the biomanufacturing programme at leiria - wordpress… · 2013-07-20 · www. cdr-sp.ipleiria.pt...

www. cdr-sp.ipleiria.pt

Paulo Jorge Bártolo

THE BIOMANUFACTURING PROGRAMME AT LEIRIA


Regional Innovation Scoreboard (2012)


The Centre is located at Marinha Grande in a two floor building of 3320 m2.

The Centre for Rapid and Sustainable Product Development was created in May, 17, 2007 by the President of the Polytechnic Institute of Leiria. It is a Research Centre of the Polytechnic Institute of Leiria (IPL) rated as excellent by the most recent evaluation of Research Units carried out by the Portuguese Foundation for Science and Technology (FCT).


FUTURE BUILDING


KEY RESEARCH THEMES:

Advanced materials – materials with novel functionalities enhancing

new manufacturing technologies and processes and the opportunities to manufacture entirely new materials-based technologies and products. SMART MATERIALS, SUSTAINABLE POLYMERS, FUNCTIONAL GRADED STRUCTURES

Emerging technologies – production technologies to exploit the potential of emerging technologies (in particular bio- and multi-scale technologies); leveraging simulation and modelling techniques to address manufacturing challenges; flexible, rapidly responsive production systems for customized manufacturing.

Sustainable manufacturing and manufacturing of green technologies


GLOBAL FUNDING

0

1000000

2000000

3000000

4000000

5000000

6000000

7000000

8000000

9000000

10000000 2013

2012 2011

2010

2009

2008

2007


Martinho, P., Bártolo, P.J., Pouzada, A., Rapid Prototyping Journal, 5, 71, 2009


Biomanufacturing - the use of additive technologies, biodegradable and biocompatible materials, cells, growth factors, etc., to produce biological structures for tissue engineering applications


BIOMEDICAL IMPLANTS I

Medical implants are devices placed either inside or on the surface of the body to accomplish some particular function, such as to replace, assist or enhance the functionality of some biological structure(s).


BIOMEDICAL IMPLANTS II

BIO

MED

ICA

L IM

PLA

NTS

Body External implants

Body internal permanent implants

Body internal temporary implants

Hip implants Knee implants

Retinal implants Dental implants

Spinal fusion constructs

Scaffolds Degradable screw and plates

Drug delivery systems Spinal fusion constructs


BIOMEDICAL IMPLANTS III


Hand

Eye

Foot

Leg

Arm


BIOMEDICAL IMPLANTS IV


Manabu OKUI – CDRSP & Tokyo Institute of Technology

WALKING INTEGRATIVE REHABILITATION DEVICE


BIOMEDICAL IMPLANTS V

Body Internal permanent implants

IBEROAMERICAN NETWORK ON BIOMANUFACTURING : MATERIALS, PROCESSES AND SIMULATION (BIOFAB) - ~200 researchers - 18 Universities and Research

Institutes - Collaborative research - Courses - Training


BIOMEDICAL IMPLANTS VI


Bucomaxillofacial prosthesis. Beyond aesthetic purposes, this prosthesis showed functional results, improving patient speech and feeding.

Implants can be functional or cosmetic. Source: CTI, Brazil


BIOMEDICAL IMPLANTS VII

Naso-orbitary reconstruction. Tumor resection - esthesioneuroblastoma

Surgery time reduced in about 40% Better quality of the surgery due to the fit and shape of the implant



BIOMEDICAL IMPLANTS VIII


BIOMEDICAL IMPLANTS IX

Biocompatibility. Both raw and processed materials should interact positively with the host environment without eliciting adverse host tissue responses. Biodegradability. Scaffolds must degrade into non-toxic products with a controlled degradation rate that matches the regeneration rate of the native tissue.


BIODEGRADABLE IMPLANTS X

Source: Holzapfel et al, Advanced Drug

Delivery Reviews, 2012

Appropriate porosity, pore size and pore shape. Generally, a high level of porosity is required (> 90%) because it increases the surface area, enabling high cell seeding efficiency, migration and proliferation, as well as neovascularisation. Pore interconnectivity (100% interconnected network of internal channels are required) is also a critical parameter in terms of cell viability and tissue regeneration, maximizing the diffusion and exchange of nutrients and the eliminations of waste.


BIODEGRADABLE IMPLANTS XI

Source: Zhang et al, Advanced Drug Delivery Reviews, 2012

Bioactive. Scaffolds should be bioactive, promoting and guiding cell proliferation, differentiation and tissue growth. This can be achieved by adding growth factors and functionalizing the scaffold with proteins or adhesion-specific peptide sequences.


