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Biolund
32
Prof. Dr. Cars
Tel.: 0351 465
werner@ipfdd
Prof. Dr. Brig
Tel.: 0351 465
logie- Mate
sten Werner
58-531
d.de
itte Voit
58-590
-insperial
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
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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
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Unter den 2013GraduierungsaPrädikat „sumDissertation „Torneal endothwei erfolgreicus dem Ausla
hre Arbeit am on der Queen
Brisbane, ausgMinister’s QueAward, und BeNational Unive
efördert durcrust, vertiefen
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Gemeinsam mMolekulare Bio
echnischen Ueptember 201
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BiomaterialienBiologie organ
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s löslichen Prr vielseitiges ltifunktionelle
hylenglykol-HyType-Additionektive Peptidfrkomponente
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
und Wechselungen von denren
oye, Dietmar Aerer
terisierung voen und deren
der oder zu anund der kompr Probensystesforderung daethode zur Moausschlusschrbei sehr häufigür sind u.a. dier Polymere u
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
-insperial
wirkungs- ndritischen
Appelhans,
n dendritischWechselwirk
nderen Molekplexen Zusammme immer wi
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
rkungen, wie zmatischen Stu
piriergesta
en kungen ülen men-eder die timmung, ie (SEC), nzen. onalität igung zu
er en cher-
n zu ungen ür die
s-bination Hierbei es se statt, üsse die eigen-
hohen öglich-
einander dieser
ungs-yko-Maltose-
von
ungen ysin-schale, ktionel-n inter-z.B. H-udien
wGKKgmmkWmA
2KBszPsdFKwdVDluneuAdBuMKIwK
rte Galtun
wurde der EinGenerationszaKonzentrationKonformationgekoppelter Dmit der AF4-Lmoleküle als konnte nachgWert die Strukmoleküle beeAggregationsv
2. AuftrennunKomponentenBei der Untersstrukturen, dizwischen biotPolypropylenisich heraus, ddispersen SysFall herrscht Komponentenwurden erstmdie einzelnen VerbindungenDabei wurde dligandenanzahund Biotin, odnisses (Avidineiner Biohybrund bivalenteAbb. 2 dargesdass in dem SBestandteile aund komplexeMolmassen voKombination mInformationenwurde auch eKonformation
renzng
nfluss der Denahl, des pH-Wn auf die Molmseigenschafte
Detektion näheLS möglich istauch Aggregaewiesen werdktur der einzeinflusst und dverhalten steu
g und Charakn eines Biokonsuchung von e auf nichtkovinylierten Glymin) und Avid
dass die Herststemen nicht meine Mischun
n vor. Diese komals mittels A
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
kterisierung denjugates: Biohybrid-valenten Bindkodendrimere
din basieren, stellung von mmöglich ist. Ing von verschieomplexen SysF4-LS separieden jeweilige
und quantifizie.a. der Biotin-
ndes zwischenularen Verhältet. Ein Beisp
ebildet aus Avitnis 1:3), ist ine nachgewiesl ungebunden
nierte Architekuren mit sehr d [2]. In r Lichtstreuunffusionskoeffizg der
möglich.
hen-
r en und 4 und . Da es el-eren, pH-r-
1].
er
ungen en (GD, stellte ono-
n jedem edenen teme ert und n ert. -n GD lt-iel idin
n der sen, ne kturen hohen
ng und zienten
f
A
s
s
W
Keywords
biopolymers
bioconjugates
guest-host-syst
field-flow-frakt
light scattering
conformation
Abb. 1:
Übersicht über d
schematischen M
strukturen der P
dendronisierten
polymere als Ein
moleküle und Ag
bei verschiedene
Werten und vers
Konzentrationen
33
tems
tionation
ie
Molekül-
Polylysin-
Glyko-
nzel-
ggregate
en pH-
chiedenen
Biolund
34
Abb. 2:
Zusammensetz
Biohybridstruk
gebildet aus Av
Glykodendrime
zwei Biotinliga
Verhältnis 1:3 b
mittels AF4-LS
logie- Mate
zung der
kturen
vidin und
eren (GD, mit
anden) im
bestimmt
S
-insperial
3. Quantifiziekapazität beiIm GegensatHybridstruktbei AufnahmGastmolekülpolymeren hhäufig eine MPolymer aufgvorliegt. Um zu quantifizieHerangehenentwickelt: (des Gast-Wirreinen Glykoten Gastmoleaufgenommemittels LS deMöglichkeit dder GastmolAbtrennung simultaner Ffiltrationseffsenkrechte LTrennkanal dtransportiertMembran kömoleküle abdetektiert bzsich dann bemenge der kersten Testmverzweigten korrelierten Methoden seAktuelle Untstoffsystemesimultane Ko
piriergesta
erung von Komi Gast-Wirt-Kotz zur Bildungturen sind die
me- oder Freislen in dendritiäufig geringe
Mischung aus genommenendie komplexi
eren, haben wsweisen mittei) über den Mort-Komplexes
opolymer die Zeküle berechnen Molekülen etektiert zu wder (ii) direkteeküle. Unser der freien Gas
Filtration. Dazekt der AF4 gLösungsmittedurch eine Met wird. Durch
önnen die kleingetrennt und
zw. quantifizieei bekannter Gkomplexierte Amessungen m
Glykopolymedie Ergebniss
ehr gut miteinersuchungen
e sollen zeigenomplexgrößen
rte Galtun
mplexierungsomplexen: von AggregatMoleküldimeetzungsstudieischen Glyko-r. Hinzu kommfreien und vo
n Gastmolekülerten Gastmo
wir zwei verschels AF4-LS olmassenzuw
s im VergleichZahl der kompnet. Ist der Anzu gering, umerden, gibt es
en QuantifizieWeg beruht astmoleküle mu wird der Ultenutzt, bei delfluss aus demembran heraudie Poren dernen, freien Gaquantitative
