This Article focuses on the fabrication of highly ordered nanotubes and some novel nanostructures of titania (TiO2) with a two-step anodization method. The first-step anodization was actually a pretreatment of the Ti foil surface and provided well-ordered imprints that served as a template for the further growth of nanotubes. As a result, the TiO2 nanotubes growing in the second-step anodization appreciably outperformed those fabricated with the conventional one-step Ti anodization in terms of size uniformity and arrangement orderliness. The parameters of the anodization were then modulated to obtain more complex structures. When the voltage in the second-step anodization was lower than that in the first-step anodization, a lotus root-shaped TiO2 nanostructure, in which each imprint contained several smaller nanopores, was achieved. When the second anodization was further divided into two stages, double-layered nanotube arrays were synthesized. They contained two distinctly separated parts, i.e., the bamboo-shaped upper one and the smooth-walled lower one. These results have demonstrated the effectiveness and controllability of the two-step anodization method in producing high-quality TiO2 nanotubes, which are believed to have potential applications in such fields as solar cells, photonic crystals, and hydrogen storage.

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A biotinylated glutathione (GSH)-responsive zinc(II) phthalocyanine has been prepared and characterized. With a 2,4-dinitrobenzenesulfonyl moiety, its fluorescence emission and singlet oxygen generation were silenced in its intact state. Upon exposure to high concentration of GSH, its photosensitizing properties were restored in phosphate buffered saline and inside tumor cells. It also showed preferential uptake on HepG2 human hepatocarcinoma cells (with higher biotin receptor expression) rather than Chinese hamster ovary (CHO) cells (with lower biotin receptor expression). Upon irradiation, it caused photocytotoxicity with an IC value down to 0.1 μm on HepG2 cells. Moreover, it can localize in the endoplasmic reticulum (ER), causing ER stress after light irradiation.

Two dual stimuli-activated photosensitizers were developed, in which two or three glutathione (GSH)-responsive 2,4dinitrobenzenesulfonate (DNBS)-substituted zinc(II) phthalocyanine units were connected via one or two cathepsin B-cleavable Gly-Phe-Leu-Gly peptide linker(s). These dimeric and trimeric phthalocyanines were fully quenched in the native form due to the photoinduced electron transfer to the DNBS substituents and the self-quenching of the phthalocyanine units. In the presence of GSH and cathepsin B, or upon internalization into A549 and HepG2 cancer cells, these probes were activated through the release of free phthalocyanine units. The intracellular fluorescence intensity was increased upon post-incubation with GSH ester or reduced upon pre-treatment with a cathepsin B inhibitor. Upon light irradiation, these photosensitizers became highly cytotoxic with IC 50 values of 0.21−0.39 μM. The photocytotoxicity was also dependent on the intracellular GSH and cathepsin B levels. The results showed that these conjugates could serve as smart photosensitizers for targeted photodynamic therapy.

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