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Antimicrobial activity of chlorophyll-based solution on Candida albicans and Enterococcus faecalis

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Abstract

Chlorophyll is a phytotherapic substance that presents curingproperties, however it is rarely used in Dentistry. Objective Objective Objective Objective Objective: to analyze the antimicrobial activity of a chlorophyll-based solution on isolates of Candida albicans and Enterococcus faecalis by agar dilution method.Material and Methods: oral isolates and Candida albicans referencestrain (ATCC 18804) and Enterococcus faecalis (ATCC 29212) wereincluded in the study. Chlorophyll extract was diluted in Sabourauddextrose or brain heart infusion (BHI) agar, according to the testingmicroorganism and poured in Petri plates, obtaining the finalconcentrations of 50%, 25%, 12.5%, 6%, 3% and 1.5%. C. albicansand E. faecalis strains were plated with the aid of a Steers’ inoculator.Plates were incubated at 37°C for 48 h for C. albicans and at 5% CO2 for E. faecalis. The experiments were performed in duplicate. Results:the results showed that the concentrations 50% and 25% chlorophyll extract were effective against C. albicans. E. faecalis grew in the presence of all the concentrations.Conclusion: chlorophyll-based solution presents effective antimicrobial activity of C. albicans but did not present any activity on E. faecalis.
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Eiko MAEKAWA, Lilian; LAMPING, Roberta; MARCACCI, Sidnei; Yasunori MAEKAWA,
Marcos; Giazzi NASSRI, Maria Renata; Yumi KOGA-ITO, Cristiane
Antimicrobial activity of chlorophyll-based solution on Candida albicans and
Enterococcus faecalis
RSBO. Revista Sul-Brasileira de Odontologia, vol. 4, núm. 2, 2007, pp. 36-40
Universidade da Região de Joinville
Brasil
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ISSN (Versión impresa): 1806-7727
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Antimicrobial activity of chlorophyll-based solution
on
Candida albicans
and
Enterococcus faecalis
Atividade antimicrobiana da solução de clorofila
sobre
Candida albicans
e
Enterococcus faecalis
Lilian Eiko MAEKAWA*
Roberta LAMPING**
Sidnei MARCACCI**
Marcos Yasunori MAEKAWA***
Maria Renata Giazzi NASSRI****
Cristiane Yumi KOGA-ITO*****
Correspondence:
Lílian Eiko Maekawa
Rua Evolução, 692 – Vila Brasilina
São Paulo – SP – CEP 04163-001
E-mail: lilian.maekawa@uol.com.br
* Post-Graduated Student – Master Restorative Dentistry Program-Dental Endodontics Especiality – School of Dentistry of São José dos
Campos – UNESP.
** Dentists.
*** Phd, Professor – Assistant Professor, Department of Dental Material and Prosthodontics – School of Dentistry of São José dos
Campos – UNESP.
**** Phd, Professor – Assistant Professor, discipline of Endodontics, School of Dentistry, University of Mogi das Cruzes.
***** Phd, Professor – Assistant Professor, department of Oral Bioscience and Diagnosis, School of Dentistry of São José dos Campos – UNESP.
Recebido em 22/8/07. Aceito em 17/10/07.
Abstract
Chlorophyll is a phytotherapic substance that presents curing
properties, however it is rarely used in Dentistry. ObjectiveObjective
ObjectiveObjective
Objective: to analyze
the antimicrobial activity of a chlorophyll-based solution on isolates
of
Candida albicans
and
Enterococcus faecalis
by agar dilution method.
Material and MethodsMaterial and Methods
Material and MethodsMaterial and Methods
Material and Methods: oral isolates and
Candida albicans
reference
strain (ATCC 18804) and
Enterococcus faecalis
(ATCC 29212) were
included in the study. Chlorophyll extract was diluted in Sabouraud
dextrose or brain heart infusion (BHI) agar, according to the testing
microorganism and poured in Petri plates, obtaining the final
concentrations of 50%, 25%, 12.5%, 6%, 3% and 1.5%.
C. albicans
and
E. faecalis
strains were plated with the aid of a Steers’ inoculator.
Plates were incubated at 37°C for 48 h for
C. albicans
and at 5% CO2
for
E. faecalis
. The experiments were performed in duplicate. Results:Results:
Results:Results:
Results:
Keywords:
chlorophyll; endodontics;
Candida albicans
;
Enterococcus faecalis
.
Artigo Original de Pesquisa
RSBO v. 4, n. 2, 2007 – 37
the results showed that the concentrations 50% and 25% chlorophyll
extract were effective against
C. albicans
.
