURE SURVEY[CONTD.]

[5]  Material  - HA-g-PDLLA nanocomposites

        Specimen preparation - Solvent casting and Compression molding
        Analysis - DSC, Shape recovery test, Invitro degradation

[6]     Material  - Co–Cr coated with Sirolimus Loaded PDLLA

        Specimen preparation - spray coating
        Analysis - SEM, FTIR, HPLC, Platelet adhesion, Histopathological Analysis,
In Vivo Examination

[7]    Material  - PDLLA and PDLLA-co-GLY(uteritic stent)

        Specimen preparation - Solvent casting 
        Analysis - Cell Adhesion, Degradation and Cytotoxicity
LITERATURE SURVEY[CONTD.]
• General survey
[8] Material  - PLA
     Specimen preparation- desktop filament extruder
      Stent  preparation -  FDM   
      Analysis -DSC,DMA, shape recovery test, FEA

[9] Material  - PCLAU/Fe3O4

      Specimen preparation- ring-opening polymerization, solution casting
      Stent  preparation -  stent prepared by rolling strip on a glass rod 
      Analysis -DSC,DMA, shape recovery test, invitro degradation

[10]  Material  -Thermoplastic PU/PCL ester–based TPU 

      Specimen preparation- magnetic stirring
      Analysis -FTIR,SEM,DSC,DMTA, shape recovery test.

[11] Material  - MM7520 polyurethane

      Specimen preparation-  Compression molding
      Stent  preparation -  dip coating on stainless steel pins
      Analysis -DSC,DMTA,Stent Crimping and Deployment
 LITERATURE SURVEY[CONTD.]

[12]  Material  - PPC and PCL (PL-25)

      Specimen preparation- Compression molding, melt processing through a twin-screw extruder    
       Analysis -DSC,DMA, shape recovery test, Creep-recovery Test, In Vitro Degradation Test, Blood
Compatibility, Platelet Adhesion, Clotting Time, Drug Loading and In Vitro Drug Release
[Metoprolol tartrate (MPT)].

[13] Material  - PCTBV

     PCTBV is synthesized  
      Analysis -NMR,GPC,DSC, shape recovery test, Tensile stress test, Cytotoxicity and biocompatibility

[14] Material  - Heparin Coating on 3D Printed Poly (l-lactic acid)

     Specimen preparation- heparinized PLA stent obtained by coating PDA, PEI, and Hep
       Stent  preparation -  3D bioprinting system (lab-made system in the Korean Institute of Machinery and
Materials)  
       Analysis - Platelet adhesion, Invitro degradation, Histopathological analysis, In vitro cell proliferation.
LITERATURE SURVEY[CONTD.]
 [15]Material  - Poly-lactic acid (PLA) and poly (D,L-lactic-co-glycolide) (PLGA)
      Specimen preparation-solution casting
       Stent  preparation - stent prepared by rolling strip on a glass rod 
       Analysis -DSC, In-vitro drug release, FTIR, Hemocompatibility, Platelet adhesion, Bacterial adhesion, Cell

proliferation assay.

[16] Material- PLA

     Stent preparation - homemade 3 d printer system

[17]  Material  - polyurethane

      Specimen preparation- Solution Mixing Method, Melt Mixing Method
   Analysis -DSC, Shape Memory Effect (SME) in Water, uniaxial tensile test, DMTA.

[18]  Material  - PEG-PCL

         PEG PCL was syntesized
         Stent  preparation -  molded in an oven 
       Analysis -DSC, shape recovery test, invitro degradation, cytotoxicity, Anticoagulant and Antiproliferation
Effect, invitro drug release
GAPS IN LITERATURE SURVEY

․Invitro degradation has not been performed on PDLLA

․ Platelet adhesion test has not been performed on PDLLA

․ Clotting time test has not been performed on PDLLA

․ Finite element analysis has not been performed on PDLLA stent model

․ PDLLA stent has not been fabricated using 3D printing

OBJECTIVE

• PDLLA procurement
• Sample preparation 
• Experimental analysis           
      
• DSC
• DMA
• Visual shape recovery
• Biodegradability
• Biocompatibility
Platelet adhesion 
Clotting time
• FEA
• Fabrication of stent
PDLLA PROCUREMENT

Poly (DL-lactide) is a biodegradable polymer for medical devices and pharmaceutical applications. It is used
to fabricate resorbable medical devices that degrade over months in physiological conditions.

