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[(times)-8(.)-366.3(SFTs)-368(were)-376.2(washed)-372.5(three)-370.8(times)-370.8(in)-371(PBS)-366.9(plus)-373(2.7)-166.9(mM)]TJ
T*
[(EDTA)-446.3(before)-446.6(an)-441.1(18-h)-447.9(wash)-444(in)-440.1(hypertonic)-445.6(buffer)-445.4(\(50)-170.3(mM)]TJ
0 -1.1174 TD
[(Tris;)-294.7(1.5)-166.9(M)-295.7(NaCl)-289.8([Sigma];)-299.2(pH)-291(7.6\).)-292.4(After)-289.8(three)-295.9(30-min)-295.9(PBS)]TJ
0 -1.1116 TD
[(washes)-9.4(,)-245.3(SFTs)-252.8(were)-249.5(sterilized)-250.9(using)-253.1(peracetic)-252.1(acid)-248.8(\(0.1%)-250.8(v/v,)]TJ
T*
[(pH)-291.1(6;)-284.6(Sigma\))-285.4(for)-285.5(3)-168.1(h)0(.)-281(Finally)-291.3(the)-286.8(SFTs)-281.5(were)-289.8(washed)-286.1(in)-290.3(PBS)]TJ
0 -1.1174 TD
[(three)-238.3(times)-238.3(for)-233.6(30)-163.5(min,)-238.5(twice)-233.6(for)-239.4(60)-163.5(h,)-234.9(and)-234.8(once)-237.1(for)-233.6(120)-164.6(h)-237.2(and)]TJ
0 -1.1116 TD
[(stored)-336(refrigerated)-340.5(in)-336.4(PBS.)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 313.0582 566.1353 Tm
[(Histology)-341.4(and)-329.4(immunohistochemistry)]TJ
/F4 1 Tf
9.843 0 0 9.843 323.0361 549.751 Tm
[(Six)-544.7(decellularized)-552.8(SFTs)-546.5(were)-549(analyzed)-549.2(and)-545.9(compared)]TJ
-1.0137 -1.1174 TD
[(with)-714(native)-717.6(tissue)-710.3(\()]TJ
/F5 1 Tf
8.9909 0 TD
(n)Tj
/F6 1 Tf
.6681 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
[(6\).)-708.3(Transverse)-717.9(and)-712.9(longitudinal)]TJ
-10.379 -1.1116 TD
[(sampl)-7.8(es)-250.8(were)-249.5(taken)-255.6(from)-247.5(the)-252.3(toe)-252.3(insertion)-253.1(point)-247.7(\(toe)-253.3(region\),)]TJ
T*
[(middle,)-502.3(and)-494(ankle)-497.5(insertion)-500.8(point)-495.3(\(ankle)-498.6(region\))-498.5(of)-497.5(each)]TJ
0 -1.1174 TD
[(tendon)-491.7(and)-488.3(xed)-489.2(in)-491.9(10%)-488.2(\(v/v\))-489.4(neutral)-488.3(buffered)-489.1(formalin)]TJ
0 -1.1116 TD
[(\(NBF;)-387.5(Genta)-383.2(Medical\))-382.1(for)-383.4(histochemical)-389.2(staining)-378.8(and)-384.6(zinc)]TJ
T*
[(xative)-292.4(\(0.1)-168(M)-284.2(tris;)-287.9(3.2)-166.9(mM)-286.6(calcium)-289.1(acetate)-290.2([Thermo)-287.7(Fisher)]TJ
0 -1.1174 TD
[(scientic],)-282.9(27)-169.2(mM)-280.8(zinc)-277.6(acetate)-284.5([Sigma)-279.6(Aldrich],)-279.4(and)-280.9(37)-169.2(mM)]TJ
0 -1.1116 TD
[(zinc)-369.7(chloride)-368.5([Fluka])-365.9(pH)-365.9(7.2\))-363.8(for)-366.1(4)-364(days)-362.5(for)-366.1(immunohisto-)]TJ
T*
[(chem)-8(ical)-439(analysis.)-440.7(The)-443.4(samples)-443(were)-439.5(processed)-445(automati-)]TJ
0 -1.1174 TD
[(cally)-595.6(\(Leica)-593.1(11020)-587.2(Tissue)-590.4(processor\))-594.6(and)-591.9(embedded)-589.5(in)]TJ
0 -1.1116 TD
[(parafn)-413(wax)-408.5(using)-402.9(standard)-408.5(techniques.)-409.7(For)-406.3(histochemical)]TJ
T*
[(staini)-8(ng,)-253.3(sections)-264.4(\(6)-255.6(and)-257.9(10)]TJ
/F7 1 Tf
11.1393 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(m\))-256.8(were)-261(taken)-261.4(from)-259(the)-258(surface)]TJ
-11.6922 -1.1116 TD
[(throug)-6.7(h)-283.3(t)0(o)-290.3(the)-292.6(central)-287.9(region)-290.1(of)-290.1(each)-290.1(block)-289(and)-292.4(stained)-292.3(with)]TJ
0 -1.1174 TD
[(hema)-8(toxylin)-520.7(\(Biostain)-530.6(ready)-526.2(reagent\))-525(and)-528.6(eosin)-525(\()-81.7(Merck\))]TJ
0 -1.1116 TD
[(\(H&E\),)-386.6(4)]TJ
/F6 1 Tf
3.9166 0 TD
()Tj
/F4 1 Tf
.2477 0 TD
[(,6-diamidino-2-phen)-9.8(ylindole)-384.7(\(DAPI;)-382.6(Sigma\),)-381(Al-)]TJ
-4.1643 -1.1116 TD
[(cian)-381.2(blue)-374.3(\(Atom)-370.6(Scientic\),)-380.8(and)-373.1(Sirius)-375.1(red)-373.1(\(WVR)-377.2(Interna-)]TJ
0 -1.1174 TD
[(tional\)/Mil)-9.2(lers)-333.8(elastin)-332.8(stains)-335.9(\(Raymond)-340.5(A)-330.3(Lamb\).)]TJ
1.0137 -1.1116 TD
[(For)-342.9(immunohistochemistry)-11(,)-337.5(sections)-350.8(were)-347.4(taken)-342(at)-343.3(6)]TJ
/F7 1 Tf
21.6968 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
(m.)Tj
-23.2634 -1.1116 TD
[(The)-391.6(Ultra)-387.9(Vision)-387.7(system-HRP)-389.2(kit)-383.7(\(Thermo)-385.6(Scientic\))-388.9(was)]TJ
0 -1.1174 TD
[(used)-397(for)-394.9(antibody)-396.1(detection.)-397.4(Sections)-396.9(were)-399.2(analyzed)-399.5(using)]TJ
0 -1.1116 TD
[(monocloncal)-362.7(antibodies)-353.4(to)-353.7(collagen)-358.1(I,)-355.8(collagen)-358.1(II,)-351.1(collagen)]TJ
T*
[(III)-337.3(\(all)-339.9(at)-337.6(1:50)-332.8(dilution;)-338.9(Chemicon\),)-340.6(and)-338.5(tenascin-C)-338.1(\(1:100;)]TJ
T*
[(Leica)-246.5(Microsystems\))-243(and)-240.6(polyclonal)-244.1(antibodies)-243.9(to)-238.5(galactose-)]TJ
/F7 1 Tf
0 -1.1174 TD
(a)Tj
/F4 1 Tf
.5529 0 TD
[(-1,3-galactose)-437.1(\()]TJ
/F7 1 Tf
6.4221 0 TD
(a)Tj
/F4 1 Tf
.5529 0 TD
[(-Gal;)-428.2(0.37)-168(mg/mL;)-431(Enzo)-433(Life)-427.3(Sciences\).)]TJ
-7.5279 -1.1116 TD
[(Omission)-339.1(of)-336.2(the)-332.9(primary)-338.5(antibody)-338.5(and)-332.8(isotype)-337.2(control)-337.4(anti-)]TJ
T*
[(bodies)-491.5(\(mouse)-492.6(IgG1;)-490.3(rabbit)-486.1(IgM\))-489.1(were)-491.4(used)-489.2(as)-492.7(controls.)]TJ
0 -1.1174 TD
[(Sections)-270.2(of)-261.4(native)-268.4(and)-263.6(decellularized)-270.6(SFTs)-258.5(were)-266.8(also)-264.7(treated)]TJ
0 -1.1116 TD
(with)Tj
/F7 1 Tf
2.1541 0 TD
(a)Tj
/F4 1 Tf
.5472 0 TD
[(-galactosidase)-378.3(for)-366.1(2024)-166.8(h)-369.7(a)0(t)-366.4(3)0(7)]TJ
/F6 1 Tf
13.2243 0 TD
()Tj
/F4 1 Tf
.3917 0 TD
[(C)-370.7(\()0(5)-169.2(U/mL;)-366.2(Sigma\))]TJ
-16.3172 -1.1116 TD
[(to)-273(remove)-275.1(the)]TJ
/F7 1 Tf
5.8403 0 TD
(a)Tj
/F4 1 Tf
.5529 0 TD
[(-Gal)-271.5(epitope)-275.2(from)-270.5(the)-275.3(tissue)-272.6(as)-273.9(an)-274.1(additional)]TJ
-6.3933 -1.1174 TD
[(negat)-8(ive)-361.7(control)-360.4(for)-360.3(determinat)-10.3(ion)-360.5(of)]TJ
/F7 1 Tf
