Odborna praâce ORTODONCIE

Transcription

1 Vliv kombinace beta-glukanu a laktaâ tu hliniteâ ho na tvorbu a slozï enõâ biofilmu u ortodontickyâ ch pacientuê. Pilotnõ placebem kontrolovanaâ studie. The effect of beta-glucan and aluminium lactate combination on the formation and composition of biofilm in orthodontic patients. Pilot placebo-controlled study. *Doc. Ing. KaterÏina ValentovaÂ, Ph.D., **MUDr. Jan Palas, ***RNDr. Jana PetrzÏelovaÂ, **doc. MUDr. MilosÏ SÏ pidlen, Ph.D., *prof. MUDr. VilõÂm SÏ imaâ nek, DrSc. *U stav leâ karïskeâ chemie a biochemie LF UP v Olomouci *Department of Medical Chemistry and Biochemistry, Medical Faculty of Palacky University, Olomouc **Ortodonticke oddeï lenõâ, Klinika zubnõâho leâ karïstvõâ LF UP v Olomouci **Department of Orthodontics, Institute of Oral Medicine, Medical Faculty of Palacky University, Olomouc ***U stav leâ karïskeâ mikrobiologie LF UP v Olomouci ***Department of Medical Microbiology, Medical Faculty of Palacky University, Olomouc Souhrn U vod: LeÂcÏ ba fixnõâm ortodontickyâ m aparaâ tem klade veïtsïõânaâ roky na uâ stnõâ hygienu a je u pacientuê sesï patnou hygienou nebo nespraâ vnou technikou cï isï teï nõâ zubuê spojena se zvyâ sï enyâ m usazovaâ nõâm zubnõâho plaku (biofilmu) v mõâstech huê rïe prïõâstupnyâ ch. Tyto prostory mohou byâ t predilekcï nõâm mõâstem vzniku zubnõâho kazu, gingivitid cï i parodontopatiõâ. CõÂl: CõÂlem studie bylo sledovaâ nõâ tvorby a slozï enõâ biofilmu na skupineï pacientuê s fixnõâm ortodontickyâ m aparaâ tem a diagnostikovanou gingivitidou po opakovaneâ m vyplachovaânõâuâ st roztoky beta-glukanu a laktaâ tu hliniteâ ho. Metody: Ve dvojiteï slepeâ zkrïõâzï eneâ (ªcross-overª) studii byly u subjektuê (n=21) hodnoceny indexy zaâ neï tliveâ, plakem podmõâneï neâ zmeï ny gingivy (CPITN), krvaâ civost daâ snï ovyâ ch papil (PBI), tvorba plaku (PLI) a prïõâtomnost neï kteryâ ch rezidentnõâch bakteriõâ prïed a po cï tyrïdennõâm vyplachovaâ nõâ uâ st roztoky kombinace laktaâ tu hliniteâ ho s beta-glukanem, laktaâ tu hliniteâ ho a placebem bez soucï asneâ ho mechanickeâhocïisïteï nõâ zubuê a mezizubnõâch prostor. Vy sledky: PLI index vzrostl po pouzïõâvaâ nõâ vsï ech trïõâ uâ stnõâch vod, u kombinace laktaâ tu hliniteâ ho s beta-glukanem poklesl index PBI; hodnoty CPITN vzrostly u placeba, u dvou experimentaâ lnõâch vod se hodnoty indexu nezmeï nily. PrÏi pouzïõâvaânõâuâ stnõâ vody s laktaâ tem hlinityâ m dosï lo k poklesu celkoveâ ho pocï tu sledovanyâ ch bakteriõâ. ZaÂveÏ ry: U stnõâ vody obsahujõâcõâ beta-glukan a/nebo laktaâ t hlinityâ se prokaâ zaly jako biobezpecïneâ. V kombinaci s mechanickyâmcïisïteïnõâmmuêzïe beta-glukan v kombinaci s laktaâ tem hlinityâ m rozsïõârïit skupinu prïõârodnõâch chemoprofylaktik v prïõâpravcõâch uâ stnõâ hygieny (Ortodoncie 2013, 22, cï. 1, s ). Abstract Introduction: For patients with fixed orthodontic appliances oral hygiene is demanding. Insufficient oral hygiene and inappropriate toothbrushing results in plaque (biofilm) in places that are difficult to reach. These can become places of early origin of caries, gingivitis, and periodontitis. Aim: This study focused on the formation and composition of biofilm in the group of patients with fixed orthodontic appliances, who were suffering from gingivitis a following repeated mouthwash with a solution of betaglucan and aluminium lactate. 18

2 rocïnõâk22 Methods: The double-blind cross-over study on 21 patients who were evaluated using community periodontal index of treatment needs (CPITN), periodontal bleeding index (PBI), plaque formation index (PLI), and some resident bacteria before and after four days treatment with rinses of beta-glucan and aluminium lactate, aluminium lactate and placebo, without simultaneous mechanical tooth brushing or cleaning of interdental spaces. Results: PLI increased after the use of all three mouthrinses. The combination of aluminium lactate and betaglucan caused decreased PBI; CPITN increased in the case of placebo; in two mouth rinses tested CPITN values did not change. When mouth rinse with aluminium lactate was used, the overall number of bacteria monitored lowered