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Peripheral neural cell sensitivity to mTHPC-mediated photodynamic therapy in a 3D in vitro model.

Wright KE, Liniker E, Loizidou M, Moore C, Macrobert AJ, Phillips JB - Br. J. Cancer (2009)

Bottom Line: MCF-7s and satellite glia were significantly more sensitive to PDT than neurons.Importantly, 4 microg ml(-1) mTHPC-PDT caused no significant neuron death compared with untreated controls but was sufficient to elicit substantial cell death in the other cell types.Initially, treatment reduced neurite length; neurons then extended neurites equivalent to those of untreated controls.

View Article: PubMed Central - PubMed

Affiliation: Life Sciences Department, The Open University, Milton Keynes MK7 6AA, UK.

ABSTRACT

Background: The effect of photodynamic therapy (PDT) on neural cells is important when tumours are within or adjacent to the nervous system. The purpose of this study was to investigate PDT using the photosensitiser, meta-tetrahydroxyphenyl chlorin (mTHPC), on rat neurons and satellite glia, compared with human adenocarcinoma cells (MCF-7).

Methods: Fluorescence microscopy confirmed that mTHPC was incorporated into all three cell types. Sensitivity of cells exposed to mTHPC-PDT (0-10 microg ml(-1)) was determined in a novel 3-dimensional collagen gel culture system. Cell death was quantified using propidium iodide and cell types were distinguished using immunocytochemistry. In some cases, neuron survival was confirmed by measuring subsequent neurite growth in monolayer culture.

Results: MCF-7s and satellite glia were significantly more sensitive to PDT than neurons. Importantly, 4 microg ml(-1) mTHPC-PDT caused no significant neuron death compared with untreated controls but was sufficient to elicit substantial cell death in the other cell types. Initially, treatment reduced neurite length; neurons then extended neurites equivalent to those of untreated controls. The protocol was validated using hypericin (0-3 microg ml(-1)), which caused neuron death equivalent to other cell types.

Conclusion: Neurons in culture can survive mTHPC-PDT under conditions sufficient to kill tumour cells and other nervous system cells.

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Related in: MedlinePlus

Reduction in neurite length due to mTHPC-mediated PDT. Dorsal root ganglion neurons were cultured on coverslips for 3 days, then treated with various concentrations of mTHPC and light, then maintained in culture for a further 24 h before fixation and quantification of the total length of all β-III-tubulin immunoreactive neurites. Data are means±s.e.m., N=4. (**P<0.001 for treatments compared with no drug or light control, one-way ANOVA with Dunnett's multiple comparison post test).
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fig2: Reduction in neurite length due to mTHPC-mediated PDT. Dorsal root ganglion neurons were cultured on coverslips for 3 days, then treated with various concentrations of mTHPC and light, then maintained in culture for a further 24 h before fixation and quantification of the total length of all β-III-tubulin immunoreactive neurites. Data are means±s.e.m., N=4. (**P<0.001 for treatments compared with no drug or light control, one-way ANOVA with Dunnett's multiple comparison post test).

Mentions: The effect of mTHPC-mediated PDT on neurite length in DRG cultures was tested in the 2D system. After 3 days in culture, cells were treated with mTHPC and light, and the effect of this treatment on neurite length was analysed 24 h later (Figure 2 and Supplementary Figure 2). There was no statistically significant effect on the neurite length when drug or light were applied separately, or when PDT was performed using 0.1 μg ml–1 mTHPC, whereas PDT using higher concentrations of mTHPC (0.3 μg ml–1 and above) caused a significant reduction in neurite length compared with untreated controls (P<0.001, one-way ANOVA with Dunnett's post test).


Peripheral neural cell sensitivity to mTHPC-mediated photodynamic therapy in a 3D in vitro model.

Wright KE, Liniker E, Loizidou M, Moore C, Macrobert AJ, Phillips JB - Br. J. Cancer (2009)

Reduction in neurite length due to mTHPC-mediated PDT. Dorsal root ganglion neurons were cultured on coverslips for 3 days, then treated with various concentrations of mTHPC and light, then maintained in culture for a further 24 h before fixation and quantification of the total length of all β-III-tubulin immunoreactive neurites. Data are means±s.e.m., N=4. (**P<0.001 for treatments compared with no drug or light control, one-way ANOVA with Dunnett's multiple comparison post test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2736832&req=5

fig2: Reduction in neurite length due to mTHPC-mediated PDT. Dorsal root ganglion neurons were cultured on coverslips for 3 days, then treated with various concentrations of mTHPC and light, then maintained in culture for a further 24 h before fixation and quantification of the total length of all β-III-tubulin immunoreactive neurites. Data are means±s.e.m., N=4. (**P<0.001 for treatments compared with no drug or light control, one-way ANOVA with Dunnett's multiple comparison post test).
Mentions: The effect of mTHPC-mediated PDT on neurite length in DRG cultures was tested in the 2D system. After 3 days in culture, cells were treated with mTHPC and light, and the effect of this treatment on neurite length was analysed 24 h later (Figure 2 and Supplementary Figure 2). There was no statistically significant effect on the neurite length when drug or light were applied separately, or when PDT was performed using 0.1 μg ml–1 mTHPC, whereas PDT using higher concentrations of mTHPC (0.3 μg ml–1 and above) caused a significant reduction in neurite length compared with untreated controls (P<0.001, one-way ANOVA with Dunnett's post test).

Bottom Line: MCF-7s and satellite glia were significantly more sensitive to PDT than neurons.Importantly, 4 microg ml(-1) mTHPC-PDT caused no significant neuron death compared with untreated controls but was sufficient to elicit substantial cell death in the other cell types.Initially, treatment reduced neurite length; neurons then extended neurites equivalent to those of untreated controls.

View Article: PubMed Central - PubMed

Affiliation: Life Sciences Department, The Open University, Milton Keynes MK7 6AA, UK.

ABSTRACT

Background: The effect of photodynamic therapy (PDT) on neural cells is important when tumours are within or adjacent to the nervous system. The purpose of this study was to investigate PDT using the photosensitiser, meta-tetrahydroxyphenyl chlorin (mTHPC), on rat neurons and satellite glia, compared with human adenocarcinoma cells (MCF-7).

Methods: Fluorescence microscopy confirmed that mTHPC was incorporated into all three cell types. Sensitivity of cells exposed to mTHPC-PDT (0-10 microg ml(-1)) was determined in a novel 3-dimensional collagen gel culture system. Cell death was quantified using propidium iodide and cell types were distinguished using immunocytochemistry. In some cases, neuron survival was confirmed by measuring subsequent neurite growth in monolayer culture.

Results: MCF-7s and satellite glia were significantly more sensitive to PDT than neurons. Importantly, 4 microg ml(-1) mTHPC-PDT caused no significant neuron death compared with untreated controls but was sufficient to elicit substantial cell death in the other cell types. Initially, treatment reduced neurite length; neurons then extended neurites equivalent to those of untreated controls. The protocol was validated using hypericin (0-3 microg ml(-1)), which caused neuron death equivalent to other cell types.

Conclusion: Neurons in culture can survive mTHPC-PDT under conditions sufficient to kill tumour cells and other nervous system cells.

Show MeSH
Related in: MedlinePlus