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Relationship between reversible and irreversible adsorption protein layers on the surface of segmented polyetherurethane
~ol~mer 8¢[¢n¢¢ U ~ .~ R Vol. ? 7, No 6, pp. 149g- 1494, |gN~ PrmLed in Poland
0012-'1')50/85 $10.010 ~. O0 Persamon Ioumal,z L,td
RELATIONSHIP BETWEEN REVERSIBLE AND IRREVERSIBLE ADSORPTION PROTEIN LAYERS ON THE S I . ~ F A C E OF SEGMENTED POLYETHERURETHANE " A ~/'A. PI~LISHCHIIK, L, V. VLADIMIRI.W, A L. IORDANSKii, G . YE". ZAIKOV, (-) G. F,~RrUNA'rt.,V, A. V. TRE.Z.VI,VA and Yt.'. V. IL.'¥AKOV IriMIlule of Ph,¢,acal Chemr,lry, I_J S.S r
A~.':ldeln,; ,~1 Selerl~'es
(Recen ed ~ 4U!lU.S't IqX4 'The ad~orpl,Otl ,~1 hunlarl ~erunl alhurrlln ~l, Ihe ,,iJrlace ~1 p~)lvclh~crur¢lh,.lne ha,; I,een sludled a,, a hJncl~,m ol Ih~ u'onlenl ill' Ihe ,,oll ,,egmenl in IIl~' polyiner I'w AT'R F,~urler IR ,;peelr~:.;cop',, II i,, .,dl,~wn Ihal I[le c'onh,rmallonal ;,el ,,I ad,J,~rhed preHeln macr,,m,fleculc,, d,ffer;J (ram Ihal o f l h e dry and dlw.,dved form,, o[ pr,,leln The kmeli~',.;ofde,~,,rpllon o f l h e eeverstl'~le layer ,d' llle alhumm tram I11¢' ;,urla~e ,,I polvc'lherurethalle has been MLid,ed. 'The physlcochemi~.al paramelern of IIw p, dyn-ler problem sy,,lelTI,; cakulaled wHh u,.,e ol Ihe dfl' liJslon kln~ll~.' n~,~J¢'l allow I1,¢ II,r,,mbu,, re,Jl.Marll pr=~pL'rll¢'. ~1 Ih~" m,llerial,., ,~liJdied Io be e~ a hJa IL-d SII.fDY of IIle adnorpll,~n of proleln:~ ,,n a n,m.phy,,uH,q:,,iLal ,,urla*.'¢¢ I,, altraeting Ihe atlenllon o1' researchers ,n connexnon wllh Ihe problem ul poal dnrecled ,.,¢arcl'l I',~r lhrombus resnslarlt ,~yrllhelnc maler~al,,. II n'.; known Ihal Ihe adsorplnem of pla~.;ma proleun', n,~ ihe prnn-,srv act or Ihrombus Iormalure orl Ill~: p~flymer ,,urface i,i conlacl wilh blood I II However, Ihe hrlk belwccn II'le ad,a~rpllorl and .SlJb!,~quenl processe.., c~rlhnlJe ~, Io remain ob,.icure II i'~ dd~cull lot example, to an,,wer Ih¢ question why is Ihe ,nleracllon of prok-in,, wilh Ihe surl'aL'¢ ol a nahJral ve,.~nel ll,~l accompamed by adhc'nlon arid morpholo.pjcal changes in Ihe Ihrombocylcs, whlk' such elfecl,¢ are ob,,erved in presence of Ihc rl(m.plzy~.,iologleal ,,urla~e of a polymer 'The mosl convmcinL.', ver;,ion appears Io be Ihat a,.'llw.lllOn ~1 Ihe Ihromb{~L '~le,, L)n Ihe pr,,le=n aled ~.;urI,Jee I.'., Mlrrlulaled by IIm ~.qmlormall,mal cl|ar, ges and denalurmg of parl of Ihe protein ad :,~rhed on Ih~: ,,urla~:e 12, :ll II may he ¢on~,iderc'd Ihat Ihe larger Ih¢ I'ra¢llon =~1'Lonh,rmal,onally unch0nged prolelrl Ihe belier will be Ihe Ihrombus re:.,i,;lanl properlies of a given polymer malerlal. 'Therelore, mm:h imporlar,¢e altache,, Io informahon on chan!?e ,n II'le ¢ompo~Hion and ~;Iru~.'lure ol Ihe protein layer m Ihe course or ii,~; ac~.'umulahon on Ih¢ polymer ,;url'ace Earlier it was .shown 12-~1 Ihal Ihe mechani,;m of ad:,Ltrpllon e,~r,stilule.~.~ a complex mulll singe pfoLe,~!, including dll-Tu.;ion ~,1 Ihc protein globule,~ Io Ihe polymer ,,urlace and Ihe suh,~cquenl slag¢,~ ~1 Iormallon or ~.'onl',,rmalt~mally ¢hanl:,ed and nahve proleln laver';. T o eslabh,di Ihe correlahon belween Ih¢ km¢l.ic puram¢lers of adsorpllon and Ihe Ihr¢,mhu~, r~:,w,lanl properl~es ol'a polymer it ,s des,rable Io sludy rtlalerials giving a good aecounl of IItemselves in medical practice Such mal.ermls w,dely used in preparinL.,, syslems of the "axtd]clal heart" type Im. lude segmenled polyethert.uelh-
aries I~1 'l'h~ mum aim of Ih¢ pre,~enl, work ,.~ to devise a reliable melhod Ior record,rig the prnlein adsorbed on Ib¢ su.rlace of segmerlled polyelher;~elharae ,n order Io delermm¢ Ihe rallo of Ihe rever,:able and ,rrevers=ble proleln layers al any momenl of lime