BIOMEDICAL IMPLANTS XII

Cells

Signals Scaffold

Cell-scaffold interactions

Cell-cell interactions

Tissue Engineering main pillars:

Physicochemical interaction (shear, pH, O2, CO2, temperature


BIOMEDICAL IMPLANTS XIII

Hydrogels

Polymers Hydrogels

Polymers Polymer/ceramic


BIOMANUFACTURING @ CDRSP I

MATERIAL CHARACTERISATION MANUFACTURING PROCESSES

COMPUTER MODELLING & SIMULATION


BIOMANUFACTURING @ CDRSP II

COMPUTER MODELLING & SIMULATION 4D Modelling Approach


BIOMANUFACTURING @ CDRSP III


BIOMANUFACTURING @ CDRSP IV

ADDITIVE PROCESSES NON-ADDITIVE

PROCESSES

HYBRID PROCESSES

EXTRUSION-BASED PROCESSES

STEREOLITHOGRAPHIC PROCESSES

ELECTROSPINNING

PRINTING PROCESSES

PLASMA TREATMENT/COATING


BIOMANUFACTURING @ CDRSP VI


BIOMANUFACTURING @ CDRSP VII

Melt-electrospun sPCL189 at a temperature of 160 ◦C (Vapp =25 kV, EFR = 3.75±0.08 mL h−1): (a) focused deposition by ‘static’ melt-ES; (b) uniform 3D fibre collection by computer-aided melt-ES (extruder translation velocity 500 mm min−1).

SEMmicrograph of melt-electrospun scaffolds produced with Vapp =25 kV and SRS = 29.0 rpm: (a) linear PCL produced with Tproc =160 ◦C; (b) sPCL64 produced with Tproc =125 ◦C; (c) sPCL189 produced with Tproc =160 ◦C; (d) sPCL189 melt-electrospun fibres collected in a coiled form following the deposition pattern (Tproc =160 ◦C) – the arrow indicates the deposition direction. Inset images show details of the fibre surface topography obtained; scale bar 10 μm.


VAT-PHOTOPOLYMERISATION I

PEGDMA Dextran PEGDMA/HA PEGDMA/Dextran UP systems


VAT-PHOTOPOLYMERISATION II

PEG


VAT-PHOTOPOLYMERISATION III

PEG


VAT-PHOTOPOLYMERISATION IV

Ethylene glycol dimethacrylate

Triethylene glycol dimethacrylate

The HEMA was copolymerized with 1-vinyl-2-pyrrolidinone using two differents crosslinks agents

1-Vinyl-2-Pyrrolidinone HEMA


VAT-PHOTOPOLYMERISATION V

HEMA with 0.5wt% of PI

HEMA with 1wt% of PI HEMA with 2wt% of PI

HEMA with 3wt% PI


VAT-PHOTOPOLYMERISATION VI

Irradiation time (s)

No

rma

lize

d w

ate

r a

bso

rptio

n

0,0

0,2

0,4

0,6

0,8

1,0

1,2

1,4

1,6

1,8

2,0

15 25 35 45 60

T= 15 s T= 60 s

PEGDA

Dextran

Influence of the irradiation time on the water absorption of PEGDMA constructs built by STLG system (left). Macroscopic images of the constructs produced using 15s and 60s of irradiation time, after immersion into distilled water during 24 hours (right). Scale bar represents 0.5mm.


VAT-PHOTOPOLYMERISATION VII

Energy (KeV)

0 1 2 3 4 5 6 7 8

Counts

0

2000

4000

6000

8000

10000

12000

14000

16000

Energy (KeV)

0 1 2 3 4 5 6 7 8

Counts

0

2000

4000

6000

8000

10000

12000

14000

16000

Ca

Ca

P

C

O

a) b)

EDS analysis and SEM micrographs of the PEGDMA (a) and PEGDMA/HA hydrogel scaffolds (b).


VAT-PHOTOPOLYMERISATION VIII


VAT-PHOTOPOLYMERISATION IX

Curing time (s)

0 100 200 300 400 500

Fra

cti

on

al co

nvers

ion

0.0

0.2

0.4

0.6

0.8

1.0

50 wt% of styrene

60 wt% of styrene

70 wt% of styrene

37 wt% of styrene

Coral-like structure

Flake-and-pore structure

Flake-type structure

Curing time (s)

0 100 200 300 400 500

Fra

cti

on

al

co

nve

rsio

n

0.0

0.2

0.4

0.6

0.8

1.0

70 wt% of styrene

37 wt% of styrene


VAT-PHOTOPOLYMERISATION XI

COMPOSITE STEREOLITHOGRAPHY Magnetic field


VAT-PHOTOPOLYMERISATION X

Cláudio Ferreira, MSc work

a) b)

c)


VAT-PHOTOPOLYMERISATION XIII


EXTRUSION-BASED PROCESSES I


EXTRUSION-BASED PROCESSES II


EXTRUSION-BASED PROCESSES III

PCL PCL/PLA PCL/HA PCL/TCP PCL/graphene PCL/PHB PLA Alginate Alginate/aloe vera Alginate/agar Alginate/chitosane

