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I take this opportunity of expressing my sincere gratitude to my revered

and esteemed teacher and guide, DR RUPAL J. SHAH, MDS.
Professor and Head, Department of Prosthetic Dentistry, without
whose guidance, constructive criticism and valuable suggestions, this
study would not have been completed. It¶s really a proud privilege to
work under her patronage and observation. I wish to place on record,
through this acknowledgement, my deepest sense of indebtness to her
and look forward to her continued support and encouragement in the
future.
I express my sincere thanks to DR. GIRISH PARMAR, MDS., Dean,
Government Dental College and Hospital, Ahmedabad, for providing all
the necessary facilities.
I express my gratitude towards Ex-Assistant Professor and Guide,
DR. SHRUTI P. MEHTA, MDS. whose guidance and motivation helped
in initiation of this work.
I express my sincere gratitude to all the staff members of the
department of Prosthetic Dentistry for helping me whenever I needed.
I am also thankful to library staff MR. R.S. KHETRA and Mrs. V.R.
CHAYA, for their help in providing the necessary academic material.
I am grateful to Mrs. CHANDRALEKHA, Professor and Head,
Department of Microbiology, B.J. Medical College, Ahmedabad for
permitting me to carry out dissertation work at their department. I also
take the opportunity of expressing sincere thanks to Mrs. MONIKA,
Laboratory Technician, who helped me to carry out the necessary tests
in the laboratory.
I am grateful to Mr. VIJAY SHARMA, Biostatistician, N.I.O.H.,
Ahmedabad for carrying out statistical analysis as per the requirements
of my work. My sincere thanks to my colleagues Dr. Vishal, Dr.
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Kuldeep, Dr. Alaap, Dr. Gautam, Dr. Prashant, my seniors and my
junior friends, who have provided support throughout my work.
I wish to place on record that it is the encouragement, love and
warmth given by my parents, my brother, my in-laws and my beloved
husband DR. RITESH GUPTA, who helped me make this venture
successful.

3





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. INTRODUCTION 

2. REVIEW OF LITERATURE 4

3. MATERIALS AND METHOD 2

4. OBSERVATION AND RESULTS 36

5. DISCUSSION 5

6. SUMMARY AND CONCLUSION 62

7. BIBLIOGRAPHY 63
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Ê

The therapeutic procedures used in the treatment of causes do not
always eliminate all the micro-organisms in the residual tissues. The
persisting bacterial presence, together with the lack of a thoroughly
hermetic seal between the prepared tooth walls and final finished fixed
prosthesis walls allow bacterial leakage and thus may be involved in
the development of recurrent caries. One possible solution for this
service problem is to use dental luting materials with a bacteriostatic
capacity. 
Microleakage is a common, unavoidable, clinical phenomenon by
which oral fluids, ions, molecules¶, and bacteria gain access to dentinal
tubuli and pulp through the junction between a ceme nted prosthesis
and the tooth. 2
Recurrent human dental caries has been associated with the
dissolution of dental luting agents and the inherent roughness of the
prepared tooth margins or prosthesis. Breakdown is marginal. Areas
between tooth prepared and the prosthesis cemented by any of the
luting cement can provide potential pathways for reinfection.
Carcinogenic micro-organisms present in normal human flora could
easily penetrate underlying dentin through such defects. A well
designed preparation with a smooth and even margin reduces or
preferably prevents such marginal breakdown reducing the chances of
recurrent carries. 3
Several species of bacteria may be isolated from plaque associated
with carious lesions and pulpal inflammation.    is
one of the bacteria most frequently implicated in dental caries. 4


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When teeth are being prepared for fixed prosthesis, infected and
softened dentin is often found extending into the deeper layers.
Preparations for abutments, which include removal of such carious
Lesions, can often result in the creation of larger cavities.
Consequently, the mechanical strength and retention force of
abutments may be weakened. Human carious lesions can be
thoroughly sterilized by the topical application of certain antibacterial
drug combinations. These sterilized carious lesions can be left
untreated and softened because the dentin recalcifies with time. Thus
this procedure can minimize the amount of infected dentin that must be
removed from the prepared abu tments.5
Failure of fixed partial denture is most frequently caused by caries. It
was proposed that carious lesions of the prepared abutments may be
sterilized by covering them with a provisional restoration and
antibacterial provisional cement containing a mixture of antibacterial
drugs.6
Permanent and provisional restorations cemented with temporary
cement need to provide biological and mechanical protection for the
abutments. A temporary cement must be strong enough to retain the
restoration but weak enough to enable removal of the restoration
without damage to the abutment. However, provisional crown luted with
temporary cements are susceptible to cement washout, marginal
leakage, Bacterial infiltration and caries, especially when placed for a
periods longer than a few weeks. 7
Antibacterial activity, during and after setting, assumes clinical
relevance because this property may help in the elimination or
reduction of bacteria that have remained viable in the cavity walls or
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bacteria that may gain access to the cavity through micro leakage
channels. 4
The physically superior cement materials can play an important
role in caries prevention. Furthermore, it would be advantageous if
such materials possessed effective antibacterial prope rties. 3
It is therefore, the objective of this study ³ANTIBACTERIAL
PROPERTIES OF AGED DENTAL CEMENTS´ to access and compare
the incessant antibacterial activity of three luting cements .
 Glass-Ionomer Cement (Ketac- Cem
2 Zing Phosphate Cement (Harvard
3 Polycarboxylate (Poly F in 2 powder: liquid ratios (wt/wt
 Ketak-Cem 3.8: 3.5:
2 Harvard .8: .6:
3 Poly F 2.8:.8:
by undertaking the following tests:

 Direct contact test

2 Agar diffusion test. 6



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  Ê Ê in 
80 reported that the adhesion to
enamel is an important aspect of periodontal dressing retention. Details
of tensile and stress bond strengths were presented for three dressing
materials i.e. Coe Pak, Peripac, Peripac improved and a positive
polycarboxylate Control (Duralon and the data was discussed with
regard to optimum levels of retention. The dr4essing materials were
tested hr, day and week after placement, in groups of five
specimens for each test, tensile and shear.
Most shear tests yielded higher results than the corresponding tensile
tests and a level of 2MN/m2 is suggested for shear bond strength and
MN/m2 for tensile bond strtength. 8
 Ê !  " # in 
80 evaluated the
antimicrobial action of various dental cements, against common micro -
organisms frequently based within the components of the normal
microbial oral flora. The cements included in the study were
 zinc oxide eugenol
2 polycarboxylate
3 zinc phosphate
4 silicate
5 silico-phosphate
6 composite resin.
The antimicrobial power were against the following micro -organisms:
 Streptococcus mutans
2 Escherichia coli
3 Streptococcus viridians
4 Lactobacillus acidophilus

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5 Streptococcus pyogenus
The cements were listed in decreasing order of effectiveness:
 Zinc oxide eugenol
2 Silico-phosphate
3 Zinc phosphate
4 Silicate
The two cements, polycarboxylate and composite resin, exhibited no
measurable antibacterial action.
$ %
 & ' ( %
 in 
86 examined the
parameters and expected that advances in the state of fixed
prosthodontic materials and methods would mean changes in cause of
failures and length of service of fixed restorations.
 The mean length of service of all fixed restorations observed was
8.3 years.
2 Caries was the most common cause of failure, affecting 22.0%f the
units and leading to the necessity for placement of 24.3% of all units
observed.
3 Mechanical problems accounted for 6
.5% of the failed units as
opposed to 28.5% for oral disease.
4 Resin veneer metal crowns provided the longest service of all crowns
types observed (3.
years and failed most frequently because of
worn or lost veneers.
   The resin veneer metal crown also provided the longest service as a
retainer, with a mean length of service of 4.7 years. 




