Abst ract
Submitted: February 24, 2017 0RGL¿FDWLRQ$SULO Accepted: June 02, 2017
Physical propert ies and biological
effect s of m ineral t rioxide aggregat e
m ixed wit h m et hylcellulose and
calcium chloride
Obj ect ives: Met hylcellulose ( MC) is a chem ical com pound derived from cellulose. MTA m ixed wit h MC reduces set t ing t im e and increases plast icit y.
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CaCl2 as a set t ing t im e accelerat or on t he physical and biological propert ies of MTA. Mat erial and Met hods: Test m at erials were divided int o 3 groups; Group 1( cont rol) : dist illed wat er; Group 2: 1% MC/ CaCl2; Group 3: 2% MC/
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gene expression of bone sialoprot ein ( BSP) was det ect ed by RT- PCR and real-t im e PCR. The expression of alkaline phosphareal-t ase ( ALP) and m ineralizareal-t ion behavior were evaluat ed using an ALP st aining and an alizarin red st aining. Result s: Com pressive st rengt h, pH, and cell viabilit y of MTA m ixed wit h MC/
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in MTA wit h MC/ CaCl2 com pared t o t he cont rol ( p< .05) . This st udy revealed higher expression of ALP and m ineralizat ion in cells exposed t o MTA m ixed wit h wat er and MTA m ixed wit h MC/ CaCl2 com pared t o t he cont rol ( p< .05) .
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physical and biological effect of MTA. I t suggest s t hat t hese cem ent s m ay
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Ke yw or ds: Flowabilit y. Met hylcellulose. MTA. Ost eogenic different iat ion. Bin-Na LEE1
Soo-Ji CHUN1
Hoon-Sang CHANG1
Yun-Chan HWANG1
In-Nam HWANG1
Won-Mann OH1
http://dx.doi.org/10.1590/1678-7757-2017-0050
1Chonnam National University, School of Dentistry, Dental Science Research Institute, Department of Conservative Dentistry, Gwangju, Korea.
Corresponding address: Won-Mann Oh, DDS, MSD, PhD Department of Conservative Dentistry -School of Dentistry - Chonnam National University.
I nt roduct ion
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and as a root perforat ion repair in 1993, t he use of
m ineral t rioxide aggregat e ( MTA) has been expanded
t o m any clinical applicat ions including pulp capping,
pulpot om y, and apical barrier for t eet h wit h necrot ic
p u lp an d op en ap ices5 , 1 2 , 1 3 , 1 5 , 2 6 , 3 4. Th is p op u lar it y
or igin at es f r om t h e su per ior biocom pat ibilit y an d
sea l i n g a b i l i t y o f MTA t o o t h er r o o t - en d f i l l i n g
m at erials1,24,34. However, it has several disadvant ages,
i n cl u d i n g l o n g se t t i n g t i m e a n d p o o r h a n d l i n g
charact erist ics because of it s granular consist ency23,33.
There are som e st udies aim ed at im proving t he
set t ing t im e and handling charact erist ics of MTA by
using cert ain addit ives8,16,23. The addit ion of am orphous
calciu m lact at e g lu con at e b ased liq u id im p r ov es
t he set t ing t im e as well as clinical m anageabilit y16,
how ever, it decr eases t he com pr essive st r engt h of
MTA25. I n anot her st udy, t he addit ion of pr opylene
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calciu m ion r elease du r in g t h e in it ial post - m ix in g
periods, despit e increasing it s set t ing t im e11.
Met hy lcellu lose ( MC) is a ch em ical com pou n d
derived from cellulose. I t is used as an addit ive t o
im p r ov e t h e p er f or m an ce of Por t lan d cem en t in
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several food and cosm et ics, and also as a const ipat ion
t reat m ent . Cellulose, for exam ple, is not digest ible,
neit her t ox ic, nor aller genic. An adm ix of 1% MC
and 2% calcium chloride ( CaCl2) int o MTA reduced set t ing t im e and im pr oved it s m oldabilit y sim ilar ly
t o a r ein f or ced zin c ox id e- eu g en ol cem en t w it h
an ap p r ox i m at el y eq u al co m p r essi v e st r en g t h6.
