www.scielo.br/jaos Antimicrobial activity of , Camellia sinensis and chlorhexidine against the adherence of microorganisms to sutures after extraction of unerupted third molars
Raquel Lourdes FARIA1, Lincoln Marcelo Lourenço CARDOSO2, Gokithi AKISUE3, Cristiane Aparecida PEREIRA4, Juliana Campos JUNQUEIRA5, Antonio Olavo Cardoso JORGE5, Paulo Villela SANTOS JÚNIOR6
1- Undergraduate student, São José dos Campos Dental School, UNESP - Univ. Estadual Paulista, São José dos Campos, SP, Brazil. 2- MSc, Faculty of Pindamonhangaba - FAPI, Pindamonhangaba, SP, Brazil. 3- PhD, Faculty of Pindamonhangaba - FAPI, Pindamonhangaba, SP, Brazil. 4- Postgraduate student, São José dos Campos Dental School, UNESP - Univ. Estadual Paulista, São José dos Campos, SP, Brazil. 5- PhD, Department of Biosciences and Oral Diagnosis, São José dos Campos Dental School, UNESP - Univ. Estadual Paulista, São José dos Campos, SP, Brazil. 6- PhD, Department of Diagnosis and Surgery, São José dos Campos Dental School, UNESP - Univ. Estadual Paulista, São José dos Campos, SP, Brazil.
Corresponding address: Juliana Campos Junqueira - Faculdade de Odontologia de São José dos Campos - UNESP - Departamento de Biociências e Diagnóstico Bucal - Av. Francisco José Longo 777, São Dimas - São José dos Campos - SP - Brazil - 12245-000 - Phone: +55 12 39479033 - Fax: +55 12 39479010 - e-mail: [email protected]
ABSTRACT
O L., Camellia sinensis (L.) Kuntze and 0.12% chlorhexidine digluconate on the adherence of microorganisms to suture materials after extraction of unerupted third molars. Material and Methods: Eighteen patients ! " # not to use any type of antiseptic solution at the site of surgery (control group). After 15 # Calendula , Camellia sinensis $ & ! " # ' * ! + # - / 3 streptococci; mannitol agar for Staphylococcus spp.; MacConkey agar for enterobacteria and Pseudomonas spp., and Sabouraud dextrose agar containing chloramphenicol for Candida ! 46*67 8':< <"=> ? ! @ ! B # [ 0.12% chlorhexidine digluconate. Conclusions: L. and Camellia sinensis (L.) Kuntze presented antimicrobial activity against the adherence of microorganisms to [ !
Key words: Calendula officinalis L. Camellia sinensis (L.) Kuntze. Chlorhexidine. Antimicrobial activity.
J Appl Oral Sci. 476 2011;19(5):476-82 Antimicrobial activity of , Camellia sinensis and chlorhexidine against the adherence of microorganisms to sutures after extraction of unerupted third molars
INTRODUCTION demonstrated [ The sutures used in dentoalveolar surgery, such bleeding18. According to Iauk, et al.15 (2003), as extraction of unerupted teeth, represent a risk also presents antimicrobial activity against periodontopathogenic bacteria. they are prone to the adherence of pathogenic Green tea is produced from the leaves of Camellia bacteria. The accumulation of microorganisms sinensis (L.) Kuntze (family Theaceae), and has on sutures may serve as a focus of odontogenic been popularly used in Japan and China for many infections. These infections are caused by both centuries3! N aerobic and anaerobic bacteria, including species of C. sinensis possess antimicrobial activity against of the genera Fusobacterium, Peptostreptococcus, Streptococcus mutans22. In addition, in vitro studies Prevotella, Porphyromonas, Streptococcus, have demonstrated a strong antimicrobial action and Bacteroides. Recent studies suggest that of C. sinensis against periodontal pathogens such odontogenic infections and bacteremia develop at as Porphyromonas gingivalis and Fusobacterium the time of suture removal and are a possible risk nucleatum29. for bacterial endocarditis4,23. Several studies have tested the use of medicinal The use of oral antiseptics after surgery is an ! ] # !14 (2002) [ compared the effects of 0.12% chlorhexidine consequent prevention of infections10. Chlorhexidine Allium sativum L. [ and Melaleuca alternifolia Cheel. Chlorhexidine antimicrobial agent used for the reduction of and Allium sativum L. microorganisms in the oral cavity13,14,19,21,22. activity against S. mutans, but not against other B # oral microorganisms seeded onto blood agar. In number of adverse effects, such as the formation contrast, Melaleuca alternifolia Cheel presented of stains on teeth and dentures, dysgeusia, parotid strong antimicrobial activity against all oral enlargement, and desquamation of the oral mucosa. microorganisms studied. Lauten, et al.18 (2005) These factors have encouraged the search for other antimicrobial agents14. and extracts from four plant species [Melaleuca Within this line of research, the use of medicinal alternifolia Cheel, Leptospermum scoparium Forst., plants indicated for the treatment of infectious L. and Camellia sinensis (L.) processes has been investigated based on the Kuntze] in 17 patients. No differences in plaque results of ethno-pharmacobotanical studies. The use of plants for the treatment of different diseases & ! on medicine and medicinal plants