Density : 1.25 to 1.27 g /cm3

Tensile strength : 27.6 to 50  MPa
Elastic modulus : 1 to 3.45 GPa
Tg   : 50 to 60 0C
Tm : Amorphous (no melting point)

We can procure PDLLA granule from carbanio.com

SAMPLE PREPARATION

SOLVENT CASTING (strips & films):

• PDLLA granules are dissolved in a suitable solvent like dichloromethane (DCM),
tetrahydrofuran (THF), ethyl acetate or acetone by continuous stirring at room temperature until
it is completely dissolved. The solution is then poured into a mold of suitable dimensions and
allowed to dry for 24 hours. The PDLLA strips are then removed from the mold after complete
evaporation of the solvent.
SAMPLE PREPARATION[CONTD.]

FILAMENT EXTRUDER(PDLLA filament)

• A 3D desktop filament extruder is available commercially which can be used to fabricate
PDLLA in the form of wire. PDLLA granules are introduced into the extruder. The extruder
then gives out PDLLA wire of required dimensions. The extruder can be either used separately
or used as a part of the 3D printing machine
EXPERIMENTAL ANALYSIS  

Differential Scanning Calorimeter

• Differential scanning calorimeter (DSC) is a technique used to investigate the response of polymers to
heating. DSC can be used to study the melting of a  polymer or the glass transition.

Instrument setup 
• The DSC set-up is composed of a measurement chamber and a computer.
• Two pans are heated in the measurement chamber. The sample pan contains the material being investigated. 
• A second pan, which is typically empty, is used as a reference. The computer is used to monitor the
temperature and regulate the rate at which the temperature of the pans changes. A typical heating rate is
around 10 °C/min.
EXPERIMENTAL ANALYSIS [CONTD.] 

Sample preparation
• The sample should weigh between 5 and 15mg.The sample must be a suitable size and shape to fit in the
aluminum pan. The sample can be in the form of powder or liquid.

Working 
• The computer is used to monitor the temperature and regulate the rate at which the temperature of the pans
changes.
•  A typical heating rate is around 10 C/min. The rate of temperature change for a given amount of heat will
differ between the two pans. 
• This difference depends on the composition of the pan contents as well as physical changes such as phase
changes. The system varies the heat provided to one of the pans in order to keep the temperature of both pans
the same. 
• The difference in heat output of the two heaters is recorded. The result is a plot of the difference in heat (q)
versus temperature (T).
EXPERIMENTAL ANALYSIS [CONTD.] 

Dynamic Mechanical Analysis

• Dynamic Mechanical Analysis (DMA) is a technique used to measure the mechanical properties of a wide
range of materials.
 
Sample preparation
•  The sample must be in the form of a strip. The standard strip dimensions are length 5 to 20mm , width 1 to 8
mm and thickness 0.5 to 2 mm.

Working 
• The shape memory properties of PDLLA can be investigated by cyclic thermo mechanical analysis using a
DMA-800 instrument in a force control mode. The strip with required dimensions is taken. The strip should
be kept at a particular temperature for the required time. A required  preload force should be applied to the
strip.
EXPERIMENTAL ANALYSIS [CONTD.] 
Working(contd.)
• The original length L0 should be recorded. Then the strip should be heated to a particular temperature at a
required rate, kept isothermal for some time and then stretched with a  required tensile stress for a particular
time and must be cooled rapidly  and kept isothermal for some time.
• The stretched length L1 should be recorded. Then the stress must be  released and the fixed length L2 should
be recorded. The strip must be reheated at a particular temperature (recovery temperature) for the required
time and kept isothermal for some time. The recovery length L 3 should be recorded.
Shape fixity ratio
• The  shape fixity ratio describes the ability of switching segments to fix the mechanical deformation.

Shape recovery ratio

•  The shape recovery ratio describes the ability of the material to memorize its permanent shape.
EXPERIMENTAL ANALYSIS [CONTD.] 

Visual recovery 
• The shape recovery behavior  can be investigated by using a visual recovery method.  The shape recovery of
the specimen can be noted with respect to time, thus allowing us to investigate the rate of recovery.

Sample preparation 
• The strip is heated to the glass transition temperature and rolled on a steel rod to form a stent shape structure
and then quenched to retain the shape.

Instrument setup 
• A hot water bath is required to change the temperature of the specimen. A camera is required to record the
shape changes with respect to time.