considerably. Conclusion: Mouth rinses with beta-glucan and/or aluminium lactate proved safe from the biological viewpoint. Together with mechanical tooth brushing, beta-glucan with aluminium lactate could be used as a natural chemo-prophylactic agent in oral hygiene (Ortodoncie 2013, 22, No. 1, p ). KlõÂcÏ ovaâ slova: fixnõâ aparaâ t, beta-glukan, laktaâ t hlinityâ, zubnõâ plak, chemoprofylaxe Key words: fixed appliance, beta-glucane, aluminium lactate, plaque, chemoprophylaxis UÂ vod UÂ stnõâ hygiena hraje v zubnõâm leâ karïstvõâ vyâ znamnou roliv prevencivzniku posï kozenõâ tvrdyâ ch zubnõâch tkaâ nõâ azaâveï sneâ ho aparaâ tu. ZubnõÂ kaz a plakem podmõâneïnaâ gingivitida a parodontitida majõâ spolecïnyâ patogenetickyâ zaâ klad - mikrobiaâ lnõâ biofilm (zubnõâ plak), jehozï nedostatecï neâ odstranï ovaâ nõâmaâ za naâ sledek rozvoj teï chto onemocneï nõâ [1]. Biofilm je spolecï enstvo mikroorganismuê, charakterizovaneâ tõâm, zï e bunï ky, ktereâ jsou ireverzibilneï prïichyceneâ k podkladu nebo jedna k druheâ, jsou zapusïteïneâ v matrici bunï kamiprodukovanyâ ch extracelulaâ rnõâch polymernõâch laâ tek. Ty vykazujõâ odlisï nyâ fenotyp s ohledem na rychlost ruê stu a transkripci genuê [2, 3]. ZubnõÂ leâ karïstvõâ bylo prvnõâ oborem medicõâny, kde bylo mikrobiology prïijato, zï e biofilm (zubnõâ plak) je prïirozenyâ m zpuêsobem zï ivota bakteriõâ [4]. S biofilmem se setkaâ vaâ me v mnoha dalsï õâch leâ karïskyâ ch oborech, zahrnujõâcõâ povrchy cï aâ stõâ lidskeâ ho organismu (traâ vicõâ uâ strojõâ, pohlavnõâ a mocï ovyâ trakt), teï lnõâch implantaâtuê, naprï. kanyl, kloubnõâch a chlopennõâch naâ hrad. RuÊ st v biofilmu je z mnoha pohleduê pro bunï kyvyâ hodneï jsï õâ. Jsou odolneï jsï õâ vuê cï iantibiotikuê m, mechanickeâ mu posï kozenõâ, systeâ mu buneïcïneâ imunity hostitele a umozïnï uje jejich uchycenõâ se v prostrïedõâ bohateâ m na zï iviny a vyuzï itõâ kooperativnõâch vyâhod [3]. Dutina uâ stnõâ je osõâdlena prïirozenou mikrofloâ rou. Dosud bylo identifikovaânoprïiblizïneï 700 druhuê bakteriõâ [5, 6], prïicï emzï kultivovatelnyâch je jen asi60 % druhuê.cï aâstecï neâ znalostio nekultivovatelnyâ ch druzõâch se zõâskaâvajõâ zejmeâ na molekulaâ rneï biologickyâ mimetodami (naprï. PCR a Ramanova mikroskopie) [7]. ZubnõÂ plak, tvorïenyâ rezidentnõâ mikrofloâ rou, pokryâvaâ jak sklovinu puê vodnõâch, tak povrchy umeïlyâch zubuê, fixnõâch ortodontickyâch aparaâtuê a gingivy. RezidentnõÂ mikrofloâ ra braâ nõâ exogennõâm mikroorganismuê m v kolonizacizubnõâ skloviny [8]. Plak je hlavnõâ prïõâcï inou parodontopatiõâ a vzniku zubnõâho kazu. V pravidelneï Introduction Oral hygiene plays an important role in dental medicine for prevention of damage hard dental and periodontal tissues. Caries, gingivitis and periodontitis caused by plaque, have a common pathogenic basis, i.e. microbial biofilm (dental plaque), which if not removed properly, results in the problems mentioned [1]. A biofilm is basically a community of microorganisms characterized by cells that are irreversibly attached to the enamel or to other bacteria embedded in a matrix of extracellular polymeric substances produced by the cells. These cells have a different phenotype with regard to growth speed and gene transcription [2, 3]. Dentistry was the first branch of medicine where microbiologists found on that the biofilm (dental plaque) is a natural living mode for bacteria [4]. Bacterial biofilms are found on the surfaces of human organs (gastrointestinal, reproductive and urinary tract), body implants, e.g. cannulas, prosthetic joints and valves. The growth in biofilm confers biological advantage to the bacterial colonies. They are then more resistant to antibiotics, mechanical damage, host immunity, and they attach in an environment rich in nutrients and take advantage of the cooperation [3]. The buccal cavity is populated by resident microflora. So far about 700 bacterial species have been identified [5,6]; however, only about 60% of the species can be cultivated. Partial knowledge of non-cultivated species can be obtained using methods of molecular biology (e.g. PCR and Raman's microscopy) [7]. Dental plaque composed of resident microflora, covers enamel of natural as well as surface of artificial teeth, fixed orthodontic appliances, and gingiva. Resident microflora prevent exogenous microorganisms from enamel colonization [8]. Plaque is the major cause of periodontal disease and caries. Plaque, that is not regularly removed, undergoes a process of calcification [9, 10]. Pathogenic bacteria containing plaque bio- 19