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6 No apparent relationship was found between the span of prosthesis
and its length of service. 0
 " 
")"% in 
87 compared the
antibacterial properties of 4 different restorative materials, nine of
which were glass-ionomer cements. The materials were mixed
according to manufacture r¶s specifications and exposed to four types of
bacteria (      
      commonly found
in caries and plaque. Zones of bacterial in hibition were measured for all
materials in millimeters. They concluded that:
 Glass- ionomer cement materials used as liners/based and restorative
materials produced zones of inhibnition with the four bacteria used in
this study.
2 The glass- ionomer cement liner/base materials which contain zinc
oxide produced zones of inhibition larger than did those not containing
zinc oxide.
3 The composite material, FluorEver, is capable of producing zones of
microbial inhibition.
4 Dispersalloy amalgam produced zones of inhibition with all bacteria
used in this study. 
'*  %  in 
8
assessed the antibacterial
properties of dental restorative materials with ratios of test
material/culture medium volume aiming to simulate conditions around a
restoration. Antibacterial activity is determined by the reduction in




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optical density of the test culture relative to controls. The method was
used for assessment of the antibacterial activity of five dental materials
of different composition against five oral bacteria. Release of zinc and
fluoride from these materials was also measured and correlated with
antibacterial activity. There was a general trend toward greater
antibacterial activity with increased zinc release, while fluoride release
had a significant effect on only one organism. While all the materials,
when freshly mixed, were strongly toxic to three out of the five bac teria
studied, much of this activity was lost after the material had set. 2
#+&
 ) $  $

# + '' in 
8

evaluated the antibacterial activity of six glass -ionomer cements on S.
   
. These cements were Glassic, Shofu lining
cement, GC lining cement, Ever Bond, Gingiva Seal and Ketac Bond,
the empty walls swerved as control. They concluded that:
 GC lining and Ever Bond had signi ficantly a greater overall
inhibition of microbial g rowth than did other tested liners.
2 Glassic and Ketac Bond had significantly more inhibitory effect
on S.   than on 
!.3
,%+"% -, $ in 

 compared the
levels of fluoride and mutans streptococci in plaque grown on glass-
ionomer and composite restorations in vivo. From tunnels left under the
brackets bonded either with glass -ionomer or composite, 4-day-old
plaque samples were collected 4 ,28 and 42 days after bonding. For
glass-ionomer the mean counts of   in plaque were
0.5x03 x 6.7x03 and 8.8x0 3 CFU at the first, second and third



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collection, whereas for composite restorations the corresponding
values were 32.x03,4.6x03 and 20.6x0 3 CFU. For glass-ionomer
the mean concentrations of fluoride were 
,
85, 5788 and 50
ppm
at first, second and third collections of 4 -day-old plaque samples,
respectively, whereas for composite restorations the mean
concentrations of fluoride were about 200 ppm throughout the study.
The results show that the fluoride leve l in plaque growing on glass
ionomer is much higher than that on composite restorations which
seems to affect the level of mutans streptococci in dental plaque. 4
$"+ Ê% # '%  %Ê '"$$ Ê, in 

2
measured the in vitro inhibition of growth and adherence of five oral
bacteria by glass-ionomer materials. Disks were prepared from two
cavity liners and four restorative class materials, by use of Teflon
plates with circular walls, five mm wide and two mm deep. The
bacterial species included: A.  
  
  
 "

  
An ion exchange electrode was used measure fluoride
release over a 7 day period for all six glass ionomers. The 2 cavity
liners and two of the restorative materials produced the largest growth
inhibition zones by direct contact. No growth inhibition occurred when
the specimens were allowed to come into contact with the agars prior
to inoculation. All four restorative materials reduced bacterial
accumulations on enamel surfaces by over 80%. Elevations in short -
term fluoride release levels were positively corr4elated w ith growth
inhibition.




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  -"!, . in 

2 studied the eff ects
of different glass ionomers on the metabolism of  
 test slabs of freshly mixed conventional glass-ionomer, silver
glass-ionomer, composite and 2-week old glass-ionomer were fitted
into the bottom of a test tube. A plaque -like layer of 
  strain
Ingbritt was centrifuged onto the test slabs, and the samples w ere
incubated for 20 hours in  .7% (w/v sucrosed solution.
Incubation with glass-ionomeer materials led to increase in the
cellular concentration of fluoride in bacteria, but intracellular fluoride did
not correlate with the fall in pH. The lowest pH was associated with the
lowest cellular magnesium content. Ketac -Silver released large
amounts of calcium in the fluid phase, and t he cellular calcium content
was doubled in this group. The resu lts show that freshly mixed glass-
ionomers affect acid production and electrolyte metabolism of 

 in vitro. The effect of conventional glass -ionomer, however,
seems to disappear after a few weeks. The effects of calcium and
silver released by cermet glass ionomer deserve further study. 6
$" Ê  . #-  & "  In 

5 assessed the
antibacterial activity of the following three dental liners; Vitrebond,
Dycal and Life. The test was based on a modification of the agar
diffusion test in which samples were placed on agar plates previously
inoculated with    and were removed after
predetermined time periods. The materials effect on bacterial growth
was evaluated.




