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[(inc)12.7(uba)12.8(ted)-304.1(wit)12.8(h)-312.1(a)-313.3(1:5)13.8(00)-301.7(dil)13.7(uti)13.7(on)-307.5(of)-307.4(the)-304.1(pri)13.9(mar)11.6(y)-312.1(a)0(n)13.9(tib)13.7(odi)13.8(es)-308.4(to)]TJ
T*
[(col)12.7(lag)12.7(en)-354.7(typ)13.8(e)-359.4(I)-363.9(\(C)14.1(ol)-359.4(I\),)-345.4(ost)14(eoc)11.7(alc)11.5(in)-353.7(\(OC)12.1(N;)-354.5(Abc)11.9(am,)-347.9(Inc)12.8(.\))-355.8(or)]TJ
0 -1.1174 TD
[(ost)14(eop)12.8(ont)13.8(in)-434.3(\(OP)13.3(N;)-429.4(cus)13(tom)-425.1(mad)11.6(e)-434.2(fro)14(m)-440.1(Abc)17.7(am,)-428.5(Inc)12.8(.\))-430.7(ove)12.8(r-)]TJ
0 -1.1116 TD
[(nig)13.9(ht)-480.4(at)-481.5(4)]TJ
/F7 1 Tf
4.2334 0 TD
()Tj
/F3 1 Tf
.3917 0 TD
[(C.)-477.9(Aft)13(erw)17.7(ard)12.8(,)-487.2(the)-471.1(cel)11.5(ls)-480.2(wer)12(e)-480.3(inc)12.7(uba)12.8(ted)-476.9(wit)18.6(h)-484.9(a)]TJ
-4.625 -1.1116 TD
[(1:2)13.9(50)-370.8(dil)19.5(uti)13.7(on)-370.8(of)-365(the)-361.7(sec)11.9(ond)14(ary)-361.5(ant)12.7(ibo)13.9(dy)-365.1(\(FI)14.2(TC-)11.8(con)18.6(jug)13.9(ate)11.5(d)]TJ
0 -1.1174 TD
[(Af )13.3(nip)13.9(ure)-350(goa)12.8(t)-358.3(ant)12.7(i-m)12.7(ou)15.1(se)-354.5(IgG)13.1(;)-358.3(Abc)11.9(am,)-347.8(Inc)12.9(.\).)-347.8(Fin)14(all)12.6(y,)-350.1(cel)11.5(l)]TJ
0 -1.1116 TD
[(nuc)12.8(lei)-310(wer)12(e)-313.3(sta)12.9(ine)12.7(d)-317.9(wit)12.8(h)-312.1(DAP)12.4(I.)-309.8(The)-310.9(sam)17.5(ple)12.7(s)-317.7(wer)12(e)-313.3(obs)14.2(erv)12.8(ed)]TJ
T*
[(usi)14(ng)-428.4(CL)12.8(SM.)-421.1(The)-420.4(sta)12.9(ini)13.7(ng)-422.7(int)13.7(ens)13(iti)13.6(es)-423.6(of)-422.6(Col)-419.2(I,)-425(OCN)11.2(,)-423.9(and)]TJ
T*
[(OPN)-459.9(wer)12(e)-468.8(ana)11.6(lyz)12.7(ed)-464.1(by)-463(Im)13.9(age)-460.6(Pro)-458.1(Plu)14(s)-467.4(Sof)14.2(twa)11.8(re)-464.1(\()-75.9(Med)11.9(ia)]TJ
0 -1.1174 TD
[(Cyb)12.9(ern)12.9(eti)12.6(cs\))13(.)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 313.0582 446.3432 Tm
[(Real-time)-339.8(quantitative)-334.1(PCR)-332.3(analysis)]TJ
/F3 1 Tf
9.843 0 0 9.843 323.0361 429.9023 Tm
[(Af)20(te)19.6(r)-335.1(7)0(,)-327.1(1)0(4)20.8(,)-337.5(an)25.4(d)-340.9(2)0(1)-324.7(d)0(a)19.7(y)0(s)-324.6(o)0(f)-324.7(c)0(u)19.7(l)0(t)20.6(u)0(r)20.9(e)0(,)-322.5(c)0(e)18.5(l)0(l)20.6(s)-335(we)18.8(re)-325.9(ly)20.7(se)19.9(d)-335.2(w)0(i)19.8(t)0(h)]TJ
-1.0137 -1.1174 TD
[(Tr)19.7(iz)19.6(ol)-388.2(Re)18.6(ag)19.7(en)25.4(t)-404.4(\()0(A)20(m)15(bi)20.7(on)20.8(\))-398.5(a)0(n)19.7(d)-398.5(to)20.7(ta)19.5(l)-398.6(R)0(N)18.9(A)-399.4(wa)18.8(s)-398.3(e)0(x)19.7(t)0(r)20.7(a)15(ct)19.5(ed)-389.3(ac)18.5(-)]TJ
0 -1.1116 TD
[(co)19.7(rd)20.9(in)20.7(g)-254.5(t)0(o)-244.2(t)0(h)20.7(e)-249.9(ma)18.4(nu)20.8(fa)19.7(ct)19.5(ur)26.6(er)19.7(s)-243.9(in)20.7(st)20.9(ru)20.9(ct)19.6(io)20.7(ns)21(.)-251.1(F)0(i)20.9(r)0(s)21.1(t)0(-)20.7(s)0(t)20.9(r)0(a)19.7(n)0(d)-244.1(c)15(DN)19.1(A)]TJ
T*
[(wa)18.8(s)-450.2(s)0(y)21(n)0(t)20.7(h)0(e)25.4(s)0(i)20.9(z)0(e)18.5(d)-450.4(wi)25.6(th)-440.1(th)20.7(e)-445.8(R)0(e)18.6(v)0(e)19.7(r)0(T)19.7(r)16.2(a)-451.5(A)0(c)18.8(e)-445.8(qP)21(CR)-436.2(RT)-442.2(M)15.3(a)0(s)19.9(t)0(e)19.6(r)]TJ
