A Prospective, Double-Blind, Randomized, Controlled Clinical Trial in the Gingivitis Prevention with an Oligomeric Proanthocyanidin Nutritional Supplement

Clinical S udy A P ospec i e, Double-Blind, Randomized, Con olled Clinical T ial in he Gingi i is P e en ion wi h an Oligome ic P oan hocyanidin Nu i ional Supplemen R. M. Díaz Sánchez, G. Cas illo-Dalí, A. Fe nández-Ola a ía, R. Mosque a-Pé ez, J. M. Delgado-Muñoz, J. L. Gu ié ez-Pé ez, and D. To es-Laga es O al Su ge y Depa men , Den al School, Uni e si y o Se ille, Se ille, Spain Co espondence should be add essed o D. To es-Laga es; [email p o ec ed] Recei ed 20 Ma ch 2017; Re ised 10 Augus 2017; Accep ed 13 Sep embe 2017; Published 10 Decembe 2017 Academic Edi o : Giuseppe Valacchi Copy igh © 2017 R. M. Díaz Sánchez e al. This is an open access a icle dis ibu ed unde he C ea i e Commons A ibu ion License, which pe mi s un es ic ed use, dis ibu ion, and ep oduc ion in any medium, p o ided he o iginal wo k is p ope ly ci ed. Aim. To e alua e he effec i eness on issue esponse o he new nu i ional supplemen made o oligome ic p oan hocyanidins in induced gingi i is a e 21 days o use. Ma e ial and Me hods. A p ospec i e, double-blind, andomized, con olled clinical ial was ca ied ou on 20 pa ien s; i is di ided in o an expe imen al g oup and a con ol g oup a e ulfilling he selec ion c i e ia. Pa ien s had o come 4 imes du ing he s udy o egis e he Silness and Löe index, he gingi al bleeding index, he plaque index, he inflamma o y c e icula fluid s udy (IL6), and he changes in he b igh ness o he gingi a. No complemen a y hygiene me hods we e allowed du ing he 21 days. Resul s. The Silness and Löe index was highe in he con ol g oup han in he expe imen al g oup, eaching a wo old diffe ence be ween he g oups (p<00001). The gingi al bleeding index also suppo s his ac , since he bleeding was lowe in he expe imen al g oup (p<0005). Howe e , he den al plaque on he oo h su ace acco ding o he plaque index was 33% highe in he expe imen al g oup (p<0006). Some diffe ences in he IL-6 we e ound in he c e icula fluid (p<00001). Conclusion. Oligome ic p oan hocyanidins ha e an effec on he pe iodon al issue’s heal h. No effec s on he accumula ion o plaque on he oo h su ace we e ound, so u he s udies a e needed o de e mine he na u e o he plaque. 1. In oduc ion Gingi i is is an inflamma o y disease o den al issue sup- po ha in he fi s ins ance affec s he gingi a. Gingi i is could mo e on o mo e ad anced s ages, leading o he de elopmen o pe iodon i is, in which he inflamma ion and bac e ial in ec ion can p oduce he des uc ion o he suppo ing issues o he ee h, gingi a, pe iodon al ligamen , and al eola bone [1–5]. Gingi i is is due o he long- e m effec s o plaque deposi s on he ee h, which is composed o bac e ia, loca ed on he su aces o he ee h and in he gingi al sulcus. I pla- que is no p e en ed o emo ed, i u ns in o a ha d deposi called a a (o calculus) ha becomes apped a he base o he oo h. Plaque and calculus i i a e and p oduce gingi al swelling. Bac e ia and he oxins p oduce in ec ion and mo e swelling gingi a [1–5]. C e icula fluid is no mal plasma exuded flowing h ough he gingi al sulcus. A pa ien wi h gingi i is de elops an inflamma o y esponse wi h a la ge numbe o media o s in ol ed, such as he cy okines IL-6 and IL-8 [1–6]. In e leukin 6 (IL-6) is a mul i unc ional cy okine ha is p oduced by a ange o cells, and i is in ol ed in he B-cell diffe en ia ion, p oli e a ion, and diffe en ia ion o T cells; in he immunoglobulin, s imula ion by he B-cell sec e ion [7] can also induce bone eabso p ion [8]. IL6 is a use ul diagnos ic indica o o de e mine he p og ession o he gin- gi i is o a pe iodon al disease [9]. The ea men