* Vysokomol soyed. A27 No.h, 1~27-1~1, Ig8'~ 1489
1490
A. YA. POtJSHCHLIK ('! (71.
PI-WSICOCH.I~MICAI, PR(1P[,R TIES AND 5LIR FACE .~'TRLIC'T'LIRE, OF $.~J~'L,['$ (IF, S['OME'NTED POLY1E'THER LIRE,'THANE Sample A B
rr., M P a 4t hO
ep o, 700 ')(X)
E, M P a II 4 '~
D~nao
Z')n~,,a,~
~'h ~"h
I h '2 h
'T'¢) rec~rd Ihe surl'ace c~mcenlral.~n o1' ads(~rbed prolein Ihe melhod ~1' ATR IR ,~pe¢'lr¢)s~.'()pyi,~ r)l'len used However, prehniiriarv invesllgahons ~d' Ihe A'T'R speelra showed Ihal in Ihe ca.~e o1' (he ~yMem.s considered Ihe hands amide I and anude II m()sl ~.'hara~.'lerlslic and inlcnse in IIie speelrum ;~1" proleln are masked hy ahsr)rpllrm o1' II'ie p;dym'elharie malrix. 'Tl'.s dltficully could be ten'loved by u,,,ir=~.~, A'T'R comhmed wilb Ih¢ lechmque q)l' ditTereulial IR F'~lurier '.,pe,.'lro.~,cqlpy (IR FS) alhl~,mg one al'ler digilal deduclion (~1 Ihe speclrum el' Ihe pulvmer ~uppor( Io is~)lale rehal')lv Ihe lar les~ mlense ~p¢clrum o1' adsorhed pr,~lein E';.u'lier Ihe A'T'R IR-FS melh¢)d wa~ used h) invesllgale the ~,pec. Ira (d' prulein ad~lrbed on Ihe surl'ace ~d' quarlz 171, germanium and PVL' IX, 91 In the pres~'nl W~Lirk we inve~,lit.:aled samples el polyelherurelhaue.~ synlhe,,ized on Ih¢ ba~,js o f Ihe copulymer o1' p~dyl'Lu'lle wilh oxvpropvlerie, diphenyl m¢lhari¢ dnsocy,auale and elhylene dnamlne wh(ch conlained 27 (.~ample A) arid ,.I,.I (.,~ample B) II,) hard urelhane uJ'ea segmenls ['he physlco mechanical pn)perlie~ :rod surluce ~,(ruclcLre of Ihe sampfe,~ are indlealed in Ihe Table The su.rl'ace ~lruelure or Ih¢ .,.,aniples was evalualed by Ihe A T R IR melh,,J I'nm~ Ibe rahu el Ihe ophcal deusilies o r l h e ahs~rplir)n band I I I ( t c m a ~.'hara~.'lvrlzmg Ihe relallve c~mccnlrahr)n ed' Ihe esler F.~oups ed' Ihe ~(')1'1.segnieuL on Ihe surl'ace add I1i¢ hand Ih4LI c m = chu.racler~z.ing (he relallve c~m~.'enlration ~.d'Ihe urea t,.,q'uups (d' Ihe hard .se~menl h) IIie ~lplical den;~ily (d' Ihe inlerrial :,laDdard - Ihe band 77U em" (-) II belong, jng Io Ihc - C - C - 1..~()up. In Ihe wnrk we used human serum album,n or Reanal (Hungary); p m l e m ~'(mlenl (~n c(~nver~non Io dry mailer was 9'~",'~ 'The IR speclra of Ihe pure and proleiualed .~amples o1' p~lvelherurelhaue (SPLr), dry and dis,~nlved prulem ( b u f f e r - 'T'yrade, pH 7.4(.)) were recorded wilh Ihe DIGIL, AB F"T'S I~C IR F'oLLrier spectrorneler wllh re~olulioa ()1' 2 and 4 c m - a A sali.sl'a,.'lc)ry .'~H..,,;nal.lonoi~e ralin was acl'neved by a large nunil'~r c~l' ,,canmn~,. To measure Ihe A'T'R speclra we u~.,ed an allachmenl wilh Iwo paralelt mirror~ and cryslals ur K R $ '; wJlh Ihe angle.~ 4.~ and 60". 'To elimmale speclrul di.,,Iorlions Ihe A'T'R spectra were ~)blamed at angles ~d' incidence o1' radnalion H ' hl.)' well exceedin/~ Ihe crilical (H,,,.'-42"). All Ihe~,peclra w e r e o h l a i n e d al rr)um lemperalare The kinehc expernmenls were bused (m Ihe lechnJque pr~'~p('~sed in rel'erenee I%, IOI The A'TR IR F'S !,pcclrum or albumin a d ~ r b e d Ir~rn hul.[er ~,ululi~m on II'ie ,,urlacc ed' SPL/ is .,~hown nn F"i~. I (curve /) and Fag 2 (curve /). This ~pec'lrum i,~ ~hlained by deduc. ling Ihe speclral conlr~bulion (d' SPI.I (Fig. I, curve 2) I'rom II'=e ..,,hd'~ul ~peclrum ol Ihe :w',lem SPl.J-ad.,,,)rbed albumin (Fit.~. I, curve 3). F';~r comparls~m Fig 2 gave!, Ihe ,,wclra ed' albumin powder ()n a cryslnl ~d' KRS ¢i (curve 2) and albumin dH:;olved in bufl"er (curve i'). As Fig 2 show:,, Ihe relalive mlen'~il(e:; and h~rrri of Ibe conlLtars ~1 Ihe m~Jsl inlen,,e burid,~ or Ihe speclra of Ihe adsnrbed, pure and dls.,,dved I'~rm:~ ul' prolein in Ihe main mah:li. AI Ihe .,~ame lime a!, compared wllh Ihe speclrum (d" Ihe alhumm powder Ihe I'requencnes o1' Iheband.,, am~de.I and annde II in Ihe ~,pe,.'lrum ~d" pn'~lein adsorbed ~m Ihe surl'ace o1' SPLI .~hil'l In h.lwer frequencJe,~' Ih,.l(I-~ IhTh (am~de I ) a n d I~ 1 ' ~ 1'~2h (arrud¢ U ) c m a. II i,~ known I I I I Ik)r proleins wdh an ¢-hehcal conformahrm ~d' the chains (l'(~r example, I'or albumin 1121)Ihal ¢hanp,e in Ihe conl'ormalional siruclu.re I'rr~m ~.~ heh~ Io Ihe '~lall~llC~.ll cull and layered //slruclure may lead Io rn~,e in I he hlw I'requency slnfl of Ihe.~e hands We w~uld uls¢~ m)le olher possible reason,,~ I'or Ihe shd'l rd' Ihe hands amide.I and amnde.II - Ihe dilferem'e in Ihe rel'rachve radices o1" prr~lem (as pr~wder and lave r ol ad~.orbcd macrorried¢,:ule~) and al'~o Ihe appearance or an addHnonal pr~.~le=n-SPLI interlace.
R.clal=on.~h=p helween reversible and irrevers=ble adsurpli,~n prolein layer~
14~)1
On the ,~.!,therhand, compaJ'ahv¢ analys=,s of the' A'T'R I R-FS ,~peclra ~.d'Ihe bulTer .,.,olul,on uf albumin and Ihc albumin puwder (F',g. 2,curve~ .1and 2) eslabhshed Ih¢ hip,b Irequcncy shdt o1" Ihc bands arrHde I and am=de II in the spectrum o1' file pru[cm solulion amounl,ng re,specl,vely Io I and ~ ~.'m A .~in)ilar tendency =n Ihe behav=~.~urof Ihe.,;e band;., was n~le~..|earlier m rel'erence [I t J where Ihe I R F'S melh~d ,~h~wed Ihal ~.m passa~,e ~1' Ihe globular pr~leirl,~ Ir,~m file .~,~lld ;dale I,~ an aque~u;~ ~ l u l t ~ n Ihe Irequeney ¢d' amide I remain.'~ prachcally unchanged while Ihe band am=de II undergL~e', a I'HI~I'~ I'rc:quency .~htl'l Jv = b-g c m ~ 'The rea.,,,,m I'~r Ih~, rc,:,ull ~, Ihal Ihe inl-luencu ¢~1 Ihe weak ¢md~)rn'=a lional ~.'hang,e~,in Ihe syslem ~r) Ihc p~.~,iliem ~Jl' Ihe hand arrude II esxenltallv hehml.,,mg I~ Ihe v=hra It(m.'.; ~'d'Ihe N H ~ fp'~'~up.',,i~, ~.'~n.~td~'rahly le.'.,,.,lhan Ihe a¢ll~rl excrled ~m il by in,.'rca'.,e ~n Ihe c~i~ cnlrJ Item and slrenglh ~1' Ihe h,vdrc~gen b~md~ =n Ihe n'=¢~le~.'ule'.,~d' d=.,.,s~'~lwd pr,qein The band amrde I rel'ers m Ihe main h~ IIle valence vlbrah~,ns ~1 Ihe (." = (_) I~r~tJp~'~ and v,'cal,.Iy rc,J~.'l,, h~ ~.hange,, in Ihc ,~ec,md=trv ,,IrLt,.'lure ¢~1' Ihe pr,~lem mu~.rc.m-=¢decuh:.
Dv~apb un ~ v o e b un
I
2
I
20
18
16 F"I~, I
lq
12.
v, lO-~cm.~
I
I
1
I
|
3D
18
16
ILl
12
I0
v,10"z, cm "1 F'r~ '?