0 -1.1174 TD
[(Mi)19.8(x)-404.3(\()0(T)19.7(o)0(y)20.8(o)0(b)26.6(o)-404.3(Co)19.8(.,)-392.8(Lt)19.5(d.)24.3(\).)-390.4(Re)18.6(al)19.5(-t)20.7(im)25.2(e)-405.4(q)0(u)20.8(a)0(n)19.7(t)0(i)20.6(t)0(a)19.6(t)16(iv)20.7(e)-405.4(p)0(o)20.8(l)0(y)20.7(m)15(er)19.7(as)19.9(e)]TJ
0 -1.1116 TD
[(ch)19.7(ai)19.5(n)-346.7(r)0(e)25.5(a)0(c)18.5(t)0(i)20.6(o)0(n)-330.5(\()0(R)19.8(T)0(-)19.7(q)0(P)21(C)0(R)18.8(\))-340.9(us)21(in)20.7(g)-346.7(F)16.4(as)19.9(t-)20.7(St)20.9(ar)19.7(t)-346.8(U)15.3(ni)20.7(ve)19.7(rs)21.1(al)-331.8(SY)20.1(BY)]TJ
T*
[(Gr)20(ee)18.5(n)-254.5(M)0(a)18.8(s)0(t)20.9(e)0(r)-245.2(\()0(R)19.8(o)0(x)20.8(;)-260.4(R)15.2(o)0(c)19.7(h)0(e)-245.3(L)0(t)19.5(d)0(.)18.5(\))-254.5(wa)18.8(s)-254.3(p)0(e)19.7(r)0(f)20.9(o)0(r)20.9(m)0(e)18.4(d)-254.5(us)21(in)20.7(g)-254.5(a)-255.7(75)20.8(00)]TJ
T*
[(Re)18.6(al)19.5(-T)19.7(im)19.5(e)-411.2(P)0(C)20(R)-416.8(Sy)21(st)20.9(em)-402(\(L)19.7(if)20.7(e)-417(T)0(e)18.5(c)15(hn)20.8(ol)20.7(og)20.8(ie)19.5(s,)-401.8(In)20.9(c.)23.1(\).)-407.7(Re)18.6(la)25.3(ti)20.6(ve)]TJ
0 -1.1174 TD
[(qu)20.8(an)19.7(ti)20.6( c)19.8(at)19.5(io)26.5(n)-335.2(w)0(a)24.6(s)-335(ca)24.3(rr)20.9(ie)19.6(d)-329.4(o)0(u)20.8(t)-329.5(us)21(in)20.7(g)-329.4(t)0(h)20.7(e)]TJ
/F10 1 Tf
7.3823 0 0 6.5614 472.7054 346.5637 Tm
(DD)Tj
/F3 1 Tf
9.843 0 0 9.843 482.3432 342.1983 Tm
[(Ct)-325.9(m)15(e)0(t)19.5(h)0(o)20.8(d)0(.)-321.3(T)15(hr)20.9(ee)]TJ
-17.1985 -1.1116 TD
[(se)19.9(pa)19.7(ra)19.7(te)-251.2(ex)19.7(pe)19.7(ri)20.7(m)15(e)0(n)19.7(t)0(s)-255.6(w)15.3(er)19.7(e)-267.2(p)16.2(er)19.7(fo)20.9(rm)19.6(ed)19.7(.)-262.6(T)15(he)-256.8(se)25.6(qu)20.8(en)19.7(ce)18.5(s)-260.1(o)0(f)-255.6(g)16.2(en)19.7(e)]TJ
T*
[(pr)20.9(im)19.5(er)19.7(s)-323.4(u)0(s)21(e)0(d)-308.6(f)0(o)20.9(r)-323.6(RT)18.6(-q)20.9(PC)25.7(R)-324.7(a)0(r)19.7(e)-324.8(li)26.3(st)20.9(ed)-314.4(in)-313.4(Ta)24.2(bl)20.7(e)-324.8(1)0(.)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 313.0582 298.3748 Tm
[(Western)-336(blot)-338.1(analysis)]TJ
/F3 1 Tf
9.843 0 0 9.843 323.0361 281.9338 Tm
[(On)-348.7(the)-350.2(third)-345.6(day)-350(after)-347.7(seeding,)-347.5(total)-352.6(cellular)-346.7(protein)-348.9(was)]TJ
-1.0137 -1.1116 TD
[(extracted)-256.7(as)-256.6(previously)-254.1(described.)]TJ
7.3823 0 0 6.5614 446.9101 275.3574 Tm
(14)Tj
9.843 0 0 9.843 456.7747 270.9921 Tm
[(Brie y,)-254.1(cells)-255.6(were)-255.2(lysed)]TJ
-14.6009 -1.1174 TD
[(with)-443.3(ice-cold)-438.7(RIPA)-438.1(\(Thermo)-443.2(Fisher)-439.5(Scienti c\))-440.8(containing)]TJ
0 -1.1116 TD
[(phosph)-6.4(atase)-262.4(inhibitor)-258.1(cocktail)-266.1(\(Applygen\).)-264.2(For)-262.3(each)-261.3(sample,)]TJ
T*
(25)Tj
/F10 1 Tf
1.1692 0 TD
(m)Tj
/F3 1 Tf
.5529 0 TD
[(g)-456.1(o)0(f)-457.2(protein)-452.6(was)-460.2(resolved)-460.3(on)-451.5(sodium)-457(dodecyl)-459.4(sulfate)]TJ
ET
62.022 195.931 490.167 .22676 re
f
62.022 68.485 490.167 .28348 re
f
62.022 176.939 490.167 .28348 re
f
BT
/F6 1 Tf
9.845 0 0 9.845 226.6582 201.5999 Tm
(Table)Tj
