organ printingdownloads.deusm.com/designnews/25419-organ_printing_how...organ printing: how to print...
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Organ Printing:How to print a human organ
Vladimir Mironov MD, PhDAssociate Professor & Director ofAdvanced Tissue Biofabrication Center (ATBC)Department of Regenerative Medicine & Cell BiologyMedical University of South CarolinaCharleston, SC 29425, USA
ATBC
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Question 1:
How new technologiesevolve?
Answer: from preexistingtechnologies
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1. Francis Bacon (1607)(old linear model)2. Terence Kealey (1996) (radical parallel model)3. Brian Arthur (2009) (new integrated model)
New technologies are emerging from preexisting technologies and not from basic science,
ut emerging technologies must explore and incorporate new phenomena discovered by basic scientists
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Question 2:
Is South Carolina an emerging high tech
manufacturing state?
Answer: yes
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South Carolina is a High Tech State
North Charleston, SC
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South Carolina is a High Tech State
Spartanburg, SC
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South Carolina is a High Tech State Rock Hill, SC
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South Carolina is a High Tech State
Charleston, SC
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Question 3:
Is tissue engineeringscience or technology?
Answer: it is both science and technology
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"Scientists discover the world that exists;
engineers create the world that never was.”
Theodore Von Kármán
Tissue engineers are trying to “recreate” human tissues
& organs that exist
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Question 4:
What is Synthetic Anatomy?
Answer:Anatomy of XXI century
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Biofabrication, 2009
From analytical to synthetic anatomy
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Question 5:
What is a status of industrial tissue
engineering today?
Answer: it could be better
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Organogenesis, USA
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Genzyme, USA
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Cytograft Tissue Engineering, USA
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Tengion, USA
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Question 6:
Why do we need bioprinted functional
human organ constructs?
A d
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OrganPrinting
$25 000 000market
only for TEkidney
$75 000annualcost ofdialysis
Huge Market
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Question 7:
What is an organ printing?
Answer: Biomedicalapplication of RP or AM
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Organ Printingis a computer-aided
robotic layer by layeradditive biofabrication
of functional livinghuman organ
constructs
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Bioimaging
Blueprint
CAD
Bioink
Bioprinter
Biopaper
Maturogens
Bioreactor
Biomonitoring
I. Pre-processing
II. Processing
III. Post-processing
Components ofOrgan PrintingTechnology
3 Steps in Bioprinting
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Question 8:What is a status
of biomedical applicationof rapid prototyping oradditive manufacturingin tissue engineering?Answer: it is growing
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RP
RP in TE
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RP
RP in TE
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1. Title: Mechanical properties and cell cultural response of polycaprolactone scaffolds designed and fabricated via fused deposition modelingAuthor(s): Hutmacher DW, Schantz T, Zein I, et al.Source: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH Volume: 55 Issue: 2 Pages: 203-216 Published: MAY 2001 36 44 35 53 35 262 29.112. Title: Fused deposition modeling of novel scaffold architectures for tissue engineering applicationsAuthor(s): Zein I, Hutmacher DW, Tan KC, et al.Source: BIOMATERIALS Volume: 23 Issue: 4 Pages: 1169-1185 Published: FEB 2002 32 42 41 42 20 222 27.753. Title: The design of scaffolds for use in tissue engineering. Part II. Rapid prototyping techniquesAuthor(s): Yang SF, Leong KF, Du ZH, et al.Source: TISSUE ENGINEERING Volume: 8 Issue: 1 Pages: 1-11 Published: FEB 2002 29 31 22 37 14 169 21.124. Title: Solid freeform fabrication of three-dimensional scaffolds for engineering replacement tissues and organsAuthor(s): Leong KF, Cheah CM, Chua CKSource: BIOMATERIALS Volume: 24 Issue: 13 Pages: 2363-2378 Published: JUN 2003 18 27 31 50 27 161 23.005. Title: Organ printing: computer-aided jet-based 3D tissue engineeringAuthor(s): Mironov V, Boland T, Trusk T, et al.Source: TRENDS IN BIOTECHNOLOGY Volume: 21 Issue: 4 Pages: 157-161 Published: APR 2003 18 25 18 40 28 154 22.006. Title: Scaffold-based tissue engineering: rationale for computer-aided design andsolid free-form fabrication systemsAuthor(s): Hutmacher DW, Sittinger M, Risbud MVSource: TRENDS IN BIOTECHNOLOGY Volume: 22 Issue: 7 Pages: 354-362 Published: JUL 2004 13 25 34 35 27 136 22.677. Title: Bone tissue engineering using polycaprolactone scaffolds fabricated via selective laser sinteringAuthor(s): Williams JM, Adewunmi A, Schek RM, et al.Source: BIOMATERIALS Volume: 26 Issue: 23 Pages: 4817-4827 Published: AUG 2005 2 18 34 42 22 118 23.608. Title: Rapid prototyping of scaffolds derived from thermoreversible hydrogels and tailored for applications in tissue engineeringAuthor(s): Landers R, Hubner U, Schmelzeisen R, et al.Source: BIOMATERIALS Volume: 23 Issue: 23 Pages: 4437-4447 Published: DEC 2002 20 22 13 25 17 117 14.629. Title: Fabrication of PLGA scaffolds using soft lithography and microsyringe depositionAuthor(s): Vozzi G, Flaim C, Ahluwalia A, et al.Source: BIOMATERIALS Volume: 24 Issue: 14 Pages: 2533-2540 Published: JUN 2003 17 23 16 27 10 108 15.4310. Title: Novel collagen scaffolds with predefined internal morphology made by solid freeform fabricationAuthor(s): Sachlos E, Reis N, Ainsley C, et al.Source: BIOMATERIALS Volume: 24 Issue: 8 Pages: 1487-1497 Published: APR 200313 16 24 15 15 92 13.14
TOP 10
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Research clusters (RP in TE)
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Question 9:
What is a “bioink” ?