E. faecalis
grew in the
presence of all the concentrations. Conclusion:Conclusion:
Conclusion:Conclusion:
Conclusion: chlorophyll-based
solution
presents effective antimicrobial activity of
C. albicans
but did
not present any activity on
E. faecalis
.
Resumo
A clorofila é uma substância fitoterápica que possui propriedades
curativas, apesar de ainda ser pouco utilizada na odontologia. O
presente trabalho objetivou analisar a atividade antimicrobiana do
extrato de clorofila sobre amostras de C
andida albicans
e
Enterococcus faecalis
, utilizando o método de diluição em ágar.
Foram incluídos no estudo isolados bucais e amostra-padrão de
Candida albicans
(ATTC 18804) e
Enterococcus faecalis
(ATCC
29212). O extrato de clorofila foi diluído em ágar Sabouraud
ou
Triptic soy, de acordo com o microrganismo a ser testado, e vertido
em placas de Petri, obtendo concentrações finais de 50%, 25%, 12,5%,
6%, 3% e 1,5%. As amostras de
Candida albicans
e
Enterococcus
faecalis
foram semeadas nas placas com auxílio de replicador de
Steers. Em seguida, as placas de Petri com
C. albicans
foram
incubadas a 37°C por 48 horas; para as de
E. faecalis
procedeu-se
do mesmo modo, em 5% de CO2. Os experimentos foram realizados
em duplicata. Os resultados mostraram que as concentrações de
50% e 25% do extrato de clorofila foram efetivas nas amostras de
Candida albicans
, entretanto nas concentrações de 12,5%, 6%, 3% e
1,5% observou-se crescimento.
Enterococcus faecalis
desenvolveu-
se em todas as concentrações. Foi possível concluir que o extrato de
clorofila, diluído em até 25%, tem efetiva ação antimicrobiana sobre
Candida albicans
, não apresentando atividade sobre
Enterococcus
faecalis
nas concentrações testadas.
Palavras-chave:
clorofila; endodontia;
Candida albicans
;
Enterococcus faecalis
.
Introduction
The aim of chemical and surgical preparation
is to clean and to disinfect the root canal, aiming to
maintain the conditions obtained by the use of
irrigation solutions. Also, intracanal medicaments
and obturing pastes are important to difficult the
metabolic changes between the oral milieu, root canal
and support tissue, blocking the recontamination
by the microorganisms presents in the oral
microbiota, that is related to treatment failures.
Endodontic infections are considered
polimicrobial and more than 150 bacterial species
are usually found in combinations of 3 to 6 species
in each canal [19]. Also, microorganisms such as
yeasts may be commonly found in root canals with
pulp necrosis [6, 12].
Najzar-Fleger
et al
. (1992) [13], studying the
prevalence of
Candida
genus in different sites of the
oral cavity, verified that 55% of the root canals
presented these microorganisms. Maekawa
et al
.
(2006) [9] analyzed the microbiota from the root
canals of teeth with pulp necrosis and showed that
in 15.3% of the cases
Candida albicans
was
identified.
Enterococcus faecalis
is also frequently
isolated from root canals in cases of pulp infection
and also recalcitrant infections after endodontic
treatment [22, 8].
Waltimo
et al
. (2000) [24] studied the penetration
of
Candida albicans
in dentine compared with
Enterococcus faecalis
through an
in vitro
model in
human dentine. An effective and deep penetration was
observed in the specimens infected with
E. faecalis
.
The authors concluded that, although
C. albicans
penetration is possible, that is less significant in
relation to the penetration ability of
E. faecalis
.
Rodrigues (2001) [15] related that enteric
bacteria, mainly
E. faecalis
, and yeasts, in particular
of the
Candida
genus, are commonly isolated from
recalcitrant endodontic infections. This microbiota
may characterize monoinfection, with the
predominance of Gram-positive microorganisms
Maekawa et al.
Antimicrobial activity of chlorophyll-based solution on Candida albicans and Enterococcus faecalis
38
particularly facultative anaerobes. These are more
resistant to antimicrobial procedures performed
during the endodontic treatment and can establish
themselves without other microorganisms.
Several studies have been developed about the
antimicrobial activity of irrigation solutions in
Endodontics. Due to several properties, such as
ability to solve organic material and neutralize toxic
content, sodium hypochlorite is the most frequently
employed substance during the instrumentation and
irrigation of root canals. It is commercially available
at the concentrations of 0.5%, 1%, 2.5% and 5.25%
[23, 11, 10, 2, 14, 4, 21, 20]. Cardoso (2003) [1],
studying the dentinal permeability of the cervical,
medium and apical thirds of the root canals after
the rotatoty instrumentation with different auxiliary
chemical substances, concluded that the use of
Endo-PTC associated to Dakin’s solution increased
the dentinal permeability when compared with EDTA
17% and chlorhexidine 2%. Despite of its advantages,
the use of high sodium hypochlorite concentrations
may cause injuries in periapical tissues [7, 21], and
alternative solutions have been proposed aiming the
association between antimicrobial effectiveness and
biocompatibility [14].
