Different concentrations of fetal bovine serum affect cytokine modulation in Lipopolysaccharideactivated apical papilla cells in vitro*Fetal bovine serum (FBS) is the most used supplement in culture media; however, it may interfere with in vitro assays via effects on cell proliferation and cytokine production. The ideal FBS concentration for assays using apical papilla cells (APCs) remains unknown. Therefore, this study aimed to evaluate the effects of FBS on APC activation, cell viability/proliferation, and cytokine production. Methodology: Human APCs were cultured, plated, and maintained in media containing increasing concentrations of FBS for 24 h, 48 h, 72 h, 7 days, and 14 days in the presence of Lipopolysaccharide (LPS -1 µg/mL).At each time point, the cells were subjected to the MTT assay. The cytokines transforming growth factor (TGF)-β1, osteoprotegerin (OPG), and interleukin (IL)-6, along with the chemokine CCL2, were quantified using the enzymelinked immunosorbent assay at the 24-h time-point. Statistical analysis was performed using two-way analysis of variance (ANOVA) followed by Tukey's post-hoc test (p<0.05). Results: In general, APCs exhibited increasing metabolic activity in an FBS concentration-dependent fashion, regardless of the presence of LPS. In contrast, FBS interfered with the production of all the cytokines evaluated in this study, affecting the response induced by the presence of LPS. Conclusion: FBS increased APC metabolism in a concentration-dependent manner and differentially affected the production of TGF-β1, OPG, IL-6, and CCL2 by APCs in vitro.

Background Elevated levels of troponin I (hsTnT) and B-type natriuretic peptide (BNP) have been related with poor prognosis in patients with LFLG-AS. Biomarkers are less expensive, more practical and more accessible than imaging tests, so their use can be an alternative to imaging in the evaluation of patients with LFLG-AS. Purpose The aim of the present study is to assess multimodality imaging findings according to systemic biomarkers (i.e. hsTnT and BNP) in Low-Flow, Low-Gradient Aortic Stenosis (LFLG-AS) and reduced left ventricular ejection fraction (LVEF) patients. Methods Prospective study with LFLG-AS patients (LVEF &lt;50%, aortic valve area ≤1,0 cm2 and mean gradient &lt;40 mmHg) that underwent hsTNnT, BNP, cardiac magnetic resonance (CMR) with T1 mapping and 2 dimensional echocardiogram (2DEcho). All patients also underwent dobutamine stress echocardiogram to define aortic stenosis severity. Patients were divided into 3 groups according to BNP and hsTnT levels: Group 1: BNP and hsTnT levels below median (BNP &lt;395 pg/ml and TnI-Ultra &lt;0.042 ng/ml); Group 2: BNP or hsTnT higher than median; and Group 3: both hsTnT and BNP higher than median. Results 49 patients with LFLG-AS were included (Group 1: 17 patients, Group 2: 14 patients and Group 3: 18 patients). Clinical characteristics (including risk scores) were not able to stratify these groups. Patients with elevation of both biomarkers had lower valvuloarterial impedance (P=0.03), lower LVEF (P=0.02), less moderate/severe mitral (P=0.01) and tricuspid regurgitation (P&lt;0.01) by 2DEcho. CMR identified a progressive increase (from Group 1 to 3) of right and left chamber volumes; reduction in right and left ejection fraction and a marked increase in myocardial fibrosis assessed by extracellular volume (ECV) and indexed extracellular volume (iECV) (Figure 1). Conclusion Higher levels of BNP and hsTnT in LFLG-AS patients were associated with worse multi-modality imaging parameters and can be a surrogate of cardiac remodeling. FUNDunding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): No funding