Material characterisation: DSC, FTIR, NMR, GPC, AFM


EXTRUSION-BASED PROCESSES IV

FIBROBLASTS


EXTRUSION-BASED PROCESSES V

Saos-2 cells


EXTRUSION-BASED PROCESSES VI

Human mesenchymal stem cells (hMSCs)


EXTRUSION-BASED PROCESSES VII

Tendogenesis/

LigamentogenesisMarrow Stroma

Transitory

FibroblastTransitory

Stromal Cell

Osteogenesis Chondrogenesis Myogenesis

MSC Proliferation

Transitory

Osteoblast

Transitory

Chondrocyte Myoblast

Mesenchymal Stem Cell (MSC)

Myoblast Fusion

Unique

Micro-niche

ChondrocyteOsteoblast

Other

Bo

ne

Ma

rro

w/P

eri

os

teu

mM

es

en

ch

ym

al

Tis

su

e

Proliferation

Commitment

Lineage

Progression

Differentiation

Maturation

OsteocyteHypertrophic

ChondrocyteMyotube

Stromal

Cells

T/L

Fibroblast

Adipocytes,

Dermal and

Other Cells

BONE CARTILAGE MUSCLE MARROWTENDON/

LIGAMENT

CONNECTIVE

TISSUE

THE MESENGENIC PROCESS


EXTRUSION-BASED PROCESSES VIII

Extruded PCL scaffolds were plasma modified with a C2H4/N2 deposition (plasma deposited ethylene: nitrogen, PdE:Ncoating) followed by H2 post treatment (PdE:N/H2 coating).


EXTRUSION-BASED PROCESSES IX

Surface Modification Plasma Argon (Ar)

Power 100 W

Time 30 s

Frequency 0,6

Graft polymerization with Acrylic Acid (Aac)

PCL/Collagen


EXTRUSION-BASED PROCESSES X

PCL/Collagen Incorporation of collagen

0

0,01

0,02

0,03

0,04

0,05

0,06

0,07

0,08

Day 7 Day 14

Ab

sorb

ance

PCL scaffold 0/90º PCL-col scaffold 0/90º

Human Fibroblasts (3T3)


EXTRUSION-BASED PROCESSES XI

Scaffolds PCL/HA (75:25)

Scaffold PCL/HA Scaffold PCL

Video 2


EXTRUSION-BASED PROCESSES XII

PCL

PCL/HA

Video 3

Scaffold PCL/Bioglass (70/30)

Video 1


EXTRUSION-BASED PROCESSES XIII

HYDROZONE – BIOACTIVATED HIERARCHICAL HYDROGELS AS ZONAL IMPLANTS FOR ARTICULAR CARTILAGE REGENERATION. Global funding: ~12MEuros; CDRSP funding: ~1 MEuros


EXTRUSION-BASED PROCESSES XIV

Physiological operating temperature (4ºC-37ºC) will be ensured, preserving temperature sensitive and bioactive components. Additionally, na irradiation system composed of a light source and na optical fibre attached to the extrusion heads will be implemented. This system will work as a secondary cross-linking mechanism of the extruded hydrogels, increasing the structural stability of the constructs.


EXTRUSION-BASED PROCESSES XV


EXTRUSION-BASED PROCESSES XVI


EXTRUSION-BASED PROCESSES XVII


EXTRUSION-BASED PROCESSES XVIII


EXTRUSION-BASED PROCESSES XIX

1 - Dermal skin graft with more flexible hydrogel, entrapping fibroblasts 2 - Epidermal skin graft with stiffer hydrogel, entrapping keratinocytes

3 1

1 2


EXTRUSION-BASED PROCESSES XX

Film

Thic

kness

in

cre

ase

(%

)

0

50

100

150

200

250

300

350

AG AGA5 AGA15 AGA25

In vitro degradation – 10 weeks in SBF solution

Degradation period (week)

0 1 2 3 4 5 6 7 8 9 10

Wei

ght

loss

(%

)

0

2

4

6

8

10

12

14

16

18

20

Film AG

Film AGA5

Film AGA15

Film AGA25

a)

Degradation period (week)

0 1 2 3 4 5 6 7 8 9 10W

ater

abso

rpti

on (

%)

0

200

400

600

800

1000

1200

1400

1600

b)

Time

Water molecules

IN

Aloe vera

OUT

Cleavable bond

Carbohydrate Polymers, In Press


MULTI-MATERIAL LASER SINTERING FOR THE PRODUCTION OF FUNCTIONAL GRADED STRUCTURES


FUTURE CHALLENGES I


FUTURE CHALLENGES II


FUTURE CHALLENGES III


SkelGEN - ESTABLISHMENT OF A CROSS CONTINENT CONSORTIUM FOR ENHANCING REGENERATIVE MEDICINE IN SKELETAL TISSUES.

FUTURE CHALLENGES IV


Thank you!

The first Portuguese AM machine

the biomanufacturing programme at leiria - wordpress… · 2013-07-20 · www. cdr-sp.ipleiria.pt...

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