Characterization of apoptosis induced by photodynamic treatment with hypericin in A431 human epidermoid carcinoma cells.

Hypericin is a naturally occurring metabolite extracted from Hypericum plants and is regarded as a promising photosensitizing agent for applications in the frame of photodynamic treatment (PDT). This treatment procedure is based on the light-induced formation of reactive oxygen species and subsequent destruction of target cells. We used an in vitro model system consisting of human epidermoid carcinoma cells (A431) and hypericin as a photosensitizer to study the time- and dose-dependent characteristics of hypericin-PDT-based induction of cytotoxicity and apoptotic cell death.

Apoptosis following photodynamic tumor therapy: induction, mechanisms and detection.

As a treatment modality for malign and certain non-malignant diseases, photodynamic therapy (PDT) involves a two step protocol which consists of the (selective) uptake and accumulation of a photosensitizing agent in target cells and the subsequent irradiation with light in the visible range. Reactive oxygen species (ROS) produced during this process cause cellular damage and, depending on the treatment dose/severity of damage, lead to either cellular repair/survival, apoptotic cell death or necrosis. PDT-induced apoptosis has been focused on during the last years due to the intimate connection between ROS generation, mitochondria and apoptosis; by this PDT employs mechanisms different to those in the action of radio- and chemotherapeutics, giving rise to the chance of apoptosis induction by PDT even in cells resistant to conventional treatments.

Photodynamic treatment with fractionated light decreases production of reactive oxygen species and cytotoxicity in vitro via regeneration of glutathione.

Photodynamic therapy removes unwanted or harmful cells by overproduction of reactive oxygen species (ROS). Fractionated light delivery in photodynamic therapy may enhance the photodynamic effect in tumor areas with insufficient blood supply by enabling the reoxygenation of the treated area. This study addresses the outcome of fractionated irradiation in an in vitro photodynamic treatment (PDT) system, where deoxygenation can be neglected.

Differential effects of glucose deprivation on the cellular sensitivity towards photodynamic treatment-based production of reactive oxygen species and apoptosis-induction.

Photodynamic treatment (PDT) employs a photosensitizer and the light-induced formation of reactive oxygen species--antagonized by cellular antioxidant systems--for the removal of harmful cells. This study addresses the effect of altered carbohydrate metabolism on the cellular antioxidant glutathione system, and the subsequent responses to PDT. It is shown that glucose-deprivation of 18 h prior to PDT causes a reduced level of intracellular glutathione and an increased cytotoxicity of PDT.

Fast and reliable determination of intracellular ATP from cells cultured in 96-well microplates.

Due to its fundamental role in cellular energetics, metabolic regulation and cellular signalling, determination of intracellular adenosine-5'-triphosphate (ATP) is an irreplaceable tool in the characterization of cellular physiology. Classical protocols describing the measurement of ATP from cells usually suffer from the use of relatively high reaction volumes and the according use of cuvettes for ATP determination, thus making them impractical for processing high sample numbers. We therefore present two methods for the fast and reliable measurement of intracellular ATP levels of cells cultured in 96-well microplates.

Glucose is required to maintain high ATP-levels for the energy-utilizing steps during PDT-induced apoptosis.

Photodynamic therapy (PDT) may trigger apoptosis or necrosis in cancer cells. Several steps in the induction and execution of apoptosis require high amounts of adenosine-5'-triphosphate (ATP). Because the mitochondrial membrane potential (delta psi) decreases early in apoptosis, we raised the question about the mechanisms of maintaining a sufficiently high ATP level.