Biol und 32 Prof. Dr. Cars Tel.: 0351 465 werner@ipfdd Prof. Dr. Brig Tel.: 0351 465 [email protected]logie- Mate sten Werner 58-531 d.de itte Voit 58-590 -insp erial Lebende Org molekulare S Funktionen u sich ändernd anzupassen. dagegen bish Regulationsm ein biorespo die Blutgerin kontrollierte effektiver un Materialien, werden (Nat dx.doi.org/10 soll künftig v Medizinprod Blutkontakt bietet das 20 Zwanzig20 a RESPONSE – Implantattec Universität R Stammzellen extrazellulär miteinander eingebettet. nachvollzieh menarbeit m Universität D und Zentrum Rahmen des forschungsb to Tissues) e die von Zellk Matrix auf Ob kann. Dieses Blutstammz Menschen an schneller ve bisher dafür Die neue Me keiten zur En Stammzellen durch ihre M könnte künft genutzt werd 10.1038/NME Regenerative Neubildung v mehr und m Zu deren Akt Matrixsystem pirier gesta ganismen nutz Steuerungsm und Stoffwech den Umgebun Synthetische her noch nich möglichkeiten nsives Hydrog nnung durch s e Freisetzung v nterdrücken ka die derzeit kl ure Communi 0.1038/ncomm vor allem zur ukten genutzt zur Anwendun 013 zur Förder usgewählte B – Partnerscha chnologien (Ko Rostock) einen n sind in der N re Matrix aus verknüpften B Um derartige en zu können mit Kollegen de Dresden (Med m für Regener 2013 verlänge ereichs 655 d eine Methode e kulturen produ berflächen ve s Prinzip wurd ellen aus dem ngewandt, die rmehrt werde angewandten thodik bietet v ntschlüsselun n aus verschie Mikroumgebun tig auch für ze den (Nature M ETH2523). e Therapien s von Gewebe u ehr auf endog tivierung werd me benötigt, d rte G altun zen rückgeko echanismen, hsel-Paramet gsbedingunge e Materialien v t über solche n. Wir konnten gel entwickeln selbständig von Hemmsto ann als alle a inisch verwen ications, ms3168). Das K Beschichtung t werden, die ng kommen. H rung im Progr BMBF Projekt aft für innovat oordination n idealen Rah Natur in eine dreidimensio Biomakromol e Matrixstrukt , wurde in Zus er Technische izinische Faku ative Therapie erten Sonder- er DFG (From entwickelt, m uzierte extraz rankert werd de für die Kult m Knochenma e so etwa drei en konnten als n Kulturbeding vielfältige Mö ng von Signale edenen Geweb ng steuern las ellbasierte Th Methods, dx.do etzen für die und Organstru gene Mechani den Polymer- die sich erst im renz ng ppelte um er unter en verfügen n nun n, das offen nderen ndet Konzept g von im Hierzu ramm ive men. nal lekülen uren sam- en ultät en) im - m Cells it der elluläre en tur von rk des mal s mit gungen. glich- en, die ben ssen und erapien oi.org/ ukturen smen. m Z E E g ü d b w 10 M F S s T vo k U G P D c Z a ih vo B M A N g T F m B E G M T S L ra B B fläch ielgewebe au Ein neues, seh Erzeugung mu lycan-Polyeth ber Michael-T urch regiosel eider Polyme werden (Advan 0.1002/adma.2 Matrices stehe Forschungen z ystemen für d eit Juni 2013 i R67 der DFG on Heilungspr önnen. Unter den 2013 Graduierungsa Prädikat „sum Dissertation „T orneal endoth wei erfolgreic us dem Ausla hre Arbeit am on der Queen Brisbane, ausg Minister’s Que Award, und Be National Unive efördert durc rust, vertiefen Forschungen d modellen für T Behandlungsm Erkrankungen Gemeinsam m Molekulare Bio echnischen U eptember 201 Life mit Beiträ agender Wiss Biomaterialien Biologie organ hen- s löslichen Pr r vielseitiges ltifunktionelle hylenglykol-Hy Type-Addition ektive Peptidf rkomponente nced Materials 201300691). De en auch im Fo zu immunmod die dermale W m Sonderfors Biomaterialie rozessen real 3 abgeschloss arbeiten war d ma cum laude Tissue engine helium“ von Ju che Nachwuch and haben sich IPF fortzuset sland Univers gezeichnet mi en Elizabeth I enjamin Newla ersity of Irelan h ein Stipend n für jeweils z des Instituts z Tumorzellen s methoden für n . it B CUBE, de oingenieursw Universität Dre 13 die Konfere gen internatio enschaftler d n, Biophysik u isiert werden rekursoren bi Prinzip zur er Glycosamin ybridnetzwerk sreaktionen k funktionalisie en entwickelt s, dx.doi.org/ erartige in situ kus der dulatorischen Wundheilung, schungsbereic en zur Steueru lisiert werden senen die mit dem e“ ausgezeich ering of the h uliane Teichm hswissenscha h 2013 entschi tzen: Laura Br sity of Techno t dem Prime II Diamond Ju and von der nd, Galway, ium des Wellc zwei Jahre die u 3D-Kultur- sowie zu neue neurodegener m Zentrum fü issenschaft de esden, konnte enz Engineerin onal heraus- er Gebiete nd Synthetisc . lden. no- ke konnte rung u die ch ung hnete uman mann. aftler ieden, ray logy, bilee come e n rative ür er e im ng he
