Photothermal therapy with silver nanoplates in HeLa cells studied by in situ fluorescence microscopy.

Photothermal therapy (PTT) is a noninvasive treatment for cancer relying on the incorporation of NIR-light absorbing nanomaterials into cells, which upon illumination release heat causing thermally induced cell death. We prove that irradiation of aqueous suspensions of poly(vinylpyrrolidone)-coated silver nanoplates (PVPAgNP) or PVPAgNP in HeLa cells with red or NIR lasers causes a sizeable photothermal effect, which in cells can be visualized with the temperature sensing fluorophore Rhodamine B (RhB) using spinning disk confocal fluorescence microscopy or fluorescence lifetime imaging. Upon red-light irradiation of cells that were incubated with both, RhB and PVPAgNP at concentrations with no adverse effects on cell viability, a substantial heat release is detected. Initiation of cell death by photothermal effect is observed by positive signals of fluorescent markers for early and late apoptosis. Surprisingly, a new nanomaterial-assisted cell killing mode is operating when PVPAgNP-loaded HeLa cells are excited with moderate powers of fs-pulsed NIR light. Small roundish areas are generated with bright and fast (<1 ns) decaying emission, which expand fast and destroy the whole cell in seconds. This characteristic emission is assigned to efficient optical breakdown initiation around the strongly absorbing PVPAgNP leading to plasma formation that spreads fast through the cell.

Riboflavin-Mediated Photooxidation of Gold Nanoparticles and Its Effect on the Inactivation of Bacteria.

Photodynamic inactivation (PDI) of microorganisms, based on the ability of photosensitizers to produce reactive oxygen species (ROS) under adequate irradiation, emerges as a promising technique to face the increasing bacterial resistance to conventional antimicrobials. In this work, we analyze the combined action of Riboflavin (Rf) and pectin-coated gold nanoparticles (PecAuNP) on Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) as suitable PDI strategy. We demonstrate that gold ions can be generated upon Rf-photosensitized oxidation of PecAuNP. Transient absorption spectroscopy shows that the Rf cationic radical can accept an electron from the nanoparticles to yield Au(I) ions, which in aqueous medium is disproportionate to yield Au0 and Au(III). Microbiological assays showed that the presence of PecAuNP enhanced the antibacterial activity of photoirradiated Rf toward S. aureus and P. aeruginosa, in line with the well-known antibacterial activity of gold ions. Moreover, the irradiation of Rf solutions containing about 100 µM PecAuNP enabled the solutions to be bactericidal against both bacteria.

Photodynamic Therapy in HeLa Cells Incubated with Riboflavin and Pectin-coated Silver Nanoparticles.

Riboflavin (Rf) is an endogenous photosensitizer, which can participate in Type I and Type II processes. We have recently shown that the yield of the triplet excited states of Rf is enhanced in the presence of pectin-coated silver nanoparticles (Pec@AgNP) due to formation of a complex between Rf and Pec@AgNP (Rf-Pec@AgNP). Consequently, under aerobic conditions, the amounts of singlet molecular oxygen and superoxide radical anion generated are also larger in the presence of the nanoparticles. This result made us suspect that the nanoparticles could have a beneficial effect in Rf-based PDT. To prove this hypothesis, we here compared the photodamage in HeLa cells incubated with Rf in the presence and in the absence of Pec@AgNP applying several optical assays. We used fluorescence imaging of irradiated HeLa cells incubated with Annexin V and propidium iodide to evaluate the occurrence of apoptosis/necrosis, the reduction of the tetrazolium dye MTT to formazan and neutral red uptake to prove cell viability, as well as synchrotron infrared microscopy of single cells to evaluate possible structural changes of DNA and nuclear proteins. The enhanced photodamage observed in the presence of Pec@AgNP seems to indicate that Rf enters into the cells complexed with the nanoparticles.

Toward biomedical application of amino-functionalized silicon nanoparticles.

Silicon blue-emitting nanoparticles (NPs) are promising effectors for photodynamic therapy and radiotherapy, because of their production of reactive oxygen species (ROS) upon irradiation. RESULTS: Amino-functionalized silicon NPs (NH2SiNP) were intrinsically nontoxic below 100 µg/ml in vitro (on two tumor cell lines) and in vivo (zebrafish larvae and embryos). NH2SiNP showed a moderate effect as a photosensitizer for photodynamic therapy and reduced ROS generation in radiotherapy, which could be indicative of a ROS scavenging effect. Encapsulation of NH2SiNP into ultradeformable liposomes improved their skin penetration after topical application, reaching the viable epidermis where neoplastic events occur. CONCLUSION: Subsequent derivatizations after amino-functionalization and incorporation to nanodrug delivery systems could expand the spectrum of the biomedical application of these kind of silicon NPs.