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November 2017 -
Volume 15, Issue 9

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From the Editor

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Original contribution/Clinical Investigation
Diabetes Mellitus - Knowledge, Management and Complications: Survey report from Faisalabad-Pakistan
Ijaz Anwer, Ahmad Shahzad, Kashmira Nanji, Farah Haider, Muhammad Masood Ahmad

Alanine aminotransferase indicates excess weight and dyslipidemia
Mehmet Rami Helvaci, Orhan Ayyildiz* Mustafa Cem Algin, Yusuf Aydin, Abdulrazak Abyad, Lesley Pocock

Comparative Analysis of Antimicrobial Peptides Gene Expression in Susceptible/Resistant Mice Macrophages to Leishmania major Infection

Hamid Daneshvar, Iraj Sharifi, Alireza Kyhani, Amir Tavakoli Kareshk, Arash Asadi

Does socio-economic status of the patients have effect on clinical outcomes after coronary artery bypass grafting surgery?
Forough Razmjooei, Afshin Mansourian, Saeed Kouhpyma

Comparison of the uterine artery Doppler indices during pregnancy between gestational diabetes and diabetes mellitus and healthy pregnant women
Nazanin Farshchian, Farhad Naleini, Amir Masoud Jaafarnejhad,
Parisa Bahrami Kamangar

Survey single dose Gentamicin in treatment of UTI in children with range of 1 month to 13 years old in Jahrom during 2015
Ehsan Rahmanian, Farideh Mogharab,
Vahid Mogharab

Evaluation of control of bleeding by electro cauterization of bleeding points of amplatz sheath tract after percutaneous nephrolithotomy (PCNL) in Jahrom Peymanieh hospital during year 2015-2016
Ali Reza Yousefi , Reza Inaloo

Comparison of the three-finger tracheal palpation technique with triple ID formula to determine endotracheal tube depth in children 2-8 years in 2016-2017
Anahid Maleki, Alireza Ebrahim Soltani, Alireza Takzare, Ebrahim Espahbodi,
Mehrdad Goodarzi , Roya Noori

Effect of Sevoflurane and Propofol on pulmonary arterial pressure during cardiac catheterization in children with congenital heart diseases
Faranak Behnaz, Mahshid Ghasemi , Gholamreza Mohseni, Azim Zaraki
Population and Community Studies

Prevalence and risk factors of obesity in children aged 2-12 years in the Abu Dhabi Islands
Eideh Al-Shehhi, Hessa Al-Dhefairi, Kholoud Abuasi, Noora Al Ali, Mona Al Tunaiji, Ebtihal Darwish

Study and comparison of psychological disorders in normal students and students with multiple sclerosis in Shahrekord
Neda Ardestani-Samani, Mohammad Rabiei, Mohammad Ghasemi-Pirbalooti, Asghar Bayati, Saeid Heidari-Soureshjani

Comparative study of self-concept, physical self-concept, and time perspective between the students with multiple sclerosis and healthy students in Shahrekord
Neda Ardestani-Samani, Mohammad Rabiei, Mohammad Ghasemi-Pirbalooti, Asghar Bayati, Saeid Heidari-Soureshjani

Relationship between Coping Styles and Religious Orientation with Mental Health in the Students of the Nursing-Midwifery Faculty of Zabol
Nasim Dastras, Mohsen Heidari Mokarrar, Majid Dastras, Shirzad Arianmehr

Tuberculosis in Abadan, Iran (2012-2016): An Epidemiological Study
Ali-Asghar ValiPour, Azimeh Karimyan, Mahmood Banarimehr, Marzieh Ghassemi, Maryam Robeyhavi, Rahil Hojjati,
Parvin Gholizadeh

Family Stability and Conflict of Spiritual Beliefs and Superstitions among Yazdi People in Iran: A Qualitative Study
Zahra Pourmovahed , Seyed Saied Mazloomy Mahmoodabad ; Hassan Zareei Mahmoodabadi ; Hossein Tavangar ; Seyed Mojtaba Yassini Ardekani ; Ali Akbar Vaezi

A comparative study of the self-actualization in psychology and Islam
Simin Afrasibi, Zakieh Fattahi

The effectiveness of cognitive - behavioral therapy in reducing the post-traumatic stress symptoms in male students survivors of earthquake in the central district of Varzeghan
Sakineh Salamat, Dr.Ahad Ahangar, Robab Farajzadeh

Reviews

Effects and mechanisms of medicinal plants on stress hormone (cortisol): A systematic review
Kamal Solati, Saeid Heidari-Soureshjani, Lesley Pocock

Comparing Traditional and medical treatments for constipation : A Review Article
Mohammad Yaqub Rajput