Chlorhexidine 2% is also largely employed. Many
studies proved its effectiveness on several
microorganisms such as
Fusobacterium nucleatum
,
Prevotella nigrescens
,
Staphylococcus aureus
, and
E. faecalis
[23, 10, 3].
Chlorophyll is a phytotherapic substance that
presents curing properties but is scarcely known and
studied in Dentistry. Santiago
et al.
(2002) [17],
testing chlorophyll solutions, concluded that it might
increase the dentinal permeability, an important
characteristics in the endodontic therapy. Guillén-
Burgos
et al
. (2004) [5] evaluated the cytotoxicity of
chlorophyll extract and verified that it can be well
tolerated in cell culture by fibroblasts of periodontal
ligament, in short and long experimental periods.
Saeki
et al
. (1993) [16] evaluated the
antimicrobial effects of the green tea, chlorophyll and
sodium copper chlorophyll on oral bacteria and
concluded that all the tested substances were effective
but each of them presented different specificities.
Chlorophyll inhibited the growth of the tested oral
bacteria, particularly
Porphyromonas gingivalis
and
Fusobacterium nucleatum
. Copper chlorophyll did
not pesent inhibitory activity on those bacteria.
Perin (2001) [14] tested a lyophilized
phytotherapic solution obtained from the extract of
Arctium lappa
as an auxiliary substance for the canal
preparation and concluded that the tested
substances were effective against
C. albicans
but did
not present any effect on
E. faecalis
,
Pseudomonas
aeruginosa
and
Staphylococcus aureus
. The
phytotherapy substance presented antifungal activity
against
C. albicans
, while that other cepas had
remained viable.
In Endodontics, chlorophyll can be indicated as
an auxiliary antimicrobial solution to assist in the
chemical and surgical preparation and to increase
the permeability of the dental tubules. Knowing the
importance of the removal of the smear layer,
originated during the instrumentation due to the
action of the instruments cut and being able still to
be contaminated, it seems interesting to use a
solution provided with antimicrobial effect in the
removal of smear layer.
Based on the findings from the literature and the
lack of data regarding the chlorophyll and its uses in
Dentistry, the present study aimed to evaluate
in vitro
the antimicrobial activity of chlorophyll-based solution
on
Candida albicans
and
Enterococcus faecalis
.
Material and methods
The solution tested in this study was composed
by sodium copper chlorophyllin (Ervas Vida, Vitória-
ES, Brazil). Thirty-two oral isolates and reference
strain (ATCC 18804) of
C. albicans
and
E. faecalis
ATCC 29212 were included in the study.
Firsty, the isolates were plated on Sabouraud
dextrose agar (Difco, Detroit, USA) or brain heart
infusion agar (BHI, Difco, Detroit, USA) and incubated
at 37°C for 24 hours, and 37°C/48h/5% CO2 for
E.
faecalis
. Then, standardized cell suspension
containing 106 cells per milliliter was turbidimetrically
obtained in sterile saline solution (NaCl 0.85%).
Plates containing serial dilutions (50%-1.5%)
of the testing solution in Sabouraud dextrose agar
(for
C. albicans
tests) and BHI agar (for
E. faecalis
)
were obtained.
Isolates were plated on each plate with the aid
of a Steers’ inoculator (Figure 1). Then, plates were
incubated at 37°C for 48 hours (
C. albicans
) and
37°C/48h/5% CO2 for
E. faecalis
. All the experiments
were performed in duplicate. After the incubation
period, readings were based on the growth of the
isolates in the presence of the dilutions tested.
Aiming at characterizing the chlorophyll-based
solution tested, it was submitted to pH measurement
(CG710, Schott Mainz, Switzerland).
Results
The concentrations of 50 and 25% of the tested
chlorophyll-based solution tested inhibited the
growth of
C. albicans
isolates. The concentrations
RSBO v. 4, n. 2, 2007 – 39
of 1.5% to 12.5% did not show any antifungal activity.
No acitivity was observed on
E. faecalis
. The pH of
the tested solution was of 5.4.
Discussion
Several studies on the antimicrobial activity of
irrigation solutions in Endodontics, such as 0.5%,
1%, 2.5% and 5% sodium hypochlorite and 2%
chlorexidine are found in the literature [23, 14, 4,
21]. On the other hand, the lack of studies on
phytotherapic substances such as chlorophyll,
propolis and castor bean detergent do not permit
more objective conclusions about their use [14, 3].