3 neodstranï ovaneâ m plaku probõâhaâ proces kalcifikace [9, 10]. Biofilm zubnõâho plaku, pokud obsahuje patogennõâ bakterie, muê zï e infikovat i jineâ povrchy v lidskeâ m teï le, naprï. ceâ vnõâ katetry, implantaâ ty, umeï leâ nebo posï kozeneâ srdecï nõâ chlopneï, ale inaâ hrady kloubuê, a zpuê sobit zaâ vazïneâ zdravotnõâ komplikace. Do krevnõâho obeï hu se bakterie dostaâ vajõâ jak prïi invazivnõâch vyâ konech, ale iprïi drobnyâ ch traumatizacõâch zpuê sobenyâ ch pouhyâ m cï isï teïnõâm zubuê [4, 11]. SlozÏenõ uâ stnõâ mikrofloâ ry kazïdeâhocï loveï ka je rozdõâlneâ a je podmõâneï no jeho konstitucõâ, veï kem s prïihleâ dnutõâm k pohlavõâ (gravidita, klimakterium), dispozicõâ k tvorbeï zubnõâho kazu a kvalitou uâ stnõâ hygieny. DuÊ lezï itou roli maâ slozï enõâ dennõâ diety (neï ktereâ mikronutrienty), onemocneï nõâ (naprï. diabetes mellitus), mnozï stvõâ a slozï enõâ slin, snõâzï eneâ samoocï isï t'ovaâ nõâ podmõâneï neâ naprï. steï snaânõâm zubuê, chybnyâmivyâ plneï mi, protetickyâ minaâ hradami a v neposlednõâ rïadeï ortodontickyâ miaparaâ ty. SlozÏ enõâ mikrobiaâ lnõâho biofilmu se takeâ li sï õâ na ruê znyâ ch povrsï õâch dutiny uâ stnõâ, odlisï naâ prostrïedõâ pro mikrobiaâ lnõâ kolonizacitak poskytujõâ naprï. rty, bukaâ lnõâ povrchy a meï kkeâ patro, jazyk, supragingivaâ lnõâ a subgingivaâ lnõâ plochy zubu, celkoveâ zastoupenõâ jednotlivyâ ch mikroorganismuê v jizï vytvorïeneâ m biofilmu vsï ak byâvaâ pomeï rneï konstantnõâ po celyâ zï ivot [4]. Mechanicke cï i sïteï nõâ zubuê a mezizubnõâch prostor zubnõâmia mezizubnõâmikartaâ cï ky za pouzï itõâ zubnõâ pasty, dentaâ lnõâch nitõâ, mezizubnõâmi stimulaâ tory, prïõâpadneï dentaâ lnõâ sprchou maâ klõâcï ovou roliv odstranï ovaâ nõâ biofilmu [1]. U cï inneïjsï õâho odstranï ovaâ nõâ plaku je mozï no dosaâ hnout pouzï itõâm chemickyâ ch inhibitoruê zpomalujõâcõâch tvorbu biofirmu a inhibujõâcõâch aktivitu oraâ lnõâch patogenuê.uâ cï inek syntetickyâ ch/ prïõârodnõâch chemoprofylaktik by nemeïlbyâtdaâ n jen uâcï inkem antimikrobiaâ lnõâm (antiseptickyâ m), ale takeâ antiadhezivnõâm. Tyto pozï adavky splnï uje neï kolik antiseptik. Prioritnõ postavenõâ maâ chlorhexidin diglukonaâ t [12] a esenciaâ lnõâ oleje thymol, eukalyptol, mentol a methylsalicylaâ t. Ty jsou aktivnõâ slozï kou ve vyârobcõâch rïady Listerine Ò [13]. Antiplakove uâcï inky vsï ak vykazuje takeâ celaâ rïada sekundaâ rnõâch metabolituê rostlin naprï. kvarternõâ benzofenantridinoveâ alkaloidy obsazï eneâ naprï. v extraktu nadzemnõâ cï aâ sti Macleaya cordata [14, 15], proanthokyanidiny typu A2 v plodech klikvy velkoplodeâ (Vaccinium macrocarpon) [16-19], polyfenoly v plodech Vitis vinifera [20]. AktivnõÂmi slozïkamiprïõâpravkuê uâ stnõâ hygieny jsou takeâ prïõârodnõâ laâ tky s protizaâ neï tlivyâ mi, antioxidacï nõâmicï iadstringentnõâmi uâcï inky, jako naprï. alantoin, fenoloveâ kyseliny, obsahoveâ laâ tky zeleneâ ho cï aje, prïõârodnõâ oleje, probatickeâ bakterie, vitaminy B, C, D, E, biomineraâ ly Ca, Mg, Zn [15, 21-25]. Beta-glukany, polysacharidy z buneïcï neâ steï ny kvasinek (Sacharomyces cerevisie), hub, neï kteryâ ch bakteriõâ a rostlin majõâ vyâ razneâ imunomodulacï nõâ uâ cï inky [26] film may infect other surfaces of the human body, e.g. vascular catheters, implants, artificial or damaged heart valves, joint substitutes, and result in severe health conditions. The invasive treatment and even minor injuries due to tooth brushing cause the bacteria to escape into the bloodstream [4, 11]. The composition of oral microflora is specific for each individual depending on their constitution, age and gender (pregnancy, menopause), tendency to caries and quality of oral hygiene. The daily diet (some micronutrients) plays an important role, as well as diseases (e.g. diabetes mellitus), amount and composition of saliva, lower self-cleaning due to e.g. teeth crowding, faulty fillings, prosthetics, and orthodontic appliances. The bacterial biofilm composition differs according to the type of oral cavity surface. There are different environments in the oral cavity - e.g. lips, buccal surfaces and soft palate, tongue, supragingival and subgingival tooth surface. However, overall representation of individual microorganisms in already formed biofilm remains stable during the life of an individual [4]. Tooth brushes and interdental brushes with toothpaste, dental thread, interdental stimulators, and dental showers (irrigators) play a key role in biofilm removal [1]. More effective elimination is ensured using chemical inhibitors that slow down biofilm formation and inhibit oral pathogens. Effective synthetic/natural chemoprophylaxis should combine antimicrobial (antiseptic) and antiadhesive effects. These requirements are met by several antiseptics. Chlorhexidine digluconate [12] and essential oils - thymol, eucalyptol, menthol and methylsalicylate are the most effective. The latter are active components of Listerine Ò products [13]. However, a number of secondary plant metabolites, e.g. quarternary benzophenantridine alcaloids found in e.g. extract of Macleya cordata [14, 15], proanthocyanidines of A2 types found in fruits of cranberry (Vaccinium macrocarpon) [16-19], polyphenols found in fruits of Vitis vinifera [20] display antiplaque effects. Natural substances with anti-inflammatory, antioxidant and adstringent effects, such as allantoin, phenolic acids, green tea components, natural oils, probiotic bacteria, vitamins B, C, D, E, biominerals Ca, Mg, Zn are used as active components of oral hygiene products as well [15, 21-25]. Beta-glucans, polysaccharides of yeast cell wall (Sacharomyces cerevisie), fungi, some bacteria and plants have significant immunomodulation effects [26], and are recommended to enhance healing process before and after surgery in the oral cavity [15]. The effect of beta-glucans on experimental gingivitis was recently studied in rats [27] and human volunteers [28]. Aluminium lactate is adstringent, it also stops bleeding, and helps in elimination of halitosis [29]. Therefore, it is often 20

4 rocïnõâk22 a jsou doporucï ovaâ ny pro lepsï õâ hojivost prïed a po chirurgickyâch zaâ krocõâch v dutineï uâ stnõâ [15]. Jejich vliv na experimentaâ lnõâ gingivitis byl nedaâ vno studovaâ n na potkanech [27] ilidskyâ ch dobrovolnõâcõâch [28]. Lakta t hlinityâ maâ uâ cï inky stahujõâcõâ (adstringentnõâ) a krvaâ cenõâ zastavujõâcõâ, pomaâhaâ takeâ odstranï ovat halitoâ zu [29]. DõÂky svyâ m vlastnostem byâvaâ prïidaâ vaâ n do prïõâpravkuê uâ stnõâ hygieny, cï asto v kombinaci s chlorhexidindiglukonaâtem[30]. NedostatecÏ neâ odstranï ovaâ nõâ zubnõâho plaku je pozorovaâ no zejmeâ na u adolescentnõâch ortodontickyâch pacientuê s fixnõâm aparaâ tem [12], a proto jsme se na neï v nasï õâ studii zameïrïili. CõÂlem teâ to praâ ce bylo zhodnocenõâ vlivu uâ stnõâch vod s obsahem b-glukanuê a/nebo laktaâtu hliniteâ ho na tvorbu zubnõâho plaku u pacientuê leâ cï enyâch fixnõâm ortodontickyâm aparaâ tem ve dvojiteï slepeâ studii na dobrovolnõâcõâch. Materia l a metodika Studie byla provedena se vzorky uâ stnõâch vod 1) placebem a s obsahem, 2) kombinace laktaâ tu hliniteâ ho s beta-glukanem a 3) laktaâ tu hliniteâ ho. Protokol studie byl schvaâ len Etickou komisõâ Fakultnõ nemocnice Olomouc a Le karïskeâ fakulty Univerzity Palacke ho dne pod cï. j. 14/11. Byl sestaven soubor 21 pacientuê veveï ku (16.0 ± 3.8) let, s kompletnõâm chrupem, celkoveï zdravyâ ch, bez medikace, nekurïaâ kuê, leâcï enyâch fixnõâm ortodontickyâm aparaâ tem a s vyâraznyâmi projevy zaâneï tu gingivy. Tab. 1. SlozÏ enõâ uâ stnõâch vod (v %) Tab. 1. Mouth wash composition (v %) Vzorek 1 Sample 1 Vzorek 2 Sample 2 Vzorek 3 Sample 3 Aluminium lactate b-1,3/1,6-glucan Zinc sulphate Potassium sorbate Saccharin Denat. ethanol Pastadent Royal Tagat CH Glycerine Purified water ad 100% ad 100% ad 100% Pro hodnocenõâ vlivu uâ stnõâch vod byl pouzï it dvojiteï slepyâ zkrïõâzï