arose in China $4!7j ! and Egypt11. Medicinal plants are valuable natural herbal products frequently used for the treatment of various diseases5 and represent an important Calendula officinalis L. (Asteraceae), Camellia 11,24. sinensis (L.) Kuntze (Theaceae) and 0.12% The antimicrobial compounds found in plants inhibit chlorhexidine digluconate on the adherence of microorganisms to suture materials after extraction that differ from those underlying the activity of of unerupted third molars. commonly used antimicrobial agents and have a [ 7,12,30. MATERIAL AND METHODS L. is an annual plant of the N # \ / Q ! U \ preparations. This plant is native to Central Europe approved by the Ethics Committee of the São José / dos Campos Dental School (FOSJC), São Paulo State locations throughout North America and Europe27. University (UNESP) (protocol 062/2007 - PH/CEP). X # \ # Preparation of the mouthwashes # # \ # carotenoids, xanthophylls, phenolic acids, steroids, study: mucilage, tocopherol, and calenduline28. The Oral solution of 0.12% chlorhexidine digluconate extract produced from (Becker, São José dos Campos, SP, Brazil). used in Europe since the 12th century as a topical A 1% ?! \ ! In vivo studies employing B ?&# #
J Appl Oral Sci. 477 2011;19(5):476-82 FARIA RL, CARDOSO LML, AKISUE G, PEREIRA CA, JUNQUEIRA JC, JORGE AOC, SANTOS P V Jr glycerine, and Nipagim® (Becker). 3 ! / 4j > * & \ $ $ ! of Camellia sinensis (L.) Kuntze (5 g), glycerine (4 3-0 silk sutures (Ethicon, Johnson & Johnson, São g), 70% sorbitol (3.5 g), sodium saccharin (0.5 g), < # 3# \ sodium lauryl sulfate (0.6 g), lauryl polyglucoside (1 repositioned in its original location. # !8 # The patients received 500 mg amoxicillin (Aché, 7!$ ! # São Paulo, SP, Brazil) and 50 mg sodium diclofenac \ + # 3 # 3# # ! instructed to brush their teeth using toothpaste Laboratory of Pharmacognosy and Medicinal Plants only in the non-operated areas. (LAFAPLAM), Faculty of Pindamonhangaba (FAPI). Control group Patient selection N [ # ! !# < # Oral and Maxillofacial Surgery and Traumatology, type of oral antiseptic solution. "Q3 <# =+3! clinical and radiographic evaluation in order to Experimental groups Fifteen days after the first surgery, the infectious process. Eighteen patients ranging in $' 8 ! B # ! These patients presented good health conditions $! medications or any oral antiseptic solution. The 18 package (250 mL) accompanied by a measuring 8 ! & " $! $ ? 8 according to the method of Metin, et al.20 (2006), Surgery for unerupted maxillary third molar extraction for 7 days. N the patients before surgery for planning of the Suture removal ! '! N # X [ # * standardized into a size of 15 mm using a sterile # ! [ transferred aseptically to a test tube containing the bottom of the buccal sulcus at the height of the 2 mL sterile 0.1 M phosphate-buffered saline [ !j + < # B '!4! mucosa close to the tooth to be extracted. Next, the Laboratory of Microbiology, FOSJC, UNESP, for microbiological analysis. the alveolar bone crest from the curvature of the tuberosity to the mid-portion of the distal side of Microbiological analysis the second molar, complemented by an oblique incision in the direction of the bottom of the sulcus for 2 min (Fanem, São Paulo, SP, Brazil) in order involving the distal side of the interdental papilla of to obtain a homogenous suspension. Decimal [ ! N \ dilutions (10-1, 10-2 and 10-3 # from this suspension in sterile saline. Aliquots of 0.1 ! ? &
Number of patients Extraction of unerupted third molar Left Right 6 Control Chlorhexidine 6 Control 6 Control Camellia sinensis
Figure 1- Distribution of the patients according to experimental group
J Appl Oral Sci. 478 2011;19(5):476-82 Antimicrobial activity of , Camellia sinensis and chlorhexidine against the adherence of microorganisms to sutures after extraction of unerupted third molars seeded in duplicate onto Petri dishes containing / # U !# 3 < # N# =3N # [ j! # # /U# =3N j RESULTS total count of aerobic and facultative anaerobic microorganisms; Mitis Salivarius bacitracin sucrose The mean number of CFU/mL (log10) of aerobic /33 / 3 !4 U=> ? blood agar obtained for the control and experimental $j " 4! 8 streptococci- of Staphylococcus spp.; MacConkey agar (Difco) for ! B # Pseudomonas spp., [ !$4j and Sabouraud dextrose agar (Difco) containing 0.1 chlorhexidine digluconate (p=0.008). mg/mL chloramphenicol (União Química, São Paulo, 8 3# Candida spp. of Candida spp. on Sabouraud agar (Figure 3). The blood agar, mannitol agar, MacConkey agar B # [
3 8' C CFU/mL (log10 46*67 ! /33 control and chlorhexidine groups (p=0.048). at 37 C for 48-72 h in a 5% CO2 atmosphere. After $*8 <"=> ? the time of incubation, plates containing 30 to 300 (log10 /33# / * and MacConkey agar obtained for the control and <"=> ? log transformed (log10). 25% C. sinensis \ # $j tincture and 0.12% chlorhexidine digluconate. No Statistical analysis [ 3 R * / !
