Dermatologica 152 (Suppl. 1): 147-153 (1976)
Electron-Microscopic Evaluation of Collagen Fibrils after Topical Corticosteroid Therