bioimpresión 3d de piel: tecnología, aplicaciones y futuro · bioimpresión 3d de piel:...
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Bioimpresión 3D de Piel: Tecnología, Aplicaciones y
Futuro
José L. Jorcano
Dept. of Bioengineering. University Carlos III (Madrid)
Division of Epithelial Biomedicine. CIEMAT (Madrid)Catedrático Ramón Areces
COSMETORIUMBarcelona, 3-4 de Octubre 2017
The Need for Artificial Skin
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• In the US, the number of burn injuries receiving medical treatment per year are 486000 and the number of hospitalizations per year for acute burn injuries are 40000, per the 2016 report
• The 2016 global wound management market is expected to hit $15 billion and forecasted to be worth over $22 billion in 2024
• The global tissue engineered skin substitutes market was valued at USD 958.8 million in 2014 and is projected to reach USD 3873.5 million by 2023
• There are two compelling reasons as to why the world is in needof artificial skins:(i) wound healing and skin regeneration—especially for
burn victims, and (ii) drug and skin care products (cosmetics) testing.
From: S. Vijayavenkataraman et al., Biofabrication, 2016
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2ª Conferencia Internacional sobre Ensayos Clínicos y Vigilancia Terapéutica de Medicamentos
In Vitro Toxicology Market
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Distribution of the global in vitro toxicology testing market in2016, by region (in billion U.S. dollars)
https://www.statista.com/statistics/679963/global-in-vitro-toxicology-testing-market-share-by-region/
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New technologies are needed that canreliably mimic human skin and can beindustrialized:
3D Bioprinting and Tissues-on-Chips
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Types of 3D printers:Stereolithography(SLA)Digital Light Processing(DLP)Fused deposition modeling (FDM)Selective Laser Sintering (SLS)Selective laser melting (SLM)Electronic Beam Melting (EBM)Laminated object manufacturing (LOM)
Additive Manufacturing (AM) is a term to describe set of technologies that create 3D objects by adding layer-upon-layer of material. Materials can vary from technology to technology
Where are 3D printing technologies
applied in medicine?
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External prosthesis (orthosis)
Lightweight exoskeleton for
Emma
Arthrogryposis multiplex congenita
(Delawere Hospital & Stratasys)
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Dentistry
Prosthesis
But we are talking about
How to Build Tissues and Organs:
3D BIOprinting
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The components of bioinks
Artificial scaffolds
-Natural polymers (collagen, fibrin)-Synthetic polymers
-Materials Science
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Chitosan-based scaffolds for bone tissue engineering
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SCANNING ELECTRON MICROSCOPE:
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Dermal fibroblasts and Col I matrix
2 m²
2.000 million cells
Renewal rate: 70
million/day
What we would like to mimic…..
……What we can mimic
Dermis
Epidermis
St. corneum
St. basale
St. granulosum
St. spinosum
Basal membrane
SKIN: A COMPLEX STRUCTURE
Technology: Bioprinting Methods
From: S. V. Murphy and A. Atala, Nature Biotech., 2014
Continous extrusion
Technology: Bioprinting Methods
(Continous extrusion)
From: S. V. Murphy and A. Atala, Nature Biotech., 2014
Scheme of a Bioprinter
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Bioinks: the Key to Organ Bioprinting
From: H-W Kang et al., Nature Biotech., 2017
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Table 1. Preparation of the cell-laden composite hydrogels for 3D bioprinted tissue constructs
Composite hydrogel
Gelatin Fibrinogen HA Glycerol Cell type & density
Bone (type I) 35 mg/ml 20 mg/ml 3 mg/ml
10 % v/v Human AFSCs, 5x106 cells/ml
Cartilage (Type I) 45 mg/ml 30 mg/ml 3 mg/ml
10 % v/v Rabbit ear chondrocytes, 40x106 cells/ml
Skeletal muscle (type II) 35 mg/ml 20 mg/ml 3 mg/ml
10 % v/v Mouse C2C12 myoblasts, 3x106
cells/ml
A 3D bioprinting system to produce human-scale tissue constructus with structual integrity. Hyun-Wook Kang, Sang Jin Lee, In Kap Ko, Carlos Kengla, James J Yoo & Anthony Atala. Nature biotechnology , 34 (3) pag. 312. DOI: 10.1038/nbt.3413, Mar2016
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Marlborough, an international fund rising firm, places BioDan together withOrganovo as the sector’s leaders (Inflation driving up costs? Your solution lies in Asia;
May 2017; Sally Macdonald).
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The 3D printing Hype Cycle by Gartner: What does the 2017 edition say?
What were we doing?
Manual production of bioengineered skin
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De aquí obtuvimos nuestras biotintas, la clave de nuestro éxito.