Fi*= I. A'T'R IR-PS .,,,peclra of Ihe hy.,.;lem SPLI-serum aibumm I - dtlTerenhal .,.,peclrum ~d' !,erum albumin (dvtTerence m speclra (,r-3)x2'~); ,?-.,,,pcclJ'Um ol' polyefherurelhane malrtx, 'l-D~h)bal specfrum ol ,.;y.slem SPLJ-albumm (De,, ,L~phcal den~tly) FiG. 2 A T R IR-FS spectJ'a ,cd"human ~¢rum albumin: I -dilTerenhal speclrum cd' alburrun adscPrbed ~.t,n ~.,url'ac~ ~d" SPLI; 3 - spc'ctrum ,)1" a l b u m m apphed Io KRS-~ ~."l'y.,.;lal; ~-dtlTerenhal speclrum ,Ld' a l h u m m de,strived in 'T'yrade buffer (speclrum of s,=.dulicm-speclrc, m ~~1";,olvenl). F'rL)m all Ihi~ we eqms=der Ihal the ~h,lt (o Ih¢ h)w Irequency re 'Luon , ol Ihe absL~rphon band amtde I ~.t,I adsorbed a l b u m m =~ primaJ'dy due Io oplteal et:T¢cls a~soctaled wilh use or ihc A'TR melhod I-he d=fl'erence in Ihe ,~hdl ~~1 Ih¢ bands am,de-I and amid¢-II poml,~ Io change ,n Ihe ~..~mlormal=4.mal ,.;el of Ihe adsorbed prol¢in molecules us compared wilh Ih¢ nalive, lyophiliz.ed prepaJrall~,n By recording the. ~pect]'um ol prolem adsorbed on SPLI al dtfferenl moments of lime it ts p~.~,.~stble Io oblarn Ihe. k,nel,¢ curves of accumulalmu and rer'nuval of Ihe macromol¢cule.'.., Irom Ihe ,~url'ac¢
1492
A. YA, POLISHCJEUI[ e l M.
to the malerial and hence to give the quanlilaUve chata~teriRt,cs of the adsorption process. We in. vestigaled the kinehc curve.,; of d¢.,;orplion or albumin (Ftg. 3). The results ,nd,cale Iirslly the existence o f a heavily adsorbed prolein layer Ihe size or which is cbaraclerized by Ihe, I,mJt of Ib¢ kiaellc de.~orpt,on curve; secondly, Ihere exists a reversibly adsorbed albumin layer removed rrom Ihe surlace rm replacing Ihe surTou.nd,ng prole,a soluHoa by butter soluhon In addtlion, Ihe dilTereal conlenl of the .~q~l( segnienl ,n poly¢lherurelhane ,nflueaces Ihe I,milmg valu¢~ ~~1'Ihe k,net,c curves and al.,,,.~ Ihe rain ol desorpli,m. The q u a n m a l i v e characleri,,,hc;~ or the de,~,~rplion curves were calculaled wilh Ihe aid ul" the previ¢~u'qy pr~p,)sed dll-]'U'~l,~n klnellc m~)del ]~-~, IOJ acc¢~rdlng h'~ wlll¢'h Ihe desorplion pr,~cess include~ Iwo ,~erlal paraJlel ,~lage~ - de,~c~rptlon o1' prolein propel' Irum Ihe ,~urlacc and diffu,~lon transfer ~d" Ihe macromnlecule,~ Ihruugh a layer ~ Ih=ck rain Ihe pure snlvenl. II m~ne ol Ihese ~lage,s i~ hrruling ~ditTu,.ion klnelic fegl,,n) Ihen Inr Ihe limes t . ~'~"I'D Ihe dv;,~,rpl.~n curve I', a p p r o x l m a l c l y described by II-ie e q u a l . m (.,,.,
A,,, f",'. Nt'?:l (' /,,,,C';: +/~,,., e~,p
'~ ,'_.'_/'r, t t I,~ + ~ . ]
(1)
where (?,,z t,~ Ihe c,mcerilr.',l,~m ,)l' Ihe rev¢'r.,,tble layer al lhe mumeril ~1 I=me ;, (?~=~,,, Ihv cuncenlraliun ~1' pr~dein iii ,,()hJh()n beh~re the ~la.l'l or Ihe de.~urpl.m experlmenl. (~,.,'~ i;~ {he concer=lrahOn ~)1 Ihe heavdy ,,.~rbed pr()lem layer, k,.. ix Ibe rale ¢,'~rl;;lanl ~.~l ad;,.~rpl.m ¢~1 Ihe rever,,tble layer, /~,,z 1,, (he rule ¢on,,lanl Id' des~)rpl[nn (d' Ihe rever;,ihlc, layer, N ix Ihc,, rriean nLirr~ber ~)1'acliv¢, cerilre~ ,)1' r¢ verstble a d s ~ r p l . m per m~deeule underg~.ng c~ml'mmaliunal ~.'hange,,, I~ ~ = 2l)/,~ ~ ( D i,; Ihe ddTu~.on co,:tT]cwril for prl~leirl =r= !,,.dul.m).
c~,
rO , k g / m z
.= .,. I
~6
i
--
1[
1E
=
=
I
I
1"8
a,
I
I
I
30 f. lO'~-~ec
FIG. 1, Kinelic de.~orpli,m curves of ;,erum albumin I'rr)m surl'm:c ~1' $PI.I I - ,;ample A; 2 - sa topic. B.
F r o m Ihe kinel,c curves pre.~enled in Fig. !, a curve, may be described by equalion (I) corresponding Io a 27 % c o n l e n t or Ihe ht~d se~,~"nentin Ihe polymer molecule. 'The ~aphie` soluhon of Ihis equalion in Ihe semi h')gaJ'ilhmJc coordmale~; (In ( ( . . s d ( . s , ) - t ) , n hne, wilh Ihe procedure described In reference II01 yields Ihe rain con.,;lan! ur desorpl,on or the revers,bin layer ka~. = U . ] 2 ~: IO- = sec" and help.s lu delerm,ne Ihe ral~o q~l' Ihe crmcenlralion.,J o1' the rever.s,bly and irreversibly bound prr~lem al Ihe irtlial moment ur lime C'~(U)/C,~'~' = 2. T o lhis ralio c o r r e s p o n d Ihe paramelers o1' Ihe syslem albumin-SPl.I A given below N A ,.,. cm~/g sec
2 4(.K)
1.
4
s
70
40
I i,
The be.~l malch of the exper,menlal and calculated craves ,s observed al N = 3, k~, = 70 ¢m ~] l g ' .,,ec, allhough Ihe 20% random errur ,n delermmmg C',s~ does nol perm,| ¢xclu.Nion of other var,anls of Ih¢ con~tanls prc,~ntcd above.
Relalionsh,p between reversible and irreversible adsorptlun prole,n layers
149.1
Wilh increase in Ihe conlenl of |he hard ~ g m e n l or polyelherurelhane Ih¢ rale of de.,,orpl,on ri~,~ and the proces,> of passage of Ihe reversible prolein layer mlu .~nluhon shins Io Ihe dilTuslon regmn. The k=n¢lic curve nf ad~orphon in Ihr, case i'~ well described by Ihe I'nllowing se! of peru. milers. N = 1,,/~,~ = 40 cm~/g net', k:., =U .1 s ~ A,~ a whole, Ihe unaly,~i,, of Ihe desorplion km¢lic curves made shows fhal wilh mcrca.se in the Iract..m o1' IIle sl~il segmenl in Ihe chemical slruffure ol SPU fhe Iolal amounl nl irreversibly adsorbed protein deLrea'~es and Ihe reaction of h~rmall~m ~1 the rever,~ible I.'lyer shifts h~ adNnrpllon. It IS mo,J likely Ihal Ihe number ol reversible blm:hng cenlrcs dries rlnf depend im Ihe ral,o r~f Ihe hard arid ,J,~ll segmerll,; in Ihe pulyrrler L,e, u,, now h.d, at the re,,iull,, in relali,}n h'l Ihe ,,earth I'or Ihrombu,,-re,;i;danl maleflab; |1 P, k n o w . Ihal increase m Ihe ,;hare i~1 Ihe ,,,d'l ,~¢gmenl un Ihe surface of polyefherurelhan¢,, =mproves the haemi~c==mpaldd¢ pr~perlw,, ol the producI,J oH' Ih,,~ cla,;,, 1141 ()n¢ esplanalion or Ihl,~ may be ll'i¢, fall ublamed I'ly us m Ihe am~iJnl id albumm undergo,ng on Ihe surrac¢ c o n l o r m a l i o n a l chanLkmS wilh relahve mcrea,;¢ =it Ihe e;,ler fra~-.,,menl m Ih¢ polymer 'The corTelah~m found in parhcularly importanl since albuillin I,, Ihe rrialn comp~menl In fhe t,Halll~, i~l i~rolelns adsorbed Irom the plasmu on Ihe surlac¢ ol SPU I1'~, Ihl In adddi~m, Ihe w¢)rsening of Ihe Ihrombus re,~lslanl propcrlie,~ of Ihe polymer chine=des wHh ~lllfl ill equdibrium m IIie reacliori of fnrmahon ~lr the rever,;ible layer towards de,,orpllon 'This pulrll,~ h~ the po~,lll~,¢ r~le played I'rom the po~nl cff view ~l haem~cnmpalibilily by Ihe deacllvahon of Ih¢ rever~ll.~l¢ ad~i~rpl,m ccrilres h)rmed ,in the mnlecule undergoing conformahonal c!iangcs. 'Thus, sludv of Ihe desorplion of albumin Irnm Ihe surface of SPU has ,~hown Ihal Ihe A r R IR FS m¢lhod chosen arid Ihe use ul Ihe ddrusion kmehc m~del makes ~1 possible In e:;labhsh a ciirrelahon belween Ihe ad,,orpl,m and Ihrombu,;.res~lam properl=es .I Ihe inhomngeneous poly ¢lhcrurelhane surla~'e ']'he developmenl ol the approach proposed may be linked with study OI the adsorption of other plasma prol¢,n.s on amide contammt., polymers, Ihe mechanism =~f ~.'ompchhve adsorph~n amJ pre dlchori ~1 Ihe hac.m~t'oinpalibl¢ pruperhes of synlhelic rrialc.rials 'Ft,mslated hi, A C'm.iz','
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4 A. Ya. P()LISHCHtlK, A. L. I O R D A N S K I l a n d G . Ye. ZAIKOV, Khim fizika I: 1268, 1982 % A. L. I(.)RDANSKII, A. ¥a. P O L I S C H U K and G. E. ZAII(OV, J Macrnnlolec Sci. Ray Macromolec. (."hem Phys. 23: 1'I, 198 I, b Polirnery medtlsinskogo naz.nacl'ieniya (Polvmer,~ for Medical Llses)(ediled by Senoo M a n a b u ) 248 pp, Medllsma, Moscow, 1081 7 R. M. GENDREALJ, AppI Spectroscopy 36: 47, It~82 8 R. M. GENDREAU, S. WINTERS, R.'~. L,ENINGER, D. FINK, C'. R. HASSLER and R. J. J A C O B S E N , Appl Speclro~copy .L~i: ht~, 1981 '9 R. M. G E N D R E A U a n d R . J . J A C O B S E N , J Biomed Maler Res. 13:89t, Iq7q 10. A. Ya. P O U S H C H U K , A. L. IORDANSKI! and G. Ye. Z,AIgOV, Vysokomol. soved A ~ : 1008, 1984 (Translaled in Polymer Sc, U S.S.R. A26: h, 1191, It)84) II J. L. KOEN1G and D. L. TABB, In. Analvlical applications of FI'.IR In moletular and bin logical ~w~lem,~ (ediled by During, J R ) p. 2,41, Pro( Nat Advanced Studies In,dilulc. N 'l' ,
1980
14q4,
V.A.
SAr, IKISYAN t't U/
12. M. E. SODERQLIIS'r and A. G. WALTON, J. C o l h , d In/ S~l 75: 1Nh, IqX0 I ~. M. M. COLEMAN ,',nd P. C. PAINTER, J Macromnk'c. Sol , R~v Ma~rmn~dcc. Chem Phys Ib: Iq7, IqT7-TN 14 M. D. LELAH, L. K. L A M B R E ( ' H T , B. R. YOLING and S. L. COOPER, I Bn~rr~,(J M~]ler. R~',,. 17: I, IqN~ I). S. I~OCHWA, R. S. L,I'rWMg, R. E. ROSENFIELD arid E. F. LEONARD, Ann. N Y. Acad S~i 283: ~7, Iq77 ih M . L E M M ~md V. L I N G E R , In' Adg;Jri~c,, 111 Bi~m]aler,al,, (~d,lcd by W l h w , I }, N Y , I: ~1),4, I "}NIl
Pul*, m¢'r 5~ lenl 1' I i ~ I'~ ~ V,d. 21, N,~ h, pp 14'J4- 14'.~8, 19N"~ PllriH'd Ill Pld,.llld
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ORIENTATION OF MACROMOLECLILES AT THE IN'rERFACE OF POLYMER AND FILLERS* V A. SAI~KI~'~I'ArIq, M (3. ANV,AIYI~%, A A. MkilirArY~,N, K KH K,~TR[IZHY,~N alld A
K
DADIVANYAN