12
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ogie- inspirierte Grenzflächen- und Materialgestaltung · 2014-04-04 · Biol und 32 Prof. Dr. Cars Tel.: 0351 465 werner@ipfdd Prof. Dr. Brig Tel.: 0351 465 [email protected] ogie-Mate
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Lebende Orgmolekulare SFunktionen usich änderndanzupassen.dagegen bishRegulationsmein biorespodie Blutgerinkontrollierteeffektiver unMaterialien, werden (Natdx.doi.org/10soll künftig vMedizinprodBlutkontakt bietet das 20Zwanzig20 aRESPONSE –ImplantattecUniversität RStammzellenextrazellulärmiteinander eingebettet. nachvollziehmenarbeit mUniversität Dund ZentrumRahmen desforschungsbto Tissues) edie von ZellkMatrix auf Obkann. DiesesBlutstammzMenschen anschneller vebisher dafür Die neue Mekeiten zur EnStammzellendurch ihre Mkönnte künftgenutzt werd10.1038/NME RegenerativeNeubildung vmehr und mZu deren AktMatrixsystem
piriergesta
ganismen nutzSteuerungsmund Stoffwechden Umgebun Synthetischeher noch nichmöglichkeitennsives Hydrog
nnung durch se Freisetzung vnterdrücken ka
die derzeit klure Communi
0.1038/ncommvor allem zur ukten genutztzur Anwendun
013 zur Förderusgewählte B– Partnerschachnologien (KoRostock) einenn sind in der Nre Matrix aus verknüpften BUm derartigeen zu können
mit Kollegen deDresden (Medm für Regener
2013 verlängeereichs 655 d
eine Methode ekulturen produberflächen ves Prinzip wurdellen aus demngewandt, diermehrt werdeangewandtenthodik bietet vntschlüsselunn aus verschie
Mikroumgebuntig auch für zeden (Nature METH2523).
e Therapien svon Gewebe uehr auf endogtivierung werdme benötigt, d
rte Galtun
zen rückgekoechanismen,
hsel-Parametgsbedingunge
e Materialien vt über solche
n. Wir konntengel entwickelnselbständig von Hemmstoann als alle ainisch verwenications,
ms3168). Das KBeschichtungt werden, die ng kommen. Hrung im Progr
BMBF Projekt aft für innovatoordination n idealen RahNatur in eine dreidimensioBiomakromol
e Matrixstrukt, wurde in Zuser Technischeizinische Fakuative Therapieerten Sonder-er DFG (Fromentwickelt, muzierte extrazrankert werd
de für die Kultm Knochenmae so etwa dreien konnten alsn Kulturbedingvielfältige Mö
ng von Signaleedenen Gewebng steuern lasellbasierte Th
Methods, dx.do
etzen für die und Organstrugene Mechaniden Polymer-die sich erst im
renzng
ppelte um er unter en verfügen
n nun n, das
offen nderen
ndet
Konzept g von im Hierzu ramm
ive
men.
nal lekülen uren sam- en ultät en) im -
m Cells it der elluläre en tur von rk des mal s mit gungen. glich-
nced Materials201300691). Deen auch im Fozu immunmoddie dermale Wm SonderforsBiomaterialierozessen real
3 abgeschlossarbeiten war dma cum laude
Tissue enginehelium“ von Juche Nachwuchand haben sich
IPF fortzusetsland Univers
gezeichnet mien Elizabeth I
enjamin Newlaersity of Irelanh ein Stipendn für jeweils zdes Instituts zTumorzellen smethoden für n
.
it B CUBE, deoingenieursw
Universität Dre13 die Konferegen internatioenschaftler d
n, Biophysik uisiert werden
rekursoren biPrinzip zur er Glycosaminybridnetzwerksreaktionen kfunktionalisie
en entwickelt s, dx.doi.org/ erartige in situkus der
dulatorischen Wundheilung, schungsbereicen zur Steuerulisiert werden
senen die mit dem e“ ausgezeichering of the huliane Teichmhswissenschah 2013 entschitzen: Laura Brsity of Technot dem Prime II Diamond Juand von der nd, Galway, ium des Wellc
zwei Jahre dieu 3D-Kultur-
sowie zu neueneurodegener
m Zentrum füissenschaft deesden, konnteenz Engineerinonal heraus-er Gebiete nd Synthetisc.
lden.