15.6434 0 TD
(a)Tj
/F4 1 Tf
.5529 0 TD
[(-Gal.)-361.3(Sections)-368.1(were)]TJ
-16.1963 -1.1116 TD
[(blocked)-263.6(with)-264.7(dual)-259.1(endogenous)-266.6(enzyme)-259(block)-266(\(Ultra)-262.3(Vision\).)]TJ
T*
[(Antibodies)-347.3(were)-347.4(diluted)-349.1(in)-342.2(Tris-buffered)-345.9(saline)-348.8(\(TBS\))-347.3(con-)]TJ
0 -1.1174 TD
[(taining)-539.2(0.1%)-531.9(\(w/v\))-536.3(bovine)-537.8(serum)-537.6(albumin)-534.5(\(Sigma\).)-537.6(The)]TJ
0 -1.1116 TD
[(sections)-310.5(were)-307.1(incubated)-307.4(with)-310.8(the)-304.1(diluted)-308.8(antibodies,)-305(isotype)]TJ
T*
[(controls,)-474.2(or)-462.9(antibody)-471(diluent)-464.3(only)-471.1(for)-464(1)-168.1(h)0(,)-465.3(TBS)-466.1(and)-471(TBS)]TJ
T*
[(conta)-8(ining)-357.1(0.05%)-366(\(v/v\))-362.7(Tween)-367(\(Sigma\))-361.4(were)-364.7(used)-362.5(as)-366(wash)]TJ
0 -1.1174 TD
[(buffers)-296.6(throughout.)-291.1(The)-293.6(sections)-293.2(were)-295.6(labeled)-299.4(with)-287.8(polymer)]TJ
0 -1.1116 TD
[(horseradish)-417.3(peroxidase)-417.6(\(HRP\),)-413.8(followed)-414.3(by)-411.1(DAB)-409.3(chroma-)]TJ
T*
[(gen)-338.5(from)-333.9(the)-332.9(Ultra)-336.1(Vision)-335.8(kit.)]TJ
1.0137 -1.1174 TD
[(The)-339.7(presence)-335.8(of)-336.2(the)]TJ
/F7 1 Tf
8.4725 0 TD
(a)Tj
/F4 1 Tf
.5529 0 TD
[(-Gal)-334.8(epitope)-338.6(in)-336.4(tissue)-335.9(sections)-333.6(was)]TJ
-10.0392 -1.1116 TD
[(also)-690.9(analyzed)-687.5(using)-685.1(biotinylated)-690.1(GSL-1)-687(lectin)-689(\(Vector)]TJ
T*
[(Labs\).)-443.1(Sections)-448.8(were)-439.6(blocked)-442.1(with)-443.3(streptavidin/biotin)-450.3(and)]TJ
0 -1.1174 TD
[(carbo-free)-274.7(blocking)-269.4(solution)-268.2(\(Vector)-275.9(Labs\).)-270.3(Lectin)-269.6(was)-270.1(used)]TJ
0 -1.1116 TD
[(at)-280(5)-162.4(mg/mL)-280(in)-273(TBS.)-273.8(The)-276.4(streptavidin)-280.8(polymer)-275.1(and)-275.2(IMMpact)]TJ
T*
[(DAB)-253.7(chromagen)-256.5(\(Vector)-252.9(Labs\))-255.3(were)-255.2(used)-253(to)-250(visualize)-255.5(lectin)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 62.022 749.6502 Tm
(2)Tj
47.719 0 TD
[(JONES)-334.9(ET)-338.7(AL.)]TJ
ET
endstream
endobj
72 0 obj
<>stream
/GS1 gs
BT
/F4 1 Tf
9.843 0 0 9.843 59.7543 725.1588 Tm
0 0 0 rg
0 Tc
0 Tw
[(binding.)-489.5(Lectin)-494.2(\(5)-163.4(mg/mL\))-488.4(absorbed)-494.7(with)-489.3(200)-164.6(mM)-488.1(galac-)]TJ
0 -1.1174 TD
[(tose,)-584.9(omission)-585.9(of)-583.9(the)-586.3(lectin,)-583(and)]TJ
/F7 1 Tf
14.981 0 TD
(a)Tj
/F4 1 Tf
.5529 0 TD
(-galactosidase-treated)Tj
-15.5339 -1.1116 TD
[(sections)-391.1(were)-393.5(used)-391.3(as)-389.1(negative)-386.9(controls.)-393.6(All)-390.3(sections)-391.1(were)]TJ
T*
[(viewed)-459.2(using)-460.5(an)-458.4(Olympus)-459(BX51)-455.6(microscope)-464.9(and)-453.7(imaged)]TJ
T*
[(using)-356.8(an)-348.9(Olympus)-355.3(XC50)-351.9(digital)-349.2(camera)-358(controlled)-353.5(through)]TJ
0 -1.1174 TD
[(Cell)-335.2(B)-336.2(software)-340.2(\(Olympus)]TJ
/F6 1 Tf
7.3823 0 0 6.5614 165.9968 674.7021 Tm
()Tj
/F4 1 Tf
9.843 0 0 9.843 171.8362 670.3369 Tm
[(,)-337.5(United)-331.4(Kingdom\).)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 59.7543 648.3967 Tm
[(Determination)-337.6(of)-336.7(total)-329(DNA)-338.7(content)-334.1(of)-330.4(native)]TJ
0 -1.2203 TD
[(and)-335.7(decellularized)-336.1(SFTs)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 621.014 Tm
[(Th)19.6(e)-290.2(t)0(o)20.7(t)0(a)19.6(l)-289.2(DN)19.1(A)-289.9(c)0(o)19.7(n)0(t)15(e)0(n)19.7(t)-289.2(of)-278.6(si)20.9(x)-289.1(d)0(e)19.7(c)0(e)18.5(l)0(l)20.6(u)0(l)20.7(a)0(r)19.7(i)0(z)13.8(e)0(d)-279.8(S)0(F)21.2(T)0(s)-279.7(w)0(a)18.8(s)-288.9(de)19.7(-)]TJ
-1.0137 -1.1116 TD
[(te)19.6(rm)13.9(in)20.7(ed)-187.7(an)19.7(d)-196.9(c)0(o)19.7(m)0(p)19.6(a)0(r)19.7(e)0(d)-187.7(w)0(i)19.8(t)0(h)-192.4(t)0(h)20.7(e)-198.1(DN)19.1(A)-197.8(c)0(o)19.7(n)0(t)20.7(e)0(n)19.7(t)-197.1(of)-192.2(na)19.7(ti)20.6(ve)-187.7(ti)20.6(ss)21.2(ue)13.9(.)]TJ
0 -1.1174 TD
[(Ti)19.5(ss)15.5(ue)-366.2(sa)19.9(mp)19.6(le)19.5(s)-375.3(w)0(e)18.8(r)0(e)-366.2(t)0(a)13.8(k)0(e)19.7(n)-375.5(fr)20.9(om)-366.3(th)20.7(e)-376.6(t)0(o)20.7(e)0(,)-368.6(m)0(i)19.5(d)0(d)20.8(l)0(e)19.6(,)-377.8(an)19.7(d)-375.5(a)0(n)19.7(k)0(l)20.7(e)]TJ
0 -1.1116 TD
[(re)14(gi)20.7(on)20.8(s)-455.9(o)0(f)-451.4(e)0(a)18.5(c)0(h)-446.9(t)0(e)19.5(n)0(d)20.8(o)0(n)20.8(,)-458.4(co)19.7(mb)19.6(in)15(ed)19.7(,)-452.7(a)0(n)13.9(d)-456.1(ma)18.4(ce)18.5(ra)19.7(te)19.6(d)-456.1(u)0(s)15.3(i)0(n)20.7(g)-456.1(a)]TJ
T*
[(sc)14.1(al)19.5(pe)19.7(l)-323.8(b)0(l)20.7(a)0(d)19.7(e)0(.)-316.7(T)0(h)19.6(e)-324.8(sa)14.1(m)15(p)0(l)14.9(e)0(s)-314.2(w)0(e)18.8(r)0(e)-314.4(w)0(e)18.8(i)0(g)15(h)0(e)19.7(d)-323.6(\(c)19.7(ir)20.7(ca)-315.5(25)-146.2(mg)-320.2(fr)20.9(es)19.9(h)]TJ
T*
[(ti)14.8(ss)21.2(ue)-274.1(an)19.7(d)-283.3(2)0(5)20.8(0)-156.6(mg)-274.1(de)19.7(ce)18.5(ll)20.6(ul)20.7(ar)14(iz)19.6(ed)-274.1(ti)20.6(ss)21.2(ue)19.7(\))-283.3(a)0(n)19.7(d)-283.3(fr)15.1(ee)18.5(ze)18.5(-d)20.9(ri)20.7(ed)-274.1(to)-273(a)]TJ
0 -1.1174 TD
[(co)19.7(ns)15.3(ta)19.6(nt)-434.3(we)18.8(ig)20.7(ht)20.7(.)-441.2(T)0(h)19.6(e)-445.8(DN)19.1(A)-439.7(w)0(a)18.8(s)-444.4(ex)19.7(tr)20.7(ac)18.5(te)13.8(d)-438.8(u)0(s)21(i)0(n)15(g)-438.8(a)-445.8(D)0(N)19.1(e)0(a)18.5(s)0(y)]TJ
0 -1.1116 TD
[(bl)14.9(oo)20.8(d)-410(a)0(n)19.7(d)-415.8(ti)20.6(ss)21.2(ue)-406.5(ki)20.7(t)-410.2(\()0(Q)14.2(i)0(a)19.6(g)0(e)19.7(n)0(\))-399.6(f)0(o)15.1(l)0(l)20.6(o)0(w)20(i)0(n)20.7(g)-415.8(th)20.7(e)-411.2(m)0(a)18.4(n)0(u)20.8(f)0(a)19.7(c)0(t)19.6(u)0(r)15.1(e)0(r)19.7()0(s)]TJ
T*
[(in)15(st)20.9(ru)20.9(ct)19.5(io)20.7(ns)21(.)-182(T)15(he)-170.4(ex)19.7(tr)20.7(ac)18.5(te)19.6(d)-179.6(D)0(N)19.1(A)-174.8(wa)18.8(s)-179.5(t)0(h)20.7(e)0(n)-170.4(q)0(u)20.8(a)0(n)19.7(t)0(i)20.6()0(e)19.8(d)-179.6(at)-164.8(26)20.8(0)-156.6(n)0(m)]TJ