goal o gingi i is is o educe inflamma ion by cleaning he ee h, using diffe en ins umen s o emo e he den al plaque deposi s acco ding o he case [6]. I is impo an o educa e he pa ien and p esc ibe special oo h- pas e, mou hwash, gel, and so o h which con ain a wide spec um o an isep ic and an i-inflamma o y p ope ies. Hindawi Media o s o Inflamma ion Volume 2017, A icle ID 7460780, 7 pages h ps://doi.o g/10.1155/2017/7460780 Howe e , he den is has he esponsibili y o explain he co - ec oo h b ushing echnique o achie e good esul s [2, 3]. The oligome ic p oan hocyanidins (OPCs) a e one o he mos abundan polyphenolic subs ances in he plan king- dom. These subs ances exhibi a ange o a he su p ising physical and chemical p ope ies which, once applied o li - ing o ganisms, a e ansla ed in o a mul i ude o biological ac i i ies [10]. The OPCs ha e been ecen ly in es iga ed, and, apa om i s powe ul an ioxidan ac i i y, hey ha e been shown o ha e an icance , an i-inflamma o y, an imi- c obial, and asodila o y p ope ies, e ealing o be a po en- ially aluable he apeu ic ool o he ea men o many illnesses [11–15]. The a ailable esea ch has demons a ed ha he acidic en i onmen o he human s omach does no easily deg ade he p oan hocyanidins; he e o e, he a es o abso p ion in he uppe gas oin es inal ac a e no high [16]. Howe e , i seems ha e en he low le els obse ed in u ine a e an o al dose (gene ally less han 250/0 o doses/ o iginal) a e sufficien o significan ly inc ease he an ioxi- dan capaci y in plasma/se um. When OPCs each he colon, hey suffe an ex ensi e deg ada ion due o colonic flo a [16]. The me aboli es and biological p ope ies o his p ocess ha e no been in es iga ed ye ; i has been sugges ed ha hey may ha e p o ec i e effec s on he an ioxidan and sys emic illness [11–16]. Rega ding he o he componen included in he OPCs, i amin C, we know ha i is a good associa i e op ion o bil- be y. Vi amin C gene ally p o ides p ope ies ha con ib- u e o he o ma ion o collagen, which helps o main ain unc ionali y o ee h and gums and o he p ope ies such as ein o cemen o he immune sys em and po en an ioxi- dan opposi ion o oxida i e s ess [17–21]. Al hough in i o s udies published abou he in e ac- ion be ween he OPCs and pa hogens in he o al ca i y ha e had e y good esul s, he e a e no publica ions wi h he app op ia e design o assess he clinical efficacy o OPCs in gingi al disease in human, excep an a icle pub- lished in 2015 compa ing he chlo hexidine and he c an- be y mou hwash [22–25]. The aim o his s udy is o e alua e he effec i eness o he new nu i ional supplemen made o oligome ic p oan ho- cyanidins (OPCs) on issue esponse o p e en gingi i is. We hypo hesise a heal hy s a e o he pe iodon al issue in he s udy g oup e sus gingi i is in he con ol g oup. 2. Ma e ial and Me hods 2.1. S udy Design and Popula ion. A andomized, double- blind, placebo-con olled, clinical ial o 21 days o du a ion was conduc ed among 20 olun ee s uden s o he School o Den is y o he Uni e si y o Se ille. The du a ion o 21 days was chosen as ha ime allows he pa ame e s udy on he gingi a and was enough o s udy he inflamma ion o he gin- gi a wi hou causing any i e e sible p oblem in i . No p e i- ous s udies ha e been ound in he li e a u e, so 20 subjec s we e de e mined o he pilo s udy o es ou hypo hesis. The E hics Commi ee o he Uni e si y o Se ille app o ed he s udy p o ocol. P io o pa icipa ion, he pu pose and p ocedu es we e ully explained o all heal hy olun ee s and all pa icipan s ga e w i en in o med con- sen in acco dance wi h he Decla a ion o Helsinki. The s udy was designed, conduc ed, analyzed, and epo ed acco ding o he guidelines o Good Clinical P ac ice. The s udy was app o ed by he h p://ClinicalT