Introduction: Photodynamic therapy (PDT) shows potential as an effective treatment for prostate cancer.  Clinical observations indicate that this approach causes fewer nerve damage related side-effects than conventional treatments.  The aim here is to investigate the effect of PDT on nerve tissue using engineered 3-dimensional cell culture models.  Initial experiments focussed on establishing photosensitiser localisation in neurones and Schwann cells, then developing a model for simulating nerve PDT in culture

Brown, R.A.

Liniker, E

MacRobert, A.J.

Phillips, J.B.

Saffrey, M.J.

Wright, K.E.

English

Open Research Online (The Open University)

Open Research OnlineThe Open University’s repository of research publicationsand other research outputsInvestigating the effect of photodynamic therapy onnerves using tissue engineered culture modelsConference or Workshop ItemHow to cite:Wright, K.E.; Liniker, E; MacRobert, A.J.; Brown, R.A.; Saffrey, M.J. and Phillips, J.B. (2006). Investigatingthe effect of photodynamic therapy on nerves using tissue engineered culture models. In: Tissue & Cell EngineeringSociety 2006, 3-4 Jul 2006, Sheffield, UK.For guidance on citations see FAQs.c© [not recorded]Version: [not recorded]Copyright and Moral Rights for the articles on this site are retained by the individual authors and/or other copyrightowners. For more information on Open Research Online’s data policy on reuse of materials please consult the policiespage.oro.open.ac.ukInvestigating the effect of photodynamic therapy on nerves using tissue engineered culture models K E Wright1, E Liniker2, A J MacRobert2, R A Brown2, M J Saffrey1 & J B Phillips1 1Biological Sciences Department, The Open University, Milton Keynes, U.K.  2 National Medical Laser Centre, UCL, London, U.K. 3Institute of Orthopaedics and Musculoskeletal Science, UCL, London, U.K.  INTRODUCTION: Photodynamic therapy (PDT) shows potential as an effective treatment for prostate cancer [1]. Clinical observations indicate that this approach causes fewer nerve damage related side effects than conventional treatments. The aim here is to investigate the effect of PDT on nerve tissue using engineered 3-dimensional cell culture models. Initial experiments focussed on establishing photosensitiser localisation in neurones and Schwann cells, then developing a model for simulating nerve PDT in culture. Schwann mTHPCSchwann controlNeurone cell body mTHPCNeurone cell body controlNeurone process mTHPCNeurone process control0255075Mean pixel fluorescenceintensityMETHODS: Neurones and Schwann cells were cultured from the dorsal root ganglia and sciatic nerves of 200g rats. Tissues were dissociated using collagenase then seeded onto glass coverslips for localisation experiments using 4 µg/ml meso tetra hydroxyl phenyl chlorine (mTHPC) for 4 hours. Intracellular localisation of mTHPC fluorescence was visualised using fluorescence and confocal microscopy and quantified using digital image analysis. Schwann cells and neurones were identified using immunoreactivity for S100 and βIII tubulin respectively. A 3-dimensional (3D) cell culture system was developed to enable PDT to be directed against distinct parts of neurones. DRG explants were embedded within tethered aligned Schwann cell-populated type I collagen gels [2].  RESULTS: Localisation experiments revealed photosensitiser fluorescence within Schwann cells and the cell bodies of neurones, but not in neuronal cell processes (fig 1 & 2).  Fig. 1: Neurone in culture (left) with mTHPC localisation in cell body and underlying Schwann cells but not in neuronal cell process (right). Fig 2: Quantifiaction of mTHPC localisation in neurons and Schwann cells DRG explants embedded within aligned Schwann cell populated collagen gels extended linear neuronal processes (fig 3), forming an effective model with which to investigate PDT in distinct parts of neurones.   mTHPC βIII tubulin Fig 3: DRG explant extending processes in 3D culture model  βIII tubulin   DISCUSSION & CONCLUSIONS: This ongoing investigation into nerve sparing indicates that mTHPC may not be localised within neuronal processes. A 3D model has been created to investigate whether this protects the neuron from PDT damage when light is directed at the axon.  REFERENCES: 1 C.M. Moore, T.R. Nathan, W.R. Lees et al. (2006) Lasers Surg Med (in press). 2 J.B. Phillips, S.C.J. Bunting, S.M. Hall et al. (2005) Tiss Eng 11:1611-17. 