enyâ (cross-over) design (Obr. 1) [31]. VsÏ ichni uâ cï astnõâcistudie tak postupneï, v naâ hodneâ m porïadõâ, uzï õâvalivsï echny testovaneâ uâ stnõâ vody. Kontrolnõ skupinou bylititõâzï pacienti pouzï õâvajõâcõâ placebo (Vzorek 1). U pacientuê byly prïikazïdeâ naâvsïteïveï hodnoceny: stav parodontu pomocõâ indexu pro potrïebu parodontolo- added to oral hygiene products, very often in combination with chlorhexidine digluconate [30]. Insufficient elimination of dental plaque is frequently observed especially in adolescent orthodontic patients with fixed appliance [12], for this reason, we focused on this group in our study. The aim of our work was to evaluate the impact of mouth rinses with b-glucanes and/or aluminium lactate on plaque formation in patients treated with fixed orthodontic appliances using a double-blind study and volunteers. Material and method Mouth rinses containing 1) placebo, and 2) combination of aluminium lactate and b-glucan, and 3) aluminium lactate were used. The study protocol was approved by the Ethics Committee of the University Hospital and Faculty of Medicine and Dentistry, Palacky University in Olomouc on February 14, 2011 (ref. No. 14/11). The sample was of 21 patients between the ages of 12 and 28 (16.0 ± 3.8), with complete dentition, healthy, nonsmokers, treated with fixed orthodontic appliances. All had significant manifestations of gingivitis. To assess the impact of mouth rinses we used double-blind cross-over design (Fig. 1) [31]. The volunteers gradually used all mouth rinses tested in random order. The control group included the same patients using placebo (Sample 1). During each check-up we evaluated the following parameters: periodontium condition by means of CPITN and PBI, the level of oral hygiene by means of plaque index PLI [32]. In the given intervals (6x) from each volunteer a sample of plaque swab was taken and sent for microbiological examination. Samples were cultivated in blood agar, Sabouraud agar, and MacConkey agar. They were evaluated semiquantitatively starting with ¹rareª and ending with ¹+++ª. The evaluation was translated into the scale 0-3 for individual microorganisms, and in each individual the amount of microbes at the beginning and at the end of the testing period was calculated. During the preliminary examination we removed tartar and plaque from all patients and gave them a sample of mouth rinse. For 4 days, the patients abandoned common oral hygiene (tooth brush and tooth paste) and 5 times a day (after they woke up, after breakfast, lunch, dinner, and before going to bed) they intensively rinsed their mouths (around teeth and gums) with 10 ml experimental mouthwash/placebo for 1 min. On day 4 there was a check-up, subjective impressions related to the use of a mouthwash were recorded, and patients were instructed on how to care for dentition according to Stillman. Over 14 days (i.e. wash-out period) the patients used tooth brush and interdental brush with a tooth paste containing no inhibitors of dental plaque with the exception of sodium fluoride, for 5 min in the morning and in the evening. 21

5 gickeâ ho osï etrïenõâ (CPITN) a indexu krvaâ civosti daâsnï o- vyâch papil (PBI) a uâ rovenï hygieny uâ stnõâ dutiny plakovyâ m indexem (PLI) [32]. KazÏ deâ mu dobrovolnõâku byl v danyâ ch intervalech 6x odebraâ n vzorek steï ru z plaku a zaslaâ n na mikrobiologickeâ vysï etrïenõâ. Zde byly vzorky kultivovaâ ny na krevnõâm agaru, SabouraudoveÏ agaru a MacConkeyho puêdeï a hodnoceny semikvantitativneï od hodnoty ¹ojedineÏ leª azï po ¹+++ª. Hodnocenõ bylo prïevedeno na cï õâselneâ vyjaâdrïenõâ 0-3 pro jednotliveâ mikroorganismy a u kazïdeâ ho jedince bylo spocïõâtaâ no celkoveâ mnozïstvõâ mikrobuê na zacïaâ tku a koncikazïdeâ ho testovacõâho obdobõâ. PrÏivstupnõÂm vysï etrïenõâ byl kazïdeâ mu pacientovi odstraneï n zubnõâ kaâ men a plak a obdrzï el vzorek uâ stnõâ vody. Po dobu 4 dnuê vynechal beïzï nou uâ stnõâ hygienu zubnõâm kartaâcï kem a mõâsto toho si5x denneï (po probuzenõâ, po snõâdani, obeï deï, vecï erïia prïed spaâ nkem) intenzivneï vyplachoval uâ sta (okolo zubuê a daâ sneï ) experimentaâ lnõâ uâ stnõâ vodou/placebem v mnozï stvõâ 10 ml po dobu 1 min. CÏ tvrtyâ den naâ sledovala kontrola, byly zaznamenaâ ny subjektivnõâ pocity prïipouzïõâvaâ nõâ uâ stnõâ vody a provedena instruktaâzï domaâcõâ peâcï e o chrup metodou dle Stillmana. Ve 14 dennõâm intervalu mezi testovacõâmi obdobõâmisipacienticï istili chrup 5 min. raâ no a vecï er zubnõâm a mezizubnõâm kartaâ cï kem za pouzï itõâ zubnõâ pasty, kteraâ neobsahovala s vyâ jimkou fluoriduê zï aâ dneâ inhibitory tvorby zubnõâho plaku. Ke statistickeâ mu zpracovaâ nõâ vyâ sledkuê byl pouzï i t program SPSS v. 15, SPSS Inc. Chicago, USA na hladineï vyâ znamnosti0,05. U kazï deâ ho pacienta byly jednotliveâ parametry hodnoceny vzïdy prïed a po testovacõâm obdobõâ s jednotlivyâmiuâ stnõâmivodami, bez ohledu na porïadõâ, paâ rovyâ mitesty (t-test, prïõâpadneï WilcoxonuÊ v paâ rovyâ test) a naâ sledneï analyâ zou rozptylu, resp. FriedmanovyÂm testem. Pro kazïdou uâ stnõâ vodu se vyhodnocoval vyâslednyâ pocï et n = 20 pacientuê. Vy sledky Testova nõâ se zuâ cï astnilo celkem 21 dobrovolnõâkuê, z nichzï jeden studii nedokoncï il pro neprïõâjemneâ subjektivnõâ pocity. DobrovolnõÂci pouheâ vyplachovaânõâ uâ stnõâmi vodamihodnotilivesmeï s jako nedostatecï nou naâ hradu cï i sïteï nõâ zubuê kartaâcï kem, neï kterïõâ dobrovolnõâcisisteï zïovalina pocit nedostatecïneâuâ stnõâ hygieny, nebyly vsïak pozorovaâ ny zïaâ dneâ negativnõâ vedlejsï õâ uâ cï inky. Pa rovyâ mitesty byl prokaâ zaâ n statisticky vyâ znamnyâ pokles indexu PBI po pouzïõâvaânõâuâ stnõâ vody s obsahem kombinace laktaâ tu hliniteâ ho s beta-glukanem; u indexu CPITN byl prokaâzaân naâruê st hodnot po pouzïõâvaâ nõâ placeba (p = 0,033), u testovanyâch uâ stnõâch vod se hodnoty beï hem testovacõâch obdobõâ vyâ znamneï nezmeï nily (p = 0,017, Obr. 2.). Plakovy index PLI statisticky vyâznamneï vzrostl po pouzï õâvaâ nõâ vsï ech vod. Analy zou rozptylu vsï ak nebyly prokaâ zaâ ny statisticky vyâ znamneâ roz- Obr. 1. Sche ma studie Fig. 1. Study scheme Statistical data were processed with software SPSS v.15, SPSS Inc., Chicago, USA, the level of significance In each patient individual parameters were always evaluated before and after a testing period with individual mouth rinses using paired tests (t-test, Wilcoxon paired test), and with ANOVA and Friedman test, respectively. For each mouth rinse, final n = 20 was evaluated. Results One volunteer dropped out due to subjective discomfort. The care that was limited only to the use of mouth rinse was considered insufficient substitute for tooth brushing; some volunteers reported the feeling of insufficient oral hygiene. However, no negative side effects were found. Paired tests confirmed statistically significant decrease in PBI after the use of mouth rinse containing the combination of aluminium lactate and beta-glucan. For CPITN, the values increased after the use of placebo (p=0.033), but they did not change significantly for tested mouth rinses (p = 0.017, Fig.2). The plaque index PLI increased significantly after the use of each mouth rinses tested. Nevertheless, ANOVA did not 22

6 rocïnõâk22 Obr. 2. Vliv testovanyâch uâ stnõâch vod na index krvaâ civosti daâsnï ovyâch papil (PBI), index pro potrïebu parodontologickeâhoosï etrïenõâ (CPITN), plakovyâ index (PLI) a celkoveâ pocï ty mikroorganismuê v plaku (N) u pacientuê prïed a po pouzïõâvaâ nõâ placeba (bõâlou), kombinace laktaâ tu hliniteâ ho s beta-glukanem (zï lutou) a laktaâ tu hliniteâ ho (zelenou, n= 20). VyÂsledky jsou prezentovaâ ny jako box grafy (vodorovnaâ cï aâ ra v boxu znaâ zornï uje hodnotu mediaâ nu, dolnõâ a hornõâ hrana 1. kvartil, resp. 3. kvartil, anteâ nky ukazujõâ maximaâ lnõâ a minimaâ lnõâ nameï rïeneâ hodnoty; pokud byly v souboru nalezeny odlehleâ a extreâ mnõâ hodnoty, jsou zakresleny krouzï ky a hveï zdicï kami). Fig. 2. The impact of mouth rinses on PBI, CPITN, PLI, and overall number of microorganisms in plaque (N) in patients before and after the use of placebo (white), combination of aluminium lactate and beta-glucan (yellow), and aluminium lactate (green, n=20). The results are presented in the form of box graphs (horizontal line in the box represents median, bottom and top edge 1st quartile and 3rd quartile, respectively, antennae show maximum and minimum values measured; if there were recorded distant and extreme values, they are depicted with rings and stars). dõâly meziefektem trïõâ uâ stnõâch vod na indexy PBI, CPITN cï ipli. Ve vzorcõâch zubnõâho plaku uâcï astnõâkuê studie byla nalezena sï irokaâ sï kaâ la mikroorganismuê, prïicï emzï neïktereâ se vyskytovaly opakovaneï, jineâ jen vyâ jimecï neï. PrÏevaÂzÏneÏ byla vykultivovaâna beï zïnaâ uâ stnõâ floâ ra, naprï.: viridujõâcõâ streptokoky, koagulaâ za negativnõâ stafylokoky, uâ stnõâ neiserie a koliformnõâ bakterie, v zaâ vislosti na stavu chrupu pacienta a jeho aktuaâ lnõâ straveï. Ve 12 odbeï rech byly prokaâ zaâ ny kvasinky, ktereâ prïetrvaâvaly po celou dobu studie. Wilcoxonovy m testem byl prokaâ zaâ n statisticky vyâ znamnyâ uâ bytek pocï tu bakteriõâ po pouzïõâvaânõâ uâ stnõâ vody s obsahem laktaâ tu hliniteâho (p = 0,025, Obr. 2.); FriedmanovyÂm testem vsï ak nebyly prokaâ zaâ ny statisticky vyâ znamneâ rozdõâly v rozdõâlech pocï tu mikroorganismuê mezijednotlivyâmiuâ stnõâmivodami, p = 0,252. proved statistically significant differences between the effectiveness of three mouth rinses related to PBI, CPITN, and PLI. In the dental plaque we found a wide range of microorganisms; some occurred repeatedly, some only rarely. Mostly common oral flora was cultivated. Viridans streptococci, coagulase-negative streptococci, oral neiseria, and coliform bacteria, depending on the patient's dental condition, and his/her food were identified. In 12 swabs yeasts were found that persisted during the whole study. The Wilcoxon paired test showed a statistically significant reduction of bacteria after the use of mouth rinse containing aluminium lactate (p = 0.025, Fig. 2); however, Friedman test did not show statistically any significant difference in number of microorganisms between individual mouth rinses (p = 0.252). 23

7 Diskuse Studie ukaâ zala, zïe naâ hrada mechanickeâ ho cï isï teï nõâ zubuê pouhyâ m intenzivnõâm vyplachovaâ nõâm uâ st experimentaâ lnõâmiuâ stnõâmivodamivedla ke zvyâsï eneâ akumulaciplaku, hodnoceneâ plakovyâ m indexem. U placeba byl zjisïteïn naâruê st indexu CPITN. Tyto vyâsledky jsou v souladu se zaâveï ry podobneâ studie, provedeneâ suâ stnõâ vodou obsahujõâcõâ polyfenoly chmele [31]. PouzÏõÂvaÂnõÂu stnõâ vody s obsahem b-glukanu se projevilo ve snõâzïenõâ indexu PBI. V nedaâ vneâ studii na potkanech b-glukan podaâ vanyâ v pitneâ vodeï signifikantneï snõâzï il ztraâ ty kostnõâ tkaâ neï v modelu experimentaâ lnõâ parodontitis [27]. Podobny uâcï inek byl pozorovaânprïipou- zï õâvaâ nõâ uâ stnõâ vody s b-glukanem u zdravyâ ch muzï uê, kteryâ m byla indukovaâ na experimentaâ lnõâ gingivitis. Ve srovnaâ nõâ s kontrolnõâ skupinou nebyl ovlivneï n PLI ani gingivaâ lnõâ index, ktereâ u vsï ech dobrovolnõâkuê v pruê beï hu experimenty vzrostly. U skupiny, kteraâ proplachovala uâ sta prïõâpravkem s b-glukanem a naâ sledneï jej spolkla, dosï lo k signifikantnõâmu zvyâsï enõâ mnozï stvõâ gingivaâ lnõâ tekutiny [28]. PouzÏõÂvaÂnõ uâ stnõâ vody s laktaâ tem hlinityâm se v nasïõâ studii projevilo snõâzï enõâm celkoveâ ho pocï tu mikroorganismuê v dentaâ lnõâm plaku. Takovy efekt nebyl pro tuto laâ tku dosud popsaâ n. Lakta t hlinityâ je v prïõâpravcõâch uâ stnõâ hygieny pouzïõâvaâ n zejmeâ na pro sveâ adstringentnõâ uâcï inky. PrÏedpoklaÂda se, zïe na povrchu daâ snõâ a krcïkuê zubuê iniciuje tvorbu koagulacï nõâ membraâ ny, kteraâ kraâtkou dobu chraânõâdaâ sneï /dentin prïed vneïjsï õâm podraâzïdeïnõâm. To vede k zastavenõâ zaâneï tuauâ tlumu bolesti. Optima lnõâho uâ cï inku laktaâ tu hliniteâ ho bylo dosazï eno prïi koncentraci0,05 % a prïetrvaâ val 3 azï 4 hodiny. KoagulacÏ nõâ uâ cï inek laktaâ tu hliniteâ ho na bõâlkovinneâ slozïky v dentinovyâ ch kanaâ lcõâch snizï uje jejich funkci ve vedenõâ podraâ zï deï nõâ, cozï prïispõâvaâ k desenzibilizaci obnazï enyâch zubnõâch krcï kuê. Lakta t hlinityâ takeâ pomaâhaâ odstranï ovat zaâ pach z uâ st (halitoâ zu) [29]. ZaÂveÏr Vy sledkem studie bylo zjisï teï nõâ, zï e prïõârodnõâ laâ tky b- glukan a