Amyloid fibrils are fibrillar polypeptide aggregates from several degenerative human conditions, including Alzheimer's and Creutzfeldt-Jakob diseases. Analysis of amyloid fibrils derived from various human diseases (AA, ATTR, A␤2M, AL , and AL amyloidosis) shows that these are associated with a common lipid component that has a conserved chemical composition and that is specifically rich in cholesterol and sphingolipids, the major components of cellular lipid rafts. This pattern is not notably affected by the purification procedure, and no tight lipid interactions can be detected when preformed fibrils are mixed with lipids. By contrast, the early and prefibrillar aggregates formed in an AA amyloidproducing cell system interact with the raft marker ganglioside-1, and amyloid formation is impaired by addition of cholesterolreducing agents. These data suggest the existence of common cellular mechanisms in the generation of different types of clinical amyloid deposits.protein folding ͉ prion ͉ lipid rafts

Cyclodextrins (CDs) are cyclic oligosaccharides that are able to form inclusion complexes with a variety of substances. For pharmaceutical applications, CD‐based drug formulations offer important advantages compared with uncomplexed drugs. These include improved water solubility of lipophilic drug molecules, increased chemical stability, as well as enhanced bioavailability and absorption rate. Also, a number of topical formulations for dermal and transdermal drug delivery contain CDs. However, the most frequently used CDs –β‐CD and MβCD – are known to extract cholesterol from plasma membranes and thus to cause cellular damage and cell death. In the present study, the influence of various CDs and CD derivatives on the human keratinocyte cell line HaCaT was assessed. We found that β‐CD and MβCD induce apoptosis via the activator caspase‐8, which subsequently activates the effector caspases‐3/‐7. Furthermore, β‐CD‐induced apoptosis is accompanied by mitochondrial cytochrome c release. A significant shift from mitochondria into the cytosol was found. These findings may provide further rationale to the use of CDs in topical formulations for dermal and transdermal drug delivery or as raw material in order to functionalize textiles for medical applications.

The two photosensitizers, chlorin e6 and pheophorbide a, were tested in an in vitro model of topical photodynamic therapy (PDT). Both dyes accumulate in HaCaT keratinocytes as verified by fluorescence measurement but pheophorbide a is enriched fivefold more strongly than chlorin e6 after 24 h. HaCaT cells are susceptible to PDT with both dyes. The phototoxicity measured by ATP bioluminescence is caused by necrosis and apoptosis depending on the photosensitizer used and the treatment modality. Chlorin e6 shows higher toxic potential because it elicits nearly 90% cell mortality 24 h after PDT comparable to pheophorbide a but with a fivefold lower rate of accumulation. These results implicate caution with topical PDT of oncologic diseases due to the risk of serious side effects on healthy skin in the course of topical photodynamic treatment. But the lack of dark toxicity and the time-dependent enrichment of both dyes in HaCaT cells are arguments for the application of these sensitizers in topical PDT of non-malign skin disorders. Further studies are necessary to discover appropriate lower doses and mechanisms of action of topical PDT with both compounds.