/F3 1 Tf
3.0002 0 TD
(1.)Tj
/F6 1 Tf
1.2496 0 TD
[(Primers)-336(Used)-332.7(for)-332.2(RT-qPCR)]TJ
/F8 1 Tf
-20.9726 -1.86 TD
[(Gene)-22678.5(Sequence)-335.1(\(5)]TJ
/F7 1 Tf
29.7486 0 TD
()Tj
/F8 1 Tf
.2476 0 TD
(-3)Tj
/F7 1 Tf
.835 0 TD
()Tj
/F8 1 Tf
.2476 0 TD
[(\))-13988.5(No.)]TJ
-31.0789 -1.9118 TD
(ALP)Tj
/F3 1 Tf
10.3827 0 TD
[(Forward)-7005.4(TCAGCTTCCTCCTCTTTGCG)-8551.5(XM_002716044.1)]TJ
0 -1.0077 TD
[(Reverse)-7163.4(CGGGTCCACAGTTCAGGAAA)]TJ
/F8 1 Tf
-10.3827 -1.0135 TD
(BMP2)Tj
/F3 1 Tf
10.3827 0 TD
[(Forward)-7005.4(ACTGCCAGAAACAAGTGGGA)-7719.5(NM_001082650.1)]TJ
0 -1.0135 TD
[(Reverse)-7163.4(TGATGGAAACCGCTGTCGTC)]TJ
/F8 1 Tf
-10.3827 -1.0077 TD
(OPN)Tj
/F3 1 Tf
10.3827 0 TD
[(Forward)-7005.4(TGGCTAAACCCTGACCCAT)-8801.5(NM_001082194.1)]TJ
0 -1.0135 TD
[(Reverse)-7163.4(TATCCACGTGGTCATCGTCC)]TJ
/F8 1 Tf
-10.3827 -1.0135 TD
(BSP)Tj
/F3 1 Tf
10.3827 0 TD
[(Forward)-7005.4(ACGCTTTCTTTCACAACACCTG)-7011.5(XM_002717020.1)]TJ
0 -1.0135 TD
[(Reverse)-7163.4(GCTGGTGCCATTGATGCTCTG)]TJ
/F8 1 Tf
-10.3827 -1.0077 TD
(GAPDH)Tj
/F3 1 Tf
10.3827 0 TD
[(Forward)-7005.4(GTATGATTCCACCCACGGCA)-8083.5(NM_001082253.1)]TJ
0 -1.0135 TD
[(Reverse)-7163.4(CCAGCATCACCCCACTTGAT)]TJ
8.7511 0 0 8.7511 70.0157 56.6929 Tm
[(RT-qPCR,)-333.2(real-time)-333.8(quantitative)-333.1(polymerase)-335(chain)-332.1(reaction.)]TJ
/F4 1 Tf
8.9663 0 0 8.9663 62.022 749.6502 Tm
(1812)Tj
49.2112 0 TD
[(WEI)-331.7(ET)-338.7(AL.)]TJ
ET
endstream
endobj
84 0 obj
<>stream
/GS1 gs
BT
/F3 1 Tf
9.843 0 0 9.843 59.7543 725.1588 Tm
0 0 0 1 k
0 Tc
0 Tw
[(polyacrylamide)-724.4(gels)-713.9(and)-712.9(electrotransferre)-14.2(d)-709.5(onto)-718.7(PVDF)]TJ
0 -1.1174 TD
[(membranes)-384.3(\()-87.5(Millipore)-381.1(Corporation\).)-381.9(The)-380(membranes)-384.3(were)]TJ
0 -1.1116 TD
[(blotted)-372.1(using)-368.3(5%)-371.9(Carnation)-370.6(nonfat)-370.7(dry)-366.1(milk)-368.8(and)-373.1(incubated)]TJ
T*
[(with)-299.3(anti-p-ERK)-297.7(antibody,)-290.1(anti-ERK)-301.3(antibody,)-295.9(anti-RUNX-)]TJ
T*
[(2)-352.4(antibody,)-347.7(and)-355.8(anti-GAPDH)-349.8(antibody)-355.8(\(custom)-349.8(made)-353.5(from)]TJ
0 -1.1174 TD
[(Abcam,)-353.2(Inc.\))-354.5(overnight)-351.1(at)-349.1(4)]TJ
/F7 1 Tf
11.4849 0 TD
()Tj
/F3 1 Tf
.3974 0 TD
[(C,)-351.1(respectively.)-355.5(Then,)-350(the)-350.2(pri-)]TJ
-11.8823 -1.1116 TD
[(mary)-283.2(antibodies)-284.2(were)-278.3(detected)-284.4(using)-281.9(horseradish)-284.8(peroxidase)]TJ
T*
[(\(HRP\)-conjugated)-374.4(secondary)-376(antibodies.)-368.3(To)-366.3(verify)-370.7(the)-373.2(role)]TJ
0 -1.1174 TD
[(of)-376.5(the)-379(ERK1/2)-378.5(signaling)-379.9(pathway)-379.7(in)-376.7(the)-373.2(osteogenic)-383.2(differ-)]TJ
0 -1.1116 TD
[(entiation)-485.1(of)-486(RaADSCs)-484.6(mediated)-486.1(by)-480.3(CACB,)-484.1(the)-488.4(selective)]TJ
T*
[(inhibitor)-286.9(U0126)-282.9(\(Cell)-278.6(Signaling)-287.8(Technology,)-283.1(Inc.\))-279.6(was)-287.4(used)]TJ
0 -1.1174 TD
[(at)-389.4(a)-388.2( nal)-391.4(concentration)-393.7(of)-388.1(20)]TJ
/F10 1 Tf
12.3661 0 TD
(m)Tj
/F3 1 Tf
.5529 0 TD
