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Inactivation of acyclovir-sensitive and -resistant strains of herpes simplex virus type 1 in vitro by photodynamic antimicrobial chemotherapy.

Latief MA, Chikama T, Ko JA, Kiuchi Y, Sakaguchi T, Obana A - Mol. Vis. (2015)

Bottom Line: To evaluate the efficacy of photodynamic antimicrobial chemotherapy (PACT) with the new porphyrin derivative TONS 504 and a light-emitting diode (LED) against acyclovir (ACV)-sensitive and -resistant herpes simplex virus type 1 (HSV-1).Complete eradication of both viruses was apparent at a TONS 504 concentration of 10 mg/l and light energy of 10 to 30 J/cm(2) as well as at a TONS 504 concentration of 1 mg/l and light energy of 20 or 30 J/cm(2).No antiviral effect was apparent with TONS 504 in the absence of irradiation or with irradiation in the absence of TONS 504.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Science, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan ; Medical Faculty, Muhammadiyah University, Makassar, South Sulawesi, Indonesia ; Department of Ophthalmology, Hasanuddin University, Makassar, South Sulawesi, Indonesia.

ABSTRACT

Purpose: To evaluate the efficacy of photodynamic antimicrobial chemotherapy (PACT) with the new porphyrin derivative TONS 504 and a light-emitting diode (LED) against acyclovir (ACV)-sensitive and -resistant herpes simplex virus type 1 (HSV-1).

Methods: Human FL cells infected with the viral strains were subjected to PACT with TONS 504 at various concentrations (0.01 to 10 mg/l) and irradiation at various light energies (10 to 30 J/cm(2)) and were then incubated for 24 h before analysis.

Results: Immunocytofluorescence analysis with antibodies to HSV-1 revealed that PACT eliminated HSV-1 and ACV-resistant HSV-1 in a manner dependent on the TONS 504 concentration and light energy. Complete eradication of both viruses was apparent at a TONS 504 concentration of 10 mg/l and light energy of 10 to 30 J/cm(2) as well as at a TONS 504 concentration of 1 mg/l and light energy of 20 or 30 J/cm(2). No antiviral effect was apparent with TONS 504 in the absence of irradiation or with irradiation in the absence of TONS 504. Staining of cell nuclei with 4', 6-diamidino-2-phenylindole revealed no apparent cytotoxicity of the PACT system, a finding that was confirmed by the system's failure to induce the release of lactate dehydrogenase from the host cells.

Conclusions: We conclude that our PACT system based on TONS 504 and an LED is effective for eliminating HSV-1 and ACV-resistant HSV-1 without a harmful effect on host cells.

No MeSH data available.


Related in: MedlinePlus

Lack of cytotoxicity of TONS 504–PACT for host cells as evaluated with measurement of LDH release. A: Control conditions. Serum-free medium alone (vehicle only) or culture supernatants of uninfected FL cells that were either not subjected to TONS–504 photodynamic antimicrobial chemotherapy (PACT), not subjected to TONS 504–PACT but exposed to a cell lysis reagent, exposed to TONS 504 (10 mg/l) without irradiation, or subjected to irradiation (30 J/cm2) in the absence or TONS 504 were assayed for lactate dehydrogenase (LDH). B: Experimental conditions. FL cells infected with herpes simplex virus type 1 (HSV-1) or acyclovir (ACV)-resistant HSV-1 were subjected to TONS 504–PACT under the indicated conditions, after which culture supernatants were assayed for LDH. All data are presented as absorbance at 490 nm (A490) minus A595 and are means ± SD for three independent experiments.
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f2: Lack of cytotoxicity of TONS 504–PACT for host cells as evaluated with measurement of LDH release. A: Control conditions. Serum-free medium alone (vehicle only) or culture supernatants of uninfected FL cells that were either not subjected to TONS–504 photodynamic antimicrobial chemotherapy (PACT), not subjected to TONS 504–PACT but exposed to a cell lysis reagent, exposed to TONS 504 (10 mg/l) without irradiation, or subjected to irradiation (30 J/cm2) in the absence or TONS 504 were assayed for lactate dehydrogenase (LDH). B: Experimental conditions. FL cells infected with herpes simplex virus type 1 (HSV-1) or acyclovir (ACV)-resistant HSV-1 were subjected to TONS 504–PACT under the indicated conditions, after which culture supernatants were assayed for LDH. All data are presented as absorbance at 490 nm (A490) minus A595 and are means ± SD for three independent experiments.

Mentions: We first examined the possible effect of TONS 504-PACT on host cell viability by manually counting the number of apparently normal nuclei. Data for infected cells from three independent experiments revealed the lack of effect of TONS 504-PACT on host cell viability as evaluated based on nuclear morphology compared with the corresponding value for uninfected cells not subjected to TONS 504-PACT. There are no significant differences at all conditions of treatment (data not shown). PACT did not exhibit cytotoxicity at any of the TONS 504 concentrations and light energies examined. We then determined the possible effect of TONS 504-PACT on the integrity of the host cells by measuring the release of LDH into the incubation medium (Figure 2 A,B). Whereas the extent of LDH release was increased slightly after exposure of infected FL cells to the higher concentrations of TONS 504 or higher light energies, these effects were not statistically significant (p>0.05, Student t test).


