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In vitro perforation of human epithelial carcinoma cell with antibody-conjugated biodegradable microspheres illuminated by a single 80 femtosecond near-infrared laser pulse.

Terakawa M, Tsunoi Y, Mitsuhashi T - Int J Nanomedicine (2012)

Bottom Line: 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.Perforation by biodegradable spheres compared with other particles has the potential to be a much safer phototherapy and drug delivery method for patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Electronics and Electrical Engineering, Keio University, Yokohama, Kanagawa, Japan. terakawa@elec.keio.ac.jp

ABSTRACT
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.

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Conceptual diagram of dielectric sphere-mediated perforation using femtosecond (fs) laser.Notes: Biodegradable spheres are conjugated to cell membrane via antigen– antibody interaction. Femtosecond laser illumination to the spheres generates a strongly enhanced optical field under the sphere for perforation.
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f1-ijn-7-2653: Conceptual diagram of dielectric sphere-mediated perforation using femtosecond (fs) laser.Notes: Biodegradable spheres are conjugated to cell membrane via antigen– antibody interaction. Femtosecond laser illumination to the spheres generates a strongly enhanced optical field under the sphere for perforation.

Mentions: Figure 1 shows a conceptual diagram of dielectric sphere-mediated perforation using femtosecond laser. Spherical protein A conjugated PLA spheres, which have a diameter of 2000 nm, were mixed with anti-epidermal growth factor receptor (EGFR) mouse monoclonal antibody (Thermo Fisher Scientific, Fremont, CA). The anti-EGFR antibody can be employed to target overexpressed EGFR on A431 cells. The mixture was stirred for 25 minutes at room temperature. After the removal of unbound antibody by centrifugation for 10 minutes at 10,000 rpm, the conjugated PLA microspheres were resuspended in phosphate-buffered saline (PBS) and added to A431 cells. The cells were incubated for 40 minutes at 37°C and washed three times with PBS to remove the unbound microspheres. The uptake of the microspheres by the cells was not observed in the incubation time of 40 minutes in this study.


In vitro perforation of human epithelial carcinoma cell with antibody-conjugated biodegradable microspheres illuminated by a single 80 femtosecond near-infrared laser pulse.

Terakawa M, Tsunoi Y, Mitsuhashi T - Int J Nanomedicine (2012)

Conceptual diagram of dielectric sphere-mediated perforation using femtosecond (fs) laser.Notes: Biodegradable spheres are conjugated to cell membrane via antigen– antibody interaction. Femtosecond laser illumination to the spheres generates a strongly enhanced optical field under the sphere for perforation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368514&req=5

f1-ijn-7-2653: Conceptual diagram of dielectric sphere-mediated perforation using femtosecond (fs) laser.Notes: Biodegradable spheres are conjugated to cell membrane via antigen– antibody interaction. Femtosecond laser illumination to the spheres generates a strongly enhanced optical field under the sphere for perforation.
Mentions: Figure 1 shows a conceptual diagram of dielectric sphere-mediated perforation using femtosecond laser. Spherical protein A conjugated PLA spheres, which have a diameter of 2000 nm, were mixed with anti-epidermal growth factor receptor (EGFR) mouse monoclonal antibody (Thermo Fisher Scientific, Fremont, CA). The anti-EGFR antibody can be employed to target overexpressed EGFR on A431 cells. The mixture was stirred for 25 minutes at room temperature. After the removal of unbound antibody by centrifugation for 10 minutes at 10,000 rpm, the conjugated PLA microspheres were resuspended in phosphate-buffered saline (PBS) and added to A431 cells. The cells were incubated for 40 minutes at 37°C and washed three times with PBS to remove the unbound microspheres. The uptake of the microspheres by the cells was not observed in the incubation time of 40 minutes in this study.

Bottom Line: 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.Perforation by biodegradable spheres compared with other particles has the potential to be a much safer phototherapy and drug delivery method for patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Electronics and Electrical Engineering, Keio University, Yokohama, Kanagawa, Japan. terakawa@elec.keio.ac.jp

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus