Time-dependent photodynamic damage to blood vessels: correlation with serum photosensitizer levels.

We have used the technique of dynamic capillaroscopy to study the time-course of photodynamic vascular occlusion in mice injected intraperitoneally with either of two photosensitizers; hematoporphyrin esters (HPE) or meso-tetrahydroxyphenyl chlorin (mTHPC). The peak of vascular occlusion induced by HPE coincided in time with peak serum levels of this photosensitizer (about 3 h after injection). However, there was also a second peak of occlusive activity at about 12 h after injection, at which time serum HPE was falling monotonically. In the case of mTHPC, no peak of occlusive activity was seen at 3 h after injection, even though the serum levels of this photosensitizer, like those of HPE, were highest around this time. Instead, a steady rise in photosensitizing activity was observed, peaking at 11 h. This decoupling between serum drug levels and vascular photosensitization--partial for HPE and complete for mTHPC-suggests that direct photosensitization of endothelial cells is unlikely wholly to explain the vascular collapse. Instead, there must be either another compartment that accumulates photosensitizer more slowly and in which photodynamic activity has an indirect effect on the blood capillaries or a slow metabolic transformation of mTHPC into a more active sensitizer.

Dynamic capillaroscopy: a minimally invasive technique for assessing photodynamic effects in vivo.

A noninvasive method for visualizing the microvasculature in the mouse tail is described, consisting of a custom-built microscope with through-lens illumination. The microscope is fitted with a television camera and images can be recorded on videotape and displayed on a television monitor. Blood vessels are imaged as columns of red blood cells, in which flow is clearly observed. Administration of photosensitizers and illumination with the standard light source produces no observable photodynamic effect on blood flow. The combination of photosensitizer and a more intense light source (either broadband light from a filtered mercury arc or red light from a laser) causes photodynamic cessation of flow within a few minutes. The magnitude of the effect is dependent on the dose and nature of the photosensitizer, the delay after photosensitization and the match between the laser light and the absorption spectra of the photosensitizers in the red region. We conclude that the technique yields results consistent with the known photodynamic effects of the photosensitizers in tumors and propose its use as an initial screening method in vivo, as a means of conducting pharmacokinetic experiments and as an assay of prolonged cutaneous photosensitivity.