o:25464 Anti-inflammatory effect of amalgam on periapical lesion cells in culture Anti-inflamacijski efekat amalgama na ćelije iz periapikalne lezije u kulturi en ABSTRACT Background/Aim. Amalgam has been used for years in dentistry, but the controversy on its adverse effects, both on local oral/dental tissues and systemic health, still exists. When used for retrograde filling in apical surgery, amalgam comes in close contact with the periapical tissue, and it is sometimes responsible for the induction of periapical lesion (PL) or its exacerbation. Therefore, the aim of the study was to examine the effect of amalgam on cytotoxicity and pro-duction of pro-inflammatory cytokine by cells isolated from PL. Methods. Conditioned medium from freshly prepared amalgam (ACM) was performed according to the ISO 10993-12 by incubating the alloy in RPMI medium (0.2 g/mL) for 3 days at 37°C. Cells were isolated from 20 hu-man PLs after apicoectomy by collagenase/DNA-ase diges-tion and cultured with different dilutions of ACM. Cytotox-icity was determined by MTT assay (n = 7 cultures) and apoptosis/necrosis assays (n = 8 cultures), whereas cytokine production was measured by a Flow Cytomix Microbeads Assay (n = 8 cultures). Results. Undiluted (100%) and 75% ACM was cytotoxic due to induction of apoptosis of PL cells. Non-cytotoxic concentrations of ACM (50% and 25%) inhibited the production of pro-inflammatory cyto-kines (TNF-α, IL-1β, IL-6, and IL-8), concentration-dependently. Conclusion. For the first time, our results showed an unexpected anti-inflammatory property of amal-gam on PL cells, which could be beneficial for PL healing after apicoectomy. SAŽETAK Uvod/Cilj. Amalgam se godinama koristi u stomatologiji, ali i dalje postoje kontroverze o njegovim neželjenim efektima na lokalno oralno/dentalno tkivo i sistemsko zdravlje. Kada se koristi za retrogradno punjenje u apikalnoj hirurgiji, amal-gam dolazi u blizak kontakt sa periapeksnim tkivom, što je ponekad povezano sa indukcijom periapeksne lezije (PL) ili njenom egzacerbacijom. Zato je cilj ovog rada bio da se ispita efekat amalgama na citotoksičnost i produkciju pro-inflamacijskih citokina od strane ćelija izolovanih iz PL. Metode. Od sveže napravljenog amalgama pripremljen je kondicionirani medijum (ACM) inkubiranjem legure na 37°C u RPMI medijumu u toku 3 dana (0.2 g/mL) kako je pred-loženo standardom ISO 10993-12. Ćelije su izolovane iz 20 humanih PL nakon apikoektomije, digestijom tkiva pomoću kolagenaze/DNA-aze, a zatim su korišćene za kulturu u pri-sustvu različitih razblaženja ACM. Citotoksičnost je ispitivana pomoću MTT testa (n = 7 kultura) i detekcijom apop-toze/nekroze (n = 8), dok je nivo produkovanih citokina me-ren simultano pomoću eseja sa mikrokuglicama uz pomoć protočne citometrije (n = 8). Rezultati. Nerazblažen ACM (100%) i onaj od 75% pokazali su citotoksični efekat, in-dukujući apoptozu PL ćelija. Necitotoksične koncentracije ACM (50% i 25%) inhibirale su produkciju pro-inflamacijskih citokina (TNF-α, IL-1β, IL-6 i IL-8) na dozno-zavisan način. Zaključak. Naši rezultati po prvi put pokazuju neočekivano antiinflamacijsko svojstvo amalgama na PL ćelije, što može biti korisno za zarastanje lezije nakon apikoektomije. dental amalgam, periapical tissue, cytokines, cytotoxicity, immunologic, inflammation, apicoectomy amalgam stomatološki, periapeksno tkivo, citokini, citotoksičnost, imunološka, zapaljenje, apikoektomija 1552099 10.2298/VSP190225043E 2022-05-18T10:42:35.661Z 44 no 46 Mile Eraković Military Medical Academy, Clinic for Stomatology, Belgrade, Serbia Miodrag Čolić Faculty of Medicine of the Military Medical Academy, Belgrade, Serbia akademik person Dragana Vučević Faculty of Medicine of the Military Medical Academy, Belgrade, Serbia person Sergej Tomić University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia doktor bioloških nauka person 0000-0003-2570-1295 Marijana Milanović Faculty of Medicine of the Military Medical Academy, Belgrade, Serbia person Marina Bekić University of Belgrade, Institute for the Application of Nuclear Energy INEP, Serbia person 0000-0002-6945-4241 Miloš Duka Military Medical Academy, Clinic for Stomatology, Belgrade, Serbia person application/pdf 577989 https://phaidrabg.bg.ac.rs/o:25464 no yes 21 70 92000001 11A29 11A2904 11A32 Vojnosanitetski pregled 78 3 289 295 2021