Investigating the effect of photodynamic therapy on nerves using tissue engineered culture models

Photodynamic therapy (PDT)

shows potential as an effective treatment for

prostate cancer. Clinical observations indicate

that this approach causes fewer nerve damage

related side effects than conventional treatments.

The aim here is to investigate the effect of PDT on

nerve tissue using engineered 3-dimensional cell

culture models. Initial experiments focussed on

establishing photosensitiser localisation in

neurones and Schwann cells, then developing a

model for simulating nerve PDT in culture

Wright, KE

MacRobert, AJ

Brown, RA

Saffrey, MJ

Phillips, JB

UCL Discovery

European Cells and Materials Vol. 11. Suppl. 3, 2006 (page 51)                                                             ISSN 1473-2262  Investigating the effect of photodynamic therapy on nerves using tissue engineered culture models K E Wright1, E Liniker2, A J MacRobert2, R A Brown2, M J Saffrey1 & J B Phillips1 1Biological Sciences Department, The Open University, Milton Keynes, U.K.  2 National Medical Laser Centre, UCL, London, U.K. 3Institute of Orthopaedics and Musculoskeletal Science, UCL, London, U.K.  INTRODUCTION: Photodynamic therapy (PDT) shows potential as an effective treatment for prostate cancer [1]. Clinical observations indicate that this approach causes fewer nerve damage related side effects than conventional treatments. The aim here is to investigate the effect of PDT on nerve tissue using engineered 3-dimensional cell culture models. Initial experiments focussed on establishing photosensitiser localisation in neurones and Schwann cells, then developing a model for simulating nerve PDT in culture. METHODS: Neurones and Schwann cells were cultured from the dorsal root ganglia and sciatic nerves of 200g rats. Tissues were dissociated using collagenase then seeded onto glass coverslips for localisation experiments using 4 µg/ml meso tetra hydroxyl phenyl chlorine (mTHPC) for 4 hours. Intracellular localisation of mTHPC fluorescence was visualised using fluorescence and confocal microscopy and quantified using digital image analysis. Schwann cells and neurones were identified using immunoreactivity for S100 and βIII tubulin respectively. A 3-dimensional (3D) cell culture system was developed to enable PDT to be directed against distinct parts of neurones. DRG explants were embedded within tethered aligned Schwann cell-populated type I collagen gels [2].  RESULTS: Localisation experiments revealed photosensitiser fluorescence within Schwann cells and the cell bodies of neurones, but not in neuronal cell processes (fig 1 & 2).  Fig. 1: Neurone in culture (left) with mTHPC localisation in cell body and underlying Schwann cells but not in neuronal cell process (right). Fig 2: Quantifiaction of mTHPC localisation in neurons and Schwann cells DRG explants embedded within aligned Schwann cell populated collagen gels extended linear neuronal processes (fig 3), forming an effective model with which to investigate PDT in distinct parts of neurones.   Fig 3: DRG explant extending processes in 3D culture model    DISCUSSION & CONCLUSIONS: This ongoing investigation into nerve sparing indicates that mTHPC may not be localised within neuronal processes. A 3D model has been created to investigate whether this protects the neuron from PDT damage when light is directed at the axon.  REFERENCES: 1 C.M. Moore, T.R. Nathan, W.R. Lees et al. (2006) Lasers Surg Med (in press). 2 J.B. Phillips, S.C.J. Bunting, S.M. Hall et al. (2005) Tiss Eng 11:1611-17. Schwann mTHPCSchwann controlNeurone cell body mTHPCNeurone cell body controlNeurone process mTHPCNeurone process control0255075Mean pixel fluorescenceintensityβIII tubulin mTHPC βIII tubulin 

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Investigating the effect of photodynamic therapy on nerves using tissue engineered culture models

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