K lr;ivJi~,Jll r'cddl],i~,,i~' lll'.,lll,ile ( Rcc,~'med 17 ,Iml.s! IqX,..I) A Hiq'lliod i,, l,rop~),,cd l'~r d~'l~rrillrllilL ~ lhe ~rlellHJfli~n i~rdt.'rlnl. ~, l;ieh~r and fh~' llil, knc,.,,; ,~I lhl.' ~)rll:i|l{'d l:i~'cl .Jr fli~' llll~rl,J('¢~ (~I fli~' l),d~ni~'r v,zilh ~iri,~lh~r pha,;e hy l[IVe,,ll!,.'illlllZ lhe riBJi,Jl d~..p~lll]~.,it.'~' ()I' hli'~'irlrll-~,~'ll('~ • appVilrli|~;, ,J( lli{: Inl~'l'l'dt't.' ~l' lh~.' p~ilvn'icl Wll!i hph~rl~.',iI ilr L'Vlllldi'll .ii hllcr pilrflcle,, l'q r l l [ L-~)llrld,.iry layer,, ul' gla~.;~,y polymL~r ,, ~n llle l]dl mlerla~'L ~ wllh ~lllcr l~lia,,t% lh~ru appear'~ ~ponl,ilB.'~pu,~ hlrclrlnl, cnL'e II-(~J lh~~ Vdlllt.~ ipl ~ l l l r h I,, pr,lp~rll,~ilal h~ ih,~ i~l~liL ,Jl ,Irll.;~driIpV ~i" lhc iri,.il.'r,)ini,lq:(lJl¢ 'TIIL' ihiCkll~:~,,., ~pl file ~r;cnIL~d laver .'illd dt'I..~¢l~ of ~ri~.=;ilaiB)n ~Ii' iJi~' ma~'r~m~decule,; 111 l{ ,tfl~ d~ll;'FmlrB'd IIv IflV~.,ll!~HllflIg p()lyITWl hlm~ ~d' dliJL'rcn| lhiulqne:~,,~,, [hl. ()rlL'flidiB~n ('}I' lhe p,~ lymer i', al!.;~ ~b,,er~ud r~lallve I,~ lli~ ~ hller ,,,llrl'a~'e 171 ~,VIIIII)IJI 12.~1111~IIIll) IhL' rllL'l h.illl,~lll ;)I' I~)rmdill;H| ~)I f i l l ~ (IrlCIllL~d hwcr c,m,~d~rud in a o u n l b c r ol MIJdi~':.; IN-11~I we :.,b~ill ;.,lli~W lllill lhc IIIIL'I~Du.~, ~d' lhe orlenled lay
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N ~,~ Ihe rlurilber of rnolecule.~ p~r unil vulume, ~z is II'le mean ;el'rachvc ,nd~x ,~1 Ihe sample.. 'To find Ihe pola.rlzahlhly c~l IIlC m a c r o m u l e c u l e m Ihe l a b o r a l o ; y :~yslem ~d' c o o r d m a l e s ~ I,, Icl u~ expres~ it in lhe sy.~lem ~, i',:~ h n k e d w i l h ihe cylmd.rlcal ,~r !~pherlcal .~,url'.~ce. Lel u~ denola
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IqX5
0012-'1')50/85 $10.010 ~. O0 Persamon Ioumal,z L,td
RELATIONSHIP BETWEEN REVERSIBLE AND IRREVERSIBLE ADSORPTION PROTEIN LAYERS ON THE S I . ~ F A C E OF SEGMENTED POLYETHERURETHANE " A ~/'A. PI~LISHCHIIK, L, V. VLADIMIRI.W, A L. IORDANSKii, G . YE". ZAIKOV, (-) G. F,~RrUNA'rt.,V, A. V. TRE.Z.VI,VA and Yt.'. V. IL.'¥AKOV IriMIlule of Ph,¢,acal Chemr,lry, I_J S.S r
A~.':ldeln,; ,~1 Selerl~'es
(Recen ed ~ 4U!lU.S't IqX4 'The ad~orpl,Otl ,~1 hunlarl ~erunl alhurrlln ~l, Ihe ,,iJrlace ~1 p~)lvclh~crur¢lh,.lne ha,; I,een sludled a,, a hJncl~,m ol Ih~ u'onlenl ill' Ihe ,,oll ,,egmenl in IIl~' polyiner I'w AT'R F,~urler IR ,;peelr~:.;cop',, II i,, .,dl,~wn Ihal I[le c'onh,rmallonal ;,el ,,I ad,J,~rhed preHeln macr,,m,fleculc,, d,ffer;J (ram Ihal o f l h e dry and dlw.,dved form,, o[ pr,,leln The kmeli~',.;ofde,~,,rpllon o f l h e eeverstl'~le layer ,d' llle alhumm tram I11¢' ;,urla~e ,,I polvc'lherurethalle has been MLid,ed. 'The physlcochemi~.al paramelern of IIw p, dyn-ler problem sy,,lelTI,; cakulaled wHh u,.,e ol Ihe dfl' liJslon kln~ll~.' n~,~J¢'l allow I1,¢ II,r,,mbu,, re,Jl.Marll pr=~pL'rll¢'. ~1 Ih~" m,llerial,., ,~liJdied Io be e~ a hJa IL-d SII.fDY of IIle adnorpll,~n of proleln:~ ,,n a n,m.phy,,uH,q:,,iLal ,,urla*.'¢¢ I,, altraeting Ihe atlenllon o1' researchers ,n connexnon wllh Ihe problem ul poal dnrecled ,.,¢arcl'l I',~r lhrombus resnslarlt ,~yrllhelnc maler~al,,. II n'.; known Ihal Ihe adsorplnem of pla~.;ma proleun', n,~ ihe prnn-,srv act or Ihrombus Iormalure orl Ill~: p~flymer ,,urface i,i conlacl wilh blood I II However, Ihe hrlk belwccn II'le ad,a~rpllorl and .SlJb!,~quenl processe.., c~rlhnlJe ~, Io remain ob,.icure II i'~ dd~cull lot example, to an,,wer Ih¢ question why is Ihe ,nleracllon of prok-in,, wilh Ihe surl'aL'¢ ol a nahJral ve,.~nel ll,~l accompamed by adhc'nlon arid morpholo.pjcal changes in Ihe Ihrombocylcs, whlk' such elfecl,¢ are ob,,erved in presence of Ihc rl(m.plzy~.,iologleal ,,urla~e of a polymer 'The mosl convmcinL.', ver;,ion appears Io be Ihat a,.'llw.lllOn ~1 Ihe Ihromb{~L '~le,, L)n Ihe pr,,le=n aled ~.;urI,Jee I.'., Mlrrlulaled by IIm ~.qmlormall,mal cl|ar, ges and denalurmg of parl of Ihe protein ad :,~rhed on Ih~: ,,urla~:e 12, :ll II may he ¢on~,iderc'd Ihat Ihe larger Ih¢ I'ra¢llon =~1'Lonh,rmal,onally unch0nged prolelrl Ihe belier will be Ihe Ihrombus re:.,i,;lanl properlies of a given polymer malerlal. 'Therelore, mm:h imporlar,¢e altache,, Io informahon on chan!?e ,n II'le ¢ompo~Hion and ~;Iru~.'lure ol Ihe protein layer m Ihe course or ii,~; ac~.'umulahon on Ih¢ polymer ,;url'ace Earlier it was .shown 12-~1 Ihal Ihe mechani,;m of ad:,Ltrpllon e,~r,stilule.~.~ a complex mulll singe pfoLe,~!, including dll-Tu.;ion ~,1 Ihc protein globule,~ Io Ihe polymer ,,urlace and Ihe suh,~cquenl slag¢,~ ~1 Iormallon or ~.'onl',,rmalt~mally ¢hanl:,ed and nahve proleln laver';. T o eslabh,di Ihe correlahon belween Ih¢ km¢l.ic puram¢lers of adsorpllon and Ihe Ihr¢,mhu~, r~:,w,lanl properl~es ol'a polymer it ,s des,rable Io sludy rtlalerials giving a good aecounl of IItemselves in medical practice Such mal.ermls w,dely used in preparinL.,, syslems of the "axtd]clal heart" type Im. lude segmenled polyethert.uelh-
aries I~1 'l'h~ mum aim of Ih¢ pre,~enl, work ,.~ to devise a reliable melhod Ior record,rig the prnlein adsorbed on Ib¢ su.rlace of segmerlled polyelher;~elharae ,n order Io delermm¢ Ihe rallo of Ihe rever,:able and ,rrevers=ble proleln layers al any momenl of lime