0 -1.1174 TD
[(us)15.3(in)20.7(g)-266(a)-273(na)19.7(no)20.8(dr)15.1(op)-255.6(sp)15.3(ec)18.5(tr)20.7(op)20.8(ho)20.8(to)20.7(me)18.4(te)19.6(r)-271.8(\()0(N)20(D)0(-)20(5)0(0)20.8(0)-271.8(La)18.5(bt)20.7(ec)18.5(h\))15.1(.)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 59.7543 500.4282 Tm
[(Determination)-337.6(of)-336.7(collagen)-331.7(content)]TJ
0 -1.2203 TD
[(of)-336.7(decellularized)-336.1(SFTs)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 473.0456 Tm
[(The)-478(total)-479.4(collagen)-479.1(content)-476.8(of)-480.2(native)-475.7(\()]TJ
/F5 1 Tf
16.2021 0 TD
(n)Tj
/F6 1 Tf
.6681 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
[(6\))-474.5(and)-476.8(decel-)]TJ
-18.6038 -1.1116 TD
[(lularized)-572.4(\()]TJ
/F5 1 Tf
4.4235 0 TD
(n)Tj
/F6 1 Tf
.6681 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
[(6\))-566.6(SFTs)-569.5(was)-569.6(determined)-574.7(using)-569.9(the)-569(hydro-)]TJ
-5.8115 -1.1174 TD
[(xyproline)-714(assay.)-707.8(Samples)-713.7(of)-704.8(native)-711.9(and)-712.9(decellularized)]TJ
0 -1.1116 TD
[(tendons)-331.4(were)-335.9(freeze-dried)-335.8(to)-330.6(a)-330.6(constant)-333.7(weight,)-334.9(hydrolyzed)]TJ
T*
[(in)-520.7(6)-162.4(M)-520.4(HCl)-518(for)-515.9(4)-168.1(h)-513.7(at)-521.9(120)]TJ
/F6 1 Tf
11.6634 0 TD
()Tj
/F4 1 Tf
.3974 0 TD
[(C,)-518.2(and)-517.1(neutralized)-518.2(with)-518.2(6)-168.1(M)]TJ
-12.0608 -1.1116 TD
[(NaOH.)-374.5(Test)-369.6(samples)-373.9(\(50)]TJ
/F7 1 Tf
10.4193 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(L\))-372(were)-370.4(added)-370.7(to)-365.2(96-well)-372.8(plates)]TJ
-10.9722 -1.1174 TD
[(with)-506.6(100)]TJ
/F7 1 Tf
3.9512 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(L)-503.4(o)0(f)-497.5(chloramine-T)-508.9(solution)-504.3(\(chloramine-T)-504.2(hy-)]TJ
-4.5041 -1.1116 TD
[(drate;)-510.3(Sigma\))-504.3(for)-498.6(5)-168.1(min)-505.8(with)-500.9(agitation.)-505.8(Ehrlichs)-507.6(reagent)]TJ
T*
(\(100)Tj
/F7 1 Tf
2.0044 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(L;)-539.2(Sigma\))-538.8(was)-540.8(added)-543.5(to)-538(each)-543.5(well)-542.4(and)-540.1(the)-540.2(wells)]TJ
-2.5573 -1.1174 TD
[(covered)-351(and)-344.3(incubated)-347.7(at)-343.3(60)]TJ
/F6 1 Tf
11.6404 0 TD
()Tj
/F4 1 Tf
.3917 0 TD
[(C)-347.7(for)-343.1(45)-163.5(min.)-347.9(The)-345.5(absorbance)]TJ
-12.032 -1.1116 TD
[(was)-523.5(read)-519.3(at)-521.9(570)-170.3(nm.)-519.5(The)-518.3(hydroxyproline)-523.6(content)-522.9(of)-514.8(test)]TJ
T*
[(samples)-443(was)-437.1(determined)-442.2(by)-439.9(interpolation)-437.7(from)-443.3(a)-434.2(standard)]TJ
0 -1.1174 TD
[(curve)-336.1(of)-336.2(trans-4-hydroxy-)]TJ
/F10 1 Tf
10.5115 0 TD
(l)Tj
/F4 1 Tf
.5011 0 TD
[(-proline)-338.5(\(Sigma\).)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 59.7543 319.5779 Tm
[(Determination)-337.6(of)-336.7(the)-328.5(glycosaminoglycan)-335.3(content)]TJ
0 -1.2203 TD
[(of)-336.7(decellularized)-336.1(SFTs)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 292.1952 Tm
[(The)-385.8(glycosaminoglycan)-387.7(\(GAG\))-382.4(content)-384.7(of)-382.3(native)-383.6(\()]TJ
/F5 1 Tf
21.0632 0 TD
(n)Tj
/F6 1 Tf
.6624 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
(6\))Tj
-23.4593 -1.1116 TD
[(and)-442.2(decellularized)-443.3(\()]TJ
/F5 1 Tf
8.3055 0 TD
(n)Tj
/F6 1 Tf
.6681 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
[(6\))-439.9(SFTs)-437.1(was)-442.9(determined)-442.2(using)-437.4(the)]TJ
-9.6936 -1.1116 TD
[(dimethylene)-455(blue)-443.4(assay.)-448.6(Native)-447.8(and)-448(decellularized)-454.9(tendon)]TJ
0 -1.1174 TD
[(samples)-276(were)-272.5(freeze-dried)-278.2(to)-273(a)-267.2(constant)-276.1(weight)-273.9(before)-273.8(being)]TJ
0 -1.1116 TD
[(digested)-270.4(in)-261.5(papain)-268.3(\(1250)-264.7(units)-259([Sigma])-269.2(in)-261.5(25)-169.2(mL)-263.8(of)-261.4(5)-168.1(m)0(M)]TJ
/F10 1 Tf
23.4593 0 TD
(l)Tj
/F4 1 Tf
.5011 0 TD
(-)Tj
-23.9604 -1.1116 TD
[(cysteine)-386.7(hydrochloride)-379.7(plus)-378.7(5)-168.1(m)0(M)-378.7(N)0(a)]TJ
7.3823 0 0 6.5614 213.2787 235.7858 Tm
(2)Tj
9.843 0 0 9.843 217.0204 237.4299 Tm
[(EDTA)-377.2([WVR)-382.9(Inter-)]TJ
-15.9774 -1.1174 TD
[(national]\))-388(for)-377.6(26)-169.2(h)-375.5(a)0(t)-383.6(6)0(0)]TJ
/F6 1 Tf
9.9873 0 TD
()Tj
/F4 1 Tf
.3974 0 TD
[(C)-382.3(with)-379.9(agitation.)-379.1(Standard)-385.5(or)-382.3(test)]TJ
-10.3848 -1.1116 TD
[(solutions)-274.9(\(40)]TJ
/F7 1 Tf
5.3853 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(L\))-268.3(were)-272.5(added)-267(to)-273(wells)-266.8(of)-272.9(96-well)-269.2(plates)-273.9(with)]TJ
-5.9383 -1.1116 TD
(250)Tj
/F7 1 Tf
1.6703 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(L)-393.9(1,9-dimethylene)-398.3(blue)-397.4(solution)-400.7(\(DMB\).)-397.6(Absorbance)]TJ
-2.2232 -1.1174 TD
[(was)-362.2(read)-352.3(at)-360.6(525)-164.6(nm)-354.8(after)-359.2(2)-168.1(min)-356(of)-353.5(incubation.)-357(The)-357(sulfated)]TJ
0 -1.1116 TD
[(sugar)-363.5(content)-361.6(of)-359.3(the)-355.9(test)-362.7(samples)-362.4(was)-362.2(determined)-361.5(by)-359.3(inter-)]TJ
T*
[(polation)-430.8(from)-420.3(the)-430.8(standard)-425.7(curve)-428.3(of)-422.6(chondroitin)-428.3(sulfate)-424.8(B)]TJ
T*
(\()Tj
/F7 1 Tf
.3341 0 TD
(m)Tj
/F4 1 Tf
.5529 0 TD
[(g/mL;)-336.5(Sigma\))-331.5(versus)-335.7(OD)-338(525)-164.6(nm.)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 59.7543 138.7842 Tm
[(Semiquantitative)-341.3(analysis)-334.5(of)]TJ
/F12 1 Tf
12.7912 0 TD
(a)Tj
/F11 1 Tf
.4995 0 TD
[(-Gal)-335.3(content)]TJ
-13.2908 -1.2266 TD
[(of)-336.7(decellularized)-336.1(SFT)]TJ
/F4 1 Tf
9.843 0 0 9.843 69.7323 111.3448 Tm