The inclusion of
C. albicans
and
E. faecalis
in
this study was based on the literature that relates
these microorganisms to pulp infections, mainly in
recalcitrant infections after endodontic treatment
[13, 22, 6, 24, 15, 9, 8].
Chlorophyll is cited as a phytotherapic substance
provided of several benefits to our organism such as
the prevention of anemia, detoxifying effect and
improvement of blood circulation (Ervas Vida, Vitória-
ES, Brazil). Considering these benefits already studied
in the medical area, the interest on chlorophyll is now
spreading to Dentisty. However, few studies are still
found in the literature.
The interest on this substance is based on these
already described properties. Moreover, relates on
an increased dentinal permeability stimulated the
study of this substance. The increase in the
permeability of the dentinal canals may be
considered a very interesting characteristic and it is
related to better cleaning activity and adaptation of
the obturing material.
Santiago
et al.
(2002) [17] observed
in vitro
an
increase in the permeability of dentinal canals when
compared chlorophyll with Clorox and Dakin’
solution and it was indicated to be used during the
chemical-surgical preparation of root canal.
Also, Guillén-Burgos
et al
. (2004) [5], in
experiments of biocompatibility with chlorophyll,
concluded that this extract is biocompatible in
human periodontal lligament fibroblast culture at
short and long periods.
The methodology of this study followed the
standard established for agar dilution tests [18]. This
method permits more objective growth control and
the use of Steer’s replicator makes the testing of
several samples possible at the same time.
The results obtained in this study showed that
the chlorophyll-based tested solution was effective
against
C. albicans
at the concentration of 25%,
however no activity was observed against
E. faecalis
.
Considering that
C. albicans
is a common
microorganism in pulp infections, this study suggests
that in certain concentrations, chlorophyll might be
indicated as a good choice as irrigation solution. On
the other hand, the results obtained for
Enterococcus
faecalis
were not so promising. Bacterial growth was
observed in all the concentrations, however more
studies on this subject are necessary.
Another fact to be considered is the value of pH,
since it is related in the literature that an alkaline
permits a better infection control interfering on
microbial growth and survival [7]. The pH observed
for chlorophyll was 5.6 that is contrary to this
affirmative. On the other hand, this result corroborates
the study of Guillén-Burgos
et al.
(2004) [5], which
states that this pH value shows that chlorophyll has
low damage potential for periapical tissues when
compared to 0.5% sodium hypochlorite (pH 11.8), 24%
EDTA (pH 7.2) and 15% EDTA (pH 7.4).
Considering the results obtained and the
characteristics cited before, the data suggest that
chlorophyll extract may have a clinical applicability
as irrigation solution of the root canal if used in
concentrations that might compensate its low
antimicrobial potential and might preserve the good
cell tolerance and capacity of permeate the dentinal
canals. More studies for searching this formulation
are necessary.
Conclusions
The chlorophyll-based tested solution presented
antifungal activity against
C. albicans
at the
concentration of 25%. No activity on
E. faecalis
was
observed.
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... Enterococcus faecalis is also frequently isolated from root canals in cases of pulp infection and also recalcitrant infections after endodontic treatment. 9 Propolis is a natural flavonoid-rich resinous product of honeybees, which is known for its biological properties, including antimicrobial, antifungal and healing properties. 10 It was suggested that flavonoids from propolis may stimulate reparative dentine formation and may delay pulp inflammation by stimulating production of transforming growth factor (TGF)-b1 and synthesis of collagen by dental pulp cells. ...
... 17 The inclusion of C. albicans and E. faecalis in this study was based on the literature that relates these microorganisms to pulp infections. 9 The agar diffusion inhibitory test (ADT), or Lawn's technique has been used for long time as the standard test for antibacterial activity of dental materials. 18 The agar diffusion test does not distinguish microbiostatic and microbicidal properties of dental materials neither does it provide any information about the microorganisms viability after the test. ...