Moreover, t he addit ion of 10% CaCl2 t o MTA did not alt er it s biologic propert ies regarding t he form at ion
of a m ineralized bar r ier aft er pulpot om y8 and MTA
m ix ed w it h calciu m com pou n ds sh ow ed a sim ilar
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in an in vivo st udy29. However, t here are few st udies on
t he physical propert ies and biological effect of MC on
MTA. I n DGGLWLRQLQWHUDFWLRQEHWZHHQURRWHQG¿OOLQJ
m at erials and periradicular t issues is very im port ant
at t he st art and developm ent of healing. Thus, t he aim
of t his st udy was t o invest igat eWKHLQÀXHQFHRI0&
and CaCl2 ( MC/ CaCl2) on t he physical and biological propert ies of MTA.
Mat erial and m et hods
Preparat ion of t est m at erials
To prepare t he addit ive solut ions, CaCl2 ( Sigm a, St
Louis, MO, USA) wit h 2% of sam ple weight was added
t o dist illed wat er and m ixed int o a solut ion. This CaCl2
solut ion was placed on a hot plat e whose t em perat ure
was raised t o 80° C. MC ( Sigm a) was added t o t he
warm solut ion t o obt ain t he concent rat ions t o be t est ed
( 1% or 2% ) and was st irred unt il all t he m at erials were
m ixed. Then, t he solut ion was st ored at 0° C for 20
m inut es t o m ake it t hicker and st irred wit h a m agnet ic
st irrer for 30 m inut es t o creat e a hom ogenous gel.
Sim ilar t o t he m anufact urer ’s recom m endat ions for
MTA ( ProRoot MTA; Dent sply Tulsa Dent al, Tulsa, OK,
USA) , t hese solut ions were used in a 3: 1 powder–liquid
rat io and assigned t o t he following t est groups. Group
1( cont rol) : dist illed wat er; Group 2: 1% MC/ CaCl2;
Group 3: 2% MC/ CaCl2.
Com pressive st rengt h
Th e co m p r essi v e st r en g t h s o f t h e m a t er i a l s
under t est were det erm ined according t o t he m et hod
recom m ended by t he I SO 9917- 120. To prepare t he
specim en s, poly et hy len e m olds w it h 4 m m in n er
diam et er and 6 m m height were used. The specim ens
w er e r em ov ed f r om t h e m old s an d a sear ch f or
any air- voids or chipped edges was conduct ed. All
defect ive specim ens were discarded. Six sam ples were
select ed t o undergo m at erial t est ing, and prepared for
each m at erial t est at each t im e int erval ( n= 6) . The
specim ens were im m ersed in dist illed wat er for 1 day,
3 d, and 7 d and m aint ained at 37° C. The com pressive
st r en gt h s w er e t h en m easu r ed u sin g a u n iv er sal
t est ing m achine ( RB Model 302 ML, R&B I nc., Korea)
at a crosshead speed of 1.0 m m per m . The m axim um
load needed t o fract ure each specim en was m easured,
and t he com pressive st rengt h ( C) was calculat ed in
m egapascals according t o t he form ula:
& 3Ⱥ'2
where P is t he m axim um force applied in newt ons,
an d D is t h e m ean diam et er of t h e specim en in
m illim et ers.
pH
The pH was m easured by using a pH m et er ( ORI ON
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elect rode for solid specim ens ( I nLab Surface; Met t ler
t est , t he apparat us was calibrat ed at t he st andard pH
solut ions of 4.0, 7.0 and 11.0. Six specim ens for each
group were prepared by loading t he m at erial under t est
int o acrylic m olds wit h 10.0 m m inner diam et er and
5.0 m m height ( n= 6) . The readings were t aken at t he
end of t he m ixing and aft er 30 m , 1 h, 3 h, 24 h, 48 h
and 72 h. Bet ween each m easurem ent , t he elect rode
was washed wit h dist illed wat er and blot dried.
Flowabilit y
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as recom m ended by I SO 687619. I m m ediat ely aft er
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glass slab ( 40x40x5 m m , 20 g weight ) . The second
glass slab was posit ioned on t he m at erial followed by
t he addit ion of 100 g of weight aft er 180 s from t he
st art of m ixing. The weight was rem oved aft er 10 m
of m ix ing. The m ax im um and m inim um diam et er s
of t he circle form ed by t he m at erial were m easured.
The m ean diam et er is considered as a m easurem ent
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m axim um and m inim um diam et ers is wit hin 1 m m .
Six t est s were carried out for each m at erial ( n= 6) .
Cell cult ure
Mouse per iodont al ligam ent ( m PDL) cells w er e
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Eagle’s m edium ( DMEM; I nv it r ogen, Car lsbad, CA,
USA) , supplem ent ed w it h 10% fet al bov ine ser um
( FBS; I nvit rogen) and 1% ant ibiot ics ( 100 U/ m L of
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CO2. Cells used in t hese experim ent s were bet ween
passages 4 and 7.
Mat erial ext ract s
MTA sam ples were m ixed wit h dist illed wat er, 1%
MC/ CaCl2 and 2% MC/ CaCl2 in a 3: 1 powder–liquid
rat io according t o t he m anufact urers’ inst ruct ions. The
m at erials were placed int o a cylindrical polyet hylene
t ube ( 5 m m in diam et er and 3 m m in height ) . To at t ain
com plet e set t ing, t he sam ples were kept for 6 h at
37° C and 95% relat ive hum idit y. Aft er set t ing, discs
were exposed t o ult raviolet light for 1 h for st erilizat ion
and t ransferred int o 24- well cult ure plat es. Discs were
incubat ed in 1.5 m L DMEM cont aining 2% or 10% FBS
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wit h 5% CO2 for 24 h. The ext ract s were collect ed and
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St edim Biot ech, Goet t ingen, Germ any) .
Cell viabilit y assay
The m PDL cells were seeded int o 96- well cult ure
p l at es at a d en si t y o f 1 x 1 04 cel l s p er w el l an d
incubat ed in a growt h m edium ( DMEM cont aining 10%
FBS and 1% ant ibiot ics) for 24 h for adhesion. The
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of experim ent al groups and incubat ed for 24 h. The
m PDL cells and t he grow t h m edium w ere used for
cont rol. To com pare dose- response relat ionships, t he
m at erial ext ract s were gradually dilut ed in t he growt h
m edium t o obt ain 5 concent rat ions ( 1, 1/ 2, 1/ 4, 1/ 10,
and 1/ 50) . Ten m icrolit er of t he WST reagent (
EZ-CyTox; Daeil Lab Service Co., Seoul, Korea) was added
t o each well and incubat ed at 37° C for 3 h. Opt ical
densit ies were m easured at 420 nm using a m ult iwell
spect r ophot om et er ( VERSAm ax Mult iplat e Reader ;
Molecular Devices, Sunnyvale, CA, USA) . The relat ive
cell viabilit y was calculat ed for each t est m at erial as
m ean percent age of t he cont rol.
Rev er se- t ranscr ipt ion PCR and quant it at iv e
real- t im e RCR
The m PDL cells were seeded in 6- well plat es at
a densit y of 2x105 cells per well and incubat ed in a
growt h m edium for 24 h. The growt h m edium was
replaced wit h a m edium cont aining 1/ 4 concent rat ion
of m at erial ext ract s. The unt reat ed cells were used for
cont rol. Aft er 1, 3, and 5 d in cult ure, t he t ot al RNA
was ext ract ed using a TRI zol reagent ( I nvit rogen) .