* Vysokomol soyed. A27 No.h, 1~27-1~1, Ig8'~ 1489
1490
A. YA. POtJSHCHLIK ('! (71.
PI-WSICOCH.I~MICAI, PR(1P[,R TIES AND 5LIR FACE .~'TRLIC'T'LIRE, OF $.~J~'L,['$ (IF, S['OME'NTED POLY1E'THER LIRE,'THANE Sample A B
rr., M P a 4t hO
ep o, 700 ')(X)
E, M P a II 4 '~
D~nao
Z')n~,,a,~
~'h ~"h
I h '2 h
'T'¢) rec~rd Ihe surl'ace c~mcenlral.~n o1' ads(~rbed prolein Ihe melhod ~1' ATR IR ,~pe¢'lr¢)s~.'()pyi,~ r)l'len used However, prehniiriarv invesllgahons ~d' Ihe A'T'R speelra showed Ihal in Ihe ca.~e o1' (he ~yMem.s considered Ihe hands amide I and anude II m()sl ~.'hara~.'lerlslic and inlcnse in IIie speelrum ;~1" proleln are masked hy ahsr)rpllrm o1' II'ie p;dym'elharie malrix. 'Tl'.s dltficully could be ten'loved by u,,,ir=~.~, A'T'R comhmed wilb Ih¢ lechmque q)l' ditTereulial IR F'~lurier '.,pe,.'lro.~,cqlpy (IR FS) alhl~,mg one al'ler digilal deduclion (~1 Ihe speclrum el' Ihe pulvmer ~uppor( Io is~)lale rehal')lv Ihe lar les~ mlense ~p¢clrum o1' adsorhed pr,~lein E';.u'lier Ihe A'T'R IR-FS melh¢)d wa~ used h) invesllgale the ~,pec. Ira (d' prulein ad~lrbed on Ihe surl'ace ~d' quarlz 171, germanium and PVL' IX, 91 In the pres~'nl W~Lirk we inve~,lit.:aled samples el polyelherurelhaue.~ synlhe,,ized on Ih¢ ba~,js o f Ihe copulymer o1' p~dyl'Lu'lle wilh oxvpropvlerie, diphenyl m¢lhari¢ dnsocy,auale and elhylene dnamlne wh(ch conlained 27 (.~ample A) arid ,.I,.I (.,~ample B) II,) hard urelhane uJ'ea segmenls ['he physlco mechanical pn)perlie~ :rod surluce ~,(ruclcLre of Ihe sampfe,~ are indlealed in Ihe Table The su.rl'ace ~lruelure or Ih¢ .,.,aniples was evalualed by Ihe A T R IR melh,,J I'nm~ Ibe rahu el Ihe ophcal deusilies o r l h e ahs~rplir)n band I I I ( t c m a ~.'hara~.'lvrlzmg Ihe relallve c~mccnlrahr)n ed' Ihe esler F.~oups ed' Ihe ~(')1'1.segnieuL on Ihe surl'ace add I1i¢ hand Ih4LI c m = chu.racler~z.ing (he relallve c~m~.'enlration ~.d'Ihe urea t,.,q'uups (d' Ihe hard .se~menl h) IIie ~lplical den;~ily (d' Ihe inlerrial :,laDdard - Ihe band 77U em" (-) II belong, jng Io Ihc - C - C - 1..~()up. In Ihe wnrk we used human serum album,n or Reanal (Hungary); p m l e m ~'(mlenl (~n c(~nver~non Io dry mailer was 9'~",'~ 'The IR speclra of Ihe pure and proleiualed .~amples o1' p~lvelherurelhaue (SPLr), dry and dis,~nlved prulem ( b u f f e r - 'T'yrade, pH 7.4(.)) were recorded wilh Ihe DIGIL, AB F"T'S I~C IR F'oLLrier spectrorneler wllh re~olulioa ()1' 2 and 4 c m - a A sali.sl'a,.'lc)ry .'~H..,,;nal.lonoi~e ralin was acl'neved by a large nunil'~r c~l' ,,canmn~,. To measure Ihe A'T'R speclra we u~.,ed an allachmenl wilh Iwo paralelt mirror~ and cryslals ur K R $ '; wJlh Ihe angle.~ 4.~ and 60". 'To elimmale speclrul di.,,Iorlions Ihe A'T'R spectra were ~)blamed at angles ~d' incidence o1' radnalion H ' hl.)' well exceedin/~ Ihe crilical (H,,,.'-42"). All Ihe~,peclra w e r e o h l a i n e d al rr)um lemperalare The kinehc expernmenls were bused (m Ihe lechnJque pr~'~p('~sed in rel'erenee I%, IOI The A'TR IR F'S !,pcclrum or albumin a d ~ r b e d Ir~rn hul.[er ~,ululi~m on II'ie ,,urlacc ed' SPL/ is .,~hown nn F"i~. I (curve /) and Fag 2 (curve /). This ~pec'lrum i,~ ~hlained by deduc. ling Ihe speclral conlr~bulion (d' SPI.I (Fig. I, curve 2) I'rom II'=e ..,,hd'~ul ~peclrum ol Ihe :w',lem SPl.J-ad.,,,)rbed albumin (Fit.~. I, curve 3). F';~r comparls~m Fig 2 gave!, Ihe ,,wclra ed' albumin powder ()n a cryslnl ~d' KRS ¢i (curve 2) and albumin dH:;olved in bufl"er (curve i'). As Fig 2 show:,, Ihe relalive mlen'~il(e:; and h~rrri of Ibe conlLtars ~1 Ihe m~Jsl inlen,,e burid,~ or Ihe speclra of Ihe adsnrbed, pure and dls.,,dved I'~rm:~ ul' prolein in Ihe main mah:li. AI Ihe .,~ame lime a!, compared wllh Ihe speclrum (d" Ihe alhumm powder Ihe I'requencnes o1' Iheband.,, am~de.I and annde II in Ihe ~,pe,.'lrum ~d" pn'~lein adsorbed ~m Ihe surl'ace o1' SPLI .~hil'l In h.lwer frequencJe,~' Ih,.l(I-~ IhTh (am~de I ) a n d I~ 1 ' ~ 1'~2h (arrud¢ U ) c m a. II i,~ known I I I I Ik)r proleins wdh an ¢-hehcal conformahrm ~d' the chains (l'(~r example, I'or albumin 1121)Ihal ¢hanp,e in Ihe conl'ormalional siruclu.re I'rr~m ~.~ heh~ Io Ihe '~lall~llC~.ll cull and layered //slruclure may lead Io rn~,e in I he hlw I'requency slnfl of Ihe.~e hands We w~uld uls¢~ m)le olher possible reason,,~ I'or Ihe shd'l rd' Ihe hands amide.I and amnde.II - Ihe dilferem'e in Ihe rel'rachve radices o1" prr~lem (as pr~wder and lave r ol ad~.orbcd macrorried¢,:ule~) and al'~o Ihe appearance or an addHnonal pr~.~le=n-SPLI interlace.
R.clal=on.~h=p helween reversible and irrevers=ble adsurpli,~n prolein layer~
14~)1
On the ,~.!,therhand, compaJ'ahv¢ analys=,s of the' A'T'R I R-FS ,~peclra ~.d'Ihe bulTer .,.,olul,on uf albumin and Ihc albumin puwder (F',g. 2,curve~ .1and 2) eslabhshed Ih¢ hip,b Irequcncy shdt o1" Ihc bands arrHde I and am=de II in the spectrum o1' file pru[cm solulion amounl,ng re,specl,vely Io I and ~ ~.'m A .~in)ilar tendency =n Ihe behav=~.~urof Ihe.,;e band;., was n~le~..|earlier m rel'erence [I t J where Ihe I R F'S melh~d ,~h~wed Ihal ~.m passa~,e ~1' Ihe globular pr~leirl,~ Ir,~m file .~,~lld ;dale I,~ an aque~u;~ ~ l u l t ~ n Ihe Irequeney ¢d' amide I remain.'~ prachcally unchanged while Ihe band am=de II undergL~e', a I'HI~I'~ I'rc:quency .~htl'l Jv = b-g c m ~ 'The rea.,,,,m I'~r Ih~, rc,:,ull ~, Ihal Ihe inl-luencu ¢~1 Ihe weak ¢md~)rn'=a lional ~.'hang,e~,in Ihe syslem ~r) Ihc p~.~,iliem ~Jl' Ihe hand arrude II esxenltallv hehml.,,mg I~ Ihe v=hra It(m.'.; ~'d'Ihe N H ~ fp'~'~up.',,i~, ~.'~n.~td~'rahly le.'.,,.,lhan Ihe a¢ll~rl excrled ~m il by in,.'rca'.,e ~n Ihe c~i~ cnlrJ Item and slrenglh ~1' Ihe h,vdrc~gen b~md~ =n Ihe n'=¢~le~.'ule'.,~d' d=.,.,s~'~lwd pr,qein The band amrde I rel'ers m Ihe main h~ IIle valence vlbrah~,ns ~1 Ihe (." = (_) I~r~tJp~'~ and v,'cal,.Iy rc,J~.'l,, h~ ~.hange,, in Ihc ,~ec,md=trv ,,IrLt,.'lure ¢~1' Ihe pr,~lem mu~.rc.m-=¢decuh:.