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Aim To evaluate and compare antimicrobial effect of various root canal medicaments against Enterococcus faecalis , Staphylococcus aureus and Candida albicans . Materials and methods Six root canal medicaments: 2% sodium hypochlorite (NaOCl), 2% chlorhexidine (CHX) Calcium hydroxide (Ca(OH) 2 ), EDTA, MTAD and propolis and three microorganisms: Staphylococcus aureus , Enterococcus faecalis and Candida albicans were used. These strains were inoculated in brain heart infusion (BHI) and incubated at 37 degrees C for 24 hours. For the agar diffusion test (ADT), petri plates with 20 ml of BHI agar were inoculated with 0.1 ml of the microbial suspensions, using sterile swabs that were spread on the medium, obtaining growth injunction. Paper disks were immersed in the experimental solutions for 1 minute. Subsequently, four papers disks containing one of the substances were placed on the BHI agar surface in each agar plate. The plates were incubated at 37°C for 48 hours. The diameter of microbial inhibition was measured around the papers disks containing the substances. One way ANOVA followed by Tukey's post-hoc test were used. p-value <0.05 was considered statistically significant. Results Propolis and other irrigants were found to be effective on C. albicans , S. aureus and E. faecalis . CHX and MTAD were found to be most effective amongst all the materials tested followed by propolis. Conclusion Propolis showed antimicrobial activity against E. faecalis , S. aureus , C. albicans . It appears that propolis is an effective intracanal irrigant in eradicating E. faecalis and C. albicans . Clinical significance Propolis is an effective intracanal irrigant in eradicating E. faecalis and C. albicans . It could be used as an alternative intracanal medicament. How to cite this article Mattigatti S, Jain D, Ratnakar P, Moturi S, Varma S, Rairam S. Antimicrobial Effect of Conventional Root Canal Medicaments vs Propolis Against Enterococcus faecalis , Staphylococcus aureus and Candida albicans . J Contemp Dent Pract 2012;13(3):305-309.
... Candida albicans is the most common opportunistic organism in oral cavity and it is also seen in infected root canal. 5 In this dynamic microbial environment, selection of an effective antimicrobial agent to treat infection is critical. Moringa, Laxmitaru, Mullatha, and Communist paccha are medicinal plants which are used to treat various health problems. ...
... algarbiensis and C.suffruticosa subsp. lusitanica, respectively), but these low yields may probably due to high concentration of chlorophyll, which is responsible by the dark green colouration of the extracts (Maekawa, Lamping, Maekawa, Nassri & Koga-Ito, 2007). Since this was a comparative study between the Calendula taxa, the chromatographic conditions chosen were the same for each extract. ...
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Calendula officinalis is an edible and medicinal plant with recognised economic value. What about the other Calendula species? This work aims to provide in-depth information about the UHPLC-MS/MS profile of other Portuguese Calendula species and in doing so contribute to answer that question. The knowledge on the metabolic machinery from C. arvensis, C. suffruticosa subsp. algarbiensis and lusitanica growing in Portugal is herein broadened using UHPLC-MS/MS. Phenolic acids, flavonoids and saponins were identified, some being firstly reported both in the species and genus. The antimicrobial potential of these species was assessed against several fungus and bacteria, strong activity against dermatophytes was observed. C. suffruticosa subsp. algarbiensis showed the best results especially against the two strains of Microsporum. The demonstrated antimicrobial activity and the richness in phenolic compounds reinforces that other Calendula species can be regarded as interesting ingredients to be included on the formulation of smart foods and/or natural medicines.
... Leaves of the henna are strikingly most effective against the spectrum of bacterial isolates tested. This may be attributed to the added presence of chlorophyll, henna leaves contain up to 5% by weight of the compound (2-hydroxy-1,4-naphthoquinone), chemically active constituents such as quinines (O. A. Habbal, Al-Jabri, El-Hag, Al-Mahrooqi, & Al-Hashmi, 2005;Maekawa et al., 2007). ...
... The level of total serum protein and bilirubin were estimated by Biuret method [15] and according to the developed protocol of Maekawa et al. [11]. ...
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This study was aimed to evaluate the pharmacological potential of various extracts (hexane, chloroform, methanol and aqueous) of dried shoots of Triticum aestivum (wheat grass) in terms of antiproliferative and hepatoprotective potential of T. aestivum. The total chlorophyll content in dried shoots of T. aestivum was 0.54±0.016 g/L (chlorophyll-a: 0.288±0.05 g/L; and chlorophyll-b; 0.305±0.05 g/L), while total carotene content was 0.42±0.066 g/L. In addition, the chloroform extract of dried shoots of T. aestivum (250 (tg/mL) exhibited 87.23% inhibitory effect with potent cytotoxicity against human hepatocellular carcinoma (HepG2) cancer cell line. Moreover, chloroform and methanol extracts significantly reduced the levels of SGOT, and SGPT enzymes, as well as total bilirubin content, while raised the level of total protein in a concentration-gradient manner, confirming the potent hepatoprotective effect of T. aestivum. A possible mechanism of apoptosis of the chloroform extract of dried shoots of T. aestivum in terms of its potent antiproliferative activity against HepG2 cancer cell line can also be proposed in this study. Our findings clearly demonstrate that T. aestivum has a significant pharmacological potential that night be used for antiproliferative and hepatoprotective purposes.
... Jayavanth et al. 19 reported the antibacterial activity of chlorophyll in pure form against Gramnegative bacteria. Lilian et al. 20 analyzed the antimicrobial activity of a chlorophyll-based solution on isolates of Candida albicans and Enterococcus faecalis and concluded that the chlorophyll-based solution possessed potent antimicrobial activity. ...
