The samples were sputtered with platinum for 50 s (15 nm, SCD 004, BAL-TEC, Wetzlar, Germany) and field emission scanning electron microscopy (FE-SEM, ZEISS Merlin VP compact, Carl Zeiss, Oberkochen, Germany) observations were taken with an acceleration voltage of 5 kV, a working range of 5
May 21, 2021
The samples were sputtered with platinum for 50 s (15 nm, SCD 004, BAL-TEC, Wetzlar, Germany) and field emission scanning electron microscopy (FE-SEM, ZEISS Merlin VP compact, Carl Zeiss, Oberkochen, Germany) observations were taken with an acceleration voltage of 5 kV, a working range of 5.6 mm and a high efficiency secondary electron detector (InlenseDuo for 2,000x, HE-SE2 detector for 100, 500x, and 5,000x). Circularity The circularity of the cells after 24 h could be evaluated with Photoshop CC 2017 using fluorescence microscopic images of fluoro-3-acetoxymethyl ester (fluo-3) stained cells ( 3 indie experiments 15 cells). of cell activity (membrane integrity, viability, proliferation, calcium mobilization) were observed on surfaces with a highly positive zeta potential (+50 mV). This systematic study shows that cells do not prefer positive charges in general, merely moderately positive ones. The cell behavior of MG-63s could be correlated with the materials zeta potential; but not with water contact angle or surface free energy. Our findings present fresh insights and provide an essential knowledge for long term applications in dental care and orthopedic surgery. = 3, three pairs of samples). Surface Wettability Surface free energies of the substrate/air flow interface and WCA were determined by the sessile drop method using the Drop Shape AnalyzerDSA25 (Krss, Hamburg, Germany) (Staehlke et al., 2018). One l drops of distilled water and diiodomethane (Sigma-Aldrich Chemie, Taufkirchen, Germany) were deposited within the sample surface (= 3 at 3 drops each). WCA ideals were calculated with the supplied software (ADVANCE, V.126.96.36.199, Krss, Hamburg, Germany) via the Youngs equation and the SFE according to Owens, Wendt, Rabel und Kaelble (OWRK). Coating Thickness Coating thickness was measured with null ellipsometry (Multiskop; Optrel GbR, Sinzing, Germany) as explained before (M?rke et al., 2017). Each sample was modeled by 6 slabs in order to account for oxide layers (Si, SiO2, Ti, TiO2, coating of interest and ambient air flow). The thickness of the Ti and TiO2 coating was identified independently prior to deposition of the coating of interest. Additionally, uncoated samples were measured and served like a research. Each measurement was carried out at several angles of incidence (50?80 in 1 methods) in two different ellipsometric zones (Nestler et al., 2012). Chemical Composition The elemental composition of the revised surfaces was analyzed by using an AXIS Supra X-ray photoelectron spectrometer from Kratos Analytical Ltd. (Manchester, United Kingdom). Measurements were performed using a monochromatic Al Ka X-ray resource (1486.6 eV) operated at 150 W. Survey and core-level spectra were acquired using a pass energy of 80 eV. For each sample, an area of 250 m was analyzed in duplicates on three different positions. During the analysis, the integrated charge neutralization system was triggered for charge compensation. XPS measurements for PEI and PPI-G4 were performed having a PHI 5700 (Physical Electronics, Bryostatin 1 United States). Here, survey and core-level spectra were acquired using pass energies of 117C187 eV. Charge neutralization was not necessary. Cell Biological Investigations Cell Tradition Human being osteoblast-like MG-63 cells (American Type Tradition Collection ATCC?, CRL1427TM, Bethesda, MD, United States) were used; this cell collection has been successfully applied like a model for studying cell-material relationships (Staehlke et al., 2019) and offers similar characteristics to primary human being osteoblasts (Clover and Gowen, 1994; Czekanska et al., LASS4 antibody 2014). The cells were cultured in Dulbeccos Modified Eagle Medium (DMEM, 31966-021, Existence Systems Limited, Paisley, United Kingdom), with 10% fetal calf serum (FCS, Biochrom FCS Superior, Merck, Darmstadt, Germany) and 1% antibiotics (gentamicin, Ratiopharm, Ulm, Germany) at 37C with 5% CO2/95% air flow atmosphere (Staehlke et al., 2018). All experiments were performed in passages 5C30, as the MG-63 cell physiology is known to remain stable over the entire range of these passages (Staehlke et al., 2019). Cell Morphology Scanning electron microscopy (SEM) MG-63s (15,000 cells/cm2) were cultured for 24 h within the Ti arrays, washed three times with phosphate buffered saline (PBS, Sigma-Aldrich Chemie, Taufkirchen, Germany), fixed with 2.5% glutardialdehyde (Merck, Darmstadt, Germany) at 4C over-night and rinsed with 0.1% sodium phosphate buffer (relating to S?rensen, Merck, Darmstadt, Germany). The samples were then dehydrated through an ethanol series of 30, 50, Bryostatin 1 75, 90 and twice 100% (for 5, 5, 15, 10, and 10 min, respectively) and dried in a critical point dryer (K 850, EMITECH, Bryostatin 1 Taunusstein, Germany). The samples were sputtered with gold for 50.