Aim To evaluate the effect of Biodentine eluate on cytotoxicity and production of pro‐ and anti‐inflammatory cytokines and osteodestructive/osteoprotective cytokines in cultures of human periapical lesion cells. Methodology Conditioned Biodentine Medium (CBM) was prepared according to ISO 10993‐12, by incubating Biodentine in RPMI medium (0.2 g mL−1) for 3 days at 37 °C. CBM contained both released microparticles and leachable soluble components. Inflammatory cells were isolated from 22 human periapical lesions after apicectomy or tooth extraction, by collagenase/DNase digestion, and cultured in several dilutions of CBM. The composition of periapical lesion cells was determined by morphological criteria, cytotoxicity was quantified by MTT and flow cytometric apoptosis/necrosis assays, whereas the levels of produced cytokines in cell culture supernatants were measured by flow cytometry and ELISA. Student t‐test and Friedman test with Dunn's post‐test were used for comparison of parametric and nonparametric variables, respectively. Results Undiluted (100%), 75% and 50% CBM were cytotoxic for periapical lesion cells due to induction of both necrosis (100% CBM) and apoptosis (75% and 50% CBM). Noncytotoxic concentrations of CBM (25%) inhibited the production of pro‐inflammatory cytokines: TNF‐α, (P < 0.005); IL‐1β (P < 0.01); IL‐6 (P < 0.005) and chemokines IL‐8: (P < 0.005); MCP‐1 (P < 0.005), stimulated the production of anti‐inflammatory cytokine (IL‐10; P < 0.005), Th2 cytokines: IL‐4, IL‐5 and IL‐33 (all P < 0.01), and IL‐17A (P < 0.01). The concentration of CBM (12.5%) inhibited the production of IL‐6 (P < 0.05), IL‐8 (P < 0.01) and MCP‐1 (P < 0.005) and augmented the production of IL‐10 (P < 0.05). No significant effects on Th1‐related cytokines (IFN‐γ and IL‐12) and IL‐23 were detected with 25% and 12.5% CBM concentrations. Both CBM concentrations inhibited the production of osteolytic receptor activator of nuclear factor kappa‐Β ligand (RANKL), dose dependently (P < 0.005 and P < 0.01, respectively). Higher CBM concentrations decreased RANKL/osteoprotegerin (OPG) ratio (P < 0.05), without significant influence on the levels of osteoprotective OPG. Conclusion Biodentine possesses immunomodulatory properties by suppressing pro‐inflammatory and augmenting anti‐inflammatory cytokines. Together with the reduction of osteodestructive mediators, this novel root‐end filling cement could be beneficial for healing and bone reparation after the surgical treatment of periapical lesions.

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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-α, RANTES, TLR-2, HIF-1α, OPG, MMP-3, SDF-1, HGF and IP-10 in P-GMSCs, whereas only IL-6, MCP-1 and GRO-α 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-β, 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.

Placental growth factor (PlGF) is crucial during placental development in early pregnancy. Several studies in pregnancies with complications such as preeclampsia or small for gestational age neonates find that PlGF levels are significantly lower in the first trimester, which implies that the concentration of PlGF could be used as an early screening biomarker for these conditions. This study aimed to compare the performance of chemiluminescence immunoassay (CLIA) and enzyme-linked immunosorbent assay (ELISA) for the quantification of human PlGF in serum. This is a comparative study on 88 pregnant women in the first trimester subjected to measurement of PlGF in serum using two commercially available kits: Human PlGF Quantikine HS ELISA (R&amp;D Systems) and PlGF CLIA (Snibe). The overall coefficient of correlation between the tests was 0.93. When the cut-off value of 40 pg/mL was applied, it dropped significantly to 0.50 towards the lower values, while remaining an excellent 0.91 in the group with higher concentrations of PlGF. While R&amp;D Systems’s ELISA seems to have better sensitivity, it is not very convenient to use for a small number of samples. Snibe’s CLIA automated method is user-friendly, fast and powerful. Both tests show excellent performance when indicating risk-free pregnancies.

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