Working 
• The stent shaped specimen is now introduced into the hot water bath set at the glass transition temperature.
The specimen then returns to the original shape (strip). This change in the shape of the stent to its original
shape(strip) is noted with the help of a digital camera.
EXPERIMENTAL ANALYSIS [CONTD.] 

In Vitro biodegradability 
• Biodegradability is another important consideration in the design of a SMP that is highly dependent on the
intended permanence of the final product. For products that are only required to serve a temporary function
such as a vascular stent
• In this work, biodegradability will allow for a transient existence that eliminates the needs for surgical
removal after their functional term.

Instrument setup
• The instrument setup includes the required specimen in the form of strip, phosphate buffered saline(PBS) and
shaker.
EXPERIMENTAL ANALYSIS [CONTD.] 

Working
• Numerous specimens should be immersed into an airtight bottle containing PBS solution which is then placed
in a shaker at a predetermined condition. After certain time intervals each specimen was removed and washed
and cleaned thoroughly with distilled water and then dried.
• Before immersing the specimen into the PBS solution their masses were calculated and after the
predetermined time interval the reduction in their mass were compared. The mass loss with respect to the
degradation time can be plotted in a graph.

Mass loss percentage

• Mass loss = (1 - Md/Mo) × 100% 
where Md and Mo denote the dry weight after degradation and the original weight of the sample, respectively.
EXPERIMENTAL ANALYSIS [CONTD.] 

Platelet adhesion 
• Platelet adhesion is an essential function in response to vascular injury and is generally viewed as the first
step during which single platelets bind through specific membrane receptors to cellular and extracellular
matrix constituents of the vessel wall and tissues.
• Platelet adhesion to biomaterial surfaces or an injured vessel wall is the primary and most important step
leading to the spreading and activation of platelets to form a thrombus. 

Instrument setup
• In this test, human blood plasma was used to test the blood compatibility of the samples. Healthy fresh human
blood was collected from a volunteer using vacuum tubes containing sodium citrate as an anticoagulant.
• The concentration of sodium citrate should be 3.8 wt.%, and the ratio of the anticoagulant to blood should be
1:9. The blood should be centrifuged at 1000 rpm for 15 min to obtain platelet-rich plasma (PRP). The
plasma should be stored at 20 °C until use.
EXPERIMENTAL ANALYSIS [CONTD.] 

Working 
• To eliminate the effects of the other components, such as erythrocytes and leukocytes, in the blood, PRP
must be used for investigating platelet adhesion. The specimens must be immersed in PBS and equilibrated
at 37 °C for 1 h.
• Then the PBS solution must be removed and PRP should be introduced. After the specimens are incubated
with PRP at 37 °C for 2 hr., the PRP is decanted off and the specimens must be rinsed with PBS.
• Finally the specimens are treated with 2.5 wt.% glutaraldehyde in PBS at 4 °C for 1 day.
• The prepared specimens should be then washed with PBS and subjected to a drying process by passing
them through a series of graded alcohol-PBS solutions (30, 50, 70, 80, 90, 95 and 100%) and isoamyl
acetate-alcohol solutions (30, 50, 70,80, 90, 95 and 100%) for 15 min each time.
• Platelet adhesion can be observed using a scanning electron microscope. The adherent platelets on the
surface of the specimens can be counted from the SEM images.
EXPERIMENTAL ANALYSIS [CONTD.] 

Clotting time
• Clotting time is the time required for a sample of blood to coagulate in vitro under standard conditions

Instrument setup
• In this test, human blood plasma was used to test the blood compatibility of the samples. Healthy fresh human
blood was collected from a volunteer using vacuum tubes containing sodium citrate as an anticoagulant.
• The concentration of sodium citrate should be 3.8 wt%, and the ratio of the anticoagulant to blood should be
1:9. The blood should be centrifuged at 4000 rpm for 15 min to obtain platelet-poor plasma (PPP). The
plasma was stored at 20 °C until use.
EXPERIMENTAL ANALYSIS [CONTD.] 