no- ke konnte rung
u
die ch ung
hnete uman
mann. aftler ieden, ray logy,
bilee
come e
n rative
ür er
e im ng
he
Biolund
Separation untersuchuBiopolymer Susanne BoAlbena Lede Die CharaktBiopolymeruntereinandstellt aufgrusetzung dereine HerausStandardmedie Größenagelangt dabGründe dafüdendritischeadsorptivenstationären innerhalb dkräfte, die bdendritischeAggregatenDegradationführen. AusAuftrennungFeldflussframit Lichtstrfindet die Auschmalen Kindem durcPolymere hschaften seDurch ihre VTrennbereickeit, sowohlAggregate ozu separiereStelle möchHerangehenuntersuchupolymeren (hülle) erläu1. AuftrennuEinzelmolekGegenstandsind hochmDendronen die aufgrunlen GruppenmolekulareBrücken ne
logie- Mate
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terisierung voen und deren
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n WechselwirkPhase. Weite
er Trennsäulebei der Untersen Biopolyme oder Biohybr
n der nichtkov diesem Grung die Asymmeaktionierung (reudetektion (uftrennung im
Kanals ohne sth verschiedeninsichtlich ihrpariert werdeVielseitigkeit ch bietet die Al Einzelmolekoder Hybridstren und zu ana
hten wir drei vnsweisen bei Wngen von den(verzweigte Ptern:
ung und Charakülen und Aggd dieser AF4-Lolekulare Polund einer dicd der hohen An sehr stark zn Wechselwirigen. In syste
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Appelhans,
n dendritischWechselwirk
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ar. Besonders olmassenbestromatographig an ihre Grene Multifunktiound deren Neikungen mit deerhin herrschee sehr hohe Ssuchung von ren und deren
ridstrukturen valenten Bindnd haben wir fetrische FlussAF4) in KombLS) gewählt. H
m Inneren einetationäre Phan wirkende Flürer Diffusionsen. und den sehr
AF4-LS die Möküle als auch rukturen vonealysieren. An dverschiedene Wechselwirkudritischen Glyolymere mit M
akterisierung gregaten: LS Untersuchlymere mit Lyhten Maltoses
Anzahl an funkur Bildung vo
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renzng
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Detektion näheLS möglich istauch Aggregaewiesen werdktur der einzeinflusst und dverhalten steu
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dass die Herststemen nicht meine Mischun
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Bestandteile n zugeordnet uder Einfluss uhl, des Abstaner des molek: GD) betrachidstruktur, gem GD (Verhälttellt. Es wurd
System sowohals auch define Nanostruktuorhanden sindmit statischern über den Difine Aufklärunsverteilung m
fläch
ndronen-Wertes und dermassen, Radieen mittels AF4er betrachtet., sowohl Einz
ate zu detektieden, dass der elnen Polymerdamit das uert (Abb. 1) [1
Anticoagulato different Manfred MaClaudia RenMikhail Tsu Activation ocontacting dvenous cathmedicine. Vstrategies inimmobilizathave been echallenges, Exploring anrecently devwhich releaheparin in rcoagulation(Fig. 1)[1,2]. achieved bycrosslinkedglycol) (starcleaved by tCleavage of liquid phaseenhanced acleavable hyapplied hepoptions for rpatient and,clinical app
Fig. 1:
Feedback cont
responsive hep
logie- Mate
ant hydrogels coagulation e
aitz, Jan Zitzmnneberg, Uwe rkan, Carsten
f the blood codevices such aheters is a perarious surfacncluding surfation of anticoaexplored to ad however, witn entirely newveloped a biorses the coagu
response to th process wheAnticoagulan
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rol loop obtained
parin-releasing h
-insperial
s with respons enzymes
mann, Freudenberg
n Werner
oagulation by bas vascular strsisting problee modificationace passivatioagulant molecdress the resh limited succ
w concept, we responsive coulation inhibitohe activation on exposed to t feedback coconsisting of
armed poly(ether peptides spon enzyme thrleases heparierts a massiveactivity. The thmpetes clinicand may offe
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piriergesta
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,
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of the blood ntrol is heparin
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rombin. n to the ely hrombin-cally r exciting he cts in the ucts.
-
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Thrombin, theof this hydrogagulation cascheparin is relethe coagulatioNumerical simprocess were assumption thtion inhibitor meffect of the gverification, hynetwork propeheparin, starPeither cleavedcoagulation fafactor Xa (FXahigher rate coestablished, Tfast, Thr-slowmediated gel buffered soluttion factors ateach (Fig. 2). Tconsidering ththe respectiveby the less abadjusted to exensure the reamounts of hecross-reactivibe specificallyfactor each.