The purit y and quant it y of t ot al RNA were det erm ined
using a spect rophot om et er ( Nanodrop 100; Therm o
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DNA was synt hesized using t he Maxim e RT PreMix
Kit ( iNt RON Biot echnology, Seongnam , Korea) . Each
react ion consist ed of an init ial denat urat ion at 95° C for
1 m followed by a t hree- st ep cycling: denat urat ion at
95° C for 30 s, annealing at 55° C for 30 s, and ext ension
at 72° C for 30 s. Aft er 25–30 cycles, t he react ions
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prim er sequences were t he following: bone sialoprot ein
( BSP) , forward 5’-ACACTTACCGAGCTTATGAG- 3’ and
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f o r w a r d 5 ’ - TGGATGGCTACGTACATGGCTGGG- 3 ’
an d r ev er se 5 ’-TTCTTTGCAGCTCCTTCGTTGCCG- 3 ’.
Each react ion was analyzed wit h 1.5% agarose gel
elect rophoresis and visualized wit h et hidium brom ide
st aining. Quant it at ive real- t im e PCR was perform ed
by using t he Quant iTect SYBR Green PCR kit ( Qiagen,
from t riplicat e m easurem ent s were used t o det erm ine
t he relat ive level of expression of t he t arget gene wit h
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relat ive change in gene expression was analyzed by
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Alkaline phosphat ase ( ALP) st aining
The m PDL cells were seeded in 24- well plat es at a
densit y of 2x104 cells per well wit h growt h m edium .
Aft er 24 h, t he gr ow t h m edium was changed t o a
m ediu m con t ain in g 1 / 4 con cen t r at ion of m at er ial
ext ract s and cult ured for 5 d. The cult ured cells were
washed w it h phosphat e buffer ed saline ( PBS) and
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t he cells were washed 3 t im es wit h deionized wat er
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USA) was applied per well under dark condit ions for
15 m . The st ains w er e ex t ract ed w it h 10% ( w / v )
cept ylpyridinium chloride ( Sigm a) in 10 m M sodium
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by r ecor din g t h e absor ban ce at 5 6 2 n m u sin g a
spect r oph ot om et er ( VERSAm ax m u lt iplat e r eader,
Molecular Devices) .
Alizarin red st aining
Alizarin red st aining was used t o assess m ineralized
deposit form at ion. The m PDL cells were cult ured wit h
growt h m edium cont aining 1/ 4 dilut ions of m at erial
ext ract s for 14 d. The cells were rinsed wit h PBS, and
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were st ained wit h 0.5% alizarin red ( pH= 4.2) for 60
m at r oom t em perat ur e w it h gent le agit at ion. The
cells were t hen rinsed wit h deionized wat er 5 t im es,
rinsed wit h PBS for 15 m and air- dried. 10% ( w/ v)
cept ylpyridinium chloride ( Sigm a) in 10 m M sodium
phosphat e w as applied t o t he st ained nodule and
t he absor bance of t he super nat ant s w as r ecor ded
at 540 nm using a spect rophot om et er ( VERSAm ax
m ult iplat e reader, Molecular Devices) for quant it at ive
assessm ent .
St at ist ical analysis
Each experim ent , cont aining t riplicat e independent
sam ples, was repeat ed at least t wice, and qualit at ively
ident ical result s were obt ained. One- way analysis of
variance ( ANOVA) was usd follow ed by Tukey post
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ALP act ivit y and alizarin red st aining. For com pressive
st rengt h, pH and cell viabilit y, t he t wo- way ANOVA
and t he Duncan t est s w er e used. Dat a fr om r
eal-t im e PCR was analyzed wieal-t h eal-t he Kruskal- Wallis and
Mann- Whit ney U t est s. The result s were considered
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Result s
Com pressive st rengt h
The com pr essiv e st r engt hs of MTA m ix ed w it h
1% and 2% MC/ CaCl2ZHUHQRWVLJQL¿FDQWO\GLIIHUHQW
com pared t o MTA m ixed wit h dist illed wat er ( Table 1) .