[(M.)-391.3(The)-385.8(protein)-389.2(bands)-392.4(were)]TJ
-12.919 -1.1116 TD
[(visualized)-815.3(using)-806(the)-811(ECL)-807.3(chemiluminesce)-12.4(nce)-806.2(reagent)]TJ
T*
[(\(Thermo)-523.9(Fisher)-520.1(Scienti c\))-527.2(and)-517.1(were)-520.2(quanti ed)-525(using)-518(an)]TJ
T*
[(image)-337.5(analysis)-339.3(system)-331.3(\(Quantity)-340.6(One)-333.6(software;)-335.7(Bio-Rad\).)]TJ
/F1 1 Tf
8.9663 0 0 8.9663 59.7543 548.3337 Tm
[(In)-336.7(vivo)]TJ
/F9 1 Tf
3.2753 0 TD
[(osteogenesis)-333.5(of)-330.4(CACB)-336.6(combined)-336.4(with)-330.2(ADSCs)]TJ
/F1 1 Tf
-2.1624 -1.8336 TD
[(Preparation)-320(of)-311.4(implantation)-317(composites.)]TJ
/F3 1 Tf
9.843 0 0 9.843 236.5795 531.8928 Tm
[(About)-317.5(1.5)]TJ
/F7 1 Tf
4.2334 0 TD
()Tj
/F3 1 Tf
.72 0 TD
(10)Tj
7.3823 0 0 6.5614 295.1999 536.2581 Tm
(6)Tj
9.843 0 0 9.843 59.7543 520.8943 Tm
[(cells)-359.2(of)-359.3(the)-355.9(third)-357.1(passage)-362.2(from)-356.9(each)-359.2(rabbit)-353.6(were)-364.7(seeded)-355.5(on)]TJ
0 -1.1116 TD
[(0.25)-168(mg)-464.2(CACB,)-466.8(which)-462.7(had)-459.5(been)-467.5(ground)-461.6(into)-465.4(particles)-462.8(of)]TJ
/F10 1 Tf
T*
(*)Tj
/F3 1 Tf
1.0022 0 TD
[(1.02.0)-167.9(mm)-361.8(in)-365.2(diameter,)-362.8(and)-361.6(then)-368.6(cultured)-362.7(for)-360.3(7)-364(days)-362.5(in)]TJ
-1.0022 -1.1174 TD
[(preparation)-338.3(for)-337.3(the)]TJ
/F8 1 Tf
7.9714 0 TD
[(in)-330.6(vivo)]TJ
/F3 1 Tf
3.1218 0 TD
(study.)Tj
/F1 1 Tf
8.9663 0 0 8.9663 69.7323 466.129 Tm
[(Surgical)-449.4(procedures.)]TJ
/F3 1 Tf
9.843 0 0 9.843 162.085 466.129 Tm
[(Bone)-444.3(defects)-443.1(in)-440.1(each)-439.9(side)-443.2(of)-434.1(the)]TJ
-10.3963 -1.1174 TD
[(mandibles)-445.5(of)-439.9(12)-439.9(rabbits)-440.9(were)-445.3(created)-444.4(as)-440.9(described)-439.5(in)-440.1(our)]TJ
0 -1.1116 TD
[(previous)-557.1(report)]TJ
7.3823 0 0 6.5614 122.7401 448.5542 Tm
(16)Tj
9.843 0 0 9.843 135.6094 444.1889 Tm
[(and)-551.6(were)-554.7(randomly)-558.5(divided)-550.5(into)-557.6(three)]TJ
-7.7065 -1.1116 TD
[(groups:)-243.8(\(1\))-239.4(CACB)-238.8(group:)-237.1(eight)-243.1(defects)-241.5(for)-239.4(implantation)-242.1(with)]TJ
T*
[(0.25)-168(mg)-475.7(CACB)-480.7(granules;)-484.6(\(2\))-475.5(CACB/ADSCs)-484.1(group:)-479(eight)]TJ
0 -1.1174 TD
[(defects)-414.3(for)-412.2(implantation)-409.1(with)-414.5(CACB)-411.6(seeded)-413.1(with)-408.7(ADSCs;)]TJ
0 -1.1116 TD
[(and)-269.4(\(3\))-262.4(Untreated)-273.7(group:)-265.9(eight)-266.1(defects)-264.5(were)-272.5(left)-265(untreated)-267.1(as)]TJ
T*
[(controls.)-526.1(Rabbits)-528.3(from)-523.9(each)-526.3(group)-523.8(were)-525.9(sacri ced)-532.7(using)]TJ
0 -1.1174 TD
[(lethal)-424(intravenous)-423.4(administrations)-429.3(of)-416.9(sodium)-422.5(pentobarbital)]TJ
0 -1.1116 TD
[(at)-337.6(4)-329.4(and)-338.5(12)-330.5(weeks)-336.8(postimplantation.)]TJ
/F10 1 Tf
8.9663 0 0 8.9663 69.7323 343.559 Tm
(m)Tj
/F1 1 Tf
.5501 0 TD
[(-CT)-345.7(analysis.)]TJ
/F3 1 Tf
9.843 0 0 9.843 136.4031 343.559 Tm
[(At)-343(12)-342(weeks)-348.3(postimplantation,)-347.8(the)-344.4(man-)]TJ