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Results showed that Vitrebnond had a strong antibacterial effect
that was evident after  min of direct contact with the inoculated
7
bacteria. It was significantly more effective that Dycal or Life.
Ê( # "& 
 )"*  " 

7 compared
the short-term tissue responses to two potential root-end filling
materials, light-cured glass ionomer cement and a reinforced zinc oxide
eugenol cement with that to amalgam.
In the 24 premolar teeth of beagle dogs, a collection of endodontic
pathogenic bacteria was first inoculated into the root canals to induce
periradicular lesions. On each root, an apicectomy was performed and
root-end cavities prepared to receive fillings of each material. The teeth
and surrounding were removed after 2 weeks (23 roots and I week (24
roots ; they were then prepared for histological examination.
Apart from amalgam in which healing was marked by the presence of
a localized focus of inflammation adjacent to the root canal filling,
response to Vitrebond and Kalzinol. Both Vitrebond and Kalzinol have
potential as root -end filling materials, as the tissue response was
considerably more favorable than that of amalgam even in the short-
term.8
  "/+ %  )$0   $ " 

7 compared the fluoride
concentrations in plaque on  -year old resin-modified GIC, compomer
and resin composite restorations intra -individually and related to the
occurrence of caries- associated bacteria. Plaque from class III
restorations of the three restorative materials and from a proximal
enamel surface in 8 individuals was analyzed. Low fluoride levels



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were detected in all the samples, while the resin -modified GIC samples
showed significantly higher amounts. The distribution of oral
streptococci surfaces and did not correlate to the fluoride levels in the
samples. A good correlation was found between the counts of 
  in salvia and their proportions in the plaque.  
The results indicate that the fluoride concentr ations released in vivo
from -year-old restoratives are not high enough to affect the plaque
levels of the caries- associated bacteria mutans streptococci and
lactobacilli.
,"
)&," in 

7 observed the antibacterial
potential of polycarboxylate temporary cement containing a mixture of
metronidazole, ciprofloxacin and cefaclor on carious lesions of
prepared abutments that were designed to leave caries on the
abutments.
Antibacterial efficacy was estimated in vitro and in vivo b y measuring
bacterial recovery from the lesions. They concluded:
 Bacteria counts ranged from 0 4 x 07 both in vitro and in vivo in time±
zero samples, just before application of the antibacterial cement.
2 When the lesions were covered by the antibacterial tem porary cement
from  to 4 days, no bacteria were recovered in most cases. This
finding indicated that the carious lesions were sterilized by the
antibacterial temporary cement. Although a small number of bacteria
were recovered from some samples, all the bacteria recovered were
sensitive to the drug mixture.
3 From the lesions covered by the temporary cement without the
mixture of drugs, more than 0 3 CFU/mg were recovered,


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4 indicating that the temporary cement that contained tannin was not a
potent disinfectant.
5 In experiments in vivo, no bacteria were recovered from the carious
lesions of prepared abutments that were covered by temporary
restorations with the antibacterial temporary cement within 2 weeks.
6 One in vivo case suggested that care should be taken for marginal
leakage and that temporary restoration should not be left for a long
time, even if antibacterial cement is used. 20
, ' Ê"$$  &  $ in 


studied the antibacterial
activity of the Glass-Ionomer restorative cements Ketac-Fil, Ketac-
Silver, Fuji II LC and Vitremer in vitro, in conjunction with a total of 32
strains of five bacterial general that may be associated with dental
caries:       
 #      . Agar plate diffusion was the
method used for the bacterial cultures, which included a chlorhexidine
control.
All the four glass-ionomer cements were found to inhibit bacterial
growth, though with noteworthy differences in their spheres of acti on.
Vitremer was the cement determined to have the greatest antibacterial
effects, whereas Ketac-Silver presented the least inhibitory action.
In light of reported results, the use of glass -ionomer cements may be
indicated in the restorative treatment o f root surface caries. These
could even prove beneficial for patients with periodontal disease
suffering from root caries. 2
#/    & )"&& ' " 2000 reviewed and
discussed some of the techniques available to excavate demineralised
dentine clinically. These methods can be classified as mechanical and

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non-mechanical, rotary and non-rotary and include: dental hand
pieces/burs, manual excavators, air-abrasion, air-polishing,
ultrasonication, sono-abrasion, chemo-mechanical methods, laser and
enzymes.
All the techniques removed carious denture with di ffering levels of
efficiency but more importantly it was still unknown if these techniques
would discriminate bet ween the soft, outer, necrotic, highly infected
zone that needs to be excavated and the inner, reversibly damaged,
less infected zone which could be retained. If this discrimination did not
take place, this could lead to over preparation of cavities with little
control over the quality and quantity of tissue removed by individual
operators. There is, therefore, an important need to assess the effects
of these techniques for their efficiency and extent of removal of carious
dentine. 22
, ' Ê"$$  &  $ " 2000 studied the antibacterial
action of different dental resin adhesive materials (Glumma 2000,
Syntac, Prisma Universal Bond 3, Scotchbond Multi -Purpose and
Prima & Bond 2.0 glass- ionomer cements (Ketac-Cem, Ketac-Bond,
Ketac-Silver, Ketac-Fil resin-modified galss-ionomer cements (Fuji II
LC, Vitremer and Vitrebond and a Compomer (Dyract . The agar plate
diffusion method was used for the microbial cultures and a
chlorhexidine control. The growth of the caries-producing
microorganisms was effectively inhibited by the Vitremer a nd ViteBond
cements, and to a lesser extent by the Scotchbond Multi -Purpose
adhesive system. 23



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)"+-)! "* !$&$ in 200 examined
the antibacterial activities of the bonding sys tems-Syntec, EBS and
Scotchbond I, the polyacid-modified composite resins-Hytac and
Compoglass, and the composite resins-Tetric, Z00 and Scalp -it. They
were evaluated using the cariogenic bacteria  
         
 in vitro with a modified cylinder drop plate agar diffusion
assay.
All adhesives of the dentin bonding systems and the polyacid -
modified composite resins exhibited vario us degree of antibacterial
activity against all of the test bacteria. On the contrary, composite resin
did not affect bacterial growth. The data suggest that the use of these
adhesives and polyacid-modified composite resins may reduce the
consequences of microleakage owing to their antibacterial properties. 24
Ê!'"Ê&$ in 200 analyzed the long-
term clinical behavior of two dental materials applied as filling under
silver amalgam restorations: glass-ionomer cement (GIC and
composite resin with adhesive system (CR . In this study, 7 posterior
teeth (2
premolars and 88 molars were selected with carious lesions
which resulted in great loss of dentin and cusps with unsupported
enamel. After caries removal, cavities were prepared and totally filled
with GIC with. In a following visit, new cavities were prepared, leaving
the employed filling material as a base and support for the enamel,
which were then restored with silver amalgam. Restorations were
evaluated periodically aft er 6 months and up to 5 years. Both fracture
and pulpal involvements rates were low.
There was a significant association between kind of tooth and long -
term survival of the restorations; and between degree of unsupported
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enamel and the same long -term survival. The results confirmed that the
technique in which GIC or CR are used as filling material under silver
amalgam restorations is clinically acceptable.