Dv~apb un ~ v o e b un
I
2
I
20
18
16 F"I~, I
lq
12.
v, lO-~cm.~
I
I
1
I
|
3D
18
16
ILl
12
I0
v,10"z, cm "1 F'r~ '?
Fi*= I. A'T'R IR-PS .,,,peclra of Ihe hy.,.;lem SPLI-serum aibumm I - dtlTerenhal .,.,peclrum ~d' !,erum albumin (dvtTerence m speclra (,r-3)x2'~); ,?-.,,,pcclJ'Um ol' polyefherurelhane malrtx, 'l-D~h)bal specfrum ol ,.;y.slem SPLJ-albumm (De,, ,L~phcal den~tly) FiG. 2 A T R IR-FS spectJ'a ,cd"human ~¢rum albumin: I -dilTerenhal speclrum cd' alburrun adscPrbed ~.t,n ~.,url'ac~ ~d" SPLI; 3 - spc'ctrum ,)1" a l b u m m apphed Io KRS-~ ~."l'y.,.;lal; ~-dtlTerenhal speclrum ,Ld' a l h u m m de,strived in 'T'yrade buffer (speclrum of s,=.dulicm-speclrc, m ~~1";,olvenl). F'rL)m all Ihi~ we eqms=der Ihal the ~h,lt (o Ih¢ h)w Irequency re 'Luon , ol Ihe absL~rphon band amtde I ~.t,I adsorbed a l b u m m =~ primaJ'dy due Io oplteal et:T¢cls a~soctaled wilh use or ihc A'TR melhod I-he d=fl'erence in Ihe ,~hdl ~~1 Ih¢ bands am,de-I and amid¢-II poml,~ Io change ,n Ihe ~..~mlormal=4.mal ,.;el of Ihe adsorbed prol¢in molecules us compared wilh Ih¢ nalive, lyophiliz.ed prepaJrall~,n By recording the. ~pect]'um ol prolem adsorbed on SPLI al dtfferenl moments of lime it ts p~.~,.~stble Io oblarn Ihe. k,nel,¢ curves of accumulalmu and rer'nuval of Ihe macromol¢cule.'.., Irom Ihe ,~url'ac¢
1492
A. YA, POLISHCJEUI[ e l M.
to the malerial and hence to give the quanlilaUve chata~teriRt,cs of the adsorption process. We in. vestigaled the kinehc curve.,; of d¢.,;orplion or albumin (Ftg. 3). The results ,nd,cale Iirslly the existence o f a heavily adsorbed prolein layer Ihe size or which is cbaraclerized by Ihe, I,mJt of Ib¢ kiaellc de.~orpt,on curve; secondly, Ihere exists a reversibly adsorbed albumin layer removed rrom Ihe surlace rm replacing Ihe surTou.nd,ng prole,a soluHoa by butter soluhon In addtlion, Ihe dilTereal conlenl of the .~q~l( segnienl ,n poly¢lherurelhane ,nflueaces Ihe I,milmg valu¢~ ~~1'Ihe k,net,c curves and al.,,,.~ Ihe rain ol desorpli,m. The q u a n m a l i v e characleri,,,hc;~ or the de,~,~rplion curves were calculaled wilh Ihe aid ul" the previ¢~u'qy pr~p,)sed dll-]'U'~l,~n klnellc m~)del ]~-~, IOJ acc¢~rdlng h'~ wlll¢'h Ihe desorplion pr,~cess include~ Iwo ,~erlal paraJlel ,~lage~ - de,~c~rptlon o1' prolein propel' Irum Ihe ,~urlacc and diffu,~lon transfer ~d" Ihe macromnlecule,~ Ihruugh a layer ~ Ih=ck rain Ihe pure snlvenl. II m~ne ol Ihese ~lage,s i~ hrruling ~ditTu,.ion klnelic fegl,,n) Ihen Inr Ihe limes t . ~'~"I'D Ihe dv;,~,rpl.~n curve I', a p p r o x l m a l c l y described by II-ie e q u a l . m (.,,.,
A,,, f",'. Nt'?:l (' /,,,,C';: +/~,,., e~,p
'~ ,'_.'_/'r, t t I,~ + ~ . ]
(1)
where (?,,z t,~ Ihe c,mcerilr.',l,~m ,)l' Ihe rev¢'r.,,tble layer al lhe mumeril ~1 I=me ;, (?~=~,,, Ihv cuncenlraliun ~1' pr~dein iii ,,()hJh()n beh~re the ~la.l'l or Ihe de.~urpl.m experlmenl. (~,.,'~ i;~ {he concer=lrahOn ~)1 Ihe heavdy ,,.~rbed pr()lem layer, k,.. ix Ibe rale ¢,'~rl;;lanl ~.~l ad;,.~rpl.m ¢~1 Ihe rever,,tble layer, /~,,z 1,, (he rule ¢on,,lanl Id' des~)rpl[nn (d' Ihe rever;,ihlc, layer, N ix Ihc,, rriean nLirr~ber ~)1'acliv¢, cerilre~ ,)1' r¢ verstble a d s ~ r p l . m per m~deeule underg~.ng c~ml'mmaliunal ~.'hange,,, I~ ~ = 2l)/,~ ~ ( D i,; Ihe ddTu~.on co,:tT]cwril for prl~leirl =r= !,,.dul.m).
c~,
rO , k g / m z
.= .,. I
~6
i
--
1[
1E
=
=
I
I
1"8
a,
I
I
I
30 f. lO'~-~ec
FIG. 1, Kinelic de.~orpli,m curves of ;,erum albumin I'rr)m surl'm:c ~1' $PI.I I - ,;ample A; 2 - sa topic. B.
F r o m Ihe kinel,c curves pre.~enled in Fig. !, a curve, may be described by equalion (I) corresponding Io a 27 % c o n l e n t or Ihe ht~d se~,~"nentin Ihe polymer molecule. 'The ~aphie` soluhon of Ihis equalion in Ihe semi h')gaJ'ilhmJc coordmale~; (In ( ( . . s d ( . s , ) - t ) , n hne, wilh Ihe procedure described In reference II01 yields Ihe rain con.,;lan! ur desorpl,on or the revers,bin layer ka~. = U . ] 2 ~: IO- = sec" and help.s lu delerm,ne Ihe ral~o q~l' Ihe crmcenlralion.,J o1' the rever.s,bly and irreversibly bound prr~lem al Ihe irtlial moment ur lime C'~(U)/C,~'~' = 2. T o lhis ralio c o r r e s p o n d Ihe paramelers o1' Ihe syslem albumin-SPl.I A given below N A ,.,. cm~/g sec
2 4(.K)
1.
4
s
70
40
I i,
The be.~l malch of the exper,menlal and calculated craves ,s observed al N = 3, k~, = 70 ¢m ~] l g ' .,,ec, allhough Ihe 20% random errur ,n delermmmg C',s~ does nol perm,| ¢xclu.Nion of other var,anls of Ih¢ con~tanls prc,~ntcd above.