[(Ti)19.5(ss)15.5(ue)-469.9(sa)19.9(mp)19.6(le)19.6(s)-484.7(\()0(1)20.9(0)0(0)-146.2(m)0(g)19.6(\))-484.9(fr)20.9(om)-475.7(v)21(e)-486.1(d)0(i)20.7(f)0(f)20.9(e)0(r)19.7(e)0(n)19.7(t)-485(na)19.7(ti)20.6(ve)-475.7(an)19.7(d)]TJ
-1.0137 -1.1116 TD
[(de)19.7(ce)12.8(ll)20.6(ul)20.7(ar)19.7(iz)19.6(ed)-302.9(SF)15.4(T)15(s)-317.7(we)18.8(re)-302.8(ma)18.4(ce)18.5(ra)19.7(te)19.6(d)-317.9(a)0(n)19.7(d)-312.1(ad)19.7(de)19.7(d)-312.1(t)0(o)-301.8(i)0(n)15(d)0(i)20.7(v)0(i)20.7(d)0(u)20.8(a)0(l)]TJ
T*
[(1.)18.5(5-)20.9(mL)-459.6(mi)19.5(cr)19.7(oc)19.7(en)19.7(tr)15(if)20.7(ug)20.8(e)-468.8(t)0(u)20.7(b)0(e)19.7(s)0(.)]TJ
/F7 1 Tf
12.9248 0 TD
(a)Tj
/F4 1 Tf
.5414 0 TD
[(-G)20(al)19.5(ac)18.5(to)20.7(si)15.1(da)19.7(se)-458.2(\(5)-146.2(U/)19.8(mL)-459.6(in)]TJ
-13.4662 -1.1174 TD
[(50)-146.2(mM)-303.8(Tr)13.9(is)-301.6(pH)-302.6(6.)18.5(0\))-301.7(wa)18.8(s)-317.7(a)0(d)19.7(d)0(e)19.7(d)-312.1(to)-301.8(th)20.7(e)-319(s)0(a)19.9(m)0(p)19.6(l)0(e)19.6(s)-311.9(of)-301.7(na)13.9(ti)20.6(ve)-302.9(an)19.7(d)]TJ
0 -1.1116 TD
[(de)19.7(ce)12.8(ll)20.6(ul)20.7(ar)19.7(iz)19.6(ed)-389.3(ti)20.6(ss)15.5(ue)-383.5(an)19.7(d)-398.5(r)0(o)15.1(t)0(a)19.6(t)0(e)19.6(d)-398.5(at)-389.4(37)]TJ
/F6 1 Tf
15.5973 0 TD
()Tj
/F4 1 Tf
.3859 0 TD
.3962 Tc
[(Cf)396.2(o)411.3(r)-2.3(2)396.2(4)250(h\()396.2(n)417.1(e)396.2(g)415.9(a)396.2(t)415.8(i)396.2(v)416.9(e)]TJ
-15.9832 -1.1116 TD
0 Tc
[(co)19.7(nt)15(ro)20.9(l)-421.7(t)0(i)20.6(s)0(s)15.5(u)0(e)19.7(\))0(.)-413.4(N)0(a)18.8(t)0(i)20.6(v)0(e)19.7(,)-423.9(de)19.7(ce)12.8(ll)20.6(ul)20.7(ar)19.7(iz)19.6(ed)19.7(,)-423.9(a)0(n)19.7(d)-427.3(ne)19.7(ga)19.7(ti)20.6(ve)-418.1(co)19.7(nt)20.7(ro)20.9(l)]TJ
25.5097 67.9242 TD
[(ti)20.6(ss)21.2(ue)19.7(s)-323.4(w)0(e)18.8(r)0(e)-308.6(p)0(l)20.7(a)0(c)18.5(e)0(d)-308.6(i)0(n)20.7(t)0(o)-307.6(i)0(n)15(d)0(i)20.7(v)0(i)20.7(d)0(u)20.8(a)0(l)-308.8(t)0(u)20.7(b)0(e)19.7(s)-323.4(an)19.7(d)-317.9(p)0(u)20.8(r)0(i)20.7()0(e)19.8(d)-317.9(mo)19.6(no)20.8(-)]TJ
0 -1.1174 TD
[(cl)19.5(on)20.8(al)-320.3(an)19.7(ti)20.6(bo)20.8(dy)-324.7(to)]TJ
/F7 1 Tf
7.6086 0 TD
(a)Tj
/F4 1 Tf
.5414 0 TD
[(-G)20(al)-320.3(di)20.7(lu)20.7(te)19.5(d)-329.4(t)0(o)-319.1(0)0(.)18.5(3)0(7)-146.2(m)0(g)19.6(/)0(m)19.5(L)-330.6(in)-319.1(TB)18.6(S)-329.2(w)0(a)18.8(s)]TJ
-8.15 -1.1116 TD
[(ad)19.7(de)19.7(d)-277.6(t)0(o)-273(a)0(l)19.5(l)-277.7(sa)19.9(mp)19.6(le)19.6(s.)-275(An)20(ti)20.6(bo)20.8(dy)-267.1(on)15.1(ly)-267.3(\(n)20.9(o)-283.3(t)0(i)20.6(s)0(s)21.2(u)0(e)19.7(\))-283.3(wa)18.8(s)-277.4(i)0(n)20.7(c)0(l)19.5(u)0(d)20.8(e)0(d)]TJ
T*
[(as)-331.5(a)-336.3(n)0(o)-330.5(t)0(i)20.6(s)0(s)15.5(u)0(e)-331.7(c)0(o)19.7(n)0(t)20.7(r)0(o)20.9(l)0(.)-327.2(T)0(h)13.9(e)-336.3(mi)19.5(xt)20.7(ur)15.1(es)-325.7(we)13(re)-325.9(ro)15.1(ta)19.6(te)19.6(d)-340.9(a)0(t)-326(4)]TJ
/F6 1 Tf
21.7544 0 TD
()Tj
/F4 1 Tf
.3801 0 TD
.3419 Tc
[(Cf)341.9(o)362.8(r)]TJ
-22.1345 -1.1116 TD
0 Tc
[(16)-146.2(h.)-344.4(Fo)21(ll)14.8(ow)20(in)20.7(g)-352.4(i)0(n)15(c)0(u)19.7(b)0(a)19.7(t)0(i)20.6(o)0(n)20.8(,)-354.8(th)15(e)-353.6(s)0(a)19.9(m)0(p)19.6(l)0(e)19.6(s)-358(we)18.8(re)-348.9(ce)18.5(nt)20.7(ri)20.7(fu)15.1(ge)19.7(d)-352.4(a)0(t)]TJ
0 -1.1174 TD
[(60)20.8(0)-156.6(g)-185.4(fo)20.9(r)-185.4(1)0(5)-146.2(m)0(i)19.5(n)-185.4(an)19.7(d)-185.4(t)0(h)20.7(e)-186.6(su)21(pe)19.7(rn)15.1(at)19.5(an)19.7(t)-185.5(\()0(1)20.9(0)0(0)]TJ
/F7 1 Tf
16.202 0 TD
(m)Tj
/F4 1 Tf
.5414 0 TD
[(L\))-176.2(wa)18.8(s)-185.2(a)0(d)19.7(d)0(e)19.7(d)-185.4(to)-175.1(th)14.9(e)]TJ
-16.7435 -1.1116 TD
[(we)18.8(ll)20.6(s)-363.8(o)0(f)-359.3(M)0(a)18.8(x)0(i)20.7(S)0(o)21(r)0(p)-353.5(p)0(l)14.9(a)0(t)19.6(e)0(s)-354.5(\()0(N)20(u)0(n)20.8(c)0(\))-360.4(t)0(h)20.7(a)0(t)-354.8(h)0(a)19.7(d)-364(be)13.9(en)-354.7(co)19.7(at)19.5(ed)-354.7(wi)19.8(th)]TJ
T*
(50)Tj
/F7 1 Tf
1.1462 0 TD
(m)Tj
/F4 1 Tf
.5414 0 TD
(L)Tj
/F7 1 Tf
.8006 0 TD
(a)Tj
/F4 1 Tf
.5414 0 TD
[(-G)20(al)19.5(-B)19.8(SA)-181.5(\(1)20.9(0)]TJ
/F7 1 Tf
5.6388 0 TD
(m)Tj
/F4 1 Tf
.5414 0 TD
[(g/)20.7(mL)-177.4(in)-175.1(PB)20(S;)-175(De)18.8(xt)14.9(ra)19.7(\))-185.4(o)0(v)20.8(e)0(r)19.7(n)0(i)20.7(g)0(h)20.8(t)-185.5(at)-176.3(4)]TJ
/F6 1 Tf
14.0306 0 TD
()Tj
/F4 1 Tf
.3859 0 TD
(C)Tj
-23.6263 -1.1174 TD
[(an)19.7(d)-277.6(i)0(n)15(c)0(u)19.7(b)0(a)19.7(t)0(e)19.6(d)-277.6(fo)15.1(r)-277.5(3)-156.6(h)-283.3(a)0(t)-268.4(R)0(T)18.6(.)-285.6(Fo)21(ll)20.6(ow)20(in)20.7(g)-283.3(t)0(h)20.7(i)0(s)20.9(,)-279.9(50)]TJ
/F7 1 Tf
18.8688 0 TD
(m)Tj
/F4 1 Tf
.5414 0 TD
-.02 Tc
[(L)-304.5(s)-20(ec)-20(on)-20(da)-20(ry)]TJ
-19.4102 -1.1116 TD
0 Tc
[(ra)19.7(bb)20.8(it)-221.3(an)19.7(ti)20.6(-m)19.6(ou)20.8(se)-222(HR)18.9(P)-225.6(a)0(n)19.7(t)0(i)14.8(b)0(o)20.8(d)0(y)-215.3(\()0(1)20.9(:)0(1)15(0)0(0)20.8(0)0(;)-215.4(D)0(a)18.8(k)0(o)15.1(\))-225.7(wa)18.8(s)-231.3(a)0(d)19.7(d)0(e)19.7(d)-225.7(to)]TJ
T*
[(th)20.7(e)-226.9(w)0(e)13(l)0(l)20.6(s)-225.5(an)19.7(d)-231.5(i)0(n)20.7(c)0(u)19.7(b)0(a)19.7(t)0(e)19.5(d)-231.5(fo)20.9(r)-231.5(1)-156.6(h)-225.7(a)0(t)-222.4(R)0(T)18.6(.)-228.1(Th)19.6(e)-232.6(p)0(l)20.7(a)0(t)19.5(e)0(s)-216.3(w)0(e)13(r)0(e)-216.4(w)0(a)18.8(s)0(h)21(e)0(d)]TJ
0 -1.1174 TD
[(wi)19.8(th)-428.5(TB)18.6(S)-438.7(b)0(e)13.9(f)0(o)20.9(r)0(e)-429.5(a)0(d)19.7(d)0(i)20.7(n)0(g)-434.2(1)0(0)20.8(0)]TJ
/F7 1 Tf
12.153 0 TD
(m)Tj
/F4 1 Tf
.5414 0 TD
(L)Tj
/F5 1 Tf
1.054 0 TD
(o)Tj
/F4 1 Tf
.4896 0 TD
[(-p)20.9(he)19.7(ny)20.8(le)13.8(ne)19.7(di)20.7(am)18.5(in)20.7(e)-440(d)0(i)20.7(h)0(y)20.8(-)]TJ
-14.238 -1.1116 TD
[(dr)20.9(oc)19.7(hl)20.7(or)20.9(id)15(e)-198.1(\()0(O)20(P)0(D)14.4(\))-202.7(so)21(lu)20.7(ti)20.6(on)-192.3(\(S)21(ig)20.7(ma)18.4(\))-202.7(a)0(n)19.7(d)-202.7(in)20.7(cu)19.7(ba)19.7(ti)14.8(ng)-186.5(fo)15.1(r)-196.9(1)0(0)-146.2(m)0(i)19.5(n)]TJ
T*