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bet ween t he t im e point s.
pH
The m ean pH of all sam ples is in Table 2. The pH
values of MTA m ixed wit h 1% and 2% MC/ CaCl2 were
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w it h dist illed wat er. Up t o 3 h, all gr oups show ed
higher pH values com pared t o t he ot her periods, wit h
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decreasing, t he pH values st ill rem ained high.
Group 1 day 3 days 7 days
MTA + DW 18.88±4.53 22.71±3.25 20.96±4.97
07$0& 19.19±3.06 23.95±7.43 25.40±7.03
07$0& 24.19±5.46 22.13±4.46 22.24±4.59 Table 1- Means and standard deviations of the compressive strengtth of test materials at various time intervals
Group Immediately 30m 1h 3h 24h 48h 72h
MTA + DW 12.83±0.02Aa 12.90±0.04Aa 12.74±0.27Aa 11.82±1.03Aa 10.93±0.94Ab 9.82±0.33Ac 9.61±0.29Ac
07$0& 12.95±0.01Aa 12.99±0.14Aa 12.76±0.22Aa 12.21±1.05Aa 9.99±0.54Ab 9.53±0.14Ab 9.43±0.15Ab
07$0& 12.90±0.06Aa 12.97±0.04Aa 12.96±0.08Aa 12.57±0.36Aa 10.02±0.43Ab 9.61±0.19Ab 9.49±0.08Ab
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Flowabilit y
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in Figure 1. The m ean diam et er of group 1 ( dist illed
wat er ) was 12.65± 1.72 m m , of gr oup 2 ( 1% MC/
CaCl2) w as 1 1 . 7 0 ± 1 . 5 1 m m , and of gr oup 3 w as
10.17± 1.68 ( 2% MC/ CaCl27KHÀRZDELOLW\RIJURXSV
2 and 3 decreased when com pared t o group 1 and we
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group 1 and group 3.
Cell viabilit y
The cell viabilit y of MTA m ixed wit h 1% and 2%
MC/ CaCl2ZDVQRWVLJQL¿FDQWO\GLIIHUHQWFRPSDUHGWR
MTA m ixed wit h wat er ( Figure 2) . However, t here were
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The cell viabilit y of t he cont rol, and of 1/ 50 and 1/ 10
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t o 1/ 4, 1/ 2 and 1 concent rat ions. And 1/ 4 and 1/ 2
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co m p ar ed t o t h e 1 co n cen t r at i o n . Th er e w as n o
st at ist ical differ ence bet w een t he t est ed m at er ials.
Based on t his dat a, t he 1/ 4 concent rat ion was ret ained
for t he following experim ent s.
RT- PCR and quant it at ive real- t im e RCR
To invest igat e t he effect of MTA m ixed wit h MC/
CaCl2 in ost eogenic different iat ion of m PDL cells, we
evaluat ed t he expression of BSP and t he ost eoblast
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of BSP m RNA increased in all t est ed groups com pared
t o t he cont rol ( t reat ed wit h m edium only) aft er 5 d.
According t o t he result s of real- t im e PCR, t he m RNA
OHYHORI%63LQFUHDVHGVLJQL¿FDQWO\LQWKH07$PL[HG
wit h 1% and 2% MC/ CaCl2 group com pared t o t he
cont rol at day 5 ( p< .05) ( Figure 3B) .