laktaâ t hlinityâ nesignifikantneï pozitivneï ovlivnï ujõâ uâ rovenï uâ stnõâ hygieny, ale jejich samotneâ uzï itõâ nenahrazuje mechanickeâ cï isïteï nõâ zubnõâm kartaâcï kem. Jako metoda volby se tudõâzï jevõâ kombinovaneâ pouzïõâvaânõâ zubnõâho a mezizubnõâho kartaâ cï ku spolu s vyâ plachy uâ stnõâ vodou s aktivnõâmi laâ tkami. PodeÏ kovaâ nõâ: AutorÏi deï kujõâ firmeï FAVEA (KoprÏivnice) za prïõâpravu testovanyâch uâ stnõâch vod a Mgr. KaterÏineÏ LangoveÂ, Ph.D., za statistickeâ zpracovaânõâvyâ sledkuê. StrÏet zaâ jmuê: ZÏ aâ dnyâ z autoruê nenõâ s firmou FAVEA v zameï stnaneckeâmcï i jineâ m pracovnõâm vztahu. Discussion The study showed that substitution of mechanical tooth brushing with intensive rinsing of mouth with experimental mouthwashes resulted in increased accumulation of plaque (according to the plaque index PLI). When placebo was applied, then CPITN increased. The results agree with the conclusions of a similar study of a mouthwash containing hop polyphenols [31]. The use of mouthwash containing b-glucan led to PBI decrease. A recent study with rats showed that b-glucan administered in drinking water significantly reduced the loss of bone tissue in a model of experimental periodontitis [27]. Similar effects were recorded in healthy men that with experimentally induced gingivitis. In comparison with a control group the PLI and gingival index were not influenced - the values of these indexes increased in all volunteers during the experiment. A significant increase in gingival liquid was observed in the group of probands who rinsed their mouths with b-glucan containing mouthwash and then swallowed it [28]. The use of a mouthwash with aluminium lactate led to decrease in overall number of microorganisms in dental plaque. This is the first report for this substance to date. Aluminium lactate is known and used especially for its astringent effects. It is assumed that on the gum and tooth neck surface it initiates the formation of a coagulation membrane that for a short time protects gums/dentine against irritation. This stops inflammation and inhibits pain. Optimum effect of aluminium lactate was achieved in the concentration 0.05% and lasted for 3-4 hours. Coagulation action of aluminium lactate on albuminoid components in dentine ducts inhibits their ability to transmit irritation which results in desenzibilitation of exposed necks. Aluminium lactate also helps to eliminate halitosis [29]. Conclusion The study found that b-glucan and aluminium lactate positively, though insignificantly, influence oral hygiene but they cannot fully substitute for mechanical cleaning with a tooth brush. The method of option thus seems to be combined use of tooth brush and interdental brush and rinses with mouthwashes containing active substances. Acknowledgment: Authors would like to thank the company FAVEA (KoprÏivnice) for preparation of tested mouthwashes, and to Mgr. KaterÏina LangovaÂ, Ph.D. for statistical processing of the data. Conflict of interests: Authors have no engagement or other professional relationship to company FAVEA. 24

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9 31. Shinada, K.; Tagashira, M.; Watanabe, H.; Sopapornamorn, P.; Kanayama, A.; Kanda, T.; Ikeda, M.; Kawaguchi, Y.: Hop bract polyphenols reduced three-day dental plaque regrowth. J. Dent. Res. 2007, 86, cï. 9, s Loesche, W. J.: Ecology of the oral flora. In: Nisengard, R. J., Newman, M. G., eds. Oral microbiology and immunology. Philadelphia: W. B. Saunders; s Doc. Ing. KaterÏina ValentovaÂ, Ph.D., U stav leâ karïskeâ chemie a biochemie LF UP HneÏ votõânskaâ 3, Olomouc Altis Group spol. s r. o. ±vyâhradnõâ zaâstupce pro CÏ eskou republiku a Slovensko V roce 2013 pro VaÂs prïipravujeme: TermõÂn: 8.± MõÂsto konaânõâ: Mikulov TeÂma: Ortodonticko-chirurgicka leâcïba pacientuê scïelistnõâmi vadami ± zaâklady a noveâ zkusïenosti PrÏednaÂsÏejõÂcõÂ: Doc. MUDr. Rene FoltaÂn, Ph.D. MUDr. JirÏõ Petr TeÏsÏõÂsenaVaÂs kolektiv Altis Group s.r.o. Altis Group spol. s r. o., ZÏ erotõânova 901/12, BrÏeclav Tel./fax: , [email protected], Petra Karafova , Marie PõÂsarÏõÂkova ± Zelena linka: (VOLEJTE ZDARMA!) 26