Bioengineered skin for therapeutical and industrial applications

1. 3D bioprinting of functional human skin:
production and in vivo analysis
Cubo N., Garcia M, Del Cañizo JF, Velasco D,
Jorcano JL.
Biofabrication. 2016 Dec 5;9(1):015006.
May 2019

2. Why?
➔ Medical: 11 million injuries
per year worldwide
require medical attention.
265 000 lead to death.
➔ Industrial: Accurately
predict the response of
human skin to products.
2
Objective
Print bioengineered skin for therapeutical and industrial applications in an
automatized manner.

3. 1.
INTRODUCTION
Let’s start with the base
3

4. Epidermis: Protective
outermost portion of the skin
● Keratinocytes:
Keratin-producing cells.
Protect and minimize heat,
solute and water loss.
Human skin
Dermis: Provides strength
and the sense of touch.
● Fibroblasts:
Collagen-producing cells.
Critical role in wound healing.
4

5. Introduction
Autografts:
▷ Tissue transplanted from one part of the body to
another in the same individual.
Laboratory-grown skin:
▷ Fibrinogen: (blood component) used to construct human
skin.
Bioprinting:
▷ Scaffolds: hydrogels, polymers ceramics.
▷ In situ bioprinting: fibrin-collagen layers containing human
fibroblasts (hFBs) and keratinocytes (hKCs)
▷ Free-Form Fabrication
▷ Laser assisted
5

6. 2.
Materials and Methods
6

7. Bioprinter design and set-up
(a)–(c) components of the dermal compartment (hFBs,
human plasma and CaCl2). (d) hKCs
7

8. Scheme of the bioprinting
process.
8

9. 3.
Results
9

10. Analysis of printed
human skin
differentiated in
vitro
(A) ‘Handmade’ skin
(B)-(D)-(C) Printed skin
(C) Expression of Keratin
(D) Proper growth of
hFBs
10
Equivalents deposited into transwells were allowed to differentiate
for 17 days with a medium that was changed every 3 days.

11. Analysis of printed
human skin
differentiated in
vivo -
(8 weeks postgraft)
(A) Visual appearance
(B) Regenerated skin
(C) Normal human skin
11
All the characteristic of normal skin, stratum
basale, stratum spinosum, stratum granulosum
and a well-developed stratum corneum are
easily identified in the printed skin.

12. 12
Analysis of bioprinted
human skin grafted to
mice
(A) Keratin detection (green line)
(B) Collagen VII (green line)
hFBs (red colour)
(C) Suprabasal keratin K10 (red line)
(D) Filaggrin - Granular layer(green
staining)
Rete ridges (asteriks)
SMA - blood vessels (red staining)
Analysis eight weeks post grafting
using antibodies against skin markers

13. hKCs survival
analysis
Microscopic appearance
of the hKC colonies
before (A) and after (B),
the printing system.
(C) Number of
keratinocyte colonies per
microscopic field
13

14. 4.
Discussion
14

15. Discussion
2 main approaches have been published:
▷ Multi-layered deposition of fibroblast and
keratinocytes in a collagen scaffold.
▷ LaBP technique: immortal human HaCaT
keratinocytes and mouse fibroblasts.
Our model:
▷ Simultaneously deposit all the elements of
dermis and on top the hKCs to form
epidermis.
15

16. Conclusion
▷ The printed skin was very similar to normal human
skin and to manually produced equivalents.
▷ This method is fast and appropriate for commercial
and clinical use.
▷ It’s automation should reduce the cost. 16

17. Thanks!
Any questions?
17