ials.go P o ocol Regis a ion and Resul s Sys em wi h he numbe NCT02515929. The p o ocol can be accessed i necessa y in h p://ClinicalT ials.go . The s udy was ca ied ou be ween Sep embe 2013 and Janua y 2014. The ec ui men s a ed in Sep embe , and he baseline s age ook place in Oc obe 2013. A e he 21-day ollow-up da a was ea ed ob aining he esul s in Janua y 2014. The medical and den al his o ies we e done a he p e- sc eening isi , and we selec ed he 21 pa icipan s based on he inclusion and exclusion c i e ia. The inclusion c i- e ia we e he ollowing: subjec s olde han 18 yea s old, male o emale, good o e all heal h, and a minimum o 20 ee h ( ee h wi h big ca ies we e c owned o ex ensi ely es o ed, and ee h ha we e o hodon ic banded, abu men s, o hi d mola s we e no included in he oo h coun ); he olun ee s signed he w i en consen be o e he ini ia ion o he s udy. The exclusion c i e ia we e he ollowing: pe iodon al disease (defined as 4mm and/o posi i e bleeding when p obing), p egnan o b eas eeding, subjec s wi h fixed o emo able p os hesis, umou pa hology in o al ca i y, use o an ibio ics du ing a 2-mon h pe iod p io o he s a o he ial, hype sensi i i y o ed ui s in gene al, xe os omy, ac i e smoke , con agious-in ec ious pa hologies, pa hology wi h se e e sys emic epe cussions, any o he judgmen ha makes he in es iga o belie e o endange o isk he subjec pa icipan , subjec s wi h phenylke onu ia o alle gy o aspa ame, and he use o any o al hygiene p oduc du ing he s udy. A single examine p e iously calib a ed de e mined assessmen o pa ien eligibili y o he s udy and en olmen o pa ien s in o ial. Pa ien s eligible o he s udy we e indi- idually andomly assigned o oligome ic p oan hocyanidins nu i ional supplemen ea men o placebo g oups by an in o ma ics p og amme by LACER S.A. The s udy was double-blind. The examine and he pa ien did no know in o which g oup hey we e assigned. The esul s we e ea ed by ano he examine who knew which pa ien s belonged o he expe imen al o placebo g oup. The masking p ocess was es ablished by a numbe assigna ion o each pa ien in he s udy, and he ea men was he same bo h o he pa ien and he examine who we e no allowed o know o which g oup hey belonged. Pa ien s had o ake he expe imen al o placebo ea - men each nigh a e dinne and a e a inse wi h wa e . The pill was main ained in he mou h un il comple e dissolu- ion. Nei he d inking no ea ing was allowed du ing 30 minu es a e aking he ea men . The expe imen al ea - men consis ed o 90 mg exocian c an 408 (equi alen o 36mg OPCs) and 120mg o i amin C, while he placebo g oup was composed o he same o ganolep ic subs ance bu ee o ac i e ing edien s. 2 Media o s o Inflamma ion Bo h we e simila in appea ance. No complemen a y hygiene me hods we e allowed du ing he 21 days ( oo h b ushing, inses o i iga ion wi h any p oduc , o flossing), which could cons i u e a bias o he s udy. A he ial baseline s age, a a a emo al was ca ied ou in each pa ien o egula ize he ini ial si ua ion; a pa ien dia y, enough medica ion o he en i e s udy, and ins uc- ions we e gi en o i s co ec ulfillmen . The inflamma ion o he c e icula fluid (IL6) and he b igh ness o he gingi a we e egis e ed. Two e alua ion isi s we e pe o med on days 14 and 21 o he s udy o an o al clinical examina ion and o egis e he Silness and Löe index, he gingi al bleeding index, he Tu esky plaque index, he inflamma o y c e icula fluid s udy (IL6), and changes in he b igh ness o he gingi a. 2.2. Silness and Löe Index [26, 27]. Six ee h we e examined acco ding o Ram jo d c i e ia (16-21-24-36-41-44). Fou su aces o each oo h we e examined, making a o al o 24 measu emen s. These measu emen s we e pe o med wi h a pe iodon al p obe by he same examine . 