A review of anti-measles and anti-rubella antibodies in 15- 25 year old women in Jahrom City in 2011
Ehsan Rahmania , Farideh Mogharab, Vahid Mogharab

Review of percutaneous nephrolithotomy in children below 12 years old in Jahrom hospital, during 2010-2014
Ali Reza Yousefi , Reza Inaloo

Physical and mental health in Islam
Bahador Mehraki, Abdollah Gholami

International Health Affairs

The Challenges of Implementation of Professional Ethics Standards in Clinical Care from the viewpoint of Nursing Students and Nurses
Saeedeh Elhami, Kambiz Saberi, Maryam Ban, Sajedeh Mousaviasl, Nasim Hatefi Moadab, Marzieh Ghassemi

Cognitive Determinants of Physical Activity Intention among Iranian Nurses: An Application of Integrative Model of Behavior Prediction
Arsalan Ghaderi, Firoozeh Mostafavi, Behzad Mahaki, Abdorrahim Afkhamzadeh,
Yadolah Zarezadeh , Erfan Sadeghi

Effect of resilience-based intervention on occupational stress among nurses
Hossein Jafarizadeh, Ebrahim Zhiyani, Nader Aghakhani, Vahid Alinejad, Yaser Moradi

Education and Training

Calculation of Salaries and Benefits of Faculty Members in the Ministry of Health and Medical Education of Iran
Abdolreza Gilavand

The effect of education on self-care behaviors of gastrointestinal side effects on patients undergoing chemotherapy
Shokoh Varaei, Ehsan Abadi Pishe, Shadan Pedram Razie, Lila Nezam Abadi Farahani

Creating and Validating the Faith Inventory for Students at Islamic Azad University of Ahvaz
Solmaz Choheili, Reza Pasha, Gholam Hossein Maktabi, Ehsan Moheb

Creating and Validating the Adjustment Inventory for the Students of Islamic Azad University of Ahvaz
Homa Choheili, Reza Pasha, Gholam Hossein Maktabi, Ehsan Moheb

Evaluating the Quality of Educational Services from the Viewpoints of Radiology Students of Ahvaz Jundishapur University of Medical Sciences
Abdolreza Gilavand, Jafar Fatahiasl

An Investigation of Psychosocial aspect of Iranian Nursing Students' Clinical Setting
Mahsa Boozaripour , Zanyar Karimi, Sima Zohari Anbohi, Amir Almasi-Hashiani, Fariba Borhani

Clinical Research and Methods

Comparison of the Antibacterial Effects of Chlorhexidine Mouth washes with Jaftex Mouth wash on Some Common Oral Microorganisms (An in Vitro Study)
Ebrahim Babadi, Zahra Bamzadeh, Fatemeh Babadi

Study of the effect of plasma jet on Fusarium isolates with ability to produce DON toxins
Elham Galin Abbasian, Mansour Bayat, Arash chaichi Nosrati, Seyed Jamal Hashemi, Mahmood Ghoranneviss

The comparison of anti-inflammatory effect in two methods of topical dexamethasone injection and topical application of ginger alcoholic extract after removing mandibular wisdom teeth
Sahar Zandi, Seyyed Muhammadreza Alavi, Kamran Mirzaie, Ramin Seyedian, Narges Aria, Saman Jokar

The effect of curcumin on growth and adherence of major microorganisms causing tooth decay
Leila Helalat, Ahmad Zarejavid, Alireza Ekrami, Mohammd Hosein Haghighizadeh, Mehdi Shiri Nasab


Middle East Quality Improvement Program
(MEQUIP QI&CPD)

Chief Editor -
Abdulrazak Abyad MD, MPH, MBA, AGSF, AFCHSE

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Lesley Pocock
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November 2017 - Volume 15, Issue 9

The effect of curcumin on growth and adherence of major microorganisms causing tooth decay


Leila Helalat (1)
Ahmad Zarejavid
(2)
Alireza Ekrami
(3)
Mohammd Hosein Haghighizadeh
(4)
Mehdi Shiri Nasab
(5)

(1) MSc in Nutrition, School of Paramedicine, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran
(2) Phd in Nutrition, School of Paramedicine, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran
(3) Phd in Laboratory sciences, School of Paramedicine, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran
(4) MSc in Biostatistics, School of health, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran
(5) MSc in Food Industry, School of Paramedicine, Ahvaz Jundishapur University of Medical Science, Ahvaz, Iran


Corresponding Author:
Ahmad Zarejavid
School of Paramedicine, Ahvaz Jundishapur University of Medical Science,
Ahvaz, Iran

Email:
ahmadzarejavid@gmail.com

Abstract

Background and objective:
Streptococcus mutans and Lactobacillus acidophilus are bacteria producing tooth decay which by adhering to tooth surfaces contribute to its pathogenesis. By increasing bacterial resistance to antibiotics, along with other high-cost of treatment, the use of natural antibacterial agents is essential. In this regard, the role of curcumin on the adhesion process of cariogenic pathogens has been studied.