For an applicahydrogels werdose anticoag37°C. To compthe hydrogelspro-coagulantmonolayers o20 % methyl t[3]. After two
renzng
e trigger enzymel, is a ‘late’ pcade reactioneased from thon reaction is mulations of t
performed cohat an earlier may improve tel system. Foydrogels witherties were prPEG and peptid by the earlieactors kallikrea-resp) or by tompared to thThrombin-cleaw, Thr-std). Co
degradation wtions of the ret a concentratThe peptides whe plasma cone enzymes, i.eundant ‘earlie
xhibit higher tlease of pharmeparin. There ity, i.e. the pepy cleaved by o
ation-orientedre exposed to
gulated humanpare the antic were co-incut surfaces (binf 80 % carboxerminated alkhours incubat
fläch
me for the cleprotease in th. As a conseq
his gel only, wfully activatedhe coagulatioonfirming the release of coathe anticoagu
or the experim comparable repared from des, which ca
er activated ein (Kal-resp)hrombin at loe previously avable peptideoagulation enzwas verified inespective coagtion of 45 nmowere designencentration lee. peptides cleer’ enzymes wurnover ratesmacologicallywas only min
ptides were fone coagulatio
d evaluation, tfreshly drawn
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ubated with hinary self-asse
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f
a
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Keywords
hydrogel
drug release
feedback contro
anticoagulation
hemocompatibil
Fig. 2:
Cumulative hepa
release from a se
coagulation facto
responsive starP
heparin hydrogel
crosslinked by di
peptides. Gels we
exposed for three
a solution of the
coagulation facto
35
ol
lity
rin
et of
or-
PEG-
ls
ifferent
ere
e hours to
respective
ors.
Biolund
36
logie- Mate
-insperial
thrombin F1+of the plasmheparin conc(Fig. 3 A). Hydegradation kallikrein-resuppressed tstandard thr(p < 0.05), whydrogel exhThe hydrogeupstream coanticoagulanthrombin resof the releasthe kallikreingood thrombby the high obe attributedhydrogel to teffect of hepFXa responsthe heparin rbut producedApparently, hresponsive hactivated coaanticoagulanAs a valuablemay allow fuheparin expohydrogel sholonger lastinpeptide-croscoagulation eheparin-reledifferent stagmaximizing tminimizing theparin. Thepotentialitiescoagulation-
piriergesta
+2 fragment, aatic coagulati
centrations weydrogels with aby thrombin (
esponsive hydrthe coagulatio
rombin-responwhereas the slhibited less anl, which respo
oagulation factnt capacity as sponsive hydrsed heparin (Fn-responsive boprotective coverall heparind to the high sthe protease aarin on kallikive hydrogel srelease of thed a similar anheparin releas
hydrogels triggagulation factnt effects at loe advantage fo
urther minimizosure. At the sows slower cong activity. In ssslinkers cleavenzymes allow
easing hydrogges of the coathe anticoaguhe required a
ese findings fus of the introd-activated anti
rte Galtun
a quantitative on and the reere determineaccelerated (Thr-fast) androgel (Kal-reson better thannsive hydrogelow degradingnticoagulant aonds to the mtor FXa had ththe reference
rogel. QuantifiFig. 3 B) indica
hydrogel obtaharacteristicsn release, wh
sensitivity of thand a low regurein. In contrashowed only 6e reference hyticoagulant efse from bio-gered by earlior (FXa) afford
ower heparin lor applicationzation of the psame time, thonsumption ansum, choosingved by differews for customels to respond
agulation casclant effect anmount of rele
urther extend uced class of icoagulant ma
renzng
marker eleased ed
sp) n the el g ctivity. ore he same e cation
ates that ained its s mainly ich can he ulating ast, the 60 % of ydrogel ffect.
ier ds levels. , this
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nd, thus, g nt
mizing d to cade for d for
eased the
aterials.
Fi
(A
in
w
[1
[2
[3
fläch
ig. 3:
A) Blood coagulat
ncubation of vario
with whole blood.
1] M.F. MaitzM. FischerCommun.
2] M. Maitz, UBeyrich, CPolymerfoJahresber
3] C. SperlingWerner: B4456
hen-
tion and (B) hepar
ous coagulation re
z, U. Freudenbr, T. Beyrich, C2013. p. 2168
U. FreudenbeC. Werner: Leiorschung Dresricht (2011), 32g, M. Fischer,
Biomaterials 3
rin release obser
esponsive hydrog
berg, M.V. TsuC. Werner: Na
rg, M. Tsurkabniz-Institut fsden e.V.
