BiomaterialsLecture17

Surfaces:InteractionsofProteins
withSurfaces
Unlessotherwisenoted,sourceInformationforthefollowingslides:
1) B.Ratner,A.Hoffman,F.Schoen,andJ.Lemons:BiomaterialsScience,
2
nd
edition(SanDiego:ElsevierAcademicPress.2004).
2) Butt,HJ.;Graf,K.;Kappl PhysicsandChemistryofInterfaces,2
nd
Edition
(WiletVCH:Weinheim 2006).
3) Modifiedfrom:Anne Mayes3.051J/20.340JMaterialsforBiomedical
Application,MIT,http://ocw.mit.edu,
ImportanceofProteinSurfaceInteractions
Modulatecelladhesion
Triggerthebiologicalcascade resultinginforeignbody
response
Centraltodiagnosticarray/sensordevicedesign&
performance
Initiateotherbioadhesion:e.g.,marinefouling,bacterial
adhesion
FundamentalsonProteins
ProteinFunctions
ProteinFunctions(cont.)
ProteinFunctions(cont.)
ProteinStructure
Proteinshavemultiplestructurallevels.
ProteinStructure
ProteinStructure
AAsidegroups
havevariable
chemical
character
ProteinStructure
SyntheticPolymersvs.Proteins
ProteinAdsorptiononBiomaterialSurfaces
Inasshortatimeascanbemeasuredafterimplantationinalivingsystem
(<1sec),proteinsarealreadyobservedonbiomaterialsurfaces.
Secondstominutes:amonolayerofproteinabsorbstomostsurfaces
Proteinadsorptionoccurswellbeforearrivalofcells thuscellsprimarily
interactwithaproteinlayer,ratherthanactualbiomaterialsurface
Background
A) Proteinactivity
variesinadsorbed
vs.solvatedstate
Why???
ProteinAdsorptiononBiomaterialSurfaces
Background
B. Entropicforces
ProteinAdsorptiononBiomaterialSurfaces
Background
B. Entropicforces
ProteinAdsorptiononBiomaterialSurfaces
Gaininentropyprobably
isthemaindrivingforce
duringspontaneous
proteinadsorption!!
Background
C. Adsorbedproteins
initiatephysiological
responsesto
biomaterials
ProteinAdsorptiononBiomaterialSurfaces
ModelsforProteinAdsorption:LangmuirModel
ModelsforProteinAdsorption:LangmuirModel
ModelsforProteinAdsorption:LangmuirModel
ModelsforProteinAdsorption:LangmuirModel
ModelsforProteinAdsorption:Scatchard Plot
ModelsforProteinAdsorption:Scatchard Plot
SurfaceSensitive
MeasurementsNecessary
1. Ellipsometry
2. SurfacePlasmon
Spectroscopy
3. QuartzCrystal
Microbalance
Summary:ProteinAdsorption
Proadsorptionfactors
Dehydrationoftheinterface
Increaseoftheproteininteriormobility
Columbicinteractions
VanderWallinteractions
Antiadsorptionfactors
Strongwaterbindingtointerface
Proteinrigidity
Surfacemobility
Lownetcharge
BiomaterialsLecture18
Surfaces:InteractionsofProteins
withSurfaces:Part2
ProteinSurfaceInteractions:Part2
Vroman Effect
Observations
IrreversibleAdsorption
Restructuring
MultilayerFormation
MeasurementofAdsorbedProtein
1) Techniquesforquantifyingadsorbedamounts
MeasurementofAdsorbedProtein
MeasurementofAdsorbedProtein
InSituEllipsometry
2) Techniquesforquantifyingadsorbedamounts
MeasurementofAdsorbedProtein
Ellipsometry
Ellipsometry
SurfacePlasmonResonanceSpectroscopy(SPR)
SurfacePlasmonResonanceSpectroscopy(SPR)
SurfacePlasmonResonanceSpectroscopy(SPR)
20 40
0.0
0.2
0.4
0.6
0.8
1.0
R
e
f
l
e
c
t
i
v
i
t
y
IncidentAngle/Deg
Simulatedcurveswithincreasingthickness
Gold Film
Glass Slide
He-Ne Laser
632.8nm
0
Detector
Polarizer
Prism
Adsorbed Molecules
Evanescentwaveopticaltechnique
sensitivetochangesinthicknessandoptical
propertiesofthinandultrathinfilms
Angstromsensitivity
Nondestructivetosamples
Attenuatedtotalreflection(ATR)setupin
aKretschmann configuration,opticsare
awayfromthesampleandsubphase
Surfaceplasmon excitationobservedin
reflectivityangularscan
Relativelyinsensitivetoenvironmental
changes temperature,viscosity,etc.
Multiskop
SurfacePlasmonResonanceSpectroscopy(SPR)
SurfacePlasmonResonanceSpectroscopy(SPR)
QuartzCrystalMicrobalance(QCM)
Time
F
r
e
q
u
e
n
c
y
AF
BeginningofAdsorption Equilibration
Injection
Sauerbrey Equation
Personal
Computer
Frequency
Counter
Oscillating
Circuit
Piezoelectric
Quartzcrystal
Scatchard Equation
Howtomeasurebindingconstants?
( )
q q q
A F m F
2
2 A = A
o o
F F K N K c F F
b b
A = A
Sauerbrey,G.,Z.Phys. 1959, 155,206.
Sauerbrey,G.,Ann.N.Y.Acad.Sci. 1949, 51,660672.
QCMD(Dissipation)
SimultaneousmonitoringofA
frequency(AF)andA dissipation(AD)
atmultipleharmonicsofthequartz
resonator
Dissipationrevealsqualitative
informationabouttheviscoelastic
propertiesoftheadsorbedlayer
Dissipationofaviscoelasticpolymer
layeronaquartzresonatorisheavily
influencebyitsstructure.Rigidfilms
showsmallADvalueswhilesoftor
flexiblefilmsshowlargerADvalues
Modelingtheviscoelasticproperties
usingaVoight modelcanyield
quantitativeinformation
stored
dissipated
E
E
D

= A
t 2
LargerE
dissipated
yields
moreviscoelasticresponse
Dissipation
http://www.qsense.com/$2/qcmd.swf
Mustuseatechniquesensitivetothestructureoftheprotein
CircularDichroism
CircularDichroism