! ,% Ê&$
 " 2002 compared the methods and
techniques used for short and long term restoration during immediately
after endodontic treatment, and to make clinical recommendations.
Proper clinical assessments could only be obtained from well
designed studies that reflect the superiority of some of the available
materials more accurately in the actual clinical environment and in
more complex cavities.
Because of the nature of these materials, they should be used for as
short a period as possible during the course of endodontic treatment. 26
,! Ê,%& 1
' " 2003 reviewed on the methodology
involved in biocompatibility testing following by a discussion on
biocompatibility of contemporary intracanal drugs and substances used
in endodontics. Research showed that the intracanal drugs and
substances can have deleterious effects on vital tissues. Although
these substances are meant to contact non vital dentine during use,
they often come into contact with the peripical tissue. It is thus
important to consider biocompatibility when choosing and endodontic
irrigant or intracanal medicament. 27
"" !
&$ in 2003 investigated the:
 retention and microleakage of provisional crowns cemented with
temporary cements to which to which stannous fluori de (SnF2 was
added
2 solubility of these cements.
Provisional crowns were constructed of acrylic resin with shoulder
preparations for 2molars. The crowns were luted with Tempbond,
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Tempbond NE and Freegenol tempo rary cements, and also with SnF 2
added to these cements. Specimens were thermocycled 00 times,
stored for 6days, and immersed in 0.5% basic fuschin. 7days after
cementation, crown removal (retention tests were concluded. Marginal
leakage and solubility in water of cements with and wi thout SnF2was
assessed.
The results confirmed the Freegenol to be more retentive than the
other cements. The incorporation of SnF2significantly increased the
retention capacity of Freegenol and Tempbond NE but had no effect on
Tempbond. Tempbond showed significantly higher dye penetration
than the Freegenol. The addition of SnF 2 did not alter the dye
penetration of the cements. There were no significant differences in the
solubility of the cements. However, the incorporation of SnF 2increased
the solubility of Freegenol and Tempbond NE P <0.00 and Tempbond
(P<0.0 . 28
(21"&"" 2003 analyzed the amount
of viable bacteria after excavation using conventional rose -bur on the
chemo-mechanical Carsilov method, a total of 22 lesions in this open,
control and randomized study. Two samples per lesion were taken
under aseptic conditions using a rose -bur, on superficially in the
carious lesion and one after complete excavation. The samples were
incubated on blood agar, Rogosa agar and mitis salivarius agar.
For blood agar (aerobic both methods resulted in a significant
decrease in CFU, for blood agar (anaerobic and MS agar only the
Carisolv method resulted in a significant decrease in CFU and for SL
agar neither method resulted in a significant decrease in CFU.
Comparing the excavation methods, there were no significant
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differences, except in the case of blood agar (aerobic , which showed
that Carisolv excavation was more effective in reducing CFU. 2

"+  $+  &  $ in 2004 evaluated the adhesion of
  ATCC 2575 to filling materials (Ariston, Tetric,
Dyract, Compoglass, Vitremer, Aqua Ionofil, Ketac Fil, amalgam,
Galloy and ceramics as controls .   was added to
saliva- coated test specimens, and a nutrient broth permanently
supplied over a time period of 48hrs and then the weight of plaque, the
number and viability of the bacteria adhering to the materials were
determined.
The weights of artificial plaque o n all filling materials tested were
higher than those on ceramics, the h ighest values were measured on
the glass-ionomers. The amount of plaque correlates with the surface
roughness, whereas there was no correlatio n of the surface roughness
with the number of colony forming units (CFU of 
 . The
plaque of Ketac Fil contained a high number of viable bacteria. The
fluorides of glass-ionomers do not efficiently prevent the attachment
and the viability of S.mutans. 30
"+!&.)" 2004 attempted to determine
whether it exerts an antibacterial effect on human saliva bacteria, and
to determine whether it can be used in dental materials. CFUs in ml
stimulated human saliva were examined using blood agar and mitis
salivarius agar after immersion, with or without YDA filler. The
antibacterial effect wa s compared with that of Ketac-Silver.
Human saliva bacteria and mutans streptococci showed reduced
viability following exposure to YDA filler after 2hrs. Two tested strains
showed reduced viability following exposure to dental materials
containing YDA filler. Thus YDA filler may help in the d evelopment of
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antibacterial dental materials, such as composite resin, glass -inomer or
temporary cement. 3
-,"


"$  & +!(
 " 2005 studied the effect
of crude khat extract on   growth and sucrose-
dependent colonization, and on its glucosyltransferase (GTF activity
and production. Lyophilised crude aqueous khat extracts were applied
to the different assays at conce ntrations of 0-% (w/v . Sucrose-
dependent colonization was assessed as the ability of  
 UA5
to form adherent biofilm in glass culture tubes. Colony
forming units (CFUs in the planktonic phase served as a measure of
bacterial growth, while CFUs in the biofilm phase were used to quantify
viability in the biofilms. GTFs production was assayed by comparing
intgensities of GTF bands in Western blots of extract from control and
khat-containing cultures.
The khat extracts effectively inhibited biofilm formation. The extract
also inhibited synthesis of both glucan types, particularly insoluble
glucans, with significant differences among cultivars. However, khat
increased bacterial growth and at sub -MCIC also viability within
biofilms; there were no inter-cultivar differences. It is shown that
Contain water-soluble constituents that inhibit some cariogenic
properties of 
 in vitro.32
  $!   $ ' & 01 % " 2005
evaluated the antibacterial activity of six products (one conventional
glass-ionomer cement (GIC , two light-activated glass-ionomers, two
polyacid-modified resin composites and one resin composite on
Streptococcus mutans. The relationship between product acidity and
antibacterial activity was evaluated.
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All the GICs demonstrated antibacterial properties in contrast to the
polyacid-modified resin composites and resin composite which did not
show any antibacterial effects. Vitrebond GIC exhibited higher
antibacterial action, probably because of a cytotoxic photo -initiator 
diphenyliodoniumchloride. A direct relationship betw een material
acidity and growth inhibition of 
was observed. 33
"" '$&$" 2005 evaluated the antibacterial
properties of 3 dental cements using the direct -contact test and agar
diffusion test. For the direct contract test, wells of microtitre plate were
coated with the tested cements (Harvard cement, Duralon, and Ketac
cement while    suspension was placed directly
on the cements. Bacterial growth was evaluated by a tempe rature-
controlled microplate spectrophotometer. For the agar diffusion test,
triplicate specimens of freshly mixed cements were poured into u niform
wells punched in the agar plates inoculated with  
.
Compared with the control group, Duralon and Harvard cement
demonstrated antibacterial properties even after 3months will the
direct-contract test, while Ketac-Cem exhibited no antibacterial
properties. In the agar diffusion test, no antibacterial property was
observed for any of the tested cements. The different powder/liquid
ratios had a negligible effect on the antibacterial properties of the
tested cedments. 34
+" .   &  " 2006 evaluated the antibacterial,
physical, and bonding properties of glass -ionomer cements (GIC
containing chlorhexidine (CHX , and to determine optimal
concentrations for incorporation of agents to obtain antibacterial GICs
for use with the ART approach.
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All experimental GICs exhibited inhibition of Streptococcus mutans,
Lactobacillus casei and Actinomyces naeslundii, but sizes of the
inhibition zones and concentrations of CHX released were not
dependent upon CHX content. Incorporation of CHX diacetate at 2% or
greater, significantly decreased compressive strength, and bond
strength to dentin was adversely affected by addition of CHX diacetate
at 2% or more, although setting time was extended a little by addi tion
of any concentrations of CHX. 35
The present results demonstrate that incorporation of % CHX
diacetate is optimal to give appropriate physical and bonding
properties. 35
,(+$ Ê  % & "$ ' " 2006 investigated
the interaction of     with two glass- ionomer
formulations (GIC: A containing fluoride and GIC: B without fluoride
with particular reference to bacterial gro wth and changes in hardness
of the cement with respect to time. Hydroxyapatite discs (HA wer e
used as controls.
 The viable counts of 
   per mm2 on GIC: A were significantly
less than those on HA and GIC:B during the first 5days.
2 The viable counts bacteria on the surface of GIC: B were lower during
the initial 5 days when compared to HA.
3 Exposure of GIC: A and GIC: B to different medium produced softening
to the surface of the cement.
4 It is apartment that the effects of the biofilms are significantly greater
than storage in water but similar to storage in lactic acid pH5. 36