2-33 M.F. Maitz, C
30 (2009), 4447
rved at
gels
urkan, ature
n, T. für
C. 7-
Biolund
Biomoleculdetected bysurface pla Ralf ZimmeCarsten We Materials mextracelluladecisions arengineeringcontaining gparticularlythe effectiverelease of nGrowth factattributed tomolecules aelectrostatisulfate groucharged provariations ofrom the binreflected in hydrogels, adetection ofAiming at nodetection ofGAG-based electrokinecand surfacemeasurememodel protehydrogels cshaped polyheparin [5]. exhibits a sifactors and physiologicamaterial wesurfaces. Thnegatively csulfate and The excess (as determimeasuremerecently devThe heparindegree of thsimilar to thhydrogel mato heparin inmeasureme
logie- Mate
lar interactiony electrokinetsmon resona
rmann, Susanrner
mimicking the ar matrices tore instrumentg [1,2]. Biohybrglycosaminogy promising foe binding, pronumerous groor binding to o the sulfationand is, thus, lac interactions
ups along the otein domainsf the system n
nding of growtthe electroki
allowing for thf growth factoovel electrokif biomoleculahydrogel matctic (streamin
e plasmon resents [4] to anaein lysozyme wonsisting of c
y(ethylene glyLysozyme wa
ize similar to a positive exc
al pH values. Tere immobilizehe films were charged due tocarboxylic acconductivity oned by the eleents) was anaveloped meann concentratiohe hydrogel filhe values for caterials [5]. Thn the hydrogeent of the exce
-insperial
ns in hydrogetic measuremnce
nne Bartsch,
exogenous si control cell f
tal for tissue rid hydrogels lycans (GAGs)r that purposetection and suwth factors [3GAGs can be n pattern of thargely determs between ioniGAGs and opp
s. Accordinglynet charge resth factors mignetic fingerprhe electrokiner binding. netic sensorsr interactionsterials we emng potential/cuonance (SPR)
alyze the bindiwithin biohybrovalently linkcol) (starPEG)
as selected asrelevant grow
cess charge atThin films of ted on transdushown to be
o the ionizatioid groups of h
of the hydrogeectrokinetic lyzed applying
n-field-approaon and the crolms were founcomparable bhe binding of
el was analyzeess conductivi
piriergesta
els ments and
gnals of ate
) are e due to ustained 3].
hese mined by
ized positely , sulting ght be rint of etic
s for the within ployed urrent) ) ing of the rid ed star-) and
s it wth t the gel
ucer
on of the heparin. el films
g a ach [6]. osslinking nd to be ulk lysozyme
ed by ity after
ictbfpccnewrhebgOaafumpc
F
E
(
a
i
r
t
f
rte Galtun
incubating theconcentrationtaking into accby SPR. The efilms significaprotein conceconcentrationcan be attribunumber of moelectrostatic bwas further shreleased fromhydrogel filmselectrolyte sobinding procegel films in seOngoing studiand release oaffinity and sizfilms prepareusing partiallymore, the minprinciple and chip system w
Fig. 1:
Excess conductiv
(ratio starPEG/he
after incubation w
incubation with 10
regenerated usin
the molecular str
formation and pro
renzng
e films with prns between 0.1count the bindxcess conduc
antly decreasentration withi
n range (Fig. 1)uted to the decobile counter ibinding of lysohown that lyso
m the heparin s with highly clutions. The rss allows for
ensoric applices aim at expf growth factoze to starPEGd at different y desulfated hniaturization oits integration
will be investig
ity of a starPEG-h
eparin = 3) in 1 mM
with different am
00 mg/mL lysozy
ng a 1 M NaCl solu
ructure of the gel
operties see [5].
fläch
rotein solution1 and 100 mg/mding kinetics sctivity of the hyed with increan the studied ). The latter efcrease of the ions in the geozyme to hepaozyme can beby incubating concentrated eversibility ofreuse of the hations. loring the bin
ors of differen-heparin hydrmolar ratios a
heparin. Furthof the measurn into a lab-ongated.
heparin hydrogel
M KCl solution be
ounts of lysozym
yme the gel film w
ution. The inset ill
l. For details on g
hen-
ns of mL studied ydrogel
asing
ffect
l by the arin. It the
the hydro-
ding nt rogel and
her-ing n-a-
l film
efore and
me. After
was
lustrates
gel
S
Keywords
biohybrid hydro
growth factor
electrokinetics
excess conducti
biosensor
SPR
37
ogel
ivity
Biologie-inspirierte Grenzflächen- und Materialgestaltung
38
Sponsor: Sächsisches Staatsministerium für Wissenschaft und Kunst, Förderkennzeichen 4-7531.50-03-843-12/1 Teil II Cooperation: Dr. Juliane Posseckardt, Prof. Dr. Michael Mertig, Kurt-Schwabe-Institut für Mess- und Sensortechnik e.V. Meinsberg [1] M. P. Lutolf, J. A. Hubbell: Nat. Biotechnol.
23 (2005) 47-55 [2] J. J. Rice, M. M. Martino, L. De Laporte, F.