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India is a rich and varied flora of medicinal plants since the Vedic period. Present study deals with the qualitative and quantitative analysis of hexane extract of Triticum aestivum grass. A qualitative analysis by thin layer chromatography (TLC) and a quantitative analysis by standard chemical protocol of secondary metabolites in the hexane extract of Triticum aestivum grass were studied. Using TLC, different components such as chlorophyll and carotene were analyzed. The R f (0.28, 0.29, 0.37, 0.65, 0.74 and 0.93) values of the 6 developed spots in the hexane: ethyl acetate (16:4) solvent system were noted. In the quantitative analysis, chlorophyll and carotene content in T. aestivum was 0.54 ±0.016 g/l out of which chlorophyll-a, and chlorophyll-b were 0.288±0.05 and 0.305±0.05 g/l, respectively while carotene content was 0.42±0.066 g/l. The antimicrobial activity of hexane extract of Triticum aestivum grass against Salmonella Enteritidis showed potential effects with inhibitory zones in the range of 9-8 mm along with minimum inhibitory concentration (50 mg/ml) and reduction of cell count. These results may be helpful for rationale use of this plant in the modern system of health care.
... We conducted a study w it h a n induct ive approach, a comparative procedure and direct documentation technique in laboratory [9]. For in vitro antifungal assessment, Salvia officinalis (sage), Anacardium occidentale (cashew) and Malva sylvestris (mallow) tinctures (Farmácia Homeopática Homeov itae Ltd., João Pessoa, PB, Brazil) were used. ...
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Objective: To evaluate through determination of minimum inhibitory concentration (MIC) the antifungal activity of Salvia officinalis (sage), Anacardium occidentale (cashew) and Malva sylvestris (mallow) tinctures on Candida albicans (ATCC 40227), C. tropicalis (ATCC 13803) and C. krusei (ATCC 40147). Material and methods: In 96-well microplates, 100 μl of Sabouraud-Dextrose broth doubly concentrated, 100 μl of the tested tinctures and 10 μl of fungal inoculums (1.5 x 106 organisms/ml) were inserted. The products were diluted from initial concentration of 100 mg/ml until 0.78 mg/ml. MIC corresponded to the lowest dilution at which there was no visible fungal growth. Nystatin (100,000 UI/ml) was used as control. Statistical analysis was performed by Kruskal-Wallis and Dunn tests (p < 0.05). Results: S. officinalis tincture did not inhibit the growth of C. albicans and C. tropicalis; MIC was 100 mg/ml for C. krusei. For A. occidentale, MIC was 100 mg/ml for C. albicans and C. krusei, and for C. tropicalis, there was no fungal inhibition. M. sylvestris tincture presented MIC at 25 mg/ml for C. krusei and 100 mg/ml for C. albicans and C. tropicalis. The best antifungal activity was showed by M. sylvestris tincture (p < 0.05). Conclusion: M. sylvestris tincture exhibited antifungal activity against all the tested strains at lower concentrations. S. officinalis tincture inhibited the action of C. krusei and A. occidentale tincture showed activity against C. albicans and C. tropicalis.
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In this study, pectin was extracted from black mulberry fruit pulp and chlorophyll was extracted from black mulberry leaves. The extracted chlorophyll was encapsulated with carboxymethylcellulose (CMC). Black mulberry pectin/chlorophyll of black mulberry leaf encapsulated with carboxymethylcellulose/silica nanoparticles was prepared (Pec/Ch-CMC/SiO2). The physicochemical, mechanical and antimicrobial properties of the prepared films were investigated. SEM, FTIR, TGA and XRD techniques were used to study the structure of the films. According to the results, silica increased the thickness and decreased the moisture and solubility of the films. Increasing the amount of CMC in the capsules used in the film increased the thickness, moisture and solubility of the films. Silica increased the tensile strength of the film and reduced the flexibility of the film, while CMC reduced the tensile strength and increased the flexibility of the film. Chlorophyll and silica greatly increased the antioxidant properties of the film. Encapsulated chlorophyll and silica increased the antibacterial properties of the film against Escherichia coli and Staphylococcus aureus. SEM images showed that by simultaneously adding encapsulated chlorophyll and silica to the pectin structure, almost all gaps in the pectin surface were filled. According to the FTIR spectra, electrostatic interactions between pectin polymer chains with encapsulated chlorophyll and silica were confirmed. According to TGA results, the simultaneous addition of encapsulated chlorophyll and silica to the film increases the thermal stability of the film. According to the results, the pure pectin film has a semi-crystalline structure that the addition of silica and encapsulated reduces its crystalline structure.