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-
'

', 
The purpose of this study was to evaluate the antibacterial properties of
3 dental cements using the Direct -Contract test and Agar Diffusion
Test.
'

31"0"4 
 Spoon excavator ( Fig 
2 Phosphate buffered saline (Fig 2
3 Distilled water
4 Rubber dam (Fig 3
&"4 
 The test cements ( Fig 4
2 Autoclaved BHI Broth with added Bacitacin (Fig 5
3 Microtitre plate (
6 -well, flat bottom Nunclon (Fig 6
4 0-µL micro-pipette, small (Fig 6
5 235-µL micro- pipette, large (Fig 6
(&"00!"4 
 The test cements (Fig 4
2 Autoclaved BHI Agar with added Bacitacin (Fig 5

'

'
 Electronic weighing machine (Fig 7
2 Clean, sterile glass slab (Fig 4
3 Dental spatula (Fig 4
4 Vortex mixer (Fig 8
5 Auto- mixer (Shaker (Fig

6 Incubator at 37 0C (Fig 0
7 Temperature- controlled spectrophotometer (THERMOMAX
(Fig 
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', 
The cements tested under this study were Harvard (Zinc -Phosphate
Cement , Poly-F (Polycarboxylate cement and Ketac-Cem (Glass-
lonomer cement . All the cements were exposed to Streptococcus
mutans under similar conditions.

Ê

$"0 !!0"4 
All consecutive young patients making a group of 25 patients, in the
age group of
-2 years (Streptococcus mutans considered to be the
primary etiological factor , appearing for regular dental examination
participated in this study. Inclusion criteria were vital teeth without
symptoms, either molars with p rimary caries that included
approximately half the dentine thickness as judged by bitewing
radiographs, or primary buccal caries lesions extending into the dentine
in the incisors, canines or molars. The consistency of the caries lesions
was medium hard and the colour yellow to light brown, No teeth with
pulpal involvement were included. 
Rubber dam was applied and surface debris including the outermost
layer of carious dentine was removed using sterile hand instruments.
The cavity was thoroughly rinsed with sterile saline, then dried with a
cotton pellet. The caries were e xcavated with spoon excavator and the
sample was transferred to a sterile bottle containing phosphate
buffered saline (mixed in a proportion of
.
5gms in

0ml and
transferred to the laboratory (Fig  , where it was shaken for 30sec in a




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23,3

vortex mixer to dislodge the dentin sample. A clinical isolate of
mutans streptococci (ATCC 2735 that is naturally resistant to
bacitracin was grown aerobically from frozen-stock cultures in brain-
heart infusion(BHI broth containing 8ug/ml bacitr acin (Fig for 48 hours
at 37 before applying it to the specimens according to the experimental
design. 34
 "0 $04 
The test cements (Harvard, Poly F and Ketac-Cem were measured
with an electronic weighing machine in a prescribed proportion and
mixed in2 powder/liquid (P/L ratios. P/L  ratio was in accordance with
the manufacturer¶s instructions and P/L 2 was more diluted (P/L ratio
wt/wt  and 2 .
& "- &5$"2!"&65 "0 
#

 Ê'
. 4
 65 
Harvard Zinc Phosphate
 .8:
cement 2 .6:
Ketac- Glass- lonomer
 3.8:
Cem cement 2 3.5:
Poly F Polycarboxylate  2.0:
cement 2 .8:
These cements represent 3 types of luting agents, which are
commonly used in fixed prosthodontics. The more diluted P/L ratio
simulates situations in which the clinician does not follow the
manufacturers¶ instructions exactly and prepares a thin mix of cement.
The antibacterial properties of the materials were examined within 60
minutes after mixing (designated hereafter as ³fresh preparations´ .
Similar experiments were performed in which the test materials were
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allowed to age in a phosphate buffere d saline (PBS for 24 hours,
week, mont, and 3months before assaying.

