RESUMEN
Conventional photocages only respond to short wavelength light, which is a significant obstacle to developing efficient phototherapy in vivo. The development of photocages activated by near-infrared (NIR) light at wavelengths from 700 to 950â nm is important for in vivo studies but remains challenging. Herein, we describe the synthesis of a photocage based on a ruthenium (Ru) complex with NIR light-triggered photocleavage reaction. The commercial anticancer drug, tetrahydrocurcumin (THC), was coordinated to the RuII center to create the Ru-based photocage that is readily responsive to NIR light at 760â nm. The photocage inherited the anticancer properties of THC. As a proof-of-concept, we further engineered a self-assembled photocage-based nanoparticle system with amphiphilic block copolymers. Upon exposure to NIR light at 760â nm, the Ru complex-based photocages were released from the polymeric nanoparticles and efficiently inhibited tumor proliferation in vivo.
Asunto(s)
Nanopartículas , Neoplasias , Fotoquimioterapia , Rutenio , Humanos , Fototerapia , Neoplasias/tratamiento farmacológico , Polímeros/uso terapéutico , Nanopartículas/uso terapéuticoRESUMEN
The development of methods for the off-on switching of immobilization or presentation of cell-adhesive peptides and proteins during cell culture is important because such surfaces are useful for the analysis of the dynamic processes of cell adhesion and migration. This paper describes a chemically functionalized gold substrate that captures a genetically tagged extracellular matrix protein in response to light. The substrate was composed of mixed self-assembled monolayers (SAMs) of three disulfide compounds containing (i) a photocleavable poly(ethylene glycol) (PEG), (ii) nitrilotriacetic acid (NTA) and (iii) hepta(ethylene glycol) (EG7). Although the NTA group has an intrinsic high affinity for oligohistidine tag (His-tag) sequences in its Ni2+-ion complex, the interaction was suppressed by the steric hindrance of coexisting PEG on the substrate surface. Upon photoirradiation of the substrate to release the PEG chain from the surface, this interaction became possible and hence the protein was captured at the irradiated regions, while keeping the non-specific adsorption of non-His-tagged proteins blocked by the EG7 underbrush. In this way, we selectively immobilized a His-tagged fibronectin fragment (FNIII7-10) to the irradiated regions. In contrast, when bovine serum albumin-a major serum protein-was added as a non-His-tagged protein, the surface did not permit its capture, with or without irradiation. In agreement with these results, cells were selectively attached to the irradiated patterns only when a His-tagged FNIII7-10 was added to the medium. These results indicate that the present method is useful for studying the cellular behavior on the specific extracellular matrix protein in cell-culturing environments.
RESUMEN
This protocol describes a method for dynamic patterning cells on a glass coverslip. The glass substrate is first functionalized with photocleavable silane bearing 2-nitrobenzyl group, thereafter a cell-repellent polymer, poly(ethylene glycol) (PEG), is conjugated. Upon absorption of near-UV light, the PEG is cleaved from the surface, changing the surface from non-cell-adhesive to cell-adhesive. The method allows not only for spatially controlling cell attachment on the substrate (conventional patterning), but also inducing cell migration or coculturing heterotypic cells (dynamic patterning). Furthermore, it should be emphasized that the surface is compatible with fluorescence imaging in a high-resolution inverted objective setup as it is composed of a normal glass coverslip functionalized with the thin layers. In this chapter, I describe the procedure for the synthesis of the silane molecule, the preparation of the photoactivatable surface, and its application for dynamic cell patterning.