Procese de interfata

biomaterial - tesut

curs 7 | 3. Fenomene de interfata B-T

Prof. Stancu Izabela
 Cuprins
3. Fenomene de interfata B-T

3.1. Structura si functiile componentelor matricei

extracelulare; Relatia compozitie – structura – proprietati

3.2. Interactiuni cu biomolecule

3.3. Interactiunea suprafetei cu celule

3.4. Calcifiere / Mineralizare

Agenti de bioactivare
C) Alte biomolecule / specii chimice
- care au bioactivitate convenabila
-Ex. DMSO, acid ascorbic, acid retinoic – stimuleaza diferentierea celulelor spre
fenotip cardiac
-AlkP stimuleaza calcifierea biomaterialului si ulterior aderenta celulara
-Fosfatii de calciu stimuleaza adreenta osteoblastelor
-Bisfosfonatii – inhiba resorptia osului prin stimularea apoptozei osteoclastelor
Exemplul 6 – bioactivare cu fosfat de calciu cristalin ce mimeaza faza minerala osoasa

Suprafata acoperita cu Celule precursoare ale osteoclastelor umane

fosfat de calciu cristalin diferentiate pe suprafata functionalizata cu fosfat

www.corning.com/worldwide/en/products/life-sciences/products/surfaces/matrigel-matrix.html
 Exemplul 7. Functionalizare superficiala cu grupari amina si carboxil

 Acoperirea cu functiuni amina (sarcina pozitiva) sau carboxil (sarcina negativa)

stimuleaza adeziunea celulara, proliferare mai rapida fata de placile de cultura standard
 Acoperirea zonala a elementelor superficiale a placii din polistiren cu o combinatie de
functiuni ce contin oxigen (incarcate negativ) si azot (incarcate pozitiv) – stimularea
interactiunii cu celulele prin influenta asupra hidratarii suprafetei

Pentru detalii :
“Corning® PureCoat™ Amine and Carboxyl Surfaces
Corning PureCoat amine (positively charged) and carboxyl (negatively charged) surfaces provide improved
cell attachment, faster cell proliferation, and enhanced recovery post-thaw over standard TC surfaces.
These surfaces function with a broad range of primary, transfected, transformed, and fastidious cell types,
and have demonstrated utility in serum-reduced or serum-free conditions.

Corning Primaria™ Surface The Corning Primaria surface features a unique mixture of oxygen-containing
(negatively charged) and nitrogen-containing (positively charged) functional groups on the polystyrene
surface. The surface supports the growth of cells that can exhibit poor attachment or limited
differentiation potential when cultured on traditional TC surfaces, including neuronal, primary, endothelial,
and tumor cells. The surface consistency of each lot is confirmed by electron spectroscopy chemical
analysis (ESCA). Corning CellBIND® Surface The Corning CellBIND surface features a net negative surface
charge due to oxygen-containing functional groups incorporated in the polystyrene surface. The surface is
more hydrophilic, and thus more wettable, compared to standard TC surfaces, which facilitates cell
attachment and spreading.”
3.3.2. Agenti de inhibare a cresterii celulare pe suprafete
- tratamente cu anticorpi anti-RGD
- acoperirea cu hidrogel incarcat neutru, legat covalent pe suprafata polistirenului;
stimuleaza formarea de sferoizi, inhiband interactiunile celula-biomaterial si
stimuland interactiuni celula-celula
- ---

www.corning.com/worldwide/en/products/life-sciences/products/surfaces/matrigel-matrix.html
 Suprafata trebuie
adaptata si optimizata
pentru interactiunea cu
celulele !
1 2
Exemplul 1.
Mimarea mediilor in vivo utilizand matrici
ECM purificate pentru acoperirea
suprafetelor

Exemplul 2.
Suprafete avansate ECM mimetice –
pentru celule specializate
(expansiune celule stem si progenitoare)
sau
Suprafete putin aderente (low
attachement) pentru formarea sferoizi
3
Exemplul 3.
Suprafete avansate cu controlul sarcinii
electrice superficiale – pentru interactii
celulare specializate
Controlul interactiunilor celule – biomaterial prin si pentru
sisteme model
Modele utilizate, in ordinea cresterii complexitatii compozitionale si organizationale:

Culturi celulare in monostrat – sisteme 2D, fara reproducerea interactiunilor celula-

celula si celula matrice

Sferoizi celulari – sisteme celulare 3D, adesea fara celule stem sau progenitoare
relevante necesare pt sustinerea culturii celulare 3D; fara celule capabile de innoire si
diferentiere

Organoizi – structuri 3D continand mai multe tipuri de celule dezvoltate intr-o matrice
cu scopul mimarii in vitro al tesuturilor si organelor; modele in vitro mai relevante
fiziologic, care nu pot fi ; asemanare cu tesuturi si organe; mimeaza interactiuni celula-
celula si celula-biomaterial matrice; disponibili pentru culturi pe terment lung; formarea
lor depinde de auto-organizarea celulara autonoma; solutie pentru obtinere de organe
artificiale, intelegerea unor patologii sau tratamente….