 Ê '
4-

Ê-Ê
Ê6Ê 
The direct-contact test originally described by Weiss et al is based on
the turbidometeric determination of bacterial growth in
6-well
microtitre plates. The kinetics of the outgrowth in each well was
recorded continuously at 650nm at 37 0C every 30minutes, using a
temperature- controlled spectrophotometer (THERMOmax . Auto
mixing prior to each reading ensured a homog enous bacterial cell
suspension.
A microtitre plate (
6 -well, flat-bottom Nunclon was held vertically
with the plate¶s surface perpendicular to the floor, and the sidewall of 4
wells was coated evenly with a measured amount of the test material
(designated as Group A . A thin coat was achieved by using a small
flat-ended dental instrument, such as a dental spatula. The material
was allowed to set in compliance with the manufacturer¶s
recommendation. Special care was taken to avoid the materials¶ flow to
the bottom of the well, which could interfere with the light path through
the microtitre well and result in false readings.
A 0-µL bacterial suspension (approximately 0 6 bacteria was
placed on the test material, while the plate remained in a vertical
position. Wells were inserted for evaporation of the suspensions¶ liquid,
ensuring direct contact between bacteria and the test materials. This
usually occurred within  hour at 37 0 C. BHI broth supplemented with
bacitracin (235µL was added to each of the Group A well s and gently
mixed for 2 minutes; 5uL were then transferred from Group A wells,
respectively, to an adjacent set of 4 wells containing 205µ L fresh
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medium (designated as Group B (Fig 3 . This resulted in 2 sets of 4
wells for each test material containing an equal volume of liquid
medium (220µL , so that bacterial outgrowth could be monitored and
compared, both in the presence and absence of the test material. Two
sets of 4 uncoated wells in the same microtiter plate served as the
positive control. Identical bacterial inoculum was placed on the sidewall
of the uncoated wells and processed as in the experimental Group A
and B wells. The growth curves from both experimental groups were
compared with the control outgrowth A and B, respectively. The
negative control consisted of a set of 4 wells coated with the test
materials, as in experimental Group A, containing an equal volume
uninoculated fresh medium. (Fig 4 .
The kinetics of the outgrowth in each well were monitored at 650 Nm
at 370C and recorded every 30minutes, using the same temperature -
controlled microplate spectrophotometer (THERMOmax . Auto -mixing
prior to each reading ensured a homogenous bacterial cell suspension.
Data were recorded in optical density (OD units. The values of the
negative control wells were considered as the baseline, and were then
subtracted from the respective experimental sets and plotted as growth
curves. The first set of data was recorded approximately  hour after
incubation. Additional experiments were performed on set test
materials that were allowed to age in the presence of 280µL of PBS for
24 hours,  week, montgh and 3months, respectively, before being
assayed by the DCT. During the aging period, PBS was replaced twice
a week. The data were subjected to -way ANOVA and the Post-Hoc
multiple comparisons test. Parallel to the experimental design,
calibration experiments were performed to establish bacterial
outgrowth under experimental conditions in a quantitative and
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r-
reproducible manner. In a typical experiment, 0µL of bacterial
suspension (approximately 0 6 cells was placed on each sidewall of 4
wells in a
6-well microtiter plate, as in experimental design. Fresh
medium, 275µL, was added and the plate gently mixed for 2 minutes.
From each well, 55µL were transferred to an adjacent set of wells that
each contained 220µL fresh medium. The dilution transfer was
repeated 6 consecutive times. The microtiter plate was incubated in the
spectrophotometer at 37 0C, and the bacterial growth was followed for
6 hours. To overcome contamination throughout the experiment,
25mg/ml of bacitracin was included in the BHI medium and an
antibiotic resistant strain was used.
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6
For the ADT, 200µL of bacterial suspension (approximately 5x0 7
cells were spread on BHI agar plates (Fig 5 . Freshly mixed
specimens (in triplicate of each test material were placed in holes
5mm in diameter that were punched in the agar using the blunt end of
a sterile Pasteur pipette 23 (Fig 6 . After incubation at 370C for
24hours, the agar plates were examined for bacteri al lawn was
measured (mm in 2 perpendicular locations for each specimen. The
first set of data was recorded after 24hours. Additional experiments
were performed on aged specimens for which test materials had been
placed in bottles containing PBS and bacitracin for  week, month,
and 3 months before assessment by the ADT. During the aging period,
PBS was replaced twice a week. The data were subjected to  -way
ANOVA and the Post-Hoc multiple comparisons test. 34 


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#





The caliberation experiments showed that bacterial outgrowth in

microtiter wells could be monitored in a quantitative and reproducible
manner.
The gradual decrease in viable bacteria, due to serial dilutions at
time zero, had virtually no effect on the bacterial growth rate or final
density of bacteria at the stationery phase in the system.
The result of DCT for freshly mixed and aged cements in differ ent P:L
ratios are shown.
The abbreviations used in the results:

KC: Ketac Cem (GIC Cement

KC: I P: L ratio of Ketac Cem
KC2: II P: L ratio of Ketac Cem

H: Harvard (Zinc Phosphate Cement

H: I P: L ratio of Harvard
H2: II P: L ratio of Harvard

PF: Poly F (Polycarboxylate Cement

PF: I P: L ratio of Poly F
PF2: II P: L ratio of Poly F
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 Ê

Success of restoration is dependent on accuracy of each and every
step from preparation to final seating and even upon duration or
longevity of restoration. Generally the prefe rred margin of the
preparation is supragingival. Subgingival margins o f cemented
restorations are major etiologic factor in periodontal disease,
particularly when they encroach on the epithelial attachment.
The junction between a cemented restoration and the tooth is always
a potential site for recurrent caries because of dissolution of the luting
agent and inherent roughness. The more accurately the restoration is
adapted to the tooth, the lesser the changes of failure. For acceptable
margin adaptation, castings should fit within 0um and a porcelain
margin with 50µm. A well designed preparation has a smooth and even
margin as compared to rough, irregular or ³stepped´ junctions which
increase the margin length and substantially reduces the adaptation
accuracy of the restoration. 37
This is consolidated by "+   $ 6-

[ who evaluated the
adhesion of    ATCC 2575 to filing materials
(Ariston, Tetric, Dyract, Compoglass, Vitremer, Aque Ionfil, Ketac fil,
Amalgam, Galloy and ceramics as control . They found that the
! 0 $2! -$ " !0 !(, whereas
there was no correlation of the surface roughness with the colony
forming units of   . The plaque on KETAQC-FIL
contained a higher number of viable bacteria. The fluoride content of GI
do not efficiently prevent the at tachment and viability of  
.30
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 -
The cross-section configuration of the margin has been proposed to be
of various types:
 &(4
Advantage: Conservation of tooth structure
Disadvantage: Does not provide sufficient bulk
Indication: Not recommended;
2. Ê"$&(4
Advantage: Conservation of tooth structure
Disadvantage: Location of margin difficult to control
Indication: on titled teeth.
3. #1$4
Advantage: Removes unsupported enamel
Allows finishing of the metal
Disadvantage: Extends preparation into sul cus if used on apical
margins
Indication: Facial margin of maxillary partial -coverage restorations and
inlay/onlay margins
4. Ê04
Advantage: Distinct margin, adequate bulk and easier to control.
Disadvantage: Care needed to avoid unsupported lip of enamel
Indication: Cast metal restoration
Lingual margins of metal ceramic crown
5. !$&4
Advantage: Bulk of restorative material
Disadvantage: Less conservative of tooth structure
Indication: Facial margin of metal ceramic crown
Complete ceramic crowns
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6. $ &!$&4
Advantage: Bulk of restorative material, advantages of bevel
Disadvantage: Less conservative of tooth structure
Indication: Facial margin of metal ceramic crowns.
7. !$&"#1$ :
Advantage: Bulk of restorative material advantages of bevel
Disadvantage: Less conservative of tooth structure
Extends preparation apically
Indication: Facial margin of posterior metal -ceramic crown with
supragingival margins. 37
Thus if not properly prepared, restoration margins can provide a
potential pathway to leakage of cariogenic micro -organisms present in
normal human flora. Failure of fixed partial dentures is most frequently
caused by caries. Therefore, physically superior cement materials play
an important role in caries prevention and lon gevity of cemented
restoration. Furthermore, it would be advantageous such materials
possessed antibacterial properties. 34
The antibacterial activity of dental cements has been intensively
tested. The direct-contact test (DTC was developed by "$ to
evaluate antibacterial activit y of non- soluble materials. This test is
based on measuring the effect of physical direct contract between test
bacteria and the tested material on the kinetics of bacterial outgrowth.
The DCT, was used to study the antibacterial properties of endodontic
sealers and pit and fissure sealants and the luting cements used to lute
fixed prosthesis to the prepared tooth. The purpose of the present
study was to assess and compare the incessant antibacterial activity of
3 luting cements mixed in 2 powder/liquid ratios using DCT and ADT.
 c
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 [
$  depicts the Bacterial growth rate using DCT as
demonstrated by the slope of the linear portion of the growth curve .
Each number is average (x0 -2 SD (x0-3 of slope of bacterial growth
measured in 8 separate wells in same microtiter plate at different time
intervals.
Two different P:L ratios (designated as  and 2 for each cem ent i.e.
Ketac-Cem, Harvard and Poly F, have been depicted in immediate,
24hours, week,  month and 3 months duration of time.
The gradual decrease in viable bacteria, due to serial dilutions at
time zero (0 , had virtually no effect on the bacterial growth rate or final
density of bacteria at the stationary phase in the system &"(
""  '$  $34 which indicates that the different P: L
ratios had a negligible effect on the antibacterial properties of the
tested cements. This is the reason that the readings of different
dilutions for the same cement could be clubbed for the result purposes
without any deviation in the final resu lts.
The results obtained above are in agreement with the studies of