-7.7871 -1.1116 TD
[(dibles)-399.5(w)15.3(e)0(r)19.7(e)-382.4(h)16.2(a)0(r)25.5(v)0(e)25.4(s)0(t)20.9(e)15(d)-381.2(i)0(n)26.5(t)0(a)19.6(c)15(t)-381.4(a)0(n)25.4(d)-381.2( x)21(e)15(d)-381.2(in)-370.9(4)16.2(%)-382.3(pa)25.4(ra)19.7(f)16.2(o)0(r)26.6(m)0(a)24.2(l)0(d)26.5(e)0(-)]TJ
T*
-.0254 Tc
[(h)-25.4(y)-4.6(d)-9.2(e)-25.4(,)-232.7(a)-25.4(n)-5.7(d)-245.4(t)-9.3(h)-25.4(e)-236.1(s)-25.4(pe)-25.4(ci)-25.4(me)-25.4(ns)-250.9(w)-10.1(e)-25.4(re)-252.3(e)-10.4(x)-25.4(am)-25.4(in)-25.4(ed)-245.4(u)-25.4(s)-4.4(i)-9.3(n)-25.4(g)]TJ
/F10 1 Tf
18.2352 0 TD
0 Tc
(m)Tj
/F3 1 Tf
.5414 0 TD
-.0185 Tc
[(-C)-18.5(T)-230.3(s)-18.5(c)7.1(a)-18.5(n)6.9(n)-18.5(i)8(n)-18.5(g.)]TJ
-18.7766 -1.1174 TD
0 Tc
[(Fi)20.9(l)16.1(e)0(s)-452.4(w)0(e)24.6(r)0(e)-458.3(r)16.2(ec)24.3(on)26.6(st)20.9(r)16.2(u)0(c)25.4(t)0(e)25.3(d)-467.6(us)26.8(in)26.5(g)-467.6(a)-468.8(m)15(o)0(d)26.6(i)0( )26.6(e)0(d)-452.6(F)0(e)19.8(l)16.1(dk)26.6(am)24.2(p)-467.6(a)15(lg)26.5(o-)]TJ
0 -1.1116 TD
[(ri)20.7(t)16.1(h)0(m)25.4(,)-337.5(w)15.3(h)0(i)26.5(c)0(h)-325.9(w)15.3(as)-325.7(c)15(r)0(e)25.5(a)0(t)25.3(e)0(d)-325.9(u)16.2(si)20.9(n)16.2(g)]TJ
/F10 1 Tf
12.9017 0 TD
(m)Tj
/F3 1 Tf
.5414 0 TD
[(-)16.2(t)0(o)26.5(m)0(o)25.4(g)0(r)20.9(a)15(ph)26.6(ic)-326(a)15(n)0(a)25.4(l)0(y)26.5(s)0(i)20.9(s)-335(s)16.4(o)0(f)26.6(t)0(-)]TJ
-13.4432 -1.1116 TD
[(wa)24.6(re)-550.5(\()16.2(T)0(o)25.4(m)0(o)-550.6(N)15.3(T;)-544.9(Sk)21(y)16.2(s)0(c)25.6(a)0(n)25.4(\))0(.)-545.9(A)0(f)20(t)16.1(er)-544.7(th)20.7(r)16.2(e)0(e)24.3(-)0(d)26.6(i)0(m)25.2(e)0(n)19.7(s)16.4(io)26.5(na)19.7(l)-554.2(\()0(3)26.6(D)0(\))]TJ
0 -1.1174 TD
[(vi)20.7(s)16.4(u)0(a)25.4(l)0(i)26.3(z)0(a)18.5(t)16.1(io)26.5(n,)-482.6(bo)26.6(ne)-487.2(m)15(o)0(r)20.9(p)16.2(ho)26.6(me)24.2(tr)26.5(ic)-487.3(a)15(n)0(a)25.4(l)0(y)20.7(s)16.4(es)25.6(,)-498.8(i)0(n)26.5(c)0(l)25.3(u)0(d)26.6(i)0(n)26.5(g)-496.4(bo)20.8(n)16.2(e)]TJ
0 -1.1116 TD
[(co)25.4(ve)19.7(r)16.2(a)0(g)25.4(e)0(,)-253.4(b)0(o)20.8(n)16.2(e)-267.2(d)0(e)25.4(n)0(s)26.8(i)0(t)20.6(y)16.2(,)-268.4(a)15(nd)-255.6(b)16.2(o)0(n)20.8(e)-261.4(vo)20.8(l)16.1(u)0(m)25.4(e)-267.2(in)-255.8(t)16.1(h)0(e)-256.8(b)16.2(on)20.8(e)-267.2(d)16.2(ef)25.5(ec)24.3(ts)20.9(,)]TJ
T*
[(we)24.6(re)-308.6(ca)24.3(rr)20.9(i)16.1(e)0(d)-308.6(o)0(u)26.6(t)-323.8(o)16.2(n)-323.6(t)16.1(h)0(e)-308.6(r)0(e)25.5(g)0(i)20.7(o)16.2(n)-317.9(o)0(f)-307.4(i)0(n)20.7(t)16.1(er)25.5(es)19.9(t)16.1(.)]TJ
/F1 1 Tf
8.9663 0 0 8.9663 69.7323 231.9873 Tm
-.0152 Tc
[(Hi)-15.2(s)13.6(t).2(ol)-15.2(o)15(g)-.6(i)-15.2(c)13.6(al)-246.1(and)-247.2(imm)-15.2(u)10.4(nohistochem)-15.2(i)11.5(c)4.3(al)-246.1(exami)-15.2(n)15(ati)-15.2(o)8.7(ns.)]TJ
/F3 1 Tf
9.843 0 0 9.843 280.3464 231.9873 Tm
0 Tc
(Four)Tj
-22.411 -1.1116 TD
[(and)-234.9(12)-238.3(wee)10.8(ks)-238.2(pos)14.2(tsu)14(rge)12.8(ry,)-230.2(all)-235.1(rab)12.8(bit)-234(man)11.6(dib)13.8(les)-234.8(fro)14(m)-238.5(all)-235.1(thr)13.9(ee)]TJ
0 -1.1174 TD
[(gro)14(ups)-406.3(we)13(re)-412.3(sub)14.2(je)13.8(cte)11.5(d)-410(t)0(o)-411.3(tis)13.9(sue)-401.7(pro)14(ces)11.9(sin)14(g)-410(and)-401.9(sec)11.8(tio)13.7(nin)13.9(g.)]TJ
