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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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Average perforation efficiency under four different conditions.Note: A single 80 fs laser pulse was irradiated at 1.06 J/cm2 (1.29 × 1014 W/cm2 under the PLA sphere).Abbreviations: fs, femtosecond; PLA, polylactic acid.
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f6-ijn-7-2653: Average perforation efficiency under four different conditions.Note: A single 80 fs laser pulse was irradiated at 1.06 J/cm2 (1.29 × 1014 W/cm2 under the PLA sphere).Abbreviations: fs, femtosecond; PLA, polylactic acid.

Mentions: Figure 6 shows a comparison of the average perforation efficiency for the four sets of the experimental conditions. The perforation efficiency by the 80 fs laser illumination with the PLA spheres conjugated to the cell membrane was significantly higher than the three other conditions. Under the condition of the cells without antibody (ie, 80 fs laser illumination without the conjugation of the PLA microspheres to the cell membrane), the perforation was not observed. The unbound microspheres were washed out before the laser illumination because of the lack of the antigen–antibody conjugation. For the cells illuminated with 80 fs laser without the PLA spheres, no perforation was observed. For the cells without the illumination of the 80 fs laser, no perforation was observed. Therefore, combination of the 80 fs laser illumination and the conjugation of the PLA spheres to the cells by antigen–antibody reactions were found necessary for the cell perforation. Based on the calculated optical intensity, the high focused intensity is kept nearly constant for distances longer than 1 μm. Therefore, PLA spheres that are deposited on the cell surface or loosely associated with unwashed cells also work for the perforation when the pulsed laser is illuminated. Considering in-vivo therapy, PLA spheres are accumulated in the close vicinity of targeted cells by antigen–antibody reactions. Antibody works for the cell targeting, and then the laser illumination initiates the perforation. These results indicate that the present method can open a new avenue, which is considered an efficient adherent for the selective perforation of the cells which express the specific antigen on the cell membrane.


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)

Average perforation efficiency under four different conditions.Note: A single 80 fs laser pulse was irradiated at 1.06 J/cm2 (1.29 × 1014 W/cm2 under the PLA sphere).Abbreviations: fs, femtosecond; PLA, polylactic acid.
© Copyright Policy
Related In: Results  -  Collection

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

f6-ijn-7-2653: Average perforation efficiency under four different conditions.Note: A single 80 fs laser pulse was irradiated at 1.06 J/cm2 (1.29 × 1014 W/cm2 under the PLA sphere).Abbreviations: fs, femtosecond; PLA, polylactic acid.
Mentions: Figure 6 shows a comparison of the average perforation efficiency for the four sets of the experimental conditions. The perforation efficiency by the 80 fs laser illumination with the PLA spheres conjugated to the cell membrane was significantly higher than the three other conditions. Under the condition of the cells without antibody (ie, 80 fs laser illumination without the conjugation of the PLA microspheres to the cell membrane), the perforation was not observed. The unbound microspheres were washed out before the laser illumination because of the lack of the antigen–antibody conjugation. For the cells illuminated with 80 fs laser without the PLA spheres, no perforation was observed. For the cells without the illumination of the 80 fs laser, no perforation was observed. Therefore, combination of the 80 fs laser illumination and the conjugation of the PLA spheres to the cells by antigen–antibody reactions were found necessary for the cell perforation. Based on the calculated optical intensity, the high focused intensity is kept nearly constant for distances longer than 1 μm. Therefore, PLA spheres that are deposited on the cell surface or loosely associated with unwashed cells also work for the perforation when the pulsed laser is illuminated. Considering in-vivo therapy, PLA spheres are accumulated in the close vicinity of targeted cells by antigen–antibody reactions. Antibody works for the cell targeting, and then the laser illumination initiates the perforation. These results indicate that the present method can open a new avenue, which is considered an efficient adherent for the selective perforation of the cells which express the specific antigen on the cell membrane.

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