Answer: self-assembling“fluidic” tissue spheroids
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Rounding
Fusion
Enveloping
Spreading
Evidence of Tissue Spheroids Fluidit
Sorting
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Question 10:
What is a fundamental principle of organ
printing technology?
Answer: tissue fusion
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‘Virtual and Physical Prototyping’ (2009)
Tissue spheroids fusion
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Question 11:
What is a clinically relevant cell source for
organ printing?
Answer: iPS cells or ADSC
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Adipose Tissue Derived Mesenchymal Stem Cell(ADSC) is a clinically relevant stem cell source
“David after two years staying in USA” (two photos on left side) “Celution System” (Cytori Therapeutics Inc. & GE Health Care)
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Immunofluorescence analysis
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ADSCs differentiate into SMCs in a developmentally appropriate
sequence
mRN
AW
este
rn a
naly
sis
Densinometric analysis
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Quantitative Phenotypic Analysis
Proliferation
Differentiation
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3-dimensional constructs of ADSC-derived SMCs contract in response to
Angiotensin administration
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Real time RT-PCR analysis of vascular ECM gene expression
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Matrigel vasculogenesis assay
anti-hPECAM1 anti-αSMA Co-incident fluorescence
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Question 12:How to print
an intraorgan branched vascular tree ?
Answer: using 3 types ofvascular tissue spheroids
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Basic Kidney Anatomy
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Design principles of biofabrication of intraorgan branched vascular tree
from self-assembling vascular tissue spheroids
Three types of vascular tissue spheroids
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Road Map & Timeline for Organ Printing
2003
2009
2020?
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Tissue fusion without cell mixing
Tissue compaction during fusionof vascular tissue spheroids
Virtual and Physical Prototyping, 2009
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Tissue Engineering, 2008
Bioengineering Vascular Tube UsingSelf-assembling Tissue Spheriods
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Bioprinting of Vascular Tree
‘Tissue Engineering’ (2008), featured in ‘Nature News’
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Dev. Dynamics (submittted)
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Tissue engineered branched segments of vascular tree
Regenerative Medicine, 2008 & Biomaterials, 2009
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Mechanism of Lumen Formation
Kamei M, Saunders WB, Bayless KJ, Dye L, Davis GE, Weinstein BM. Endothelial tubes assemble from intracellular vacuoles in vivo.
Nature. 2006 Jul 27;442(7101):453-6
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Bioprinting of Vascular Tree
Virtual and Physical Prototyping, 2009
‘Surface Evolver’, Ken Brakke
‘Blueprint’
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A B C
D E F
G
How to print a human organ?
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Stem cells
Tissue spheroids
Differentiated cells
Tissuespheroidsin hydrogel
Functionalmature macrotissue
Microfluidic minibioreactorsand cell sorters
Microfluidic mixers,droplet generators and encapsulators
Microfluidic dispensors
and bioprinters
Macrobioreactorwith removable porous tubes
“Trends in Biotechnology’ (under review process)
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Question 13:
What is a tissue spheroids “biofabricator” ?
Answer: robotic tissue spheroids bioassembler
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Scalable Robotic Biofabrication of Tissue Spheroi
EpMotion 5070 (Eppendorf)
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Digital (Droplets) Microfluidics
Acoustic excitation based droplet generator in collaboration with Dr. Wen and Dr. Huang (Clemson University)
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Digital Microfluidics: Droplet Generator
Jennifer Lewis & David Weitz
Self-directedself-assembly
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(a)
(c)
(b)
Digital Microfluidics: Cascade Droplet Generator
David Weitz, Science, 2005
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Question 14:
What is a “bioprinter” ?
Answer: robotic dispenso& tissue bioassembler
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Bioprinters
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(a) (b)Microfluidics: Bioprinting
Continuous and digital (droplet) microfluidics-based bioprinting
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Question 15:
What is a “bioreactor” ?
