Prospects for a novel ultrashort pulsed laser technology for pathogen inactivation

Tsen, Shaw Wei D; Wu, T. C.; Kiang, Juliann G.; Tsen, Kong-Thon

doi:10.1186/1423-0127-19-62

Cited by 52 publications

(49 citation statements)

References 41 publications

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“…1 shows the experimental setup for selective photonic disinfection employed in this work. It is a diode-pumped cw mode-locked Ti-sapphire laser system, which includes A 5Ws-cw diode-pumped laser operated at a wavelength of 532 nm (Coherent Inc. Santa Clara, CA) and a Ti-sapphire laser (Del Mar Photonics Inc., San Diego, CA) [13–24]. The laser produced a continuous train of 60-fs pulses at a repetition rate of 80 MHz.…”

Section: Methodsmentioning

confidence: 99%

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Selective Photonic Disinfection of Cell Culture Using a Visible Ultrashort Pulsed Laser

Tsen

Kibler

Jacobs

et al. 2016

IEEE J. Select. Topics Quantum Electron.

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Microbial contamination of cell culture is a major problem encountered both in academic labs and in the biotechnology/pharmaceutical industries. A broad spectrum of microbes including mycoplasma, bacteria, fungi, and viruses are the causative agents of cell culture contamination. Unfortunately, the existing disinfection techniques lack selectivity and/or lead to the development of drug-resistance, and more importantly there is no universal method to address all microbes. Here, we report a novel, chemical-free visible ultrashort pulsed laser method for cell culture disinfection. The ultrashort pulsed laser technology inactivates pathogens with mechanical means, a paradigm shift from the traditional pharmaceutical and chemical approaches. We demonstrate that ultrashort pulsed laser treatment can efficiently inactivate mycoplasma, bacteria, yeast, and viruses with good preservation of mammalian cell viability. Our results indicate that this ultrashort pulsed laser technology has the potential to serve as a universal method for the disinfection of cell culture.

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Section: Methodsmentioning

confidence: 99%

“…Ultrashort pulsed (USP) laser technology has recently emerged as a novel chemical-free method for pathogen inactivation [13]. Previous studies indicate that visible USP lasers inactivate pathogens by vibrational excitation of protein structures via impulsive stimulated Raman scattering [14–16].…”

Section: Introductionmentioning

confidence: 99%

Selective Photonic Disinfection of Cell Culture Using a Visible Ultrashort Pulsed Laser

Tsen

Kibler

Jacobs

et al. 2016

IEEE J. Select. Topics Quantum Electron.

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show abstract

“…Ultrashort pulsed (USP) lasers are an attractive, chemical-free method to inactivate pathogens in pharmaceuticals and blood products 1 . Visible USP laser irradiation is a unique strategy to overcome the limitations of conventional antiviral agents due to its physical mechanism of action – the excitation of molecular vibrations within viral capsids through impulsive stimulated Raman scattering (ISRS) 2 .…”

Section: Introductionmentioning

confidence: 99%

Ultrashort pulsed laser treatment inactivates viruses by inhibiting viral replication and transcription in the host nucleus

Tsen¹,

Chapa²,

Beatty³

et al. 2014

Antiviral Research

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Ultrashort pulsed laser irradiation is a new method for virus reduction in pharmaceuticals and blood products. Current evidence suggests that ultrashort pulsed laser irradiation inactivates viruses through an impulsive stimulated Raman scattering process, resulting in aggregation of viral capsid proteins. However, the specific functional defect(s) in viruses inactivated in this manner have not been demonstrated. This information is critical for the optimization and the extension of this treatment platform to other applications. Toward this goal, we investigated whether viral internalization, replication, or gene expression in cells were altered by ultrashort pulsed laser irradiation. Murine Cytomegalovirus (MCMV), an enveloped DNA virus, was used as a model virus. Using electron and fluorescence microscopy, we found that laser-treated MCMV virions successfully internalized in cells, as evidenced by the detection of intracellular virions, which was confirmed by the detection of intracellular viral DNA via PCR. Although the viral DNA itself remained polymerase-amplifiable after laser treatment, no viral replication or gene expression was observed in cells infected with laser-treated virus. These results, along with evidence from previous studies, support a model whereby the laser treatment stabilizes the capsid, which inhibits capsid uncoating within cells. By targeting the mechanical properties of viral capsids, ultrashort pulsed laser treatment represents a unique potential strategy to overcome viral mutational escape, with implications for combatting emerging or drug-resistant pathogens.

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“…[10][11][12][13] Recently, a novel method for pathogen inactivation using visible ultrashort pulsed (USP) lasers has been reported (Ref. 14). Visible USP laser treatment inactivates viruses through a physical mechanism: the impulsive stimulated Raman scattering (ISRS) process.…”

Section: Introductionmentioning

confidence: 99%

Chemical-free inactivated whole influenza virus vaccine prepared by ultrashort pulsed laser treatment

Tsen

Donthi

et al. 2014

J. Biomed. Opt

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There is an urgent need for rapid methods to develop vaccines in response to emerging viral pathogens. Whole inactivated virus (WIV) vaccines represent an ideal strategy for this purpose; however, a universal method for producing safe and immunogenic inactivated vaccines is lacking. Conventional pathogen inactivation methods such as formalin, heat, ultraviolet light, and gamma rays cause structural alterations in vaccines that lead to reduced neutralizing antibody specificity, and in some cases, disastrous T helper type 2-mediated immune pathology. We have evaluated the potential of a visible ultrashort pulsed (USP) laser method to generate safe and immunogenic WIV vaccines without adjuvants. Specifically, we demonstrate that vaccination of mice with laser-inactivated H1N1 influenza virus at about a 10-fold lower dose than that required using conventional formalin-inactivated influenza vaccines results in protection against lethal H1N1 challenge in mice. The virus, inactivated by the USP laser irradiation, has been shown to retain its surface protein structure through hemagglutination assay. Unlike conventional inactivation methods, laser treatment did not generate carbonyl groups in protein, thereby reducing the risk of adverse vaccine-elicited T helper type 2 responses. Therefore, USP laser treatment is an attractive potential strategy to generate WIV vaccines with greater potency and safety than vaccines produced by current inactivation techniques.

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Prospects for a novel ultrashort pulsed laser technology for pathogen inactivation

Cited by 52 publications

References 41 publications

Selective Photonic Disinfection of Cell Culture Using a Visible Ultrashort Pulsed Laser

Selective Photonic Disinfection of Cell Culture Using a Visible Ultrashort Pulsed Laser

Ultrashort pulsed laser treatment inactivates viruses by inhibiting viral replication and transcription in the host nucleus

Chemical-free inactivated whole influenza virus vaccine prepared by ultrashort pulsed laser treatment

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