Relalionsh,p between reversible and irreversible adsorptlun prole,n layers
149.1
Wilh increase in Ihe conlenl of |he hard ~ g m e n l or polyelherurelhane Ih¢ rale of de.,,orpl,on ri~,~ and the proces,> of passage of Ihe reversible prolein layer mlu .~nluhon shins Io Ihe dilTuslon regmn. The k=n¢lic curve nf ad~orphon in Ihr, case i'~ well described by Ihe I'nllowing se! of peru. milers. N = 1,,/~,~ = 40 cm~/g net', k:., =U .1 s ~ A,~ a whole, Ihe unaly,~i,, of Ihe desorplion km¢lic curves made shows fhal wilh mcrca.se in the Iract..m o1' IIle sl~il segmenl in Ihe chemical slruffure ol SPU fhe Iolal amounl nl irreversibly adsorbed protein deLrea'~es and Ihe reaction of h~rmall~m ~1 the rever,~ible I.'lyer shifts h~ adNnrpllon. It IS mo,J likely Ihal Ihe number ol reversible blm:hng cenlrcs dries rlnf depend im Ihe ral,o r~f Ihe hard arid ,J,~ll segmerll,; in Ihe pulyrrler L,e, u,, now h.d, at the re,,iull,, in relali,}n h'l Ihe ,,earth I'or Ihrombu,,-re,;i;danl maleflab; |1 P, k n o w . Ihal increase m Ihe ,;hare i~1 Ihe ,,,d'l ,~¢gmenl un Ihe surface of polyefherurelhan¢,, =mproves the haemi~c==mpaldd¢ pr~perlw,, ol the producI,J oH' Ih,,~ cla,;,, 1141 ()n¢ esplanalion or Ihl,~ may be ll'i¢, fall ublamed I'ly us m Ihe am~iJnl id albumm undergo,ng on Ihe surrac¢ c o n l o r m a l i o n a l chanLkmS wilh relahve mcrea,;¢ =it Ihe e;,ler fra~-.,,menl m Ih¢ polymer 'The corTelah~m found in parhcularly importanl since albuillin I,, Ihe rrialn comp~menl In fhe t,Halll~, i~l i~rolelns adsorbed Irom the plasmu on Ihe surlac¢ ol SPU I1'~, Ihl In adddi~m, Ihe w¢)rsening of Ihe Ihrombus re,~lslanl propcrlie,~ of Ihe polymer chine=des wHh ~lllfl ill equdibrium m IIie reacliori of fnrmahon ~lr the rever,;ible layer towards de,,orpllon 'This pulrll,~ h~ the po~,lll~,¢ r~le played I'rom the po~nl cff view ~l haem~cnmpalibilily by Ihe deacllvahon of Ih¢ rever~ll.~l¢ ad~i~rpl,m ccrilres h)rmed ,in the mnlecule undergoing conformahonal c!iangcs. 'Thus, sludv of Ihe desorplion of albumin Irnm Ihe surface of SPU has ,~hown Ihal Ihe A r R IR FS m¢lhod chosen arid Ihe use ul Ihe ddrusion kmehc m~del makes ~1 possible In e:;labhsh a ciirrelahon belween Ihe ad,,orpl,m and Ihrombu,;.res~lam properl=es .I Ihe inhomngeneous poly ¢lhcrurelhane surla~'e ']'he developmenl ol the approach proposed may be linked with study OI the adsorption of other plasma prol¢,n.s on amide contammt., polymers, Ihe mechanism =~f ~.'ompchhve adsorph~n amJ pre dlchori ~1 Ihe hac.m~t'oinpalibl¢ pruperhes of synlhelic rrialc.rials 'Ft,mslated hi, A C'm.iz','
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12. M. E. SODERQLIIS'r and A. G. WALTON, J. C o l h , d In/ S~l 75: 1Nh, IqX0 I ~. M. M. COLEMAN ,',nd P. C. PAINTER, J Macromnk'c. Sol , R~v Ma~rmn~dcc. Chem Phys Ib: Iq7, IqT7-TN 14 M. D. LELAH, L. K. L A M B R E ( ' H T , B. R. YOLING and S. L. COOPER, I Bn~rr~,(J M~]ler. R~',,. 17: I, IqN~ I). S. I~OCHWA, R. S. L,I'rWMg, R. E. ROSENFIELD arid E. F. LEONARD, Ann. N Y. Acad S~i 283: ~7, Iq77 ih M . L E M M ~md V. L I N G E R , In' Adg;Jri~c,, 111 Bi~m]aler,al,, (~d,lcd by W l h w , I }, N Y , I: ~1),4, I "}NIl
Pul*, m¢'r 5~ lenl 1' I i ~ I'~ ~ V,d. 21, N,~ h, pp 14'J4- 14'.~8, 19N"~ PllriH'd Ill Pld,.llld
li~'dl,'H'l 'l, llJlX1~- iN) PL~rl;amlm h~iJrn,]l,~ Lid,
IIII]~
ORIENTATION OF MACROMOLECLILES AT THE IN'rERFACE OF POLYMER AND FILLERS* V A. SAI~KI~'~I'ArIq, M (3. ANV,AIYI~%, A A. MkilirArY~,N, K KH K,~TR[IZHY,~N alld A
K
DADIVANYAN
K lr;ivJi~,Jll r'cddl],i~,,i~' lll'.,lll,ile ( Rcc,~'med 17 ,Iml.s! IqX,..I) A Hiq'lliod i,, l,rop~),,cd l'~r d~'l~rrillrllilL ~ lhe ~rlellHJfli~n i~rdt.'rlnl. ~, l;ieh~r and fh~' llil, knc,.,,; ,~I lhl.' ~)rll:i|l{'d l:i~'cl .Jr fli~' llll~rl,J('¢~ (~I fli~' l),d~ni~'r v,zilh ~iri,~lh~r pha,;e hy l[IVe,,ll!,.'illlllZ lhe riBJi,Jl d~..p~lll]~.,it.'~' ()I' hli'~'irlrll-~,~'ll('~ • appVilrli|~;, ,J( lli{: Inl~'l'l'dt't.' ~l' lh~.' p~ilvn'icl Wll!i hph~rl~.',iI ilr L'Vlllldi'll .ii hllcr pilrflcle,, l'q r l l [ L-~)llrld,.iry layer,, ul' gla~.;~,y polymL~r ,, ~n llle l]dl mlerla~'L ~ wllh ~lllcr l~lia,,t% lh~ru appear'~ ~ponl,ilB.'~pu,~ hlrclrlnl, cnL'e II-(~J lh~~ Vdlllt.~ ipl ~ l l l r h I,, pr,lp~rll,~ilal h~ ih,~ i~l~liL ,Jl ,Irll.;~driIpV ~i" lhc iri,.il.'r,)ini,lq:(lJl¢ 'TIIL' ihiCkll~:~,,., ~pl file ~r;cnIL~d laver .'illd dt'I..~¢l~ of ~ri~.=;ilaiB)n ~Ii' iJi~' ma~'r~m~decule,; 111 l{ ,tfl~ d~ll;'FmlrB'd IIv IflV~.,ll!~HllflIg p()lyITWl hlm~ ~d' dliJL'rcn| lhiulqne:~,,~,, [hl. ()rlL'flidiB~n ('}I' lhe p,~ lymer i', al!.;~ ~b,,er~ud r~lallve I,~ lli~ ~ hller ,,,llrl'a~'e 171 ~,VIIIII)IJI 12.~1111~IIIll) IhL' rllL'l h.illl,~lll ;)I' I~)rmdill;H| ~)I f i l l ~ (IrlCIllL~d hwcr c,m,~d~rud in a o u n l b c r ol MIJdi~':.; IN-11~I we :.,b~ill ;.,lli~W lllill lhc IIIIL'I~Du.~, ~d' lhe orlenled lay
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N ~,~ Ihe rlurilber of rnolecule.~ p~r unil vulume, ~z is II'le mean ;el'rachvc ,nd~x ,~1 Ihe sample.. 'To find Ihe pola.rlzahlhly c~l IIlC m a c r o m u l e c u l e m Ihe l a b o r a l o ; y :~yslem ~d' c o o r d m a l e s ~ I,, Icl u~ expres~ it in lhe sy.~lem ~, i',:~ h n k e d w i l h ihe cylmd.rlcal ,~r !~pherlcal .~,url'.~ce. Lel u~ denola
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