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Biocompatibility of materials used in dental and biomaterials applications is very important and depends on the components of these materials. Photopolymerized materials for dental and biomaterials applications have been progressively used since the 1970s. One of the crucial components in these materials is the photoinitiator (PI) that initiates the polymerization reaction. Synthetic PIs are the most commonly used types, but owing to their drawbacks such as cytotoxicity, insolubility in water, and high cost, research on naturally derived (bio-sourced) PIs is growing, to find an alternative to these synthetic types, especially in the growing field of three-dimensional (3D) printing and bioprinting of biomaterials for tissue engineering applications. Naturally derived PIs are biocompatible, highly water-soluble, and abundant. Naturally derived PIs have been used to prepare experimental dentine bonding agents, dentine primers, photo-crosslinked hydrogels for tissue engineering applications, antibacterial coatings, guided tissue regeneration membranes, and 3D printed biomaterials. An electronic search was done using MEDLINE/PubMed and Scopus databases using the keywords naturally derived, bio-sourced, PIs, dental, biomaterials, 3D printing, and 3D bioprinting, to review potential naturally derived PIs for dental and biomaterials applications. There are a variety of naturally derived PIs with various colors and absorption spectra to choose from, according to the intended application. Most of naturally derived PIs can be used with modern conventional dental light curing units, making them applicable for experimental studies for potential dental and biomaterials applications. Due to their biocompatibility and availability it is expected that in the upcoming years, research on naturally derived PIs and their dental and biomaterials applications will increase especially in the growing field of 3D bioprinting in which cell viability is essential; thus this review was done.
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Statement of problem: Although different forms of denture adhesives are widely used by the elderly, insufficient information is available for the effect of adhesives on the growth of Candida species. Purpose: The purpose of this in vitro study was to compare the influence of 3 different forms of a commercially available denture adhesive material on the growth of Candida species by monitoring the pH and number of colonies in the growth medium at different incubation periods. Material and methods: A total of 160 specimens of acrylic resin sheets of standard dimensions were fabricated and divided into 4 groups of 40 specimens each. The first group of 40 specimens was used as a control to which no adhesives were added. To the remaining groups, powder, cream, and strip forms of secure denture adhesives were added. After the addition of standard strains of Candida, 10 specimens from each group were then incubated for a specific period of 6 hours, 24 hours, 48 hours, and 120 hours. The pH and the number of colonies in the medium were recorded for each specimen at its specific incubation period. The data obtained were statistically analyzed using ANOVA and the Tukey honest significant differences post hoc test (α=.05). Results: The specimens with different forms of adhesive showed a significant (P<.001) increase in the pH of the medium (pH 4.70 to 5.37) compared with the specimens without adhesives (pH 3.83 to 4.65). The specimens with adhesives also showed a reduction in the number of Candida colonies (216.50 to 388.10×10(3) cells/μL) compared with the specimens without adhesives (325.90 to 400.50×10(3) cells/μL). Conclusions: All forms of denture adhesives tested exhibited an antifungal effect. Prolonged antifungal effect was shown by the strip form of adhesives.
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The purposes of this study were to determine what microbial flora were present in teeth after failed root canal therapy and to establish the outcome of conservative re-treatment. Fifty-four root-filled teeth with persisting periapical lesions were selected for re-treatment. After removal of the root filling, canals were sampled by means of advanced microbiologic techniques. The teeth were then re-treated and followed for up to 5 years. The microbial flora was mainly single species of predominantly gram-positive organisms. The isolates most commonly recovered were bacteria of the species Enterococcus faecalis. The overall success rate of re-treatment was 74%. The microbial flora in canals after failed endodontic therapy differed markedly from the flora in untreated teeth. Infection at the time of root filling and size of the periapical lesion were factors that had a negative influence on the prognosis. Three of four endodontic failures were successfully managed by re-treatment.
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The choice of an irrigating solution for use in infected root canals requires previous knowledge of the microorganisms responsible for the infectious process as well as the properties of different irrigating solutions. Complex internal anatomy, host defenses and microorganism virulence are important factors in the treatment of teeth with asymptomatic apical periodontitis. Irrigating solutions must have expressive antimicrobial action and tissue dissolution capacity. Sodium hypochlorite is the most used irrigating solution in endodontics, because its mechanism of action causes biosynthetic alterations in cellular metabolism and phospholipid destruction, formation of chloramines that interfere in cellular metabolism, oxidative action with irreversible enzymatic inactivation in bacteria, and lipid and fatty acid degradation. The aim of this work is to discuss the mechanism of action of sodium hypochlorite based on its antimicrobial and physico-chemical properties.
