Measurement of cytokines in cantharidin bisters

1. Foot ulcers are a feared complication of diabetes mellitus and occur mainly
in polyneuropathy patients. We assume that in polyneuropathy patients the
inflammatory response to exogenic stimuli is decreased which predisposes to the
development of ulcers and infections.
2. Charcot osteoarthropathy is a rare but very invalidating complication of
(diabetic) polyneuropathy, often resulting in a deformed foot. We assume that
an excessive inflammatory response, in which pro-inflammatory cytokines play a
role, to exogenic stimuli is the underlying mechanism in the development of a
acute Charcot foot.

The primary goal of this study is to te verify that it is possible to measure
te concentrations of cytokines IL-1α, IL-1RA en IL-8 in the fluid of a blister
that is applied tot the skin of healthy volunteers. using cantharidin. In the
future we wish to use this knowledge for developing a method for:
1. testing the hypothesis that in polyneuropathy patients the inflammatory
response to exogenic stimuli is decreased.
2. testing the hypothesis that the acute Charcot foot develops as a result of a
excessive inflammatory response.

Pilotstudie (n=6). 25 µl of 0.1% cantharidin solution is applied to the skin of
the upper arm of healthy volunteers. Afterwards a blister will arise. At t=24h
the bister will be perforated and the blister fluid will be collected using a
pipet. The blister fluid will be kept at a temperature of -80°C and more later
concentrations of cytikines IL-1α, IL-1RA and IL-8 will be performed using
ELISA.

We don't expect any risks for the volunteers. Cantharidin is prescriped more
than 1 million times without severe side effects.