Answer: perfusion containerfor tissue maturation
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Perfusion Bioreactor for Bioprinted Organs
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Perfusion Bioreactor for Bioprinted Organs
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Microfluidics-based irrigation dripping tripled perfusion bioreactor with removable microporous tubes
In collaboration with Dr. Kasyanov, RSU, Latvia, EU & Prof. Chua Chee Kai, NTU, Singapore
12
3
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Question 16:
What is a “maturogen” ?
Answer: factor acceleratingbioprinted tissue maturation
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Methods of quantitative evaluation ofthe material properties of tissue spheroids
Tensiometry Aspiration Centrifugation
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Tissue Spheroids Enveloping Assay
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Question 17:
How robotic automated industrial production of
human organs will look like?
Answer: like assembly line
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Clinical cell sorters
TGI 1200
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Robotic Biofabrication of Tissue Spheroids
EpMotion 5070 (Eppendorf)Acoustic excitator
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(i) (k)
Neatco-1(A), Neatco-2(B), Sciperio/nScript(C)
A B C
Robotic bioprinters
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Perfusion Bioreactors for Bioprinted Organs
Lungs Kidney Liver
Organ Recovery Systems Inc.
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nScript Inc.(Sciperio), USA
Kidney Biofabrication Plant
$1 billion?
$20 million NSF grant2025?
ATBC
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Question 18:
What is a future of bioprintingtechnology?
Answer: in vivo bioprinter
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“Da Vinci” - First Generation of Robotic Surgery Tool(Intuitive Surgery Inc, CA)
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“Face/Off” (1997)
Bioprinting in situ(a science fiction)
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Bioprinting in situ (an emerging concept, 2003)
Clinical Bioprinter - Project “Michelangelo”?
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(scientific reality, 2009)Bioprinting in situ
“In Vivo High-Throughput Biological Laser Printing of Nano-Hydroxyapatite in Mice Calvaria Critical Size Defect: Preliminary Results”
Virginie Keriquel et al. (best poster)Presented in Bordeaux 6-8 July 2009 at The International Conference Bioprinting and Biofabrication in Bordeaux (3B’09)
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“There is no such thing
as a science fiction.There is only
science eventuality.”Prof. Krummel
Chair of Department of SurgeryStanford University, CA, USA
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“The best way to predict the future
is to invent it”Alan C. Kay
fellow at Apple Computer Inc., a visionary
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Question 19:
What must be done?
Answer: a lot
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We need training course“Biomedical Applications of Rapid Prototyping”Charleston, USA 2010Organizer: Vladimir Mironov
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We need to build necessary infrastructure for emerging fielda) research society,b) new journal“Biofabrication”Editor: Wei Sun,c) e-textbook “Biofabrication”Editor: Vladimir Mironov
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We need multidisciplinaryNational Research CentersOn Biomedical Applicationof Rapid Prototyping or Bioprinting and Biofabricationin every developed nation and large developing and emerging nations
ATBC
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We need virtual conferences on“Virtual Tissues” and “Digital Human” enabled by modern telecommunication center as well as virtual global community and networks and interactive website
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We need Industrial Report on:“Biomedical Applications of Rapid Prototyping” similarto world famous & well respected “Wohlers Report”
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We need annual conference:5th International Conference on Bioprinting, Biofabrication and Bioassembly (3B’10)Philadelphia, USA, October 2010Organizers: Wei Sun & Gabor Forgacs
Welcome in City of Brotherhood Love and Motherland of US democracy
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Question 20:How to build a new
biofabrication industry in South Carolina?
Answer: joined focused& integrated efforts
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ATBC
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Advanced Tissue Biofabrication Center
ATBC
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ATBC
Prototypehuman organbiofabricationassembly line
Tissue spheroidsrobotic biofabricator
Robotic bioprinter Perfusion bioreactorCell sorter
From blueprint to organ
ATBC
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Development of new curriculum and new Bachelor Program inBiomedical Rapid Prototyping
SC RAPID PROTOTYPINGTRAINING CENTERS
ATBC
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MUSC & NEATCO Inc.
ATBC
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South Carolina Robotic Biofabrication Industry Cluster
Medical Devices & Implants, Cell, Tissue & Organ
Based Therapy
Tissue-Based BiotechnologyAnimal-Free Animal Products
(meat, leather, fur)
Business, financial & legal service
Tissue-Based In Vitro Diagnostic, Drug Discovery &Toxicology High Throughput
& High Content Assays
3D System UniversityYork Technical CollegeMUSC, USC, CU
South CarolinaGovernment:
Department ofCommerce,
South Carolina Research Authority
Blueprints, Softwares
SC Stem Cells Bank
Biomaterials
Risk capital:VC, Angel Groups
Advanced TissueBiofabrication Research
& Training CenterPerfusion Media
Bioreactors
Palmetto InstituteNew Carolina
Robotic Biofabrication Industry Companies:
Existing (3D Systems), Recruited & Start-up
Bioprinters
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Acknowledgement:1. NSF FIBR grant2. NSF EPSCoR R-II3. MUSC Bioprinting
Research Center grant4. Tan Chin Tuan Fellowship
e-mail: [email protected]
ATBC