Inactivation of acyclovir-sensitive and -resistant strains of herpes simplex virus type 1 in vitro by photodynamic antimicrobial chemotherapy.

Latief MA, Chikama T, Ko JA, Kiuchi Y, Sakaguchi T, Obana A - Mol. Vis. (2015)

Lack of cytotoxicity of TONS 504–PACT for host cells as evaluated with measurement of LDH release. A: Control conditions. Serum-free medium alone (vehicle only) or culture supernatants of uninfected FL cells that were either not subjected to TONS–504 photodynamic antimicrobial chemotherapy (PACT), not subjected to TONS 504–PACT but exposed to a cell lysis reagent, exposed to TONS 504 (10 mg/l) without irradiation, or subjected to irradiation (30 J/cm2) in the absence or TONS 504 were assayed for lactate dehydrogenase (LDH). B: Experimental conditions. FL cells infected with herpes simplex virus type 1 (HSV-1) or acyclovir (ACV)-resistant HSV-1 were subjected to TONS 504–PACT under the indicated conditions, after which culture supernatants were assayed for LDH. All data are presented as absorbance at 490 nm (A490) minus A595 and are means ± SD for three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440479&req=5

f2: Lack of cytotoxicity of TONS 504–PACT for host cells as evaluated with measurement of LDH release. A: Control conditions. Serum-free medium alone (vehicle only) or culture supernatants of uninfected FL cells that were either not subjected to TONS–504 photodynamic antimicrobial chemotherapy (PACT), not subjected to TONS 504–PACT but exposed to a cell lysis reagent, exposed to TONS 504 (10 mg/l) without irradiation, or subjected to irradiation (30 J/cm2) in the absence or TONS 504 were assayed for lactate dehydrogenase (LDH). B: Experimental conditions. FL cells infected with herpes simplex virus type 1 (HSV-1) or acyclovir (ACV)-resistant HSV-1 were subjected to TONS 504–PACT under the indicated conditions, after which culture supernatants were assayed for LDH. All data are presented as absorbance at 490 nm (A490) minus A595 and are means ± SD for three independent experiments.
Mentions: We first examined the possible effect of TONS 504-PACT on host cell viability by manually counting the number of apparently normal nuclei. Data for infected cells from three independent experiments revealed the lack of effect of TONS 504-PACT on host cell viability as evaluated based on nuclear morphology compared with the corresponding value for uninfected cells not subjected to TONS 504-PACT. There are no significant differences at all conditions of treatment (data not shown). PACT did not exhibit cytotoxicity at any of the TONS 504 concentrations and light energies examined. We then determined the possible effect of TONS 504-PACT on the integrity of the host cells by measuring the release of LDH into the incubation medium (Figure 2 A,B). Whereas the extent of LDH release was increased slightly after exposure of infected FL cells to the higher concentrations of TONS 504 or higher light energies, these effects were not statistically significant (p>0.05, Student t test).

Bottom Line: To evaluate the efficacy of photodynamic antimicrobial chemotherapy (PACT) with the new porphyrin derivative TONS 504 and a light-emitting diode (LED) against acyclovir (ACV)-sensitive and -resistant herpes simplex virus type 1 (HSV-1).Complete eradication of both viruses was apparent at a TONS 504 concentration of 10 mg/l and light energy of 10 to 30 J/cm(2) as well as at a TONS 504 concentration of 1 mg/l and light energy of 20 or 30 J/cm(2).No antiviral effect was apparent with TONS 504 in the absence of irradiation or with irradiation in the absence of TONS 504.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology and Visual Science, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan ; Medical Faculty, Muhammadiyah University, Makassar, South Sulawesi, Indonesia ; Department of Ophthalmology, Hasanuddin University, Makassar, South Sulawesi, Indonesia.

ABSTRACT

Purpose: To evaluate the efficacy of photodynamic antimicrobial chemotherapy (PACT) with the new porphyrin derivative TONS 504 and a light-emitting diode (LED) against acyclovir (ACV)-sensitive and -resistant herpes simplex virus type 1 (HSV-1).

Methods: Human FL cells infected with the viral strains were subjected to PACT with TONS 504 at various concentrations (0.01 to 10 mg/l) and irradiation at various light energies (10 to 30 J/cm(2)) and were then incubated for 24 h before analysis.

Results: Immunocytofluorescence analysis with antibodies to HSV-1 revealed that PACT eliminated HSV-1 and ACV-resistant HSV-1 in a manner dependent on the TONS 504 concentration and light energy. Complete eradication of both viruses was apparent at a TONS 504 concentration of 10 mg/l and light energy of 10 to 30 J/cm(2) as well as at a TONS 504 concentration of 1 mg/l and light energy of 20 or 30 J/cm(2). No antiviral effect was apparent with TONS 504 in the absence of irradiation or with irradiation in the absence of TONS 504. Staining of cell nuclei with 4', 6-diamidino-2-phenylindole revealed no apparent cytotoxicity of the PACT system, a finding that was confirmed by the system's failure to induce the release of lactate dehydrogenase from the host cells.

Conclusions: We conclude that our PACT system based on TONS 504 and an LED is effective for eliminating HSV-1 and ACV-resistant HSV-1 without a harmful effect on host cells.

No MeSH data available.


Related in: MedlinePlus