2.3. Gingi al Bleeding Index [28]. A pe iodon al p obe was used o ake his index. The alues es ablished o his exam- ina ion a e 0—absence o inflamma ion; 1—mild inflamma- ion, sligh change in colo , no gingi al edema, and no bleeding on p obing; 2—mode a e inflamma ion, edness, edema, and gingi al hype ophy, and bleeds o p obe (a e 10 seconds); 3—se e e inflamma ion, ma ked ed- ness, and hype ophy. The e may be ulce a ions; he gingi a ends o be spon aneous in bleeding. 2.4. Tu esky Plaque Index [29, 30]. The buccal su aces o he an e io ee h we e examined using a mou hwash o basic uchsine as de eloping agen plaque, and he ollowing nume ical sco ing sys em om 0 o 5 was es ablished: 0— he e is no plaque; 1—independen s eaks o plaque in he ce ical ma gin o he oo h; 2—a hin con inuous band o plaque (up o 1 mm) a he ce ical ma gin; 3—a band g ea e han one millime e wide, bu co e s less han one- hi d o he c own; 4— he plaque co e s a hi d, bu no mo e han wo- hi ds o he c own; 5— he plaque co e s wo- hi ds o mo e o he c own. 2.5. Inflamma o y C e icula Fluid S udy (IL6) [7–9]. C e- icula fluid samples we e collec ed om in e den al a eas (lingual, buccal, mesial, and dis al) o six ee h dis ibu ed in he ou quad an s (16, 21, 24, 36, 41, and 44) using fi e s ips o p essed pape ha we e 2cm long, especially o c e icula fluid. The imp egna ion ime o each pa ien was 5 seconds, and he s ips we e immedia ely inse ed in o mic o ubes ha con ained 0.5 ml o Eppendo wi h 50 μl saline a 4 ° C o p ese a ion. The samples we e hen anspo ed o he biological labo a o y in a e ige a o and we e s o ed ozen a −80 ° C a he labo a o y un il hey we e p ocessed. Then, we p oceeded o analyze he concen a ion (pg/ml) o in e leukin 6 p esen in each sample using panels 96-well bioplex b and Luminex®. 2.6. B igh ness o he Gingi a. The b igh ness o he gingi a was aken o iden i y possible changes in he gingi al colo . The eddening o he gingi a accompanies he inflamma ion o he issue, which is a ac o ha may help o diffe en ia e inflamma o y changes a his le el. The luminosi y o he gingi a was egis e ed wi h Mic o Spec oShade™MHT Op ic Resea ch AG. This ins umen is designed o ake den al colo . Howe e , besides including colo guides, he Spec oShade has he abili y o measu e he b igh ness o any colo , no jus shades o whi e. The change o colo o he gingi a was obse ed in each pa ien , and he colo e e ence was measu ed in he same poin in each e alua ion isi , which helped us compa e he change in he colo a ion o he same poin . 2.7. S a is ical Analysis. The managemen o he da a was pe o med using he SPSS s a is ical p og am. A simila da abase o No ebook Da a Collec ion was c ea ed. The e a e a minimum ange and a maximum ange o each a i- able, and he name o he a iable and i s alues we e defined. The da a a e p esen ed as he mean ±s anda d de ia ion o he measu emen s, and a p alue less han 0.05 was consid- e ed s a is ically significan . Chi-squa e es was pe o med o quali a i e a iable and S uden ’s - es o he quan i a i e a iable, a e he Kolmogo o -Smi no o assu e he no mali y. The E hical Commi ee app o ed his s udy, and all examina ions and ea men s we e pe o med wi h he con- sen o he subjec s and acco ding o he guidelines o he Decla a ion o Helsinki. 