Tortelli, P. S. Briquez, J. A. Hubbell: Adv. Healthcare Mater. 2 (2013) 57-71
[3] I. Capila, R. J. Linhardt: Angew. Chem. Int. Ed. 41 (2002) 391-412
[4] J. Posseckardt, R. Zimmermann, C. Werner, M. Mertig: Proceedings zum 11. Dresdner Sensor-Symposium (2013) 351-354, DOI: 10.5162/11dss2013/F11
[5] U. Freudenberg, J.-U. Sommer, K. R. Levental, P. B. Welzel, A. Zieris, K. Chwalek, K. Scheider, S. Prokoph, M. Prewitz, R. Dockhorn, C. Werner: Adv. Funct. Mater. 22 (2012) 1391-1398
[6] R. Zimmermann, S. Bartsch, U. Freudenberg, C. Werner: Analytical Chemistry 84 (2012) 9592-9595
Culture of human hematopoietic stem cells on decellularized cell-derived matrices Marina Prewitz, F. Philipp Seib, Malte von Bonin, Jens Friedrichs, Aline Stißel, Martin Bornhäuser, Carsten Werner The therapeutic potential of human bone marrow mesenchymal and hematopoietic stem and progenitor cells (MSC and HSPC, respectively) offers great promise for their use in regenerative medicine. A key regulator of these cells is their extracellular matrix micro-environment. However, a major obstacle in defining the exact role of extracellular matrices (ECM) is the lack of suitable methods that recapitulate complex ECM microenviron-ments in vitro. Our current work therefore concerned a methodology for the reliable anchorage of decellularized extracellular matrices generated from cultured bone marrow MSC to mimic the bone marrow stem cell microenvironment. Cell-secreted ECM was immobilized to culture carriers via maleic anhydride copolymer layers used for covalent binding of fibronectin to the culture carrier surface. Fibronectin was selected for its properties to interact with cell-secreted extracellular matrix proteins, so that culture and decellularization of a cell monolayer on top yields in a superior anchorage of cell-secreted ECM with the opportunity to study the decellularized extracellular phenotype of various cell-derived matrices. The ECM preparations were thoroughly characterized to identify their molecular composition, supra-structural features and nano-mechanical properties. The obtained MSC-derived ECM substrates served as in vitro culture environments for human HSPC, resulting in a significant increase in proliferation of these cells over a culture period of 7 days, with up to 3-fold expansion of CD34+ cells on MSC-derived matrices without exhaustion of CD34/CD133 double positive cells, and with the potential for long-term (20 weeks) engraftment into NSG (NOD/SCID/IL2 receptor {gamma} chain (null)) mice. Taken together, we demonstrated the unique ability of cell-derived ECM scaffolds to support expansion and differentiation of bone marrow stem cells in vitro.
Keywords
extracellular matrix
mesenchymal stem cells
haematopoietic stem
cells
Biolund
Beyond thatenables declike multicosources offedepth analyexogenous c
Fig. 1:
Human haema
extracellular m
marrow mesen
Sponsor: Deutsche FoCollaborate CooperationProf. Dr. M.Universitäts [1] M. Prew
FriedricK. AnasM. BornMethod
logie- Mate
t, the establisciphering and omponent ECMering exciting
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topoietic stem ce
matrix that was d
nchymal stem cel
orschungsgem Research Ce
n: Bornhaeuser
sklinikum Dre
witz, P. F. Seibchs, A. Stißel,stassiadis, C. Wnhäuser, C. Wds, 10(8), 788-7
-insperial
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meinschaft, ntre SFB 655
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b, M. von Bon C. Niehage, KWaskow, B. H
Werner: Nature794
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Multifaceted, matrices to in
Mikhail V. TsuSilvana ProkoKandice R. Leand Carsten W
Polymer matrindependent c(biodegradatioligands and sonetwork paramof particular i[1]. Fast self-apresence of cetissue is crucistimulation ofproperties in vprinciples havforming matesuch as poly(eglycosaminogpreviously repwith respect tnetwork charaincorporating molecules. Toorthogonal syto produce anpeptide- and gconjugates (Fzation of starPresulted in PEmultiple functadhesion liganenzymaticallyunits, allowinghydrophobic sSIKVAV. The Gconverted to cmaleimide groclick reaction the above-meA multistep puultra-high purconjugates wifunctionality awith RGD peptconjugation offactor (VEGF).heparin-maledefined 3D-ne
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urkan, Karolinoph, Andrea Zievental, Uwe FWerner
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as demonstrattides and by tf vascular end. Reacting PEGimide-(peptid
etworks were
fläch
ng polymer orphogenesis
na Chwalek ,ieris, Freudenberg,
for the precismultiple biocheion of adhesiors) and physic
h size, stiffnesrrent bioenginuch matrices njection into aee-dimension-defined mate
o. Several desred to create nthetic polymeol), peptides a) [2]. However
aches were limover the polymd did not allowsignaling ese limitation
egy was devele a set of PEGlycan-peptidewise function
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cluding differeKVAV) and able cross-linementation ofides such as
block (heparinent quantities for subseque
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Keywords
hydrogel
biomaterials
growth factors
cell adhesive pe
39
eptides
Biolund
40
Fig. 1:
Synthetic sche
formation of
multifunctiona
forming starPE
hydrogels (� =
of StarPEG/hep
Fig. 2:
(A) Cell elonga
ratio, 1 corresp
round cell, > 1
ponds to an elo
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is regulated by
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ratio of StarPE
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biomolecular m
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MMP-peptide,
(B) SIKVAV pep
promotes EC m
genesis and ne
outgrowth from
chicken dorsal
ganglia, ± SD.