Materials and Methods: The bacterial strains of Streptococcus mutans (PTCC1683) and Lactobacillus acidophilus (PTCC1643) were obtained from the Iranian Research Organization for Science and Technology). The early growth of the bacteria was carried out in BHI medium, and then the concentration of microorganisms reached the half-MacFarland standard, and using different concentrations of curcumin (1, 2, 4, 8, 16, 31, 25, 52, 62, 125, 250, 500 µg/ml) obtained by serial dilution method the substances were mixed. BHI medium without bacteria was considered as negative control. Then samples were incubated for 24 hours at 37°C in anaerobic conditions for determining the minimum inhibitory concentration. Inhibitory concentrations determined in the previous step were used to determine adhesion using the amount of light absorbance determination method.

Results: The minimum inhibitory concentration of growth was determined in both bacterial strains of 250 µg/ml. The results showed a decrease in light absorption with increasing curcumin concentration (P<0.001), which indicates a high correlation (correlation coefficient of -0.93) of curcumin concentrations with reverse adhesion.

Conclusion: Adhesion is the most important factor in tooth decay and its reduction is an effective solution in preventing the disease. Considering the inhibitory role of curcumin on growth and binding of bacterial strains, this curcuminoid agent is considered as a potent anti-decay agent.

Key words: Adhesion, Curcumin, Tooth decay


INTRODUCTION

The oral cavity diseases, especially tooth decay, are common diseases with high prevalence in human societies (1). 95% of people in each society suffer from dental caries and periodontal diseases (2). However, our country’s state of health in terms of oral hygiene and the teeth of children aged 6 to 12 in the Middle East region are the best, but unfortunately we (Iranian) are the worst in the 20-25 age group among the countries of the region. There is no exact data on the rate of caries of Iranian teeth, but what is certain is that we do not have a good situation in this age group (3).

Dental caries is a multi-factorial infectious disease, mainly due to the reduction in pH following carbohydrate fermentation, due to the action of two bacterial strains of streptococci and lactobacillus (1 and 4).

The formation of dental biofilms has been associated with the onset and development of dental caries as the extracellular matrix surrounding the bacteria (7-5). The first step in the formation of dental biofilms is bacterial adhesion to the dental surfaces (8). Oral and oral diseases have a negative effect on the quality of life, general health, nutritional status and social function of individuals. The pain, infection, sleep disorders and difficulty in chewing and eating and talking are considered as adverse outcomes of oral and dental illnesses in individuals (9). Dental diseases such as dental caries and gum disease may also significantly affect the health of the individual. Dental caries are associated with the development of chronic diseases such as diabetes and heart disease (9, 27).

Considering the high cost of treatment in oral cavity diseases (1) as well as increased bacterial resistance to commonly used antibiotics for treatment, new strategies for the prevention and treatment of dental caries and the fight against pathogens are needed (10). Nutrition is an effective factor in the prevention and treatment of dental caries and primary prevention programs at community level, including nutritional control, have played an important role in reducing tooth decay (11). Today, the use of natural antimicrobial agents in the form of alternative therapies has expanded due to the low and effective nature of the treatment and even the prevention of many diseases, including oral and dental diseases (13-12). Therefore, finding these natural and low risk compounds and studying the effect of these compounds on growth and adhesion of cariogenic bacteria, as the first step in the onset of tooth decay is considered necessary (5.8).

Tumeric is a yellow spice of the family Zingiberaceae (14), which grows mainly in the southern and tropical regions of Asia, such as China, India and Malaysia. The root and stem of this plant mainly contains a yellow curcuminoid compound (15). Turmeric is one of the most popular plants with extensive therapeutic properties in traditional medicine (16).

Curcumin (diferuloylmethane), as the main constituent of the main color of yellow in turmeric (17), due to its various antioxidant, anti-inflammatory, anti-carcinogenic, antimicrobial and antiparasitic effects can be used in various parts of the body, including the oral cavity (18-20). Most studies in the field of oral cavity diseases, such as the study by Izui et al. (29), have focused on periodontal disease due to the anti-inflammatory effects of curcumin, or in the form of mouthwash containing curcumin in the study of Mali et al. (22) and in the form of a curcumin-based gel by Bhatia et al (21), curcumin has been shown to be effective on periodontal disease, but the effect of curcumin on dental caries is less marked. On the other hand, studies such as Mandroli and Bhat (23) have mainly focused on the growth and microbiological counting of decay pathogens. Therefore, studying the effect of curcumin on growth of major microorganisms of dental caries as a biological factor along with their adhesion, which is one of the main causes of their pathogenicity, is one of the goals to be considered in this study.