3. Resul s A s a is ical analysis be ween he wo s udy g oups, 10 pa ien s in each g oup, in e ms o sex, age (yea s), weigh (kg), heigh (cm), obacco use, alcohol, and use o concomi- an medica ion, was ca ied ou o ensu e he eliabili y o he esul s. We ound no significan diffe ences be ween he s udy g oup and he con ol g oup, so we can say ha he homogenei y be ween he g oups was achie ed (Table 1). Table 1: G oups cha ac e is ics. Expe imen al (n=10) Con ol (n=10)p Sex Men 2 5 0.674 Women 8 5 Yea s 24.6 ±4.27 23.5 ±1.64 0.525 Weigh (Kg) 57.04 ±9.33 66.6 ±16.43 0.420 Heigh (cm) 164.7 ±8.02 169.3 ±8.59 0.325 Smoke Yes 0 0 1.000 No 10 10 Alcohol Yes 1 1 1.000 No 9 9 Medical ea men Yes 1 (ACO) 0 0.875 No 9 10 3Media o s o Inflamma ion The medica ion pe cep ion o bo h g oups, as he ini ial fla ou , he p oduc du abili y, he appea ance, size, and effi- ciency, was he same; he e we e no s a is ically significan diffe ences be ween he da a ob ained (Table 1). No ad e se e en s we e no ified du ing he s udy. Once he s udy g oups we e analysed, he da a p ocessing was conduc ed depending on he diffe en iews held. 3.1. Pe iodon al Index o 15 Days o T ea men (In e media e Visi ). The Silness and Löe gingi al index (o 0–3 om leas o g ea es gingi al inflamma ion) (Table 2) was highe in he con ol g oup han in he expe imen al g oup, eaching a wo old diffe ence be ween he wo g oups (p<00001). The gingi al bleeding index also co obo a es his ac , as he bleeding was lowe in he expe imen al g oup e sus he con ol g oup (p<0005) (Table 2). In con as o he alues ob ained in he Silness and Löe and gingi al bleeding indexes, he amoun o den al plaque deposi ed on he su ace o he pa ien s acco ding o he Tu esky index plaque was sligh ly highe (33%) in he expe imen al g oup e sus in he con ol g oup (p<0006) (Table 2). 3.2. Pe iodon al Index o 21 Days o T ea men (Final Visi ). The esul s ob ained 21 days a e s a ing he ea men showed he endency obse ed in he in e media e isi , inc easing he diffe ence be ween he s udy g oups. Silness and Löe index was highe in he con ol g oup compa ed o he expe imen al g oup, wi h a diffe ence o 50% be ween he s udy g oups (p<00001). The gingi al bleeding index, which showed bleeding in he con ol g oup, was highe in he expe imen al g oup (p<00001). Howe e , he plaque deposi ion was highe in he expe - imen al g oup han in he con ol g oup acco ding o he Tu esky plaque index ha ing plaque deposi doubled in he pa ien s in he expe imen al g oup (p<00001) (Table 2). 3.3. Gingi al B igh ness. The gingi al b igh ness was aken in each pa ien using he Mic o Spec oShade Op ic Resea ch MHT AG. This alue was aken a he beginning, middle isi , and in he final isi . The da a ob ained om hese h ee measu emen s in each pa ien showed no s a is ically signifi- can diffe ences (p alues om 0.351 o 0.545) (Table 3). 3.4. Inflamma o y C e icula Fluid S udy (IL6). S a is ically significan diffe ences we e ound a he baseline be ween he expe imen al g oup and he con ol g oup and in he sub- sequen isi s. No s a is ically significan diffe ences be ween he fi s isi and he second isi , and he fi s and hi d isi s we e ound, al hough i was significan be ween he second isi and he hi d isi (Table 4). 4. Discussion Gingi i is is caused by deposi ion o plaque on he oo h su - ace; his deposi ion cons i u es an i i an ha igge s he gingi al inflamma ion and bleeding. Gingi i is could be e e sible i he pa ien has a cons an and good hygiene. These measu es could be ca ied ou by a p ope b ushing echnique [1–5]. The d ug used in he cu en s udy is a nu i ional supplemen based on bluebe y and ed ui ich in oligo- me ic p oan hocyanidins (OPCs) [7]. P oan hocyanidins ha e been ecen ly in es iga ed, as we sugges ed in he in o- duc ion, due o hei powe ul an ioxidan and an icance ac i i ies and due o hei an i-inflamma o y, an imic obial, and asodila o sys emical p ope ies. The beneficial effec o p oan hocyanidins is a ibu ed o i s abili y o educe oxi- da i e s ess, lipid pe oxida ion, ee adical gene a ion, and LDL oxida ion [11]. Mohana e al. published a s udy in 2015 abou he effec o he OPCs in he p e en ion and ea men o a he oscle o- sis. Oxida ion o low-densi y