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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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Time course of neuronal and satellite glial death after mTHPC-mediated PDT. Cells in 3D gels were incubated with 4 μg ml–1 mTHPC for 4 h, exposed to light (A) or used as no-light controls (B), then maintained in culture for various times before cell death analysis and immunodetection of neurons.
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fig4: Time course of neuronal and satellite glial death after mTHPC-mediated PDT. Cells in 3D gels were incubated with 4 μg ml–1 mTHPC for 4 h, exposed to light (A) or used as no-light controls (B), then maintained in culture for various times before cell death analysis and immunodetection of neurons.

Mentions: In Figure 3, the extent of cell death at 24 h after PDT was used to show the differences in sensitivity between neurons and other cells. The time course for cell death after PDT can vary according to cell type, drug/light dose and various other parameters, so it was important to establish that the 24 h time point was appropriate for this study. Therefore, parallel cultures of neurons and satellite glia were treated using 4 μg ml–1 mTHPC and maintained in culture up to 36 h. Figure 4 shows the time course of cell death after mTHPC-mediated PDT. There was no significant difference in the extent of cell death in the cultures analysed after different lengths of time.


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)

Time course of neuronal and satellite glial death after mTHPC-mediated PDT. Cells in 3D gels were incubated with 4 μg ml–1 mTHPC for 4 h, exposed to light (A) or used as no-light controls (B), then maintained in culture for various times before cell death analysis and immunodetection of neurons.
© Copyright Policy
Related In: Results  -  Collection

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

fig4: Time course of neuronal and satellite glial death after mTHPC-mediated PDT. Cells in 3D gels were incubated with 4 μg ml–1 mTHPC for 4 h, exposed to light (A) or used as no-light controls (B), then maintained in culture for various times before cell death analysis and immunodetection of neurons.
Mentions: In Figure 3, the extent of cell death at 24 h after PDT was used to show the differences in sensitivity between neurons and other cells. The time course for cell death after PDT can vary according to cell type, drug/light dose and various other parameters, so it was important to establish that the 24 h time point was appropriate for this study. Therefore, parallel cultures of neurons and satellite glia were treated using 4 μg ml–1 mTHPC and maintained in culture up to 36 h. Figure 4 shows the time course of cell death after mTHPC-mediated PDT. There was no significant difference in the extent of cell death in the cultures analysed after different lengths of time.

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