Working
•  The anticoagulant properties of the specimen  can be  analysed by using blood coagulation analyzer. The
analyzer measures activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time
(TT).
• The specimens must be placed in a 24 well cell culture and then  PPP should be used to incubate the samples
in a well . Subsequently, 50 μL of the incubated PPP must be added into a test cup, followed by the addition
of 50 μL of APTT agent.
• The resulting solution should be  incubated at 37 °C for 3 min. Thereafter, 50 μL of a 0.025 M CaCl2
solution is  added, and the APTT can be measured. For the PT test, 100 μL of the incubated PPP must be 
added into a test cup and then mixed with 100 μL of Thromborel S agent at 37 °C for 3 min before measuring
the PT.
• For the TT test, 100 μL of the incubated PPP must be  added into a test cup, followed by the addition of 50
μL of test thrombin agent, and the resulting solution should be incubated at 37 °C for 2 min.
• Then, the TT can be  measured. Finally, the clotting times for APTT, PT and TT can be  automatically
collected and printed by the equipment. The assays must be performed in triplicate for each sample, and the
results were averaged to obtain a reliable value.
FINITE ELEMENT ANALYSIS

• Finite Element Analysis or FEA is the simulation of a physical phenomenon using a numerical mathematical
technique. The finite element has emerged as a practical tool for evaluating the biomechanical performance of
endovascular stents.
• FEA is performed using stent models with cylindrical structure and radial wall displacement to cause stent
deformation.
• It aims at estimating the distribution of residual stress, plastic strain, and the expanded shape under
simulation state, Maximum Stress & Strain: From initial loading to final device deployment, Recoil and
Foreshortening after loading, Recoil and Foreshortening after deployment, Operating Stress Levels, Stent
fatigue (contraction and expansion).
• In the simulation, the stent is compressed while above Tg, then cooled below Tg such that it remains smaller
than the artery, after placement in the artery, the stent is heated again and it pushes against the artery wall.
• Inside the artery the stent undergoes  longitudinal contraction and radial expansion due to the pressure exerted
by the blood vessels and the flow of blood. The large longitudinal contraction and nonuniform radical
expansion would induce injury to vascular(blood vessels).
• In this simulation we subject the stent to the same pressure such as in the blood vessel and study the
longitudinal contraction and radial expansion.
FABRICATION OF STENT
FDM
• Filament extrusion 
• The PDLLA sample is first dried by heating at 800C  for 12 hrs. to prevent degradation of PDLLA or bubble generation
during the hot melt extrusion process.
• The extrusion can be performed using desktop filament extruder. The required extrusion temperature and rate are set.
PDLLA filaments with smooth surfaces and required diameter can be obtained.

• FABRICATION
•  The design or model selected for stent fabrication must be exported as an STL file. The STL file should be imported into
the print software along with the polymer filament ready to feed the FDM machine.
• The filament is melted into the extruder nozzle, which then deposits the melted material onto a controlled rotatory platform.
• The PDLLA comes out of the extruder nozzle in molten form and is deposited layer by layer to fabricate a stent.
LOCATION OF TEST

• DSC in material characterization lab

• DMA at IITM(status-not confirmed)
• Shape memory effect with time in materials lab
• Invitro Degradation in materials lab
• Platelet adhesion in health center and materials lab
• Clotting time in health center and material lab
• Design and FEA in open foam or other open source software
• Filament extruder at IITM(physics lab) (status-not confirmed)
• FDM at AUFRG
EXPECTED OUTCOME

DSC
• Differential Scanning Calorimetry (DSC) is used to determine the glass transition temperature (Tg) and
melting temperature (Tm) of PDLLA.
DMA
• The shape fixity ratio (Rf) and shape recovery ratio (Rr) of PDLLA can be determined by Dynamic
Mechanical analysis. High  shape fixity and shape recovery ratios  are the desirable outcomes.
Visual recovery
• The rate of recovery from temporary to permanent shape is determined by using visual recovery. The
purpose of this test is to analyze the shape memory property of PDLLA by using heat as a stimuli and fast
recovery is desirable.
In Vitro biodegradability 
• Mass loss percentage of  PDLLA is calculated using this test over a period of time(degradation time). The
rate of degradation can be determined from a plot of mass loss and degradation time.
EXPECTED OUTCOME[CONTD.]