MATERIALS AND METHODS


Analysis method
This study is an experimental study approved by the Ethics Committee of the Medical University of Ahvaz number 1395.590IR.AJUMS.REC. The curcumin used in this study was from the German Merck company with a purity of about 97%. DMSO (Dimethyl sulfoxide) solution was used to increase solubility and the final concentration of 1mg/ml solution was prepared from curcumin solution, from which was obtained a uniform solution of curcumin using shaker. A bacterial filter was used to remove contaminants.

Microorganisms
The bacterial strains of Streptococcus mutans (PTCC1683) and Lactobacillus acidophilus (PTCC1643) were obtained from the Iranian Research Organization for Science and Technology). BHIB (Brain heart infusion broth) containing 1% sucrose was used for early growth of the microorganisms. The culture media was incubated in anaerobic conditions after inoculation of bacteria for 24 hours at 37°C.

Preparation of bacterial suspension: A linear culture of the initial BHI medium was performed on a Blood Agar culture medium under sterile conditions and resumed for 24 hours at 37°C incubation. The selective harvesting of microbial colonies was performed from Blood Agar level (surface) under sterile conditions and the colonies were transferred to a culture medium (BHIB) (Brain heart infusion broth) containing 1% sucrose and then incubated for 24 hours under anaerobic conditions at 37°C. The growth concentration was adjusted to 5×106 organism/ ml by using 0.5 McFarland’s turbidity standard.

To prepare McFarland’s standard, 0.6 ml of 1% sodium chloride solution was dissolved in 100 ml of sulfuric acid in a volume of 100 ml and uniform solution of bacterial suspension was obtained by shaker.

Using an ultraviolet spectrophotometer (UV-2802 United States Unicode), the OD (optical absorption) at a wavelength of 620 nm was set at a pH of about 0.1 to achieve a concentration of 5×106 CFU (24).

The method of exposure to bacterial strains with different concentrations of curcumin
Different concentrations of curcumin solution were obtained by serial dilution method. In such a way 12 sterile tubes were considered. In all tubes, 1ml of bacterial solution was added at a concentration equivalent to the 0/5 McFarland’s turbidity standard. Then in the first tube containing 1 ml bacterial solution, 1 ml of the curcumin solution was added. After mixing well, 1 ml was transferred to the second tube, this was continued till the last (10th) tube. From the last tube 1ml of final solution was discarded. By following this serial dilution, the concentration of, 500, 250, 125, 5.62, 25.31, 16, 8, 4, 2, 1µg/ml, respectively was acheived) (23).

BHI medium without bacteria was considered as a negative control and non-curcumin bacterial suspension as a positive control, and then the test and control samples were incubated at 37°C for 24 hours under anaerobic conditions.

Determination of MIC (Minimum inhibitory concentration)
Considering the generation of turbidity from bacterial growth, examination of the transparency of tubes indicated the inhibition of growth in tubes.

The the tubes were incubated for 24 hours at 37°C after the incubation, the MIC values were determined by visual inspection of tubes. In each series of tubes, the last tube with clear supernatant was considered to be without any growth and taken as MIC value. Turbidity in the tube indicated growth of the bacteria implying that the bacteria are resistant to curcumin. All steps for each bacterial strain were repeated 2 times and the mean of measurements was expressed in 2 replicates (23.31).

Assessment of adhesion
2.5 ml of inhibitory concentrations of curcumin, which did not inhibit the growth of bacteria in the previous stage, was added to 2.5 ml of BHI medium containing 1% sucrose and 250l of bacterial suspension prepared at a concentration of 5×106 CFU. In positive control sample, instead of the curcumin solution, DMSO alone was used and in the negative control sample, BHI medium containing 1% sucrose without bacteria was considered. Then, at 37°C, for 24 hours, with Angle 30, the incubation was performed under anaerobic conditions. The tubes were examined externally, and the lowest concentration without attachment of visible cell to tubes wall was determined as Total Bacterial Adherence Inhibition (TBAI) (25). After removing the contents of the tubes containing suspended cells for isolation of the cells that were attached to the tubes, 3ml of KPB 0.5M buffer with Ph=6.8 was used and 0.25ml trypsin. The amount of cells suspended by optical density was measured by a spectrophotometer (UV -2802 Unico) at a wavelength of 490nm. All tests were repeated for 2 bacterial strains and the mean of measurements was expressed in 2 replicates (26).

Statistical analysis
In this study, data analysis and statistical analysis of data were performed using SPSS22 software. Spearman statistical test to examine the correlation between quantitative variables and Independent sample-T test to compare the mean of quantitative variables in two groups and One Way ANOVA test followed by a Sidak test to compare the mean of quantitative variables in the control group with each of the test groups were used.