Explant tisular – reproduce pe termen scurt complexitatea si caracteristicile tisulare; isi

pierde rapid specificitatea (fenotip, nu poate fi conservat…)

 Alte modele complexe …

[Cell Stem Cell 18, January 7, 2016 ª2016 Elsevier Inc.]
Controlul interactiunilor celule – biomaterial prin si pentru
sisteme model

Figure 1. Model Systems in the Life Sciences

Organisms comprise a hierarchy of systems
from the subcellular level to the whole body. In
the life sciences, many models have been
developed across this organismal hierarchy, to
address specific questions across biology and
medicine. Each model system possess unique
attributes; in
general, with increasing scale comes increasing
system complexity and challenges in cell culture
and the reduced availability of biochemical and
quantitative tools, which can limit study
insights. Organoid models provide a unique
opportunity to incorporate moderate system
complexity while still affording many tools for
probing structure and
function. When compared to tissue explants,
organoid systems can mimic similar cell-cell and
cell-matrix interactions while maintaining the
ability for long-term cultures thanks to
maintained signaling cues important for
survival. Fig 1 din Cell Stem Cell 18, January 7, 2016 ª2016 Elsevier Inc.
Schematic – organoizi intestinali – de la obtinere la aplicatie

Organoizii
intestinali
recapituleaza
caracteristici ale
intestinului in vivo;

Necesita control
spatio-temporal al
moleculelor de
semnalizare, al
repartitiei tipurilor
de celule in
matrice si al
structurarii
matricei pt
asigurarea
functionalitatii

Fig 3 din Cell Stem Cell 18, January 7, 2016 ª2016 Elsevier Inc.
Organoizi intestinali Conditii de obtinere:IntestiCult™ Organoid Growth Medium
(Mouse) + CorningR MatrigelR Matrix
• prezenta tuturor tipurilor
de celule din epiteliul
intestinal adult
• organizare specifica
intestinului, cu vilozitati,
epiteliu polarizat si lumen
functional
• instrument pentru
investigarea biologiei
intestinale, a celulelor stem
intestinale, a unor patologii
• potential pentru medicina
regenerativa – incorporati in
colonul unor soricei lezati
chimic – formeaza segmente
intestinale ce nu pot fi
deosebite morfologic de
tesutul sanatos.

Scientists Helping Scientists™ I WWW.STEMCELL.COM DOCUMENT #28223 VERSION 1.0.0 JAN 2015;
Schematic – organoizi cerebrali

Organoizii cerebrali
– pentru modelarea
dezvoltarii creierului
uman; inca multe
limitari (absenta
comunicarii cu alte
tesuturi si organe) !

Fig 4 din Cell Stem Cell 18, January 7, 2016 ª2016 Elsevier Inc.
Perspective:

•Organogeneza
•Modelare in vitro
•Transplantare

Fig 5 din Cell Stem Cell 18, January 7, 2016 ª2016 Elsevier Inc.
Surse de informare:

Strategies in Regenerative Medicine. Integrating Biology with Materials Design,

Ed. Matteo Santin, Springer, 2009
An Introduction to Biomaterials, CRC, Taylor & Francis, 2006
MEDLINE – Medical Subject Heading, National Library of Medicine, US National Institute of Health
The Williams Dictionary of Biomaterials, Liverpool University Press, 1999
Stancu I.C. Polimeri biodegradabili si biocompatibili in patologia osoasa , Teza de doctorat 2003;
Dragusin D.M., Hydrogels for Biomedical Applications, Teza de doctorat, 2012;
L. A. Heinz and W. Stephen, On biomineralization, 1989 ;
M. Stephen, Biomineralization: Principles and Concepts in Bioinorganic Materials Chemistry, 2002;
S. Astrid, S. Helmut and S. K. O. Roland, eds., Biomineralization: From Nature to Application, 2008;
S. V. Dorozhkin,"Nanosized and nanocrystalline calcium orthophosphates", Acta Biomaterialia, vol. 6,
no. 3, 2010 pp. 715-734.
si multe altele…..
Alte materiale recomandate :
http://magnesiumforlife.com/medical-application/calcification-and-its-treatment-with-magnesium-
and-sodium-thiosulfate/
http://vimeo.com/25604437
http://www.med.umich.edu/anatomy/plastinate/index.html
Surse de informare:

Societatea Europeana de Biomateriale – ESB (www.esbiomaterials.eu)

Food and Drug Administration – FDA (http://www.fda.gov/MedicalDevices)

Strategies in Regenerative Medicine. Integrating Biology with Materials Design,

Ed. Matteo Santin, Springer, 2009

ESB Consensus Conference I – Williams, D.F., Definitions in Biomaterials, Elsevier, Amsterdam, 1987
ESB Consensus Conference II – Dohery P.J., Williams R.L., Williams D.F., and Lee A.J.C., Biomaterial-Tissue
Interfaces, Elsevier, Amsterdam, 1992
Larousse Science – Larousse Dictionary of Science and Technology, Larousse, Edinburgh, 1995
Oxford Science – Oxford Concise Science Dictionary, Oxford University Press, 3rd Edition, 1996
MEDLINE – Medical Subject Heading, National Library of Medicine, US National Institute of Health
The Williams Dictionary of Biomaterials, Liverpool University Press, 1999
http://www.cbte.pratt.duke.edu/biomaterials-tutorials
Adezivi, lacuri si vopsele – B. Marculescu, E. Rusen, L.M. Butac, ISBN 978-973-558-403-0, Ars Docendi
2009 ……