Biofunctionalization of silicon substrates is important to the development of silicon-based biosensors and devices. Compared to conventional organosiloxane films on silicon oxide intermediate layers, organic monolayers directly bound to the non-oxidized silicon substrates via Si-C bonds enhance the sensitivity of detection and the stability against hydrolytic cleavage. Such monolayers presenting a high density of terminal alkynyl groups for bioconjugation via coppercatalyzed azide-alkyne 1,3-dipolar cycloaddition (CuAAC, a "click" reaction) were reported. However, yields of the CuAAC reactions on these monolayer platforms were low. Also, the nonspecific adsorption of proteins on the resultant surfaces remained a major obstacle for many potential biological applications. Herein, we report a new type of "clickable" monolayers grown by selective, photo-activated surface hydrosilylation of α,ω-alkenynes, where the alkynyl terminal is protected with a trimethylgermanyl (TMG) group, on hydrogen-terminated silicon substrates. The TMG groups on the film are readily removed in aqueous solutions in the presence of Cu(I). Significantly, the degermanylation and the subsequent CuAAC reaction with various azides could be combined into a single step in good yields. Thus, oligo(ethylene glycol) (OEG) with an azidotag was attached to the TMG-alkyne surfaces, leading to OEG-terminated surfaces that reduced the non-specific adsorption of protein (fibrinogen) by >98%. The CuAAC reaction could be performed in microarray format to generate arrays of mannose and biotin with varied densities on the protein-resistant OEG background. We also demonstrated that the monolayer platform could be functionalized with mannose for highly specific capturing of living targets (Escherichia coli expressing fimbriae) onto the silicon substrates.
In this article, we present the first report on the antibacterial activity and cytotoxicity of poly(amidoamine) (PAMAM) dendrimers immobilized on three types of titanium-based substrates with and without calcium phosphate coating. We show that the amino-terminated PAMAM dendrimers modified with various percentages (0-60%) of poly(ethylene glycol) (PEG) strongly adsorbed on the titanium-based substrates. The resultant dendrimer films effectively inhibited the colonization of the Gram-negative bacteria Pseudomonas aeruginosa (strain PAO1) and, to a lesser extent, the Gram-positive bacteria Staphylococcus aureus (SA). The antibacterial activity of the films was maintained even after storage of the samples in PBS for up to 30 days. In addition, the dendrimer films had a low cytotoxicity to human bone mesenchymal stem cells (hMSCs) and did not alter the osteoblast gene expression promoted by the calcium phosphate coating.
We show that coating of decellularized
extracellular matrix (DC-ECM)
on substrate surfaces is an efficient way to generate a platform mimicking
the native ECM environment. Moreover, the DC-ECM can be modified with
a peptide (QK) mimicking vascular endothelial growth factor without
apparently compromising its integrity. The modification was achieved
through metabolic incorporation of a “clickable” handle
to DC-ECM followed by rapid attachment of the QK peptide with an azido
tag using copper-catalyzed click reaction. The attachment of the QK
peptide on to DC-ECM in this way further enhanced the angiogenic responses
(formation of branched tubular networks) of endothelial cells.
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