lipop o eins (OxLDL) has been s ongly sugges ed as a key ac o in he pa hogenesis o a he oscle osis [11]. Due o he an ioxidan ac ion o he Table 2: Silness and Löe, gingi al bleeding, and plaque index. Tes E alua ion isi s Expe imen al Con ol p Silness and Löe index Day 14 0.65 ±0.58 1.24 ±0.66 0.0001 Day 21 0.97 ±0.54 1.48 ±0.60 0.0001 Gingi al bleeding index Day 14 0.91 ±0.84 1.33 ±0.75 0.005 Day 21 0.98 ±0.67 1.61 ±0.66 0.0001 Plaque index Day 14 1.68 ±0.79 1.25 ±0.91 0.006 Day 21 3.35 ±1.59 1.68 ±1.43 0.0001 Table 3: B igh ness o he gingi a. B igh ness o he gingi a Expe imen al Con ol p Baseline 30.75 ±4.19 31.37 ±3.55 0.351 Day 14 32.70 ±3.48 31.50 ±3.10 0.480 Day 21 32.57 ±4.72 31.22 ±2.52 0.425 Di (day 21 and day 14) −0.12 ±1.85 −0.28 ±1.92 0.545 Table 4: IL-6 in he c e icula fluid (pg/ml). IL-6 in he c e icula fluid (pg/ml) Expe imen al Con ol p Baseline 62.85 ±45.8 126.60 ±54.03 0.011 Day 14 17.95 ±13.02 47.40 ±33.61 0.019 Day 21 22.15 ±15.14 69.40 ±50.10 0.011 Di (day 14—baseline) 44.90 ±44.05 79.20 ±64.54 0.095 Di (day 21—baseline) −40.70 ±45.02 −56.70 ±70.63 0.245 Di (day 21 and day 14) −4.20 ±3.21 −22.00 ±21.03 0.016 4 Media o s o Inflamma ion p oan hocyanidins and he modula ion o mac ophage di - e en ia ion, Mohana e al. epo ed he benefi o he p oduc as p e en ion and ea men o he a he oscle osis [11]. Wang e al. ha e also es ed he an i-inflamma o y mech- anisms o OPCs [12]. They designed an in i o s udy in mouse models o in es iga e he p o ec i e effec o OPCs agains alcohol-induced li e s ea osis and inju y. The esul s showed ha OPC significan ly imp o ed alcohol-induced dyslipidemia and alle ia ed li e s ea osis by educing le els o se um alanine amino ans e ase (ALT), aspa a e amino ans e ase (AST), o al iglyce ide (TG), o al cho- les e ol (TC), low-densi y choles e ol (LDL-c), and li e malondialdehyde (MDA) and inc easing le els o se um high-densi y lipop o ein (HDL-c) and li e supe oxide dismu ase (SOD). Fu he in es iga ion indica ed ha OPC ma kedly dec eased he exp essions o lipid syn hesis genes and inflamma ion genes such as s e ol egula o y elemen binding p o ein-1c (S ebp-1c), p o ein-2 (S ebp2), in e leu- kins IL-1b and IL-6, and TNF-α10. These s udies sugges ed ha an effec in he inflamma- ion genes and media o s could be modified by he OPCs, educing he inflamma ion ob ained [11, 12]. Ou esul s do no show significan diffe ences in he IL-6 be ween he baseline and he second isi and be ween he baseline and he hi d isi . Significan diffe ences we e ound be ween he 14 h and 21s days and be ween g oups in each isi , including in he baseline s age, so he g oups we e no simila a he beginning o he s udy, which could be a bias in he esul s. Tha eason o ces us o no conside significan ly he esul s ob ained. Howe e , he da a is lowe in he expe imen al g oup in each isi and his ac sugges s ha an effec could be in he IL-6, bu we canno assu e his like in o he s udies because o ou diffe ence in he baseline g oup. Howe e , he clinical pa ame e s, he Löe and Silness index and gingi al bleeding index, showed us ha he clinical inflamma ion was highe in he con ol g oup han in he expe imen al g oup, finding s a is ically significan diffe ences. Du ing he s udy, we obse ed ha he deposi ion o pla- que, Löe and Silness index, gingi al bleeding index, and he b igh ness o he gingi a, inc eased sequen ially, so he inflamma o y ac o s mus ha e also inc eased due o he esul s ob ained in each pa ame e , al hough he e we e no diffe ences be ween he s udy g oups in he b igh ness o he gingi a. We a ibu e his ac o he mechanical i i an ha he plaque cons i u es i sel , wi hou aking in o accoun i s na u e. Besides he an i-inflamma o y effec , OPCs ha e shown an imic obial