0 -1.1116 TD
[(The)-403.1(sam)17.5(ple)12.7(s)-409.8(wer)12(e)-405.4( xe)13(d)-410(i)0(n)-399.7(10%)-401.8(for)14(ma)12.7(lin)-401(for)-394.9(24)-157.7(h)-410(and)-396.1(de-)]TJ
T*
[(min)12.6(era)11.7(liz)12.6(ed)-452.6(in)-451.6(15%)-442.1(for)14(mic)-449.3(aci)11.5(d)-456.1(for)-446.7(6)-456.1(wee)10.8(ks.)-443.2(The)-449.2(tis)13.9(sue)13(s)]TJ
T*
[(wer)12(e)-480.3(the)12.7(n)-473.4(emb)11.6(edd)12.8(ed)-469.9(in)-468.9(par)12.8(af )13(n)-479.1(and)-465.2(sec)11.9(tio)13.7(ned)-465.2(to)-474.6(a)-474.6(4)]TJ
/F10 1 Tf
22.9639 0 TD
(m)Tj
/F3 1 Tf
.5472 0 TD
(m)Tj
-23.5111 -1.1174 TD
[(thi)13.7(ckn)12.8(ess)13.2(.)-262.6(H)0(e)13(mat)11.5(oxy)14(lin)-251.2(and)-246.4(eos)13(in)-255.8(sta)12.9(ini)13.7(ng)-255.6(was)-252.8(per)12.8(for)14(me)12.7(d)-260.3(o)0(n)]TJ
0 -1.1116 TD
[(tis)13.9(sue)-488.1(sec)11.8(tio)13.7(ns)-491.6(acc)10.5(ord)14(ing)-487.2(to)-491.9(the)-488.4(man)11.6(ufa)12.8(ctu)12.7(re)14(rs)-492.6(pro)14(toc)18.4(ols)14(.)]TJ
T*
[(The)-420.4(ima)11.5(ges)-413.2(wer)12(e)-428.5(cap)11.7(tur)13.9(ed)-418.1(usi)14(ng)-422.7(lig)13.7(ht)-422.8(mic)11.5(ros)14.2(co)13.9(py)-422.7(\(CX)12.1(21;)]TJ
0 -1.1174 TD
[(Oly)13(mpu)12.7(s\).)-575.6(Th)13.9(e)-584(amo)11.6(unt)-573.6(of)-578.1(new)-575.5(bon)14(e)-584(and)-574.7(bo)15.1(ne)-579.3(mar)11.6(row)]TJ
0 -1.1116 TD
[(cav)11.7(iti)13.6(es)-383.3(wa)13(s)-386.8(ana)11.7(lyz)12.7(ed)-377.7(by)-382.3(Im)13.9(age)-380(Pro)-371.7(Plu)14(s)-386.8(Sof)14.2(twa)17.6(re)-383.5(\()-75.9(Med)11.9(ia)]TJ
T*
[(Cyb)12.9(ern)12.9(eti)12.6(cs\))13(.)]TJ
1.0137 -1.1174 TD
[(The)-316.7(num)18.5(ber)-315.5(of)-319(blo)13.8(od)-313.2(ves)13(sel)12.9(s)-323.4(a)0(t)-320.3(the)-309.9(imp)12.6(lan)12.7(tat)12.6(ion)-308.7(sit)13.9(e)-324.8(was)]TJ
-1.0137 -1.1116 TD
[(det)12.7(erm)11.6(ine)12.7(d)-531(b)0(y)-532.1(imm)11.4(un)15.1(ohi)13.8(sto)14(che)11.7(mic)11.5(al)-527.6(ana)11.7(lys)14(is.)-523.9(De-)12(par)12.8(af )13(-)]TJ
T*
[(niz)12.7(ed)-510.2(sec)11.9(tio)13.7(ns)-514.6(we)13(re)-515.9(was)12.1(he)13.9(d,)-511.4(and)-505.6(non)14(spe)13(ci )12.8(c)-514.9(end)12.8(oge)12.8(nou)14(s)]TJ
0 -1.1174 TD
[(per)12.8(oxi)13.8(das)13(e)-232.6(act)11.5(ivi)13.7(ty)-227(was)-224(que)18.6(nch)12.8(ed)-228(by)-226.8(imm)11.4(ers)13(ing)-222.3(in)-227(3%)-222.1(H)]TJ
7.3823 0 0 6.5614 281.7637 65.9905 Tm
(2)Tj
9.843 0 0 9.843 285.4487 67.5779 Tm
(O)Tj
7.3823 0 0 6.5614 292.4786 65.9905 Tm
(2)Tj
9.843 0 0 9.843 296.107 67.5779 Tm
(/)Tj
-24.0122 -1.1116 TD
[(met)11.5(han)12.8(ol)-296.1(for)-285.5(15)-157.7(min)12.6(,)-297.2(the)12.7(n)-300.6(inc)12.7(uba)12.8(te)13.8(d)-300.6(wit)12.8(h)-300.6(ant)12.7(i-C)12.9(D3)14.2(1pr)14(ima)11.5(ry)]TJ
25.5097 67.9184 TD
[(ant)12.7(ibo)13.8(dy)-249.9(\(1:)13.9(200)14(;)-254.7(Abc)11.9(am\))-247.6(for)-250.9(2)-162.4(h)0(.)-252.2(HRP)12.2(-co)12.9(nju)13.8(ga)13.9(ted)-252.3(sec)11.8(ond)14(ar)13.9(y)]TJ
0 -1.1174 TD
[(ant)12.7(ibo)13.8(dy)-336.3(wa)13(s)-340.7(app)12.8(lie)12.6(d)-340.9(t)0(o)-336.4(the)-332.9(sli)13.9(des)-332.6(for)-331.5(1)-162.4(h)-340.9(at)-337.6(roo)14(m)-342.1(tem)11.5(pe)13.9(ra-)]TJ