Aproximación desde la biomedicina, no desde la ingeniería
3-D plasma matrix
Fibroblasts
Keratinocytes
Bio-Engineered Skin Equivalent
SKIN BIOPSYBLOOD
A. Meana et al., Burns 1998
S. Llames, et al Transplantation, 2004
PATIENT
1cm² → 2m² in 3 weeksExpansion Factor: 105
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IMPLANTATION
EPIDERMOLYSISFOREARM
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• Extensive burns• Chronic vascular and diabetic ulcers• Surgical and traumatic wounds• Skin hereditary diseases
(Epidermolysis Bullosa)• Necrotizing Fasciitis• GVHD• Giant Nevus• Oral mucosa restitution• Urogenital epithelium sustitution
BIOENGINEERED SKIN – THERAPIES:
RESPONSIBLE FOR PRODUCTION AND MARKETING IN SPAINAS A “CONSOLIDATED MEDICAMENT”
UNDER LICENSE
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SkinMed Autologous Bioactive Skin is manufactured in cleanrooms under strict sterile conditions
and analytical controls.
Generation of Autologous Bioactive Skin under GMP conditions
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3-D In Vitro Skin Cultures
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H/E H/E
Col7 Invol. Fillag.
Vimentin
K14/k10
Tricromicro
Gel diferenciado in vitro 16 días
Alta reproducibilidad:Células de un único donante
En colaboración con BioDan
Colágeno humano
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Our two skin model is unique in the market: It is bilayerd and fully human
Next step: How to improve, automatize and standardize the production of skin equivalents.
A collaboration with BioDan
Biofabrication, 2016
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BIOMATERIALS 2014/2015 – S – 17/03/2014BIOMATERIALS EXPERIMENTAL DESIGN 2014/15 –S2 – 17/03/2014 – MOD 7 - 1
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Our bioprinting system
FIRST-GENERATION BIOPRINTER
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THIRD-GENERATION BIOPRINTER
Juan CañizoLucía GullónFátima PérezCristina Quílez
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IN VITRO 3D CULTURE
Hematoxylin-
Eosin Staining
VimentinK10
Suprabasal
strata
Fibroblast
cytoskeleton
E
D E
D
A
D
C
E F
B D
EBM
IN VIVO ASSAY (I)
h Vimentin
Collagen VII h K10 h Filaggrin
H/E h K5
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Conclusiones
- I. Gracias a los conocimientos científicos y tecnológicos
desarrollados, nos encaminamos a una nueva era en la
cosmética y en el testeo de sus productos (COSMETICA
AVANZADA).
- II. La bioimpresión 3D aplicada al testeo facilitará:
1. Incrementar la producción y reducir los costes
significativamente.
2. Estandarizar los métodos de producción. Para ello, necesita
madurar como tecnología de producción.
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3. Incrementar muy significativamente la reproducibilidad
de los datos frente a los obtenidos con biopsias humanas.
4. Generar in vitro piel más parecida a la real: pelo,
glándulas, hipodermis, mayor complejidad celular,
gradientes de moléculas, etc.
III. El trasplante de órganos producidos industrialmente
será una realidad. Es necesario comenzar a diseñar la
FÁBRICA DE ÓRGANOS DEL FUTURO.
ACKNOWLEDGEMENTS
Tissue Engineering: Alvaro Meana (Centro Comunitario de Sangre y Tejidos de Asturias), Marcela Del Rio (UC3M/CIBERER/Fundación Jiménez Díaz), Fernando Larcher (CIEMAT)
BIOPRINTING: Nieves Cubo Mateo (UC3M), Lucía Gullón (BioDan),
Marta García (UC3M), Diego Velasco (UC3M), Juan Cañizo (HUGM), Fátima Pérez (BioDan), Cristina Quílez (UC3M).
….and many dermatologists and plastic surgeons from Spanish and European hospitals
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THANK YOU !!
Which are the main technologies
involved in 3D printing?
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3D PRINTERS: Layer by layer
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3D Bioprinter SolutionsOrganovo U.S. Army
3DS - 3Dynamical Systems
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Andamiaje sintético biodegradable
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Ceramic (e.g. hydroxyapatite) scaffolds for bone tissue engineering
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Technology: Bioprinting Methods
(Continous extrusion)
From: S. V. Murphy and A. Atala, Nature Biotech., 2014
Also: Mixed systems
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THE PRODUCT
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Therapy: Extensive and Severe Burns
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IN VIVO ASSAY (II)
Vascularization:
A relevant issue in bioprinting.
In this case:
Mouse blood vessels invade our engineered tissue.
E
D
sc
BM
BV
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