effec , epo ed by u he s udies [10–16, 23, 25]. This p ope y also jus ifies es ing he effec i eness o he OPCs in pe iodon al disease. A e pe o ming he cu - en s udy, we can affi m ha he OPCs ha e an effec in he heal h o he pe iodon al issues, as we could see h ough he Silness and Löe and gingi al bleeding indexes, which we e significan ly lowe in he expe imen al g oup han in he con ol g oup, al hough i was no co obo a ed by he Tu esky plaque index, whe e he p esence o plaque was pa adoxically highe by 33% in he s udy g oup han in he con ol g oup (Table 1). These esul s lead us o suspec ha his supplemen ce ainly has an imic obial effec , since he gingi al bleed- ing index and Silness and Löe index we e lowe e en in he p esence o mo e den al plaque, so he na u e o he plaque mus be modified, possibly because o a change in he pH o by a diffe en bac e ial composi ion. In Ma ch 2015, Mahesh e al. published in he Con empo a y Clinical Den is y a andomized pa allel clinical ial compa ing a 0.2% chlo hexidine glucona e mou hwash e sus a 0.6% c anbe y mou hwash on S ep ococcus mu ans in 50 sub- jec s, ob aining simila esul s in he colonisa ion o his bac e ia in he plaque [25]. This s udy confi ms he esul s ob ained in ou clinical ial, in which we hypo hesised ha he plaque could be diffe en because o he mino inflamma ion o he pe iodon al issue. Un il his momen , only one s udy has been published abou he use o c anbe y mou hwash (a componen o OPCs) in pe iodon al disease in humans [25]. Howe e , in 2011, Löh e al. published an in i o s udy abou he an imi- c obial effec o he OPCs ac ing in he adhesion o Po phy - omonas gingi alis o KB cells [23]. They pos ula ed ha he OPCs had an an iadhesi e effec agains P. gingi alis in he KB cells. The esul s o his s udy showed ha OPCs exe an iadhesi e effec s agains P. gingi alis, concluding ha OPCs may be use ul o he p e en ion o P. gingi alis-asso- cia ed pe iodon al diseases [23]. Ou s udy does no include bac e ial analysis, bu ou esul s suppo i due o he ac ha in he expe imen al g oup he pa ame e s showed less inflamma ion and less cell esponse o he plaque, which could be a ibu ed o i s na u e. Due o he p e ious s udy [23], we suspec ha he an i- adhesion effec modified he plaque o he subjec s in he expe imen al g oup, making he inflamma ion lowe . Lö h e al. showed ha polyphenols in e ac wi h p o ein su ace (e.g., gingipains, p edominan ly because o i s ac i i y agains A g-gingipain) and change he unc ionali y and eac i i y. Thus, OPCs mainly in e ac wi h he ini ial adhesion p ocess o he bac e ia o he cell memb ane. This p ocess could be ex apola ed o ou s udy, gi ing us an explica ion o ou esul s. IL-6 has no shown significan diffe ences, bu a eal effec in he inflamma ion o he gingi a ook place in ou pa ien s; hus, he Löe and Silness index and gingi al bleeding index we e lowe in he expe imen al g oup. 5. Conclusion The nu i ional supplemen made o oligome ic p oan ho- cyanidins (c anbe y and i amin C) induces an imp o emen in he heal h o he pe iodon al issues, since he Silness and Löe index and gingi al bleeding index we e ma kedly lowe in he expe imen al g oup, al hough ha supplemen has no been shown o ha e effec s on he accumula ion o plaque on he oo h su ace. Fu he s udies a e needed o de e mine he na u e o he plaque. Disclosu e The abs ac was p esen ed as a pos e in he Jou nal o Clinical Pe iodon ology, in he special issue Abs ac s o 5Media o s o Inflamma ion Eu oPe io8, London, UK, 3–6 June as pe he below URL (page 48, D056): h p://onlinelib a y.wiley.com/doi/ 10.1111/jcpe.12398/epd . Con lic s o In e es The au ho s decla e ha he e a e no conflic s o in e es in his s udy. 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