Platelet adhesion 
• The  platelet adhesion property of PDLLA is determined by this test. The number of platelets adhering
to the surface of PDLLA must be minimum so that there will be no coagulation and thrombosis in the
blood vessels.
Clotting time
•  A longer clotting time is desirable so that blood coagulation and clot formation doesn’t occur in the
blood vessel.
FEA
• In this analysis the longitudinal contraction and radical expansion of the PDLLA model stent is
simulated. The desired outcome is that the stent model must have hardly any longitudinal contraction or
radical expansion under its working condition. The shape recovery ratio must be high.
REFERENCE

[1]Shape memory properties of poly(d,l-lactide)/hydroxyapatite composites Xiaotong Zheng, Shaobing Zhou,

Xiaohong Li, Jie Weng
[2]Recent progress in shape memory polymers for biomedical applications Hong-mei Chen, Lin Wang and
Shao-Bing Zhou
[3]Shape memory effect of thermal-responsive nano-hydroxyapatite reinforced poly-d-l-lactide composites with
porous structure Ling Chen, Jia Xin Wang, Chak Yin Tang, Da Zhu Chen, Wing Cheung Law
[4]In vitro degradation and drug release from polymer blends based on poly(dl-lactide), poly(l-lactide-
glycolide) and poly(e-caprolactone) Paul F. Mcdonald, John G. Lyons, Luke M. Geever, Clement L.
Higginbotham
[5]Shape memory behaviour of ha-g-pdlla nanocomposites prepared via in situ polymerization Ke Duab and
Zhihua Gan
[6]A promising approach for improving the coating stability and in vivo performance of biodegradable polymer-
coated sirolimus-eluting stent tarek M. Bedair, Sung Nam Kang, Yoon Ki Joung and Dong Keun Han
[7]Development of a biodegradable ureteric stent: surface modification and in vitro assessment Andreas brauers
M.D., Helmut thissen ph.D. ,T Oliver Pfannschmidt ph.D., Hans Bienert, ph.D., Alexandra Foerster, Doris klee,
ph.D., T Walter Michaeli, ph.D. ,Í Hartwig Höcker, ph.D. ,F and Gerhard Jakse, M.D
[8] 3D printed self-expandable vascular stents from biodegradable shape memory polymer Han Jia | Shu-Ying
Gu| Kun Chang
[9] A dual-induced self-expandable stent based on biodegradable shape memory polyurethane nanocomposites
(PCLAU/Fe3O4) triggered around body temperature Shu-Ying Gu Kun Chang, Sheng-Peng Jin
 REFERENCE [CONTD.]

[10] An experimental–numerical study on shape memory behavior of PU/PCL/ ZnO ternary blend M Abbasi-Shirsavar,
M Baghani, M Taghavimehr,M Golzar, M Nikzad, M Ansari and D George
[11] Thermomechanical Properties, Collapse Pressure, and Expansion of Shape Memory Polymer Neurovascular Stent
Prototypes Ge´ raldine M. Baer,Thomas S. Wilson,Ward Small IV, Jonathan Hartman,William J. Benett, Dennis L.
Matthews, Duncan J. Maitland
[12] Biocompatible Shape Memory Blend for Self-Expandable Stents with Potential Biomedical Applications Yu
Zheng, Ying Li, Xue Hu, Jiabin Shen, and Shaoyun Guo
[13] Biodegradable shape-memory block co-polymers for fast self-expandable stents
Liang Xue, Shiyao Dai, Zhi Li*
[14] Heparin Coating on 3D Printed Poly (l-lactic acid) Biodegradable Cardiovascular Stent via Mild Surface
Modification Approach for Coronary Artery Implantation Sang Jin Lee ,Ha Hyeon Jo , Kyung Seob Lim , Dohyung
Lim , Soojin Lee, Jun Hee Lee , Wan Doo Kima, Myung Ho Jeong , Joong Yeon Lim , Il Keun Kwon ,Youngmee
Jung , Jun-Kyu Park, Su A Park.
[15] Fabrication and characterization of shape memory polymers based bioabsorbable biomedical drug
eluting stent Vratika C. Sonawane, Mahesh P. More, Abhijeet P. Pandey, Pravin O. Patil & Prashant K. Deshmukh.
[16] Experimental Study of Polymeric Stent Fabrication Using Homemade 3D Printing System Danyang Zhao, Ruiqi
Zhou, Jianxing Sun, Hongxia Li,Yifei Jin.
 REFERENCE [CONTD.]

[17] Shape memory characterization of poly(ε-caprolactone) (PCL)/polyurethane (PU) in combined torsion-tension

loading with potential applications in cardiovascular stent Mehdi Ansari , Mohammad Golzara, Mostafa Baghani ,
Masoud Soleimani
[18] Thermo-Induced Shape-Memory PEG-PCL Copolymer as a Dual-Drug-Eluting Biodegradable Stent Chien-Shen
Yang, Hsi-Chin Wu, Jui-Sheng Sun, Hao-Ming Hsiao zu-Wei Wang