ert werden. SoGastmolekülgeAnteil berechnit einem hochr und einem Fse der beiden ander [3]. an Polymer-Wn, wie effektivnbestimmung
renzng
-
ten oder ensionen en von
mt, dass m len
oleküle hiedene
wachs zum
plexier-nteil an m s die rung uf der
mittels tra-
em der m us r ast-
o lässt esamt-nen. Bei
h Farbstoff
Wirk-v sich die g und -
qlä
KP
[1
[2
[3
fläch
uantifizierungässt.
Kooperation: Prof. L. Nilsso
1] Boye, S.; AZschoche,FormanekLederer, A(12), 4222-
2] Ennen, F.;M.; KombeAppelhans(DOI: 10.10
3] Boye, S.; PLederer, A(29), 4841-
hen-
g auf diese Sys
n University L
Appelhans, D. S.; Komber,
k, P.; Janke, AA.: Biomacrom-4235 Boye, S.; Leder, H.; Brutscs, D.: Polymer039/c3py01152fPolikarpov, N.A.: J. Chromat-4849
steme übertra
Lund
; Boyko, V.; H.; Friedel, P.
A.; Voit, B. I.; molecules 2012
derer, A.; Cernhy, B.; Voit, B.r Chemistry 20f) ; Appelhans,
togr. A 2010, 12
agen
.;
2, 13,
nescu, .; 014,
D.; 217,
Biolund
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
y a gel system with a four-a
rPEG) by linkethe coagulatiothe linker rel
e, where it exenticoagulant aydrogel outcoarin coatings reducing the d, thus, minimilication of gel
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
drug load to tzing side effe-coated produ
d with a thrombin
hydrogel
piriergesta
siveness
,
blood-tents or em in n on and cules sulting cess. have ating, or
of the blood ntrol is heparin
hylene ecifically
rombin. n to the ely hrombin-cally r exciting he cts in the ucts.
-
ToahtNpatevnhecfhefmbtectbaeacbf
Fhd3tpm2[
rte Galtun
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
n whole bloodoagulant capa
ubated with hinary self-asse
xyl-terminatedkanethioles ontion, the pro-
hen-
eavage e co-uence,
when d. n
agula-ulant mental
an be
, by ower or
e (Thr-zyme-n gula-ol/l d
evels of eaved were s to y active imal
ound to on
he n, low at acity, ghly embled d and n gold)
f
a
a
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
patient’s is
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
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Sponsor: Deutsche FoCollaborate CooperationProf. Dr. M.Universitäts [1] M. Prew
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Keywords
hydrogel
biomaterials
growth factors
cell adhesive pe
39
eptides
Biolund
40
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[1
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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
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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
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ubbell: Biomac
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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
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macroporous oped as threeof the extraceg applications
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Welzel, ffen Vogler, en Werner
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[1] P. B. WelL. NaujoxWerner: B2358 (201
[2] P. Joly, GWelzel, UPetersen
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arameters onmechanics allon of the cryolasticity and t
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Keywords
cryogel
AFM
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Green: cryogel st
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their local mecha
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extracted.
41
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of the PBS
scaffold.
truts.
d inter-
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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
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of applied
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g geometry
on).
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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
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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
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exp A2 4B
rte Galtun
t of charged eometries
Sommer
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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
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where the conspplied electrind characteriompression-e
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when the conseriodically (orelds). These f
Fig. 2.
ig. 2:
n a set time interv
may move a certai
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heir positions. Th
istances z for two
onstant (noted cy
ylinders under co
modulated cylinde
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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
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o chains of length
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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
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he chains by obse
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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),
(2013), 3007-3021