logie- Mate
me for the
al in situ
EG-heparin
molar ratio
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y cross-
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and the
modification
ls ( ± RGD, ±
± VEGF),
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independentenzymatic dedomain peptbiological funcell-instructdemonstrateprimary humchicken dors(Fig. 2). The establishconcluded todeciphering exogenous s
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[1
[2
[3
fläch
1] M. Lutolf, 2005, 23, 4
2] (a) M. P. Lmolecules(b) A. Mettmolecules(c) M. Ehrbmakers, BWeber, M.2007, 8, 30(d) S. J. BrMater. Res(e) M. KimRevzin, G. (f) T. Nie, AKiick: J. C
3] M. TsurkaZieris, K. LWerner: A
hen-
J. Hubbell: N47-55
Lutolf, J. A. Hus 2003, 4, 713; ters, J. A. Hubs 2005, 6, 290;bar, S. C. Rizz
B. San Miguel, P. Lutolf: Bio
000; ryant, K. S. Ans. 2002, 59, 63
m, J. Y. Lee, C. Tae, Biomate
A. Baldwin, Nont. Rel. 2007n, K. ChwalekLevental, U. F
Adv. Mater., 20
at. Biotechno
ubbell: Biomac
bbell: Biomacr; i, R. G. Schoe J. A. Hubbell
omacromolec
nseth: J. Biom3; N. Jones, A.
erials 2010, 31,. Yamaguchi,
7, 122, 287 k, S. Prokoph, reudenberg, C
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, 3596; K. L.
A. C. 2610
Biolund
Cryogel micforce micro Jens FriedrMilauscha GUwe Freude Polymeric mbeing develoanalogues oengineeringobtained by mixture of pzero temperformed: ice microphaseconcentratecryogel formremoved leawith intercoCompared tpores, the inin the non-fresults in deding the macryoconcentconcentratiolike cryogelhigher localbulk hydrogsame reactiHowever, thmentally, yeapplied an ananoindentamechanicalaminoglyca[1,2]. The ansections of tallows for varchitecturemicroscopy positioning struts of opmechanicalthose of reffrom the satemperaturthat cryoconstiffness of AFM-based stress-straifurthermore
logie- Mate
cromechanicsoscopy-based
ichs, Petra B.Grimmer, Stefenberg, Carste
macroporous oped as threeof the extraceg applications
cooling an aqpolymeric gel ratures. Therecrystals and
e. The gel preced in the liquidmation, the iceading to spongonnected maco bulk hydrogncreased precfrozen liquid menser polyme
acropores, whtration effect.on effect, the s can be assul stiffness tha
gel materials oion mixture at
his was not deet. Therefore, atomic force mation method properties ofn-poly(ethylen
nalytical approthe swollen cisualizing the e during nano(Fig. 1 A). Thiof the AFM tipen pores (Fig. properties oference bulk hme reaction me directly provncentration efthe cryogel ponanoindentat
in compressioe provides val
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s unraveled by nanoindenta
Welzel, ffen Vogler, en Werner
cryogel scaffoe-dimensionalllular matrix f. Cryogels are
queous reactioprecursors toeby, two phasa non-frozen cursors are d microphasee crystals cange-like structropores (Fig.
gels without mcursor-concemicrophase ofr networks suich is designa Due to the crstruts of the s
umed to exhibn the correspobtained fromt room tempemonstrated ewe developed
microscopy-bato quantify th
f biohybrid glyne glycol) cryooach uses veryryogels and thcryogel microindentation bys enables thep directly over. 1 B). Comparf cryogel struthydrogels obtamixtures at roves, for the firffects result inore walls. Comtion with uniaxon experimentluable insights
piriergesta
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olds are l for tissue e on o sub-ses are liquid
. After be ures 1 A).
macro-ntration f cryogels urround-ated as ryo-sponge-it a
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the effect of cintegral and lofor a far-goingmatch a particinduce a desir
Sponsors: Bundesminist(BMWA)/AiF Deutsche Fors
Cooperation:Dr. P. Joly, DrCharite Unive
[1] P. B. WelL. NaujoxWerner: B2358 (201
[2] P. Joly, GWelzel, UPetersen
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ryogelation paocal scaffold mg customizatiocular tissue ered cellular re
terium für Wir
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U. Freudenber: PLoS ONE 8
fläch
arameters onmechanics allon of the cryolasticity and t
esponse.
rtschaft und A
einschaft
, Prof. Dr. G. Dn Berlin
mer, C. Rennebe, U. Freudenbecules 13, 234
Schoene, P. Brg, C. Werner, , e73545 (2013
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Keywords
cryogel
AFM
Nanoindentation
Fig. 1:
(A) Confocal micr
image of the spon
microstructure o
swollen cryogel s
Green: cryogel st
White: PBS filled
connected pores.
indicate where A
nanoindentation
performed.
