RESUMO
The use of small particles has expanded the capability of ultrashort pulsed laser optoinjection technology toward simultaneous treatment of multiple cells. The microfluidic platform is one of the attractive systems that has obtained synergy with laser-based technology for cell manipulation, including optoinjection. We have demonstrated the delivery of molecules into suspended-flowing cells in a microfluidic channel by using biodegradable polymer microspheres and a near-infrared femtosecond laser pulse. The use of polylactic-co-glycolic acid microspheres realized not only a higher optoinjection ratio compared to that with polylactic acid microspheres but also avoids optical damage to the microfluidic chip, which is attributable to its higher optical intensity enhancement at the localized spot under a microsphere. Interestingly, optoinjection ratios to nucleus showed a difference for adhered cells and suspended cells. The use of biodegradable polymer microspheres provides high throughput optoinjection; i.e., multiple cells can be treated in a short time, which is promising for various applications in cell analysis, drug delivery, and ex vivo gene transfection to bone marrow cells and stem cells without concerns about residual microspheres.
Assuntos
Sistemas de Liberação de Medicamentos/métodos , Lasers , Microfluídica/métodos , Microesferas , Animais , Células Cultivadas , Sistemas de Liberação de Medicamentos/instrumentação , Microfluídica/instrumentação , PolímerosRESUMO
The parameters critically influencing the delivery rate on the biodegradable microsphere-mediated femtosecond (fs) laser perforation are investigated in detail with the aim of developing efficient molecular delivery. Cell membrane was perforated by the irradiation of weakly focused fs laser pulses to the spherical polylactic acid microspheres conjugated to the cell membrane. The delivery of fluorescein isothiocyanate-dextran and fluorescent silica particles to A431 cells is investigated in detail. The increase in the number of irradiated laser pulses had resulted in the increase of delivery rate. The delivery rate depends on the size and functionalization of fluorescent silica particles in which silica particles of 100 nm in diameter were able to be delivered into 20% of the irradiated cells, suggesting that the pore sizes are large enough for the delivery of therapeutic agents into cells. These findings contribute to the development of an efficient and safe phototherapy and drug delivery.
Assuntos
Sistemas de Liberação de Medicamentos , Lasers , Microesferas , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Fluoresceína-5-Isotiocianato/química , Proteínas de Fluorescência Verde/química , Humanos , Ácido Láctico/química , Tamanho da Partícula , Fototerapia/métodos , Poliésteres , Polímeros/química , Dióxido de Silício/química , TemperaturaRESUMO
Fabrication of a nanopore in a hollow microcapsule was demonstrated using near-infrared femtosecond laser irradiation. The shape of the irradiated microcapsules was kept spherical except for a pore in the shell owing to the nonthermal processing by a femtosecond laser. The simulation results for the near-field and far-field scattering around a microcapsule revealed that highly-enhanced optical intensity can be generated at a spot on the shell of a microcapsule, which would in turn contribute to localized ablation. To the best of our knowledge, this is the first demonstration of the nanoperforation of transparent hollow microcapsules by a near-infrared laser without any doping with absorbing metals or dyes that may cause cell toxicity. The presented method is a promising approach for safer drug delivery and the controlled release of therapeutic drugs.
Assuntos
Cápsulas/química , Cápsulas/efeitos da radiação , Lasers , Nanoporos/ultraestrutura , Raios Infravermelhos , Teste de Materiais , Porosidade/efeitos da radiaçãoRESUMO
Pulsed laser interaction with small metallic and dielectric particles has been receiving attention as a method of drug delivery to many cells. However, most of the particles are attended by many risks, which are mainly dependent upon particle size. Unlike other widely used particles, biodegradable particles have advantages of being broken down and eliminated by innate metabolic processes. In this paper, the perforation of cell membrane by a focused spot with transparent biodegradable microspheres excited by a single 800 nm, 80 fs laser pulse is demonstrated. A polylactic acid (PLA) sphere, a biodegradable polymer, was used. Fluorescein isothiocyanate (FITC)-dextran and short interfering RNA were delivered into many human epithelial carcinoma cells (A431 cells) by applying a single 80 fs laser pulse in the presence of antibody-conjugated PLA microspheres. The focused intensity was also simulated by the three-dimensional finite-difference time-domain method. Perforation by biodegradable spheres compared with other particles has the potential to be a much safer phototherapy and drug delivery method for patients. The present method can open a new avenue, which is considered an efficient adherent for the selective perforation of cells which express the specific antigen on the cell membrane.