[(at)-303(RT)-303.9(in)-301.8(th)20.7(e)-313.3(d)0(a)19.7(r)0(k)20.9(.)-314.4(Th)19.6(e)-313.3(r)0(e)19.7(a)0(c)18.5(t)0(i)20.6(o)0(n)-301.7(w)0(a)13(s)-311.9(st)20.9(op)20.8(pe)19.7(d)-312.1(w)0(i)19.8(t)0(h)-301.8(5)0(0)]TJ
/F7 1 Tf
21.2764 0 TD
(m)Tj
/F4 1 Tf
.5472 0 TD
.3133 Tc
[(L3)156.7(M)]TJ
-21.8235 -1.1116 TD
0 Tc
[(su)21(lf)20.7(ur)20.9(ic)-326(ac)18.5(id)-324.9(an)19.7(d)-335.2(t)0(h)20.7(e)-330.6(ab)19.7(so)15.3(rb)20.9(an)19.7(ce)-321.3(me)18.4(as)14.1(ur)20.9(ed)-320.1(at)-326(49)20.8(0)-156.6(n)0(m)-326(w)0(i)19.8(t)0(h)-324.9(a)]TJ
0 -1.1174 TD
[(re)19.7(fe)19.7(re)19.7(nc)19.7(e)-273()0(l)20.9(t)0(e)19.5(r)-271.8(of)-261.4(63)20.8(0)-156.6(n)0(m)19.6(.)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 310.8472 537.4487 Tm
[(Determination)-337.6(of)-330.4(the)-334.8(biocompatibility)]TJ
0 -1.2203 TD
[(of)-330.4(decellularized)-336.1(SFT)-338(tissue)]TJ
/F4 1 Tf
9.843 0 0 9.843 320.8251 510.066 Tm
(The)Tj
/F5 1 Tf
2.1945 0 TD
[(in)-635.9(vitro)]TJ
/F4 1 Tf
3.9454 0 TD
[(biocompatibility)-638.4(of)-635.7(decellularized)-644.9(SFTs)]TJ
-7.1536 -1.1116 TD
(\()Tj
/F5 1 Tf
.3341 0 TD
(n)Tj
/F6 1 Tf
.6624 0 TD
(=)Tj
/F4 1 Tf
.72 0 TD
[(6\))-439.9(was)-437.1(determined)-442.2(using)-437.4(contact)-443.4(and)-436.4(extract)-443.4(biocom-)]TJ
-1.7164 -1.1174 TD
[(patibility)-335.5(assays.)]TJ
/F2 1 Tf
8.9663 0 0 8.9663 320.8251 464.8251 Tm
[(Cell)-551.3(culture.)]TJ
/F4 1 Tf
9.843 0 0 9.843 380.3527 464.8251 Tm
[(Murine)-532.9(3T3)-534.4(broblastic)-538.6(cells)-537.8(\(European)]TJ
-7.0614 -1.1116 TD
[(Collection)-608.1(of)-601.2(Cell)-605.9(Cultures;)-611.3(ECCAC\))-605(were)-606.6(cultured)-610.4(in)]TJ
0 -1.1174 TD
[(Dulbeccos)-435.7(modied)-436.3(Eagles)-431.6(medium)-436.6(\(DMEM\))-436.7(plus)-430.6(10%)]TJ
0 -1.1116 TD
[(\(v/v\))-293.6(fetal)-296(calf)-294.8(serum)-301.4(\(FCS\),)-291.8(2)-168.1(m)0(M)]TJ
/F10 1 Tf
14.4799 0 TD
(l)Tj
/F4 1 Tf
.5011 0 TD
[(-glutamine,)-300.6(and)-292.4(100)-170.3(U/)]TJ
-14.981 -1.1116 TD
[(mL)-315.6(penicillin/streptomycin)-325.9(\(all)-316.8(from)-316.6(Lonza\).)-318.8(Baby)-317.6(hamster)]TJ
T*
[(kidney)-434.1(\(BHK;)-440.4(ECCAC\))-438(cells)-434.1(were)-439.5(cultured)-437.6(in)-434.3(Glasgows)]TJ
0 -1.1174 TD
[(minimum)-363(essential)-364.7(medium)-361.7(plus)-361.4(5%)-360.4(\(v/v\))-362.7(FCS,)-358.8(10%)-361.5(\(v/v\))]TJ
0 -1.1116 TD
[(tryptose)-384.4(phosphate)-386.6(broth)-385.7(\(29.5)-163.3(g/L;)-386(Sigma\),)-386.8(and)-384.6(100)-164.6(U/mL)]TJ
T*
[(penicillin/streptomycin.)-363.9(The)-357(cells)-359.2(were)-358.9(cultured)-362.7(at)-360.6(37)]TJ
/F6 1 Tf
22.0885 0 TD
()Tj
/F4 1 Tf
.3974 0 TD
.3592 Tc
[(Ci)359.2(n)]TJ
-22.4859 -1.1174 TD
0 Tc
[(5%)-331.6(\(v/v\))-333.9(CO)]TJ
7.3823 0 0 6.5614 363.3448 364.5353 Tm
(2)Tj
9.843 0 0 9.843 370.318 366.1794 Tm
[(in)-330.6(air)-338.6(in)-330.6(a)-336.3(humidied)-334.9(atmosphere.)]TJ
/F2 1 Tf
8.9663 0 0 8.9663 320.8251 342.8787 Tm
[(Contact)-471.6(biocompatibility)-480.7(assay.)]TJ
/F4 1 Tf
9.843 0 0 9.843 456.9448 342.8787 Tm
[(Samples)-471.8(from)-460.6(the)-465.4(toe,)]TJ
-14.8428 -1.1116 TD
[(middle,)-387.1(and)-384.6(ankle)-388.1(regions)-388.9(of)-382.3(decellularized)-391.5(SFTs)-391(were)-387.7(at-)]TJ
T*
[(tached)-252.1(to)-255.7(the)-252.3(center)-257.9(of)-255.6(wells)-255.3(of)-249.8(a)-255.7(six-well)-252.9(plate)-255.8(\(Nunc\))-256.2(using)]TJ
0 -1.1174 TD
[(Steri-Strips)-417.5(\(3)-163.4(M;)-412.2(Medisave\).)-420.8(Cyanoacrylate)-418.8(\(RS)-413.1(Systems\))]TJ
0 -1.1116 TD
[(was)-298.9(used)-293.4(as)-296.9(the)-298.3(positive)-298.2(control)-297.1(and)-298.2(Steri-Strip)-301.4(alone)-296(as)-296.9(the)]TJ
T*
[(negative)-294.8(control.)-289(BHK)-294.1(and)-292.4(3T3)-286.7(cells)-295.9(were)-295.6(seeded)-292.1(at)-291.5(5)]TJ
/F6 1 Tf
22.1979 0 TD
()Tj
/F4 1 Tf
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(10)Tj
7.3823 0 0 6.5614 546.2361 292.4787 Tm
(5)Tj
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18.2237 0 TD
()Tj
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[(C)-313.2(i)0(n)-313.3(5)0(%)-314.3(\(v/v\))]TJ
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(CO)Tj
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(2)Tj
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[(in)-278.8(air)-286.8(in)-278.8(a)-284.5(humidied)-283.1(atmosphere.)-289.8(The)-282.1(cell)-283.5(monolayers)]TJ
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[(were)-485.6(washed)-481.9(with)-483.6(PBS,)-485.5(xed)-483.5(with)-483.6(10%)-482.4(\(v/v\))-483.6(NBF,)-486.5(and)]TJ
0 -1.1116 TD
[(stained)-447.8(with)-449(Giemsa)-448.7(solution)-446.7(\(WVR)-446.3(International\))-451.3(before)]TJ
T*
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T*
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/F2 1 Tf
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[(Extract)-301.4(cytotoxicity)-301.8(assay.)]TJ
/F4 1 Tf
9.843 0 0 9.843 433.7574 188.0503 Tm
[(Samples)-310.5(from)-305.1(each)-307.4(region)-307.4(of)]TJ
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[(the)-649.7(decellularized)-656.5(SFTs)-650.2(\(toe,)-654.2(middle,)-652(and)-649.6(ankle\))-654.1(were)]TJ
T*
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T*
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()Tj
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[(C)-370.7(with)-368.4(agitation.)-367.5(BHK)-374.7(\(1)]TJ
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()Tj
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(5)Tj
9.843 0 0 9.843 534.8408 155.2252 Tm
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[(3T3)-407.7(\(2.5)]TJ
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3.7726 0 TD
()Tj