Mineralizat ion effect
We invest igat ed t he m ineralizat ion effect of MTA
m ixed wit h MC/ CaCl2 wit h ALP st aining and alizarin
red S st aining. According t o t he result s of ALP st aining
( Figur e 4A and B) , MTA m ixed w it h dist illed wat er
and 1% and 2% MC/ CaCl2JURXSVVKRZHGVLJQL¿FDQW
in cr ease in ALP act iv it y at 5 d com p ar ed t o t h e
cont rol ( p< .05) . Alizarin red st aining, used t o det ect
Figure 1-7KHÀRZDELOLW\RIWHVWPDWHULDOV7KHÀRZDELOLW\RI07$PL[HGZLWKDQG0&&D&O2 decreased when compared to MTA
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Figure 2-7KHFHOOYLDELOLW\RIP3'/FHOOVH[SRVHGWRH[WUDFWVRIWKHWHVWPDWHULDOV7KHFHOOYLDELOLW\RI07$PL[HGZLWKDQG0& CaCl2ZDVQRWVLJQL¿FDQWO\GLIIHUHQWWKDQWKDWRI07$PL[HGZLWKZDWHU7KHFHOOYLDELOLW\RIWKHFRQWURODQGRIDQGFRQFHQWUDWLRQV
m ineralized nodule form at ion, showed all experim ent al
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( Figure 4C and D) .
Discussion
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si l i cat es2 1. D esp i t e MTA h av i n g m an y f av o r ab l e
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t o m ix MTA t o an ideal consist ency, t o deliver it t o t he
operat ion sit e, and t o condense it densely. Several
research has focused on t hese lim it at ions10,17,23,25,35.
Therefore, t he obj ect iveof t his st udy was t o evaluat e
t h e p h y si ca l a n d b i o l o g i ca l ef f ect s o f MC t h a t ,
w h en ad d ed t o MTA, cou ld im p r ov e it s h an d lin g
charact erist ics.
The com pr essiv e st r engt hs of MTA m ix ed w it h
1% and 2% MC/ CaCl2ZHUHQRWVLJQL¿FDQWO\GLIIHUHQW
com pared t o MTA m ixed wit h dist illed wat er. These
values are sim ilar t o t hose report ed by a previous
st udy23. Com pressive st rengt h is an im port ant fact or t o
FRQVLGHUZKHQSODFLQJD¿OOLQJPDWHULDOLQDFDYLW\WKDW
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com pressive st rengt h of MTA, it can be used wit h MTA
IRUSHUIRUDWLRQUHSDLUDVZHOODVURRWHQG¿OOLQJV7KHUH
were som e st udies on ant im icrobial act ivit y of MTA.
They insist ed t hat alkaline pH played an im port ant role
for t his propert y2,4,14,30. I n t his st udy, t he pH values
of MTA m ixed wit h 1% and 2% MC/ CaCl2 were not
VLJQL¿FDQWO\ GLIIHUHQW FRPSDUHG WR 07$ PL[HG ZLWK
dist illed wat er. This pH st abilit y at alkaline condit ions
m ay not affect t he ant im icr obial pr oper t ies of t he
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soluble cellulose et her and incr eases v iscosit y and
dispersion resist ance30&GHFUHDVHGWKHÀRZDELOLW\
RI 07$ VLJQL¿FDQWO\ LQ WKLV VWXG\ :H ¿UVW WKRXJKW
t hat MC im proved t he handling charact erist ics of MTA.
