Bone

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BO Bioengineering

Head of Unit: Prof. L. Meseguer-Olmo, MD, PhD.

Associated: Francesca Cragnolini
Enrique Bautista
Francisco J. Gmez

Profile URL: http://www.researcherid.com/rid/A-3141-2011

PProfile: http://es.linkedin.com/pub/luis-meseguer-olmo-md-phd/34/b1/a15
Further info on Facebook (Clulas madre or Stem cells and Tissue engineering).
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BO Bioengineering

Nuestros retos clnicos

(Our Clinical challenges in orthopaedic)
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BO Bioengineering

BIOMATERIALES
(Biomaterials)

Naturales (fibroina-seda, hueso deshidratado)

(Naturals)

Sintticos (cermicas FC, CaSO4)

(Synthetic)

Sistemas liberacin Ingeniera de Tejidos

controlada molculas (Tissue Engineering)
(controlled delivery of molecules)
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BO Bioengineering

Sistemas liberacin controlada molculas

(Controlled delivery of molecules)

a b c
Local
(Local)
*
Efecto *
*
(Effect)
Sistmico
(Systemic)

Antibiticos Accin antimicrobiana

(Antibiotics) MSCs
Efecto citotxico
OBs

- Proliferacin celular
Factores crecimiento - Diferenciacin
(Growth factors) - Migracin
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BO Bioengineering

Construccin basada
El plasma rico en plaquetas (PRP) aporta los
en PRP-Yeso vs PRP-TCP-Yeso factores de crecimiento (VEGF, TGF, IGF, etc.)
(PRP-Plaster vs PRP-TCP-Plaster constructs)

VEGF release in PBS

250

200

150
pg/ml

TCP+Yeso-PBS
100 Yeso-PBS

50

0
1d 3d 6d 9d
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Construccin basada
en Yeso-Vancomicina-Gentamicina y Yeso-PRP.
Aplicacin clnica.
(Plaster-vancomycin-gentamicin and PRP-plaster constructions.
Clinical applications)

X-R plains

2 mth post-OP
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BO Bioengineering

Ingeniera de Tejidos
Bone tissue with / without bone marrow
(Tissue Engineering)

Design of synthetic scaffold

Vs
 Bone
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BO Bioengineering

Human cancellous bone

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Clulas madre adheridas al biomaterial

(Stem cells adhered to the biomaterial)
 Bone
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BO Bioengineering
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BO Bioengineering

Queratinocitos adheridos y creciendo en una malla de fibroina

(Keratinocytes attached and growing on a fibroin mesh)
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BO Bioengineering

Multilayer growth

Mesh remains uncovered

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General and specific cell cultures

Scanning electron microscopy
Transmission electron microscopy
Energy dispersive spectroscopy
Electrospinning
Animals laboratory
Histology & Histopathology techniques
Immuno-histochemical assays
rt-PCR
and other ralated techniques.
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BO Bioengineering

Instituto Bioingeniera. Universidad Miguel Hernndez (Elche, Alicante)

Instituto Murciano de Investigacin y Desarrollo Agrario Alimentario. Murcia

Dpt. Qumica Orgnica y Bioinorgnica. Universidad Complutense. Madrid

Instituto Tecnolgico de Canarias. Las Palmas de Gran Canarias

Dpt. de Materiales. E. S. Ingenieros de Caminos. U. Politcnica. Madrid

Instituto de Cermica y Vidrio. CSIC. Madrid

 Artificial Skin
Challenging the creation of a human
skin prototype in vitro.
 Support
Fibroin matrix

The support chose for this project is a silk

fibroin scaffold made using as starting material
the fibroin protein extracted from silk cocoons.
 Cultures
Two cell types from skin are now testing for the capacity to adhere
and proliferate normally (as in plastic cultures) in the same
culture media and temperature/CO2 conditions.

Culture conditions: 37C, 5% CO2, HAMs F12 medium

supplemented with 10% SBF, 1% Pen/Strep, 1% Glutamine, 5%
MelanoMax.
The medium is changed 2 time a week.

Keratinocytes: Human Epidermal Keratinocytes (HEK001,

ATCC).

Melanocytes: Human Epidermal Melanocytes (HEM, (Gentaur).

 Results
Melanocytes and Keratinocytes adhere and
proliferate on the silk fibroin matrix.

Fig. 1: Keratinocytes on the left; Melanocytes on the right.

 Results
SEM images demonstrate that cells adhere,
proliferate and after 28 days in culture cells
are confluent on a 1cm2 flat matrix (Fig. 2c).

For Keratinocytes it is also possible to

observe an ulterior organization stage; they
organize in areas giving the surface a puzzle
like aspect (Fig. 2d).
 72 h after seeding 10 days after seeding

Detail

Complete covering of the surface at 28

days
Fig. 2: Keratinocytes after seeding: a) 72h, b) 10 days, c) 28 days, d)
puzzled aspect of keratinocytes covered surface after confluence.
 Results
SEM imagines confirm that the two types of
cells adhere and proliferate each one on a
side of the matrix, as seeded, without mix.
For melanocytes is important to underline
that they are smaller and thin enough to
threaded they way trough the matrix
framework (Fig. 3).
Fig.3: Melanocyte into the silk fibroin matrix framework.
 Results
Keratinocytes and Melanocytes grow up on
multiple layers after reacing confluence.

Some fibers not yet covered Multiple layers

 Conclusions
It is possible to produce prototypes of
human skin in vitro, using keratinocytes and
melanocytes, two human skin types. In this
prototype they organize in layers like in
normal human skin. An advantage is offered
by the substrate, that is a protein matrix
totally compatible with human body and
reabsorbable.
 Future perspectives
It is possible to produce other skin
prototypes, combining other skin cells
or more than two types of cells,
resembling more the skin composition.

This model of human skin is directed at

everyone researcher interested in test
their products on human skin before
commercialize this products.