| Issue
21 |
|
 |
|
|
|
|
|
|
|
| Category |
|
Titel |
|
Author |
| Newsletter |
|
Spectroscopy
of the Udder Model After Application of Carotinoids |
|
Wolfgang
Pittermann |
The “isolated perfused bovine udder” is an in vitro model
which is widely used in dermatological and cosmetic research for absorption and
irritation studies after topical application. It is an ideal carrier of typical
mammalian skin. Continuous perfusion maintains the barrier effect and metabolism.
The starting point for this in vitro study were human studies on sun protection
based on the “inside-outside concept”. In this concept, a systemic application
is an effective addition to the standard topically applied sun protection (1).
In the present work, the isolated perfused bovine
udder model was used to study systemic carotinoid application by reflection
spectroscopy and investigated for its suitability as a UV protection model.
Carotinoids from the algae Dunaliella salina were used in the form of pH-neutral,
deep-orange red suspensions in glycerol. The active component is Betatene 30
percent as a natural carotinoid mix in soy oil. The haemoglobin- and cell-free
organ perfusion eliminates the need for the absorption spectra of haemoglobin
and carotinoids which overlap under in vivo conditions. New starting points
are thus created.
In clinical studies, the concentration of carotinoids
in the skin or blood plasma was measured noninvasively by reflection spectroscopy.
In this study, the reflection spectroscopy from the short-wave UV range to the
long-wave infrared range was used. Two different light intensities were focussed
at two skin depths: up to a depth of 0.5 mm with the epidermis, the vascular
capillary network and the papillary loops and up to a depth of 1.5 mm with the
medium sized vessels. Figure 1 shows the shallow as opposed to the deep measurement
of reflection spectroscopy schematically. These two methods of measurement were
developed for separately measuring the carotinoid concentration in the outer
skin and in the deep layers. The kinetics of carotinoids in the skin after short-term
application thus becomes clearer. In the measurement up to a depth of 0.5 mm
the maximum is already reached after four minutes and maintained for over 80
minutes. The accumulation is concentrated in the subpapillary vascular network
and the ascending capillary loops. The measurement of up to a depth of 1.5 mm
reveals a relatively low concentration in the tissue immediately after the start
of application, but a very much higher concentration 20 minutes after the end.
Focussing on two vascular networks at different
levels is significant for dermatological sun protection because the particularly
exposed capillary loops and the papillary dermal connective tissue surrounding
them need protection. These capillary loops are nearer the skin surface than
the corresponding epidermal regions (Figure 1) and represent an essential part
of the carotinoid-containing connective tissue which is specifically covered
solely by the shallow measuring variant. Other parts of the vascular system
and the connective tissue are not as affected by light because they are better
protected by the epidermis.
Under the experiment conditions of the non-physiologically
high in-use concentration (0.01 percent solution = 190µmol/l) over a period
of 10 minutes as a short term application (190 µmol) and over 130 minutes as
a long-term application (2.740 µmol), followed by washing, the carotinoid is
differently distributed in the surface and deep vascular networks as a function
of time. The subpapillary network fills up quickly. The endothelia appear to
store large amounts of carotinoids very quickly and initially delay a distinct
increase in concentration in the deeper venous vessels.
The grey region in Figure 2 reproduces the accumulation
measured in the human study after 12 weeks’ supplementation in physiological
concentrations (deep variant) and lies over the values of the shallow 0.5 mm
measurement. In addition, the carotinoid accumulation probably occurs partly
outside the vessels on account of the chronic supplementation.

Source: Pittermann, W.; Jungmann, H.; Kietzmann,
M.; Schmitt, M.; Blume, B.; Spektroskopie im UV- und sichtbaren Bereich am isoliert
perfundierten Eutermodell nach systemischer Carotinoidapplikation, Kosmetische
Medizin 19 (1998) 152-162
Literature
(1) Schweitzer, C.; Natural Mixed Carotenoids Provide
‘Inside-Outside’ Sun Protection, Drug & Cosmetic Industry 161 (1997) No.
6, 40-44; Pittermann, W.; The Isolated Perfused Bovine Udder Skin: A Natural
in vitro Skin Model, Skin Care Forum No. 15 (1996) 1-5

Dr. Wolfgang Pittermann, DVM (ECVP), was formerly head of the Laboratory
for Experimental Toxicology and Pathology. Now he is in charge of Henkel's research
platform Biochemistry of the Skin in the fields of experimental
dermatology for the penetration/irritation in the skin and mucous membrane as
well as new technologies.