Consider ing it cont ains var ious st em cells31, w e
used t he m PDL cell for several biological experim ent s
su ch as cell v iab ilit y, RT- PCR an d m in er alizat ion
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cell v iabilit y of MTA m ix ed w it h 1 % and 2 % MC/
CaCl2 ZDV QRW VLJQL¿FDQWO\ GLIIHUHQW FRPSDUHG WR
MTA m ixed wit h wat er. However, t he cell viabilit y of
experim ent al groups in higher concent rat ions ( 1/ 4,
DQGFRQFHQWUDWLRQZDVVLJQL¿FDQWO\ORZHUWKDQ
Figure 3-([SUHVVLRQSUR¿OHVRIERQHVLDORSURWHLQ%63GXULQJRVWHRJHQLFGLIIHUHQWLDWLRQE\WHVWPDWHULDOVLQP3'/FHOOVDVVD\HGYLD57 PCR and quantitative real-time PCR. (A) RT-PCR results. (B) The relative expression of BSP genes normalized against a housekeeping
t he groups in lower concent rat ions ( cont rol, 1/ 50, and
1/ 10 concent rat ion) . Kang, et al.22 ( 2013) report ed
t hat MTA m ixed wit h calcium chloride showed lower
biocom pat ibilit y t han MTA m ixed wit h wat er. Therefore,
calciu m ch lor ide added w it h MC m igh t con t r ibu t e
t o t he lower cell viabilit y of experim ent al groups in
high concent rat ions. The different iat ion of progenit or
cells int o ost eoblast - like cells is crit ical in t he healing
pr ocess, and pr om ot ing differ ent iat ion is r equir ed
ZKHQFRQVLGHULQJDELRPDWHULDODVDURRWHQG¿OOLQJ
m at erial. BSP is t he m aj or phosphorylat ed prot ein of
m am m alian bone and it s use has been suggest ed at
t he st art of m ineralizat ion7,18. Consideringit is believed
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t he init ial st age of m ineralizat ion9, incr eased BSP
ex pr ession suggest s t he differ ent iat ion of sev eral
cells int o ost eoblast s. I n t his st udy, t he expression of
BSP m RNA increased in all t est ed groups com pared Figure 4- Alkaline phosphatase (ALP) staining and alizarin red staining in mPDL cells exposed to extracts of the tested materials. (A)
3KHQRW\SHH[SUHVVLRQRI$/3GXULQJRVWHRJHQLFGLIIHUHQWLDWLRQE\WKHWHVWHGPDWHULDOVLQP3'/FHOOVDWG%4XDQWL¿HGGDWD&0DWUL[ PLQHUDOL]DWLRQRIP3'/FHOOVREVHUYHGE\VWDLQLQJFDOFLXPGHSRVLWVLQWKHH[WUDFHOOXODUPDWUL[DIWHUG'4XDQWL¿HGGDWD:HREVHUYHG DKLJKHU$/3H[SUHVVLRQDQGPLQHUDOL]DWLRQLQFHOOVH[SRVHGWRWHVWHGPDWHULDOVFRPSDUDWLYHO\ZLWKWKHFRQWUROSFRPSDUHGWRWKH
t o t he m edium - only t reat ed group in RT- PCR. These
result s suggest t hat MTA st im ulat es t he ost eogenic
different iat ion of m PDL cells and MC did not int errupt
t he biological effect of MTA. Consider ing ALP and
alizarin red st aining dat a, it seem s t hat MTA and MTA
m ixed wit h MC/ CaCl2 st im ulat e t he expression of ALP
and t he form at ion of calcium nodules in m PDL cells.
These result s also suggest t hat MTA st im ulat es t he
m ineralizat ion effect28,32 of m PDL cells and MC did not
int errupt t he m ineralizat ion effect of MTA.
7KHVH ¿QGLQJV VXSSRUW WKH K\SRWKHVLV WKDW 0& GHFUHDVHGWKHÀRZDELOLW\RI07$DQGGLGQRWLQWHUUXSW
t he physical and biological effect s of MTA. However,
t here is need for furt her st udies t o clarify t he det ailed
m echanism of how MTA and MTA m ixed wit h MC/ CaCl2
induce ost eogenic different iat ion of m PDL cells.
Conclusion
MC d ecr eased t h e f low ab ilit y of MTA an d d id
n ot in t er r u p t it s p h y sical an d b iolog ical ef f ect s,
whichsuggest s t hat t hese cem ent s can be useful as
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Acknowledgem ent s
Bin- Na Lee and Soo-Ji Chun cont ribut ed equally
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int er est int his st udy. This st udy was suppor t ed by
a grant ( CRI 16025- 1) from t he Chonnam Nat ional
Un iv er sit y Hospit al Biom edical Resear ch I n st it u t e
and t he Nat ional Research Foundat ion of Korea ( NRF)
grant funded by t he Korea governm ent ( MSI P) ( No.
2016R1C1B1012703) .
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