Прегледај по Аутор "Eraković, Mile"

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    Anti-inflammatory effect of amalgam on periapical lesion cells in culture
    (Ministry of Defance, Serbia, 2021) Eraković, Mile; Duka, Miloš; Bekić, Marina; Milanović, Marijana; Tomić, Sergej; Vučević, Dragana; Čolić, Miodrag
    Background/Aim. Amalgam h as b een u sed f or y ears 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 production 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 human PLs after apicoectomy by collagenase/DNA-ase digestion and cultured with different dilutions of ACM. Cytotoxicity 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 cytokines (TNF-α, IL-1β, IL-6, and IL-8), concentrationdependently. Conclusion. For t he f irst t ime, o ur results showed an unexpected anti-inflammatory property of amalgam on PL cells, which could be beneficial for PL healing after apicoectomy
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    Mesenchymal Stromal Cells from Healthy and Inflamed Human Gingiva Respond Differently to Porphyromonas gingivalis
    (MDPI, 2022) Bekić, Marina; Radanović, Marina; Đokić, Jelena; Tomić, Sergej; Eraković, Mile; Radojević, Dušan; Duka, Miloš; Marković, Dejan; Marković Milan; Ismaili, Bashkim; Bokonjić, Dejan; Čolić, Miodrag
    Gingiva-Derived Mesenchymal Stromal Cells (GMSCs) have been shown to play an important role in periodontitis. However, how P. gingivalis, one of the key etiological agents of the disease, affects healthy (H)- and periodontitis (P)-GMSCs is unknown. To address this problem, we established 10 H-GMSC and 12 P-GMSC lines. No significant differences in morphology, differentiation into chondroblasts and adipocytes, expression of characteristic MSCS markers, including pericyte antigens NG2 and PDGFR, were observed between H- and P-GMSC lines. However, proliferation, cell size and osteogenic potential were higher in P-GMSCs, in contrast to their lower ability to suppress mononuclear cell proliferation. P. gingivalis up-regulated the mRNA expression of IL-6, IL-8, MCP-1, GRO- , RANTES, TLR-2, HIF-1 , OPG, MMP-3, SDF-1, HGF and IP-10 in P-GMSCs, whereas only IL-6, MCP-1 and GRO- were up-regulated in H-GMSCs. The expression of MCP-1, RANTES, IP-10 and HGF was significantly higher in P-GMSCs compared to H-GMSCs, but IDO1 was lower. No significant changes in the expression of TLR-3, TLR-4, TGF- , LAP, IGFBP4 and TIMP-1 were observed in both types of GMSCs. In conclusion, our results suggest that P-GMSCs retain their pro-inflammatory properties in culture, exhibit lower immunosuppressive potential than their healthy counterparts, and impaired regeneration-associated gene induction in culture. All these functions are potentiated significantly by P. gingivalis treatment.
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    Production of Soluble Receptor Activator of Nuclear Factor Kappa-Β Ligand and Osteoprotegerin by Apical Periodontitis Cells in Culture and Their Modulation by Cytokines
    (Hindawi, 2019) Duka, Miloš; Eraković, Mile; Dolićanin, Zana; Stefanović, Dara; Čolić, Miodrag
    RANKL, a bone-destructive cytokine, and OPG, its osteoprotective counterpart, are expressed in periapical lesions (PLs), which represent hystopatological manifestations of apical periodontitis. However, their regulation in PLs has not been elucidated yet. Therefore, our aim was to study the production of RANKL and OPG and their modulation by pro- and anti-inflammatory cytokines in PL cell cultures. Isolated PL cells were cultured alone or with addition of TNF-α, IFN-ϒ, IL-17, IL-4, IL-10, and IL- 33, respectively. The levels of RANKL and OPG in supernatants were measured by ELISA. The proportion of CD3+ (T cells) and CD19+/CD138+ (B cells/plasma cells) within isolated PLs was determined by immunocytochemistry. The levels of RANKL were higher in cultures of symptomatic PLs compared to asymptomatic PLs and PLs with the dominance of T cells (T-type lesions) over B cells/plasma cells (B-type lesions). A higher proportion of osteodestructive processes (RANKL/OPG ratio > 1.0) were detected in symptomatic PLs. The production of RANKL was upregulated by IFN-ϒ and IL-17 and higher concentrations of IL-33. IL-10 and lower concentrations of IL-33 augmented the production of OPG. The addition of either RANKL or anti-RANKL antibody to the cultures did not modify significantly the production of OPG. In conclusion, this original PL cell culture model suggests that increased bone destruction through upregulated production of RANKL could be associated with exacerbation of inflammation in PLs with the predominance of Th1 and Th17 responses and increased secretion of IL-33. In contrast, IL-10 and lower levels of IL-33, through upregulation of OPG, may suppress osteolytic processes.