The binding of cell integrins to proteins adsorbed on the material surface is a highly dynamic process critical for guiding cellular responses. However, temporal dynamic regulation of adsorbed proteins to meet the spatial conformation requirement of integrins for a certain cellular response remains a great challenge. Here, an active CoFeO/poly(vinylidene fluoride-trifluoroethylene) nanocomposite film, which was demonstrated to be an obvious surface potential variation (Δ V ≈ 93 mV) in response to the applied magnetic field intensity (0-3000 Oe), was designed to harness the dynamic binding of integrin-adsorbed proteins by in situ controlling of the conformation of adsorbed proteins. Experimental investigation and molecular dynamics simulation confirmed the surface potential-induced conformational change in the adsorbed proteins. Cells cultured on nanocomposite films indicated that cellular responses in different time periods (adhesion, proliferation, and differentiation) required distinct magnetic field intensity, and synthetically programming the preferred magnetic field intensity of each time period could further enhance the osteogenic differentiation through the FAK/ERK signaling pathway. This work therefore provides a distinct concept that dynamically controllable modulation of the material surface property fitting the binding requirement of different cell time periods would be more conducive to achieving the desired osteogenic differentiation.
We describe a Salmonella biosensor that was obtained by electrochemical immobilization of a nanocomposite consisting of reduced graphene oxide (rGO) and carboxymodified multi-walled carbon nanotubes (MWCNTs) directly on the surface of a glassy carbon electrode (GCE). An aminomodified aptamer specific for Salmonella was covalently bound to the rGO-MWCNT composite via amide bonds. The morphology of the rGO-MWCNT nanocomposite was characterized by transmission electron microscopy and scanning electron microscopy. Cyclic voltammetry and electrochemical impedance spectroscopy were used to monitor all steps during assembly. When exposed to samples containing Salmonella, the anti-Salmonella aptamer on the electrode captures its target. Hence, electron transfer is blocked, and this results in a large increase in impedance. Salmonella can be quantified by this aptasensor, typically operated at a working voltage of 0.2 V (vs. Ag/AgCl), in the range from 75 to 7.5×10 5 cfu⋅mL −1 and detection limit of 25 cfu⋅mL −1 (at an S/N of 3). The method is perceived to have a wide scope in that other bacteria may be detected by analogy to this approach and with very low limits of detection by applying respective analyte-specific aptamers.
The surface electric
potential of biomaterials has been extensively
proven to play a critical role in stem cells’ fate. However,
there are ambiguous reports on the relation of stem cells’
osteogenic capacity to surface potential characteristics (potential
polarity and intensity). To address this, we adopted a surface with
a wide potential range and both positive/negative polarity in a comprehensive
view to get insight into surface potential-regulating cellular osteogenic
differentiation. Tb
x
Dy1–x
Fe2 alloy/poly(vinylidene fluoride-trifluoroethylene)
magnetoelectric films were prepared, and the film could provide controllable
surface potential characteristics with positive or negative polarity
and potential (ϕME) intensity variation from 0 to
±120 mV as well as keep the surface chemical composition and
microstructure unchanged. Cell culture results showed that osteogenic
differentiation of mesenchymal stem cells on both positive and negative
potential films was obviously upregulated when the /ϕME/ intensities were set from 0–55 mV. Differently, the highest
upregulated osteogenic differentiation on the positive potential films
corresponded to the /ϕME/ intensity from 35–55
mV and was better than that on the negative potential films whereas
the highest on the negative potential films corresponded to the /ϕME/ intensity from 0–35 mV and was better than that
on the positive potential films. This fact could illustrate why previous
reports appeared ambiguously; i.e., the comparative result in osteogenic
differentiation between the positive and negative potential films
strongly depends on the selection of surface potential intensity.
On the basis of assaying of the exposed functional sites (RGD and
PHSRN) of the adsorbed fibronectin (FN) and the expression of cellular
integrin α5 and β1 subunits, the difference in the behavior
between the positive and negative potential films was attributed to
the distinct conformation of adsorbed fibronectin (FN) and the opposite
changing trend with /ϕME/ for the two films, which
triggers the osteogenesis-related FAK/ERK signaling pathway to a different
extent. This study could provide new cognition for the in-depth understanding
of the regulation mechanism underlying surface potential characteristics
in cell behaviors.
Inonotus obliquus, a wild wood-decay fungus which grows on Betula trees in cool climates, has a variety of biological activities that the scientific community is paying more and more attention to. However, the research work is moving at a snail's pace. The methods of strain identification and the hypha microstructure have not been reported. We isolated one strain of filamentous molds from fruit body which was collected from birch wood on Changbai Mountain, cultivated mycelia on an inclined plane, and examined its micromorphology based on macroscopic examination. The strain was identified as I. obliquus by sequencing its ITS (internal transcribed spacer) domain. We subsequently investigated some of the mycelium polysaccharides' biological activities. The strain used in this study as the producers of antioxidation and anticancer polysaccharides was LNUF008. After fermentation in a 30-L fermenter, mycelia were obtained. The polysaccharides were extracted by transonic recirculation and ethanol precipitation. In order to identify the antioxidation effect, we designed an assay to test the inhibition of endogenous and Fe(2+)-Cys-induced lipid peroxidation as well as ferrous sulfate/ascorbate (Fe(2+)-VC)-induced mitochondrial swelling. The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method was used to study the antiproliferation activity of the polysaccharides on SMMC7721 hepatoma cells. The results indicate that I. obliquus polysaccharides exhibit high antitumor and antioxidation effects. The submerged culture method of growing I. obliquus will enable large-scale production of the polysaccharides.
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