0 -1.1116 TD
[(tur)13.9(e.)-224.6(Fin)14(all)12.6(y,)-223.4(the)-217.7(dia)12.7(min)12.6(obe)18.6(nzi)12.7(din)13.9(e)-232.6(\(DA)12.3(B;)-222.2(Bey)11.8(oti)13.7(me)12.7(\))-231.5(kit)-222.4(wa)13(s)]TJ
T*
[(use)13(d)-329.4(t)0(o)-324.9(d)0(e)13.9(vel)12.7(op)-324.7(the)-321.4(col)12.7(or,)-316.6(fol)13.9(low)13(ed)-325.9(by)-324.7(cou)12.8(nte)12.7(rst)14.1(ain)12.7(ing)-320.2(wi)14.1(th)]TJ
T*
[(hem)17.4(ato)12.7(xyl)13.8(in.)-247.7(A)-255.4(vas)13(cul)12.7(ar)-251(sec)11.8(tio)13.7(n)-254.5(was)-247.1(de )13(ned)-246.4(as)-250.8(a)-255.7(ves)13(sel)-252.1(wit)18.6(h)]TJ
0 -1.1174 TD
[(a)-307.5(rec)11.7(ogn)14(iza)11.5(ble)-298.3(lum)12.6(en.)-300.5(Mic)11.8(ro)-307.4(hem)17.3(orr)14(hag)12.8(es)-308.4(wer)12(e)-307.5(exc)11.6(lud)13.9(ed)-308.6(in)]TJ
0 -1.1116 TD
[(the)-373.2(ves)13(sel)-373(cou)12.8(nt.)-374.4(Thr)12.8(ee)-378.9(dif)13.9(fer)12.9(ent)-373.2(ind)13.8(ivi)13.7(dua)12.8(ls)-382.3(pe)13.9(rfo)14(rme)11.6(d)-387(the)]TJ
T*
[(eva)11.7(lu)15(ati)12.6(on)-324.7(wit)12.8(h)-329.4(sin)14(gle)-315.6(bli)13.7(nd.)]TJ
/F9 1 Tf
8.9663 0 0 8.9663 310.8472 626.5132 Tm
[(Statistical)-336.1(analysis)]TJ
/F3 1 Tf
9.843 0 0 9.843 320.8251 610.0723 Tm
[(A)15.3(l)0(l)-342.3(q)16.2(ua)19.7(nt)20.7(it)20.6(at)19.5(iv)20.7(e)-347.8(d)0(a)19.7(t)0(a)-337.5(a)0(r)19.7(e)-347.8(ex)25.4(pr)20.9(es)19.9(se)19.9(d)-346.7(a)0(s)-343(m)15(ea)18.5(n)]TJ
/F7 1 Tf
17.809 0 TD
()Tj
/F3 1 Tf
.7084 0 TD
[(st)20.9(an)19.7(d)16.2(a)0(r)19.7(d)-352.4(e)15(r)0(-)]TJ
-19.5312 -1.1116 TD
[(ro)20.9(r.)-517.1(St)20.9(at)19.5(is)20.9(ti)20.6(c)15(a)0(l)-521.9(a)0(n)19.7(a)0(l)19.5(y)0(s)21(e)15(s)-530.8(w)0(e)18.8(r)0(e)-521.7(p)0(e)19.7(r)0(f)20.9(o)16.2(rm)19.6(ed)-521.7(us)21(in)20.7(g)-525.2(S)0(P)21.2(S)0(S)-520.2(1)0(3)20.8(.)13.8(0)]TJ
0 -1.1174 TD
[(so)21(ft)20.7(wa)24.6(re)19.7(.)-354.8(T)15(he)-343.2(Sh)21(ap)19.7(ir)20.7(o)20.8(W)13.9(i)0(l)20.6(k)-352.4(te)19.5(st)-342(wa)24.6(s)-352.2(a)0(p)19.7(p)0(l)20.7(i)0(e)19.5(d)-352.4(to)-336.4(ch)19.7(ec)18.5(k)-352.4(w)0(h)20(e)15(-)]TJ
0 -1.1116 TD
[(th)20.7(er)-274(th)20.7(e)-290.2(v)0(a)25.4(l)0(u)20.7(e)0(s)-279.6(w)0(e)18.8(r)0(e)-274(n)0(o)20.8(r)0(m)19.6(a)0(l)19.5(l)0(y)-273(d)0(i)20.7(s)0(t)20.9(r)0(i)20.7(b)0(u)20.8(t)0(e)19.5(d)0(.)-275.2(O)0(n)20(e)15(-w)20(ay)-279.8(AN)19.1(OV)24.9(A)]TJ
T*
[(an)19.7(d)]TJ
/F8 1 Tf
1.7279 0 TD
[(po)20.8(st)-290.1(ho)20.8(c)]TJ
/F3 1 Tf
3.6632 0 TD
[(mu)19.6(lt)26.3(ip)20.7(le)-291.5(co)19.7(mp)19.6(ar)19.7(is)20.9(on)-290.2(te)19.5(st)20.9(s)-300.4(w)0(e)18.8(r)0(e)-291.3(u)0(s)21(e)0(d)-291.4(t)0(o)-290.3(a)0(n)19.7(a)0(l)19.5(y)0(z)19.7(e)]TJ
-5.3911 -1.1116 TD
[(th)20.7(e)-561(d)0(i)20.7(f)0(f)20.9(e)0(r)19.7(e)0(n)19.7(c)0(e)24.3(s)-565.4(of)-555.1(th)20.7(e)-560.9(r)0(e)19.7(s)0(u)21(l)0(t)20.6(s)-565.3(of)-555.1(th)26.5(e)-566.7(i)0(m)19.5(m)0(u)25.4(n)0(o)20.8( )0(u)21(o)0(r)20.9(e)0(s)19.9(c)0(e)18.5(n)0(c)25.4(e)]TJ
0 -1.1174 TD