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(10)Tj
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(5)Tj
9.843 0 0 9.843 368.6172 144.2267 Tm
[(\))-410(cells)-411.1(in)-411.3(200)]TJ
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[(L)-411.2(o)0(f)-411.1(appropriate)-413.2(culture)-413.4(me-)]TJ
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T*
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/F6 1 Tf
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()Tj
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[(C)-370.7(i)0(n)-370.9(5)0(%)-371.9(\(v/v\))-374.2(CO)]TJ
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(2)Tj
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[(in)-370.9(air.)-370.9(The)]TJ
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[(L)-255.7(o)0(f)-255.6(appropriate)-257.7(culture)-263.7(medium)]TJ
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0 -1.1116 TD
[(in)-405.5(culture)-413.4(medium)-413.5(\(200)]TJ
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10.091 0 TD
(m)Tj
/F4 1 Tf
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[(L\))-406.5(and)-413.4(cells)-411.1(cultured)-408.8(in)-411.3(medium)]TJ
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[(alone)-301.7(served)-302.5(as)-302.7(negative)-306.3(controls)-303.8(for)-297(cytotoxicity.)-307.6(The)-299.4(cells)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 59.7543 749.7069 Tm
[(ACELLULAR)-338.8(PORCINE)-339.6(TENDON)]TJ
54.1178 0 TD
(3)Tj
ET
endstream
endobj
79 0 obj
<>stream
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BT
/F4 1 Tf
9.843 0 0 9.843 62.022 725.1588 Tm
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-4.7287 -1.1116 TD
[(turers)-400.4(instructions.)-398.1(The)-397.3(luminescence)-400.5(\(counts)-400.4(per)-396.1(second\))]TJ
T*
[(in)-250(each)-255.6(well)-248.6(was)-252.8(measured)-255.2(using)-247.3(a)-255.7(TopCount)-247.3(\(C9902)-255.3(Perkin)]TJ
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[(Elmer\))-338.6(luminescence)-337.1(plate)-336.4(reader.)]TJ
/F2 1 Tf
8.9663 0 0 8.9663 62.022 645.2786 Tm
[(In)-336.7(vivo)]TJ
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3.2689 0 TD
[(biocompatibility)-341.6(of)-330.4(decellularized)-336.1(SFT)-338(tissue)]TJ
/F4 1 Tf
9.843 0 0 9.843 72 628.8376 Tm
[(The)-587.4(host)-591.8(response)-592.4(to)-589.8(fresh)-588.1(\()]TJ
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(n)Tj
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[(3\))-589.7(and)-586.2(decellularized)]TJ
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(n)Tj
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(=)Tj
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[(6\))-295.9(SFTs)-298.8(was)-298.9(assessed)-305.3(in)-296.1(a)-301.8(mouse)-295.7(subcutaneous)-304.4(implant)]TJ
-1.7222 -1.1116 TD
[(model.)-391.6(Female)-391.4(C3H)-385.3(mice,)-391.7(612)-390.3(weeks)-388.7(old)-389.3(\(Harlan\),)-389.9(were)]TJ
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[(anesthetized,)-402.8(the)-390.5(mid-dorsum)-393.5(of)-393.8(the)-396.3(mice)-394(was)-396.8(shaved)-394.6(and)]TJ
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[(aseptically)-391.5(prepared)-392.4(and)-390.4(a)-388.2(small)-388.1(incision)-390.3(was)-391(made)-388.1(in)-388.2(the)]TJ
T*
[(dorsal)-341.7(skin.)-341.8(Tissue)-342.8(samples)-339.3(\(3)-169.2(mm)]TJ
7.3823 0 0 6.5614 201.0897 578.4376 Tm
(3)Tj
9.843 0 0 9.843 204.8314 574.0723 Tm
[(\))-335.1(taken)-342(from)-339.6(the)-344.4(central)]TJ
-14.5087 -1.1174 TD
[(region)-480.2(of)-480.2(the)-482.6(midportion)-479.1(of)-480.2(each)-480.2(tendon)-480.2(were)-479.9(implanted)]TJ
0 -1.1116 TD
[(subcutaneously)-427.7(along)-421.5(the)-419.3(mid-dorsal)-428.1(line)-420.5(distant)-423.8(from)-420.3(the)]TJ
T*
[(skin)-309.6(incision.)-313.1(Mice)-313(were)-312.8(fed)-315.5(food)-309.6(and)-309.7(water)]TJ
/F5 1 Tf
18.3792 0 TD
[(ad)-313.2(libitum)]TJ
/F4 1 Tf
4.4638 0 TD
(and)Tj
-22.843 -1.1174 TD
[(left)-247.7(for)-239.4(a)-244.2(period)-244(of)-244.1(3)-243(months.)-241.6(Mice)-243.9(were)-249.5(sacriced)-244.7(according)]TJ
0 -1.1116 TD
[(to)-417(schedule)-421.2(\(1\))-417.9(and)-413.4(the)-419.3(sample)-419(and)-419.2(surrounding)-420.9(skin)-413.3(har-)]TJ
T*
[(vested)-331.4(for)-331.5(analysis.)-337(All)-332.7(animal)-333(procedures)-331(were)-335.9(carried)-333.8(out)]TJ
T*
[(in)-394(accordance)-398.1(with)-391.4(UK)-395.6(Home)-393.6(Ofce)-394.4(regulations)-394.8(and)-390.4(with)]TJ
0 -1.1174 TD
[(appropriate)-338.3(Home)-336(Ofce)-336.8(licenses.)]TJ
1.0137 -1.1116 TD
[(The)-282.1(explanted)-284.4(samples)-281.7(were)-284(xed)-276.1(in)-284.6(NBF)-281.5(for)-279.7(48)-163.5(h)-283.3(before)]TJ
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[(being)-479.1(processed)-479.5(and)-476.8(embedded)-480.1(in)-474.6(parafn)-482.1(wax.)-475.3(Sections)]TJ
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[(were)-537.5(taken)-537.9(at)-533.4(6)]TJ
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(m)Tj
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[(m,)-535.7(retaining)-536.7(one)-534.4(section)-540(in)-532.2(every)-537.7(200)]TJ
-7.6777 -1.1116 TD
[(throughout)-402.9(the)-402(full)-407.8(thickness)-403.8(of)-399.6(the)-407.8(tissues.)-403.6(Sections)-402.7(were)]TJ
T*
[(stained)-338.4(with)-333.8(H&E)-334.9(and)-332.8(viewed)-338.2(as)-331.5(described)-341.6(above.)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 62.022 406.5448 Tm
[(Tensile)-334.8(testing)-333(of)-336.7(native)-335.7(and)-329.4(decellularized)-336.1(SFTs)]TJ
/F4 1 Tf
9.843 0 0 9.843 72 390.1039 Tm
[(Six)-423.7(native)-423.9(and)-424.9(acellular)-429.6(SFTs)-425.5(were)-428(snap-frozen)-430(by)-422.7(im-)]TJ