"!+,")/"&,"!6· observed the

antibacterial potential of polycarboxylate temporary cement containing
in mixture of Metronidazole, Ciprofloxacin, and Cefaclor on carious
lesions of prepared abutments that were designed to leave caries on
the abutments. 20
,  $ who compared the antibacterial activity of various
restorative study materials with cariogenic bacteria using the ADT. The
authors found a wide range of antibacterial activity of different cement
while on 2% of Ketac cement specimens showed positive antibacterial
properties. The antibacterial activity of cement could not be attributed
 c
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to fluoride contents because not all of the fluoride containing cements
2
and antibacterial activity.
To support above study,  "/+ %   $ 6· who
compared the fluoride concentra tions in plaque on  year resin
modified GIC, compomer and resin composite restoration intra
individually and related to the occurrence of caries -associated bacteria.
Low fluoride levels were detected in all the cements, while the resin -
modified GIC cements showed significantly higher amounts. The
results indicated that the fluoride concentration released in vivo from  -
year old restoratives are not high enough to effect the plaque levels of
the caries associated bacteria, ! 
$%
In the $ , the samples (0 of the two P:L ratios of each test
cement specimen were added forming sample size of 20 for each
material (eg. KC +KC2=KC . It was illustrated that the mean value of
optical density was 0.002674 for immediate group -, 0.0005776 for
24hours group, 0.00088
37 for  week group, 0.0022723 for  month
group and 0.00266 for the 3 months group.
As shown in $  by applying the ONE WAY ANOVA test, it was
seen that there is statistically significant difference between the optical
densities of Ketac Cem, Harvard and Poly F for each given time group.
This is illustrated by the fact that the value of p¶ being <0.05 which
states that the result is significant.
$ analyzes the differences in the mean values of different test
cements from other test cements for each time group. The values are
found to be different with a wide range among themselves as is also
depicted in table III by the p¶ factor <0.05.
The results are confirmed by *$$ who examined the factors
involved in the antibact erial activity of various GICs on  
 c
c
 ‰
. The authors found that the KETAC -CEM cement exhibited low
antibacterial properties which could be attributed to low fluoride release
of the cement.
Another explanation could be the lack of zinc oxide powder in its
composition. It has been suggested that GICs containing zinc oxide
powder are more effective in microbial inhibit due to direct effect of zinc
oxide powder and cationic effect of zinc (Zn 2+ .
Also, '*  6† assessed the antibacterial properties of
dental restorative materials. Release of zinc and fluoride from these
materials was also measured and correlated with the antibacterial
activity. They found that there was a general trend towards greater
antibacterial activity with zinc release, while fluoride release had a
significant effect on only one organism. 2
However, certain authors such as #+&
 ) $ 


# & + '' 6† disagree and concluded from
their studies that all glass-ionomer cements showed significantly
greater overall inhibition of microbial growth and were more toxic
towards        
$& In other
study$"+Ê%#'%$6- measured the inhibition
of growth and adherence of five oral bacteria by glass - ionomer
materials. The two cavity liners and two of the restorative produced the
largest growth inhibition areas by Direct Contact. No growth inhibition
occurred when the specimens were allowed to come in cont act with the
agar prior to inoculation. All four restorative materials reduced bacterial
accumulation on enamel surfaces by over 80%. Elevations in short -
term release levels were positively correlated with growth inhibition. 5
And  , %+" %   $ 6 compared the levels of
fluoride and   in plaque grown on glass -ionomer
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c
 ·
and composite restoration in vivo and concluded from his study that the
fluoride level in the plaque growing on Glass -io
nomer is much higher
than that on composite restorations which seems to affect the level of
  in dental plaque. 4
$ depicts the Agar Diffusion Test in which the diameter of the
halo formed in the bacterial lawn was measure(mm in 2 perpendicular
locations for each specimen for each given time intervals.
Two different P:L ratios (designated as  and 2 for each cement i.e.
Ketac-Cem, Harvard and Poly F, have been depicted in immediate, 24
hours, week,  month and 3 months duration of time.
$ studied the short acting antibacterial effect of polycarboxylate
cement (PCC and zinc phosphate cement (ZPC and found that in the
agar diffusion test (ADT , ZPC exhibited the strongest antibacterial
properties, while PCC appeared to be the strongest against
Streptococcus mutans in vivo. In another study, the antibacterial
activity of several glass ionomer cements (GICs , dentin bonding
systems, and luting agents were investigated using the ADT and
growth-inhibition tests. Marked antibacterial activity was shown with the
GIC, while amalgam, composites, luting agents, and dentinal bonding
systems did not affect the bacterial growth. However, the antibacterial
activity of GIC ranged from nonactive to very effective (or high
activity . 34
In the $  the samples (6 of the two P:L ratios of each test
cement specimen were clubbed forming sample size of 2 for each
material (eg. KC +KC2=KC . It was illustrated that the mean value of
the diameter of the bacterial inhibition zone was 5.5536 for immediate
group, 5.48 for 24 hrs g roup, 5.364 for  week group, 5.
03 for 
month group and 5.0775 for the 3 months group. As shown in $
 c
c
 †
by applying the ONE WAY ANOVA test, it was seen that there is not
any statistically significant difference between the bacterial inhibition
zones of Ketac Cem, Harvard and Poly F for each given time group.
This is illustrated by the fact that the value of p¶ being>0.05 which
states that the result is not significant.
The lack of evidence for antibacterial activity for all the test cements
using ADT in the present study may be attributed to the characteristics
of this test. ADT is a popular test for antibacterial properties of
restorative material. However, it has several disadvantages. One
primary disadvantage is that it depends on the solubility and diffusion
properties of both the test material and the media. It is possible that the
antibacterial activity of the tested cements, which were not amply
soluble, could not be detected by ADT   $ found low
antibacterial activity for Polycarboxylat e and Zinc Phosphate, whereas
$ found high activity for these cements, For restorative materials
and cements, which are expected to have low solubility and are less
likely to diffuse, the ADT might not detect antibacterial properties.
Furthermore, the results demonstrated that DCT, which relies on direct
and close contact between the test micro-organism and the test
material, independent of the diffusion properties, may be more suitable
for testing restorative materials and cements than ADT. With respect to
caries prevention at the margins of cemented restorations, the DCT
simulates the clinical situation in which the cariogenic micro -organisms
are in contact with the cement better than the ADT. However, it must
be noted that the DCT experimental design employed is still far from
the clinical situation in terms of oral environment, margin location, and
surface area of cement exposed at restoration margins,. Furthermore,
the antibacterial effect in the present study was observed with respect
 c
c
 