Table 1- Mean number and standard deviation of colony-forming units (CFU)/mL (log10) of microorganisms isolated from suture materials and grown on Mitis Salivarius bacitracin sucrose agar obtained for the control groups and groups using Camellia sinensis
Groups Mouthwash Control p Camellia sinensis 1.49±1.57 2.19±1.36 0.098 1.83±0.74 2.60±0.41 0.054 Chlorhexidine 2.10±1.17 3.68±0.63 0.081
Figure 3- Mean number and standard deviation of Figure 2- Mean number and standard deviation of colony- colony-forming units (CFU)/mL (log10) of microorganisms forming units (CFU)/mL (log ) of microorganisms isolated 10 isolated from suture materials and grown on Sabouraud from suture materials and grown on blood agar obtained agar obtained for the control groups and groups using Camellia sinensis for the control groups and groups using , Camellia sinensis, and chlorhexidine Calendula officinalis and chlorhexidine mouthwash the mouthwash and control groups of cholorhexidine and control groups of cholorhexidine (Student's t-test) (Student's t-test)
J Appl Oral Sci. 479 2011;19(5):476-82 FARIA RL, CARDOSO LML, AKISUE G, PEREIRA CA, JUNQUEIRA JC, JORGE AOC, SANTOS P V Jr
Table 2- Mean number and standard deviation of colony-forming units (CFU)/mL (log10) of microorganisms isolated from suture materials and grown on Mannitol agar obtained for the control groups and groups using Camellia sinensis, Calendula
Groups Mouthwash Control p Camellia sinensis 0.08±0.95 1.77±1.52 0.174 0.17±0.41 0.23±0.57 0.363 Chlorhexidine 0.28±0.69 1.05±0.83 0.081
Table 3- Mean number and standard deviation of colony-forming units (CFU)/mL (log10) of microorganisms isolated from suture materials and grown on MacConkey agar obtained for the control groups and groups using Camellia sinensis,
Groups Mouthwash Control p Camellia sinensis 0.96±1.53 1.50±1.68 0.343 0.12±0.29 0.22±0.53 0.363 Chlorhexidine 0.12±0.29 0.48±0.76 0.177
DISCUSSION extremely important procedure, reducing the risks # Microorganisms of the oral microbiota can a serious predisposition to bacterial endocarditis adhere to suture materials, a fact favoring their 6. 3 * odontogenic infections and bacteremia. This oral hygiene, but studies investigating the action situation is favored or prevented, depending on the of these phytotherapeutic agents on microbial adsorption properties of the suture material and adherence to suture materials are scarce. oral hygiene care during the postoperative period23. U # 8 The results of the present study demonstrated # the adherence of a large number of microorganisms [ to the suture material after third molar extraction. !$4j According to Banche, et al.2 (2001), contamination of suture material in the oral cavity mainly originates on blood and Sabouraud agar. [ 7.5x108 microorganisms/mL. aerobic and anaerobic Gram-positive and Gram- negative bacteria and yeast. Chlorhexidine has The largest mean number of CFU/mL (log10) of microorganisms adhered to the suture material [ induces cell surface alterations that lead to the loss # /33# of osmotic balance and cytoplasmic precipitation9. In the present study, although the antimicrobial / < & # [ Staphylococcus and enterobacteria, respectively. of the C. sinensis and # that staphylococci and enteric bacilli usually are not of the number of microorganisms. The antimicrobial found in the oral cavity8,31; if present, they occur activity of C. sinensis has been attributed to its main in smaller numbers and are part of the transitory # # microbiota. bacterial plasma membrane, causing irreversible The smallest number of microorganisms adhered damage and bacterial death16. In addition to its antimicrobial activity, also possesses 3 # high topical antiinflammatory activity due to Candida spp. According \ to Appleton1 (2000), yeast of the genus Candida phytochemical composition10! * # colonize the mucosal membranes of approximately al.33 (1997) observed that extracts 75% of the healthy population and are usually \ present in small numbers. prostaglandin inhibitors. Thus, the use of an antiseptic for the control of @ microorganism adherence to suture material is an process, chlorhexidine, in a concentration-and time-
J Appl Oral Sci. 480 2011;19(5):476-82 Antimicrobial activity of , Camellia sinensis and chlorhexidine against the adherence of microorganisms to sutures after extraction of unerupted third molars
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J Appl Oral Sci. 482 2011;19(5):476-82