Медицина/6. Экпериментальная и клиническая фармакология

Candidate of Pharm. Sc. Butko Y. O.

National University of Pharmacy, Ukraine

Effect of lotion with glucocorticosteroid and ceramides on skin morphostructure in allergic contact dermatitis

High level of dermatitis diseases in Ukrainian population, increasing the number of severe clinical forms, and shortening the term of remission and low indicator of recovery make a problem of dermatitis therapy, which is the one of topical problems in sophisticated dermatology [2, 5].

Currently, the treatment of dermatoses paid much attention to the state of proliferation of skin and restore its barrier function, because sound is the use of tools that stimulate proliferation and restore skin structure, including ceramides [4].

The aim of this work was morphological study of rats skin state in treating by lotion "Mometasone with ceramides" in the course of allergic contact dermatitis (ACD).

Materials and methods. The object of study was animals skin after treating with lotion “Mometasone with Ceramides”. 18 rats were used in study, which were divided on: 1 group – intact, 2 group – positive control (control pathology), 3 group – rats, which was treated with lotion “Mometasone with Ceramides”. To develop AСВ animals were coated 5% alcoholic solution of 2,4-dinitro-chlorobenzene (DNCB) by the method of P. Zalkan [1]. Lotions were applied a thin coat once a day. After 7^th treating day animals were removed from study and all skin material were fixed in 10% formalin solution for morphological studying. Than skin examples were swamped in celloidin-paraffin and sections were painted in hematoxylin and eosin (1) [3]. We performed review micropreparations on a microscope Micros400 (Austria). Photomicrography microscopic images was performed a digital camera Nikon Col Pix 4500. Pictures were processed on a computer Pentium 2,4 GHz using Nikon View 5.

Results and discussion. As we can see in studies of intact rats, epidermis developed well, consists of 3-5 rows of cells. We differentiated in it: one row of basal cells, 1-3 rows of thorny cells, one or rarely two rows of granular cells and the stratum corneum, which consists of fluffy corneal plates. Derma is wide with compact bundles of collagen fibers with a few thin-walled blood vessels. Cell saturation dermis is mild (lymphoid cells, fibroblasts). Epidermal-dermal border is clear. Hair follicles are multiple, cut as extended and cross section (pic.1а).

а	b	c
d	f	g

Рiс. 1 – intact rats skin (а). Positive control group rats skin: b – epidermis and papillary dermis layer necrotized, demarcation shaft on the border with mesh layer of the dermis; c – regenerated epidermis thickened under the crust, there are visible signs are not sharply distinct spongiosis, fiber dermis are swollen. d – hypertrophy of the epidermis, acanthosis, inflammatory reaction in the dermis; f – inflammatory infiltrate around the hair follicle with the destruction of the wall; g – narrowing of the capillary lumen, endothelial proliferation, inflammatory reaction in the subcutaneous tissue. Hematoxylin-eosin. х 100.

On 7^th day after applying crucial dose DNCB in all control pathology rats we found a spreading skin destructions. Epidermis is deformed on this parts, often it looks like amorphous mass, which densely infiltrated a cellular detritus, macrophages. Collagen fibers are homogenized in papillary layer of the dermis, strongly eosinophilic, epithelial cells of hair follicles and sheaths are in the necrosis. There is visible demarcation shaft some places among the crust and saved part of the dermis (pic. 1b). Often crust extends far beyond the damage and covers (peels tightly or partially) thickened epithelial layer, in which we could see a mild distinct spongiosis (intercellular edema in the thorny layer), vacuolar degeneration of epidermocytes. The epithelium of hair follicles proliferates. Collagen matrix is swollen in the dermis and moderate polymorphonuclear infiltrates are visible around some hair follicles and blood vessels (pic. 1c). Probably it was the regeneration of damaged during primary-contact reactions to allergen skin in these areas at the time of the experiment.

Often epithelial layer hypertrophied on the skin, free from peel, we can incomplete keratinization, acanthosis, more expressive spongiosis. We observed in the dermis infiltrates of histiocytes, lymphocytes, macrophages, plasma cells (pic. 1d). Inflammatory cell infiltration was observed often and around hair follicles and combined with their sebaceous glands (epithelial cells of them are necrotic) and in subcutaneous tissue. We traced wall thickening by hypertrophy and hyperplasia of endothelial cells and pericapillary in mesh dermis capillaries, capillary lumen is diminished (pic. 1f,g). The morphological picture is characteristic ACD.

During the treatment lotion improved skin condition in 83.33% of the rats. Morphological characteristics AKD were not in 33.3% of these animals (pic. 2а). On other 50% of the rats was observed (mostly) small, limited both in depth and across the injury site, covered the crust that is quite easy to flake off. We traced obvious signs accelerating regeneration of the epithelium in these areas (expressive as boundary epithelization and from the epithelium of saved hair follicles). There is no or there is reduced inflammatory cell reaction in the dermis under the, the condition of the walls and the size of the lumen were normal of most blood capillaries (pic. 2b).

a	b	c

Рiс 2. Rats skin which were treated with lotion “Mometasone with Ceramides”: а – no signs of allergic dermatitis; b – limited area of damage, no inflammatory cell reaction in the dermis, the condition of capillaries is normal; c – in the dermis condition of capillaries is close to normal, inflammatory reaction precapillary and subcutaneous tissue is decreased. Hematoxylin-eosin. х 100.

Spreading peel outside reduced markedly damage zones. Crust is sequestered often, and large areas of the skin surface are clean. The thickness of the epithelial layer is clearly reduced relative to control pathology, signs of acanthosis were not found. Sometimes there was a temperate spongiosis and vacuolar dystrophy of epidermocytes. In the dermis condition of collagen matrix, arteriovenula bridge, the hair follicles are close to normal, inflammatory reaction is absent or greatly reduced (pic. 2c). Only one rat (16,7%), which was treated with lotion “Mometasone with Ceramides”, skin condition was a little different from that of the control rats.

Conclusion. Thus, we can concluded on the basis of these studies, that rats have clear allergic skin inflammation after applications with DNCB. Application of lotion “Mometasone with Ceramides” reduces the intensity of ACD (50% rats did not have destructive epidermis changes, the zone of damage is limited, reduced exudative manifestations and inflammatory reaction in the dermis, signs of accelerating epithelial; 33,3% rats had normal skin). Thus, the introduction of ceramides to the cream of mometasone furoate helps normalize skin structure that makes it worthwhile to use in the treatment of allergic dermatitis.

References

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