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[(mersion)-395.9(in)-388.2(dry)-394.9(ice)-391.7(and)-390.4(processed)-398.9(into)-390.6(dumbbell)-391.6(shapes)-394.4(of)]TJ
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[(consistent)-606.6(cross-sectional)-608.4(area)-601.1(of)-601.2(3.5)]TJ
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16.3 0 TD
()Tj
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[(5)-162.4(m)0(m)-603.7(and)-603.5(gauge)]TJ
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[(length)-451.5(of)-451.4(30)-169.2(mm.)-451.6(Specimens)-457.9(were)-451.1(then)-455(wrapped)-454.4(in)-451.6(PBS-)]TJ
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1.0137 -1.1116 TD
[(Specimens)-423.4(were)-422.3(mounted)-419.2(through)-422.5(bespoke)-424.5(cryogrips)-419.9(to)]TJ
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[(an)-314.4(Instron)-318.6(3366)-309.7(\(Instron,)-317.3(Bucks,)-316.2(United)-314.2(Kingdom\))-316.3(material)]TJ
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[(testing)-383.4(machine)-381.2(equipped)-385.7(with)-379.9(a)-376.6(1)-168.1(kN)-383.2(load)-380.1(cell.)-379(The)-380(grips)]TJ
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[(were)-531.7(manufactured)-531.8(with)-523.9(a)-532.2(void)-522.9(into)-528.8(which)-526(dry)-527.4(ice)-529.9(was)]TJ
T*
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[(temperatures.)-581.2(Once)-577.8(secured)-581.2(in)-578.3(the)-574.8(cryogrips,)-578.8(specimens)]TJ
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[(were)-312.8(tensioned)-317.6(to)-313.4(a)-307.5(preload)-315.4(of)-313.2(0.5)-166.9(N)-313(t)0(o)-307.6(ensure)-314(tautness)-316.3(was)]TJ
T*
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[(testing)-492.9(performed.)-494.9(This)-489.4(consisted)-490.2(of)-491.7(10)]TJ
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()Tj
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T*
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[(min.)-313.4(Load)-312(and)-309.7(extension)-317.6(data)-312.1(were)-312.8(recorded)-317.5(at)-308.8(a)-313.3(frequency)]TJ
25.504 52.972 TD
[(of)-416.9(10)-163.5(Hz,)-416.6(from)-408.7(which)-416.6(the)-407.8(ultimate)-416.1(tensile)-413.4(strength)-413.2(\(UTS\),)]TJ
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(e)Tj
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(FAIL)Tj
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[(\),)-344.3(and)-344.3(Youngs)-342.4(modulus)-342.9(of)-347.8(the)-338.7(collagen)]TJ
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[(phase)-335.9(\()]TJ
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(E)Tj
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(COLL)Tj
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[(\))-335.1(o)0(f)-330.5(the)-332.9(specimens)-336.9(were)-341.6(calculated.)]TJ
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[(Analysis)-340.7(of)-330.4(data)]TJ
/F4 1 Tf
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(t)Tj
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[(-t)-5.9(e)-7(s).1(t)]TJ
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12.3834 0 TD
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[(\(A)-4.3(NO)-4.3(VA)-4.3(\),)-334(follow)-4.3(ed)-331.7(by)-326(the)]TJ
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11.8304 0 TD
0 Tc
(T)Tj
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(Results)Tj
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[(The)-489.5(SFTs)-483.1(harvested)-491.4(from)-483.6(pigs)-488.2(\()]TJ
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[(1.1)-166.9(cm)-511.4(\()]TJ
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()Tj
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[(95%)-505.5(condence)-515.5(limits)-509.3([CL]\),)-512.2(a)-509.1(width)-507.7(of)]TJ
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(11.2)Tj
/F6 1 Tf
1.918 0 TD
()Tj
/F4 1 Tf
.72 0 TD
[(1.3)-166.9(mm,)-330.6(and)-332.8(a)-330.6(depth)-335.1(of)-330.5(3.9)]TJ
/F6 1 Tf
11.2602 0 TD
()Tj
/F4 1 Tf
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[(0.28)-168(mm.)-330.6(Images)-338.2(of)-330.5(the)]TJ
-14.6124 -1.1116 TD
[(porcine)-303.9(SFT)-297.9(before)-302.6(and)-298.2(after)-301.6(decellularization)-302.9(are)-299.3(shown)-299.9(in)]TJ
T*
[(Figure)-336(1.)]TJ
/F11 1 Tf
8.9663 0 0 8.9663 313.0582 494.192 Tm
[(Histological)-339.8(and)-335.7(immunohistochemical)-333.7(analysis)]TJ
0 -1.2203 TD
[(of)-336.7(native)-335.8(and)-329.4(decellularized)-336.1(SFT)]TJ
/F4 1 Tf
9.843 0 0 9.843 323.0361 466.8093 Tm
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-1.0137 -1.1174 TD
[(\(F)15.3(ig)20.7(.)-452.7(2)0(A)20(,)-446.9(C\))-446.7(re)19.7(ve)19.7(al)19.5(ed)-441.1(th)20.7(e)-451.5(p)0(r)20.9(e)0(s)14.1(e)0(n)19.7(c)0(e)-442.3(o)0(f)-439.9(c)0(e)18.5(l)0(l)20.6(s)-450.2(ne)19.7(at)13.8(ly)-440.1(al)19.5(ig)20.7(ne)19.7(d)]TJ
0 -1.1116 TD
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T*
[(ve)13.9(ss)21.2(el)19.5(s)-358(w)0(e)18.8(r)0(e)-348.9(a)0(p)13.9(p)0(a)19.7(r)0(e)19.7(n)0(t)-347.9(i)0(n)-347.9(t)0(h)20.7(e)-359.4(en)19.7(do)15.1(te)19.5(no)20.8(n)-358.2(r)0(e)19.7(g)0(i)20.7(o)0(n)20.8(s)-363.8(wh)20(er)19.7(e)-359.4(t)0(h)20.7(e)0(r)19.7(e)]TJ
T*
[(we)13(re)-285.6(la)19.5(rg)20.9(e)-301.8(a)0(g)19.7(g)0(r)20.9(e)0(g)19.7(a)0(t)19.5(i)0(o)20.7(n)0(s)-290(o)0(f)-290.1(c)0(e)18.5(l)0(l)20.6(s)0(.)-286.6(S)0(e)19.8(c)0(t)19.5(i)0(o)20.7(n)0(s)-290(s)0(t)20.9(a)0(i)19.5(n)0(e)13.9(d)-294.8(wi)19.8(th)-290.3(Si)20.9(ri)20.7(us)]TJ
0 -1.1174 TD