to   , which is the most common caries-related
micro-organism. Since restoration margins may be subgingivally
located, it would be of interest to investigate the antibacte rial effect of
these cements on the subgingival flora. The prolonged antibacterial
activity found in this study and the initial low pH of Polycarboxylate
cement and Zinc Phosphate cement could indicate that these cements
have the potential for bacterial eradication in prepared abutments and
carious dentin. 34
$ analyzes the differences in the mean values of different test
cements from other test cements for each time group. The Values are
not found to be significantly different among themselves as is also
depicted in table VII by the p¶ factor >0.05.
The results obtained are in accordance with  $! 
  $ 6-

  who used Agar Diffusion test to evaluate the
antibacterial activity of six products (one conventional glass-ionomer
cement(GIC , two light activated glass-ionomer cements, two poly acid-
modified resin composites and one resin composite on  

All The GICs demonstrated antibacterial properties in contrast
to the polyacid-modified resin composite and resin composites which
did not show any antibacterial effects. 33
However, " 
)"% disagreed by comparing
antibacterial properties of 4 different restorative materials, nine of
which were GICs. The materials were mixed according to
manufacturer¶s specifications and exposed to four types of bacteria
commonly found in caries and plaque. Zones of bacterial inhibition
were measured for all materials in millimeters and they found that GIC
materials containing zinc oxide and amalgam produced measurable
zone of inhinbition .
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The r-  (  justifies the results obtained in the tables. As is
clearly seen, the optical density of Poly F is remains zero meaning that
it has the highest antibacterial property throughout the given time
durations. The Harvard has zero optical density for immediate, 24hrs
and  week duration depicting to be possessing antibacterial property
for that duration after which the antibacterial property declines. The
Ketac-Cem did not show any antibacterial activity for any of the time
groups.
The results can be depicted also by the line graphs as in line
graphs as in $" (     the bacterial outgrowth in microtiter
wells could be monitored in a quantitative and reproducible manner.
Each point on growth curve is an average of the optical densities
measured in 4 wells at a given same time. The results of DCT for
freshly mixed and aged cements in different P:L ratios are shown. The
immediate curves demonstrate antibacterial activity of Poly F and
Harvard cement with ( (  immediate A and without 6( 
immediate B the presence of the cement specimens. Bacterial growth
was significantly reduced when compared with the control (P=0.002 .
Ketac Cem did not show antibacterial activity for any of the test groups.
Prominent antibacterial activity was evident for the aged Poly F and
Harvard cements along the test periods 6(  - . However,
Harvard cement became less antibacterial than the Poly F after 3
months. The different P: L ratios had a negligible effect on the
antibacterial properties of the tested cements. The ADT showed no
antibacterial activity of the te3sted cements for any of the test groups.
In the present study, according to ""  '$   $
POLY F and HARVARD cements maintained antibacterial properties
even after three months of aging in Phospated Buffered Saline
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‰
(PBS .The prolonged antibacterial a ctivity was measurable only in
Direct Contact between bacteria and the cements, whereas in agar
plates, no diffusion of antibacterial components from the cements was
observed. This may confirm that the cement specimens are stable with
low solubility in an aqueous environment and do not excrete sufficient
antibacterial substances to the surrounding media to effect the
bacterial growth.
The limitations of this study include the two P:L ratios of the test
cements may not be so accurately measured thus leading to not exact
proportions as mentioned in the study. The temperature of the
operatory may be an environmental factor affecting the growth of the
bacteria thus affecting some of the results of cements. Not proper
manipulative or technical procedures may in culate inaccuracies in
study affecting the result significantly.
Many studies have tested the antibacterial activity of dental cements
and demonstrated that GIC is the most active (antibacterial cement ,
followed by Zinc Phosphate Cement and PolyCarboxylate Cement. On
the contrary, in the present study using the DCT, POLY F was the most
active followed by the HARVARD cement whereas KETAC CEM
showed no antibacterial activity even after 24 hours. In addition to that
the dilution of cements did not seem to affect the antibacterial
properties of the cements. 34





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''
.
Ê
Ê

Since failure of fixed partial dentures is most frequently caused by
caries, it would be advantageous if cement possessed antibacterial
properties.
For the direct-contact test, wells (n=4 of microtiter plates were coated
with the tested cements (Harvard cement, Poly F, and Ketac -Cem
while Streptococcus mutans suspension was placed directly on the
cements. Bacterial growth was evaluated by a temperat ure-controlled
microplate spectrophotometer. Eight wells of bacterial without the
tested cements served as the positive control. Six wells of the tested
cement without bacteria served as the negative control.
For the agar diffusion test, triplicate speci mens of freshly mixed
cements were poured into uniform wells (5 mm in diameter punched in
the agar plates inoculated with Streptococcus mutans. After incubation
at 370C for 24 hours, the agar plates were examined for bacterial
growth and the diameter of the halo formed in the bacterial lawn was
measured.
In both tests, each cement was mixed in 2 different powder/ liquid
ratios. For the direct-contact test, data were initially recorded after 
hour of incubation. Additional experiments were performed on
specimens that were aged for 24 hours,  week,  month, and 3
months before assessment by either direct -contact test or agar
diffusion test. The data were subjected  -way ANOVA with the POST
HOC test (Į=.05 .
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Ê
Ê

Compared with the control group and within the limitations of this study,
the following conclusions were drawn:
 No correlation was found between the DCT and the ADT.
2 DCT was more effective in detecting antibacterial properties of
definitive dental cements than ADT.
3 Using DCT, aged Poly F and Harvard cements demonstrated
antibacterial properties even after 3 months.
4 Antibacterial properties of the tested cements were not detected by
ADT.
5 The different P/L ratios had a negligible effect on the antibacterial
properties of the tested ce ments.
6 Ketac-Cem exhibited no antibacterial activity for any of the test groups.
 c
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Ê
 Herrrera M, Castillo A, Baca P, Carrion: Antibacterial activity of Glass -
Ionomer restorative cements exposed to cavity - producing
microorganisms. Operative Dentistry 


; 24:286 -2
.
2 Eli I, Cooper Y, Ben-Amar A, Weiss E: Antibacterial activity of three
dental liners. J Of Prosthodontics 

5;4:78 -82.
3 Palenik CJ, Behnen MJ, Setcos JC,Miller CH: Inhibition of microbial
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