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To evaluate the antimicrobial effect of the ozonized sunflower oil (Oleozon) on different bacterial species isolated from different sites. The effect of Oleozon on Mycobacteria, staphylococci, streptococci, enterococci, Pseudomonas and Escherichia coli was tested. The sunflower oil was ozonized at the Centro de Investigaciones del Ozone (CENIC, Havana, Cuba) by an ozone generator. MICs were determined by the agar dilution method. For Mycobacteria, the MIC of Oleozon was determined on solid medium by a microdrop agar proportion test. Oleozon showed antimicrobial activity against all strains analysed, with an MIC ranging from 1.18 to 9.5 mg ml-1. Oleozon showed a valuable antimicrobial activity against all micro-organisms tested. Results suggest that Mycobacteria are more susceptible to Oleozon than the other bacteria tested. The wide availability of sunflower oil makes Oleozon a competitive antimicrobial agent. These results should prompt the setting up of some clinical trials to compare Oleozon with other antimicrobial agents.
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An in vitro model was developed for investigation of Candida albicans penetration into human dentinal tubules. The model consisted of a dentin disc mounted between two cuvettes that each had a circular opening facing the disc. The cuvettes were filled with Tryptic-Soy-Broth, and the pulpal side cuvette was inoculated with C. albicans and incubated at 37 degrees C in air until growth occurred in the uninoculated cuvette or up to 30 days. The system was also used with Enterococcus faecalis. Completely glue-covered dentin specimens served as negative controls. Brown & Brenn-stained histological preparations of the specimens were examined with light microscopy. The time needed before growth occurred in the uninoculated cuvette showed great variation with C. albicans, whereas E. faecalis penetrated within 1 to 5 days of incubation. Slight penetration both by hyphae and yeast cells was observed in specimens inoculated with C. albicans, whereas specimens inoculated with E. faecalis showed deep and effective penetration. This study demonstrates the penetration of dentin as a possible pathway of infection by C. albicans. However, dentin penetration by C. albicans was slow and limited in comparison with E. faecalis.
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The aim of this study was to identify a possible mechanism that would explain how E. faecalis could survive and grow within dentinal tubules and reinfect an obturated root canal. Cells of Streptococcus gordonii DL1, Streptococcus mutans NG8, or E. faecalis JH2-2 were grown in brain heart infusion broth containing various amounts of human serum for 56 days. The ability of the three species to invade dentine and bind to immobilized type I collagen in the presence of human serum was assessed by dentine invasion and microtitre well experiments. All three species remained viable over the period of the experiment when grown in human serum. Cells of all three bacteria were able to invade dentine and bind to immobilized collagen. Both of these properties were inhibited by the presence of collagen in the cell solution. Human serum inhibited dentine invasion and collagen adhesion by S. gordonii DL1 and S. mutans NG8, whilst dentine invasion by E. faecalis JH2-2 was reduced in the presence of serum, but not inhibited, and binding to collagen was enhanced. It is postulated that a virulence factor of E. faecalis in failed endodontically treated teeth may be related to the ability of E. faecalis cells to maintain the capability to invade dentinal tubules and adhere to collagen in the presence of human serum.
Soluções irrigadoras versus dor pós-endodontia em sessão única, em dentes com patologias periapicais
  • Fa Soares
  • Cas César
  • Pires Jr
  • Dr Aguiar
  • Cf Silva
  • Prm Souza
Soares FA, César CAS, Pires Jr DR, Aguiar CF, Silva PRM, Souza MSGS. Soluções irrigadoras versus dor pós-endodontia em sessão única, em dentes com patologias periapicais. Rev Assoc Paul Cir Dent. 2001;55(2):125-9.
Atividade antimicrobiana de águas sanitárias disponíveis no mercado nacional
  • Siqueira Jr
  • Jf Moraes
  • Sr Lopes
Siqueira Jr JF, Moraes SR, Lopes HP. Atividade antimicrobiana de águas sanitárias disponíveis no mercado nacional. Rev Bras Odontol. 1999;56(2):57-60.
Avaliação da atividade antibacteriana de medicação intracal – três bases fortes e pastas à base de hidróxido de cálcio e paramonoclorofenol canforado
Avaliação da atividade antibacteriana de medicação intracal – três bases fortes e pastas à base de hidróxido de cálcio e paramonoclorofenol canforado. RGO. 1996;44(5):271-4.
Avaliação sob MEV da limpeza de canais radiculares através da clorofila, clorox e clorexidina
  • Mrj Santiago
  • Lk Szmajser
  • M Martinelli
  • Ras Fidel
  • Sr Fidel
Santiago MRJ, Szmajser LK, Martinelli M, Fidel RAS, Fidel SR. Avaliação sob MEV da limpeza de canais radiculares através da clorofila, clorox e clorexidina. Pesqui Odontol Bras. 2002;16(1):38.