[(re)14(d/)20.7(Mi)19.8(ll)20.6(er)19.7(s)-237(v)0(i)20.7(e)0(w)18.8(e)0(d)-222.2(u)0(n)20.8(d)0(e)13.9(r)-231.4(po)20.8(la)19.5(ri)20.7(ze)18.5(d)-237.2(l)0(i)20.6(g)0(h)20.8(t)-237.4(\(F)21(ig)20.7(.)-233.8(2)0(E)19.6(\))-231.5(sh)15.3(ow)20(ed)-222.2(th)20.7(e)]TJ
0 -1.1116 TD
[(al)13.8(ig)20.7(ne)19.7(d)-323.6(c)0(o)19.7(l)0(l)20.6(a)0(g)19.7(e)0(n)-314.4()0(b)21(e)0(r)14(s)-323.4(an)19.7(d)-323.6(a)0(l)19.5(s)0(o)-313(d)0(e)19.7(m)0(o)19.6(n)0(s)21(t)0(r)20.7(a)0(t)13.8(e)0(d)-314.4(t)0(h)20.7(e)-324.8(cr)19.7(im)19.5(p)-323.6(f)0(o)20.9(r)0(-)]TJ
T*
[(ma)18.4(ti)14.8(on)-324.7(of)-324.7(th)20.7(e)-336.3()0(b)21(e)0(r)19.7(s)0(.)-326.9(U)0(n)20(d)0(e)19.7(r)-335.1(br)15.1(ig)20.7(ht)20.7(-)21(el)19.6(d)-335.2(i)0(l)20.6(l)0(u)14.9(m)0(i)19.5(n)0(a)19.7(t)0(i)20.6(o)0(n)20.8(,)-337.5(Mi)19.8(ll)20.6(er)19.7(s)]TJ
0 -1.1174 TD
[(st)15.1(ai)19.5(n)-346.7(d)0(i)20.7(d)-346.7(no)15.1(t)-346.8(r)0(e)19.7(v)0(e)19.7(a)0(l)-337.6(t)0(h)15(e)-347.8(pr)20.9(es)19.9(en)19.7(ce)-338.6(of)-342(an)19.7(y)-346.7(e)0(l)19.5(a)0(s)14.1(t)0(i)20.6(n)-346.7(in)-336.4(th)20.7(e)-353.6(t)0(i)20.6(s)0(s)21.2(u)0(e)]TJ
0 -1.1116 TD
[(se)14.1(ct)19.5(io)20.7(ns)-226.6(wh)20(en)-228(th)15(e)-238.4(s)0(t)20.9(a)0(i)19.5(n)-237.2(wa)18.8(s)-237(c)0(o)13.9(m)15(bi)14.9(ne)19.7(d)-237.2(w)0(i)19.8(t)0(h)-227(S)0(i)20.9(r)0(i)15(u)0(s)-226.6(r)0(e)19.7(d)0(.)-229.2(M)0(i)19.8(l)0(l)20.6(e)0(r)19.7(s)]TJ
T*
[(st)15.1(ai)19.5(ni)20.7(ng)-399.6(al)19.6(on)20.8(e)-411.2(s)0(h)21(o)0(w)20(e)0(d)-400.8(t)0(h)20.7(a)0(t)-400.9(t)0(h)20.7(e)0(r)14(e)-405.4(wa)18.8(s)-409.8(e)0(l)13.8(a)0(s)19.9(t)0(i)20.6(n)-410(pr)20.9(es)19.9(en)19.7(t)-410.2(i)0(n)-399.7(t)0(h)20.7(e)]TJ
0 -1.1174 TD
[(en)13.9(do)20.8(te)19.5(no)20.8(n)-444.6(r)0(e)19.7(g)0(i)20.7(o)0(n)15.1(s)-444.4(su)21(rr)20.9(ou)20.8(nd)20.8(in)15(g)-444.6(b)0(l)20.7(o)0(o)20.8(d)-444.6(ve)19.7(ss)15.5(el)19.5(s.)-430.5(Al)14.1(ci)19.6(an)-435.3(bl)20.7(ue)19.7(-)]TJ
0 -1.1116 TD
[(st)15.1(ai)19.5(ne)19.7(d)-427.3(t)0(i)20.6(s)0(s)15.5(u)0(e)-418.1(s)0(e)19.9(c)0(t)19.5(i)0(o)15(n)0(s)-416.7(\()0(F)21(i)0(g)15(.)-423.9(2G)14.2(\))-427.3(s)0(h)21(o)0(w)20(e)0(d)-423.8(G)0(A)19.1(G)0(s)-417.6(d)0(i)20.7(s)0(t)15.1(r)0(i)20.7(b)0(u)20.8(t)0(e)19.5(d)]TJ
T*
[(th)15(ro)20.9(ug)20.8(ho)20.8(ut)-394(th)20.7(e)-405.4(t)0(i)20.6(s)0(s)21.2(u)0(e)13.9(;)-398.6(ho)20.8(we)13(ve)19.7(r,)-390.4(th)20.7(e)-405.4(G)0(A)19.1(G)0(s)-394.5(w)0(e)18.8(r)0(e)-395(m)0(o)19.6(r)0(e)-395(c)0(o)19.7(n)0(-)]TJ
T*
[(ce)12.8(nt)20.7(ra)19.7(te)19.6(d)-323.6(a)0(r)19.7(o)0(u)20.8(n)0(d)-313.2(t)0(h)20.7(e)-324.8(va)19.7(sc)19.9(ul)14.9(ar)19.7(iz)19.5(ed)-314.4(en)19.7(do)20.8(te)19.6(no)20.8(n)-323.6(r)0(e)19.7(g)0(i)20.7(o)0(n)15.1(s)0(.)]TJ
1.0137 -1.1174 TD
[(Sections)-598.5(of)-589.7(decellularized)-598.9(tendons)-590.5(stained)-597.6(with)-593(H&E)]TJ
-1.0137 -1.1116 TD
[(\(Fig.)-359.1(2B\))-355.6(and)-350(DAPI)-357.3(\(Fig.)-359.1(2D\))-349.7(revea)-9(led)-350.2(the)-355.9(absence)-357.8(of)-359.3(any)]TJ
T*
[(cell)-410.2(nuclei)-400.9(or)-405.3(double-stranded)-405.9(DNA.)-406.8(The)-403.1(histoarchitecture)]TJ
0 -1.1174 TD
[(of)-295.9(th)20.7(e)-307.5(m)0(a)18.4(t)0(r)20.7(i)0(x)-296.1(a)0(p)19.7(p)0(e)19.7(a)0(r)19.7(e)0(d)-297.1(s)0(i)20.9(m)0(i)19.5(l)0(a)19.6(r)-306.3(to)-290.3(na)19.7(ti)20.6(ve)-297.1(ti)20.6(ss)15.5(ue)-291.3(wi)19.8(th)-296.1(ev)19.7(id)20.7(en)19.7(ce)]TJ
0 -1.1116 TD
[(of)-382.3(so)21(me)-379(lo)20.7(ss)-381.9(of)-376.5(co)19.7(ll)14.8(ag)19.7(en)-377.7(b)21(er)-377.7(cr)14(im)19.5(p.)-373.1(Se)14.1(ct)19.5(io)20.7(ns)-376.4(st)20.9(ai)13.8(ne)19.7(d)-387(w)0(i)19.8(t)0(h)]TJ
T*
[(Si)15.1(ri)20.7(us)-290(re)19.7(d/)20.7(Mi)19.8(ll)20.6(er)19.7(s)-300.4(\()0(F)21(i)0(g)20.7(.)-297.2(2F)21(\))-300.6(d)0(e)19.7(m)0(o)19.6(n)0(s)21(t)0(r)20.7(a)0(t)19.6(e)0(d)-291.3(t)0(h)20.7(e)-301.8(re)19.7(te)19.5(nt)20.7(io)20.7(n)-300.6(o)0(f)-284.4(t)0(h)20.7(e)]TJ
0 -1.1174 TD
[(te)13.8(nd)20.8(on)-272.9(co)19.7(ll)20.6(ag)19.7(en)-279.8(b)21(er)-274(al)13.8(ig)20.7(nm)19.6(en)19.7(t.)-275.4(Al)19.8(ci)19.5(an)-274.1(bl)14.9(ue)19.7(-s)21.1(ta)19.6(in)20.7(ed)-279.8(se)19.9(ct)19.5(io)20.7(ns)]TJ
0 -1.1116 TD
-.0163 Tc
[(\(Fig)5.6(.)-353.8(2)-.1(H\))-351.4(sh)5.6(owed)-351.5(a)-358.4(d)5.6(ecr)5.7(eas)5.8(e)-358.4(i)5.5(n)-351.5(t)-.3(he)-352.6(GAG)-352.3(c)-1.3(ont)5.5(ent)-351.6(t)5.5(hrou)5.6(gho)5.6(ut)]TJ
T*
-.0158 Tc
[(the)-582.5(d)6.1(ece)5(llu)6.1(l).3(ari)6(zed)-575.6(tis)6.3(sue)-582.5(c)5(omp)6.1(a)-.8(re)5(d)-581.3(w)-.5(i)6(t).3(h)-581.3(t).3(h)6.1(e)-582.5(na)5(tive)-576.7(t).2(is)6.3(sue.)]TJ
0 -1.1174 TD
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[(Scan)5.8(ned)-236.1(i)-.1(mages)-241.6(o)5.8(f)-241.8(t)0(h)5.8(e)-243(d)5.8(ecellul)5.7(a)-1.1(ri)5.7(zed)-236.1(po)5.8(rcin)5.8(e)-243(S)6(FT)-237.3(stai)5.7(ned)-236.1(w)-.8(i)5.7(t)0(h)]TJ
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BT
/F8 1 Tf
9.845 0 0 9.845 62.022 127.6724 Tm
0 Tc
[(FIG.)-336.7(1.)]TJ
/F4 1 Tf
4.1174 0 TD
[(Im)19.9(ag)19.9(e)-324.6(o)0(f)-312.9(t)0(h)20.9(e)-324.6(po)21.1(rc)19.9(in)20.9(e)-324.6(s)0(u)21.2(-)]TJ
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[(pe)19.9(r)21.2(ex)14.1(or)-312.9(te)19.7(nd)21.1(on)-312.9(\(S)21.2(FT)20.1(\))-323.5(b)0(e)19.9(f)0(o)21(r)0(e)-314.1(\()]TJ
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[(to)20.9(p)]TJ
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1.2496 0 TD
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[(an)19.9(d)-306.2(a)0(f)19.9(t)0(e)19.7(r)-306.2(\()]TJ
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[(bo)21.1(tt)20.7(om)]TJ
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[(\))-326.3(d)-20.1(ec)-20.1(el)-20.1(lu)-20.1(la)-20.1(ri)-20.1(z)-6.2(a)-20.1(ti)-20.1(on)-20.1(.)]TJ
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