(B) Schematic rep
ation of the AFM-
dentation measu
principle. The tip
cantilever is brou
contact with a cry
covered with PBS
their local mecha
properties in the
state. The applie
induces a deform
the tested mater
proportional to th
force and the stif
the tested region
the nanoindentat
experiments the
exerted on the ca
as well as cantile
movement are re
and can be plotte
force-distance cu
Shading represe
regimes before a
contact. From the
distance curve th
nical properties o
tested material c
extracted.
41
n
roscopy
nge like
of the PBS
scaffold.
truts.
d inter-
. Arrows
AFM-based
was
present-
-nanoin-
rement
p of an AFM
ught into
yogel strut
S to probe
anical
swollen
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tion
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antilever
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nts
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he mecha-
of the
can be
Biolund
42
Keywords
electrophores
electric field
molecular dyn
simulations
confinement
Fig. 1:
Schematic des
type, direction
external electr
corresponding
(in cross-sectio
logie- Mate
sis
namics
piction of the
of applied
ric field, and
g geometry
on).
-insperial
Electrophorepolymers in Sorin Nedelc Taking as anelectrolyte Dthe sequenceis. To read thinto smaller multiple timelater reassemin free solutiwith the sam(except for veoligomer ranseparate the In a first set work has beeelectrolyte cthese simula(parallel to telectric field
The transveratmosphere the net chargcondensed cobserved to resulting in sseparation. Aconsidered stapered constime-constandirection, thecavity into th
A
B2e
piriergesta
etic transport cylindrical ge
cu, Jens-Uwe
n example the DNA, it is impoe of the base he full sequenfragments whes to find overmbled togetheion long charg
me velocity irreery short fragnge), which mem in laborato
of simulationen on electrophains in slit g
ations, we usehe walls) and s (Fig. 1, top).
rse field led toaround the po
ge of the chaiounterions. Lmove faster thselective elecA second set ostraight cylindstrictions [2] (nt applied fiele chain escape next could b
exp A2 4B
rte Galtun
t of charged eometries
Sommer
model poly-ortant to knowpairs in a DNA
nce it is typicahich are sequrlapping regioer. In an electged polymers espective of thgments in the akes it difficu
ory setups.
s, the focus ophoresis of poeometries [1].
ed longitudinatransverse ap
o an asymmetolyion and mon by the numb
Longer polyionhan shorter otrophoretic of simulationsder geometrieFig. 1, bottom
ld in the axial e time from obe written as f
(EqB
renzng
w what A strand lly split enced
ons, and tric field, move
heir size
lt to
f our oly-. In l pplied
ric ion odified ber of ns were nes
s s with ). In a
one follows
q. 1)
waac
TFcthapcyswpfiF
Fi
In
m
al
th
di
co
cy
m
fie
re
It(mvacsu
fläch
where the conspplied electrind characteriompression-e
wo conclusionFirst, if the lononstant (no chhe array of core more effecolymers than ylinders. The eparation effi
when the conseriodically (orelds). These f
Fig. 2.
ig. 2:
n a set time interv
may move a certai
lways the same, o
heir positions. Th
istances z for two
onstant (noted cy
ylinders under co
modulated cylinde
elds of relative st
eported in Lennar
t indicates thameasured as talues) is obtaonstant applieeparation taknder pulsed e
hen-
stants A and Bc field, polyelestic dimensio
expansion cha
ns of our workgitudinal app
hange in direcmpression-ex
ctive in separastraight unifosecond conclciency increastant field is swr otherwise infindings are su
val, charged chain
n distance z. If th
one can identify t
e bar diagram ab
o chains of length
yl.) and variable d
onstant applied fi
ers (noted mod.) u
trength Eeff (= E|
rd-Jonnes units.
at the best septhe differenceined in tapereed fields, whiles place in ta
electric fields.
B depend on tectrolyte lengns of the
ambers [2].
k are importalied field is kection or value)xpansion chamating charged orm diameterusion is that tses even furthwitched on an pulsed applieummarized in
ns of different len
he elution order is
he chains by obse
bove compares th
h N=150 and N=70
diameter (noted m
elds E||, and simi
under applied pul
| /3). The quantiti
paration resole between theed cylinders inle the fastest pered constri.
he gth,
nt. ept ), then mbers
r the her d off ed n
ngth
s
erving
he
0 in
mod.)
lar in
lse
ies are
lution z-
n
ctions
Biologie-inspirierte Grenzflächen- und Materialgestaltung
43
Lastly, we studied were straight cylinders with wavy surfaces [3]. Here, one could explicitly measure the fluid flow inside and outside the polyelectrolyte coils, and electrolyte friction. The ultimate goal of such complex computer experiments is to find a way of individually controlling the dynamics of charged chains, by smart choices of the type of environment, properties of the buffer solution, and applied electric fields. Sponsor: Deutsche Forschungsgemeinschaft SO 277/4-1 [1] S. Nedelcu,J.-U. Sommer: J. Chem. Phys.
133, (2010), 244902 [2] S. Nedelcu,J.-U. Sommer: J. Chem. Phys.
138, (2013), 104905 [3] S. Nedelcu,J.-U. Sommer: Materials 6(7),