[(st)20.9(ai)19.6(n)16.2(i)0(n)20.7(g)0(,)-425(t)0(h)20.7(e)-434.2(RT)18.6(-q)20.9(PC)20(R)15.2(,)-435.4(an)19.7(d)-433.1(t)0(h)20.7(e)-434.2(we)18.8(st)20.9(er)19.7(n)-433.1(b)0(l)20.7(o)0(t)20.7(t)0(i)20.6(n)0(g)20.8(,)-435.4(an)25.4(d)-438.8(t)0(h)20.7(e)15(y)]TJ
0 -1.1116 TD
[(we)18.8(re)-343.2(al)19.6(so)-341.8(us)21(ed)-343.2(to)-342.1(an)19.7(al)19.5(yz)19.7(e)-353.6(t)0(h)20.7(e)-353.6(di)20.7(ff)20.9(er)25.5(en)19.7(ce)18.5(s)-352.2(i)0(n)-347.9(b)0(o)26.6(n)0(e)-348.9(c)15(ov)20.8(er)19.7(ag)19.7(e,)]TJ
T*
[(bo)20.8(ne)-216.5(de)19.7(ns)21(it)20.6(y)-231.5(a)0(n)19.7(d)-231.5(bo)26.6(ne)-222.2(vo)20.8(lu)20.7(me)18.4(,)-228.1(t)0(h)20.7(e)-232.6(am)24.2(ou)20.8(nt)-221.2(of)-221(ne)19.7(w)-232.3(b)0(o)26.6(n)0(e)19.7(,)-233.8(bo)20.8(ne)]TJ
0 -1.1174 TD
[(ma)24.2(rr)20.9(ow)-509.9(c)15(a)0(v)19.7(i)0(t)20.6(i)0(e)19.5(s)0(,)-505.4(a)0(n)19.7(d)-513.7(bl)20.7(oo)20.8(d)-513.7(v)0(e)19.7(s)0(s)21.2(e)0(l)-504.6(s)0(e)19.9(c)0(t)19.5(i)0(o)26.5(n)0(s)-508.9(i)0(n)-503.4(t)0(h)20.7(e)-514.9(an)19.7(im)19.5(a)15(l)]TJ
0 -1.1116 TD
[(ex)19.7(p)16.2(e)0(r)19.7(i)0(m)19.5(e)0(n)19.7(t)0(s)20.9(.)-539.1(Wh)18.6(en)-533.3(th)20.7(e)-543.7(v)0(a)19.7(r)0(i)20.7(a)0(n)19.7(c)15(es)-538.8(a)15(m)0(o)19.6(n)0(g)-532.1(d)0(i)20.7(f)0(f)20.9(e)0(r)19.7(e)0(n)19.7(t)-542.6(gr)20.9(ou)26.6(ps)]TJ
T*
[(we)18.8(re)-331.6(no)20.8(t)-341(e)0(q)19.7(u)0(a)19.7(l)0(,)-333(D)0(u)20(n)0(n)26.6(e)0(t)19.5(t)-346.8(T3)-331.7(te)19.6(st)-330.4(wa)18.8(s)-346.5(a)15(pp)20.8(li)20.6(ed)-337.4(t)16.1(o)-346.7(ev)19.7(al)19.5(u)16.2(a)0(t)19.6(e)-347.8(th)20.7(e)]TJ
0 -1.1174 TD
[(di)20.7(ff)20.9(e)15(r)0(e)19.7(n)0(c)19.7(e)0(s)19.9(.)-642.8(Di)19.8(ff)20.9(er)19.7(en)19.7(c)15(e)0(s)-636.7(b)0(e)19.7(t)0(w)19.8(e)15(en)-636.9(gr)20.9(ou)20.8(ps)-629.8(wh)20(er)19.7(e)-641.6(*)]TJ
/F8 1 Tf
21.2072 0 TD
(p)Tj
/F7 1 Tf
.6566 0 TD
(<)Tj
/F3 1 Tf
.7084 0 TD
[(0.)18.5(0)16.2(5)]TJ
-22.5723 -1.1116 TD
[(we)18.8(re)-233.7(co)19.7(ns)21(id)26.5(er)19.7(ed)-239.5(st)26.7(at)19.5(is)20.9(ti)20.6(ca)18.5(ll)20.6(y)-243(s)0(i)20.9(g)0(n)20.8(i)0( )20.9(c)0(a)18.5(n)16.2(t)-248.9(a)0(n)19.7(d)-243(wh)20(er)19.7(e)-244.2(*)0(*)]TJ
/F8 1 Tf
20.9711 0 TD
(p)Tj
/F7 1 Tf
.6566 0 TD
(<)Tj
/F3 1 Tf
.7085 0 TD
[(0.)24.3(01)20.8(,)]TJ
-22.3361 -1.1116 TD
[(th)20.7(e)-319(d)0(i)20.7(f)0(f)20.9(e)0(r)19.7(e)0(n)19.7(c)0(e)18.5(s)-317.7(we)18.8(re)-314.4(co)19.7(ns)21(i)16.1(d)0(e)19.7(r)0(e)19.7(d)-323.6(hi)20.7(gh)20.8(l)16(y)-323.6(si)20.9(gn)20.8(i )20.9(ca)18.5(nt)20.7(.)]TJ
/F4 1 Tf
8.9663 0 0 8.9663 310.8472 429.2786 Tm
(Results)Tj
/F9 1 Tf
0 -1.8336 TD
[(Characterization)-335.2(of)-330.4(the)-334.8(CACB)]TJ
/F3 1 Tf
9.843 0 0 9.843 320.8251 396.3968 Tm
-.0217 Tc
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T*
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[(RaADSCs)-461.6(in)-463.1(CACB)-463.4(increased)-463.7(with)-460.5(prolonged)-460.4(incubation)]TJ
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[(Immuno uorescence)-285.8(staining)-286.6(of)-278.6(Col)-286.7(I,)-281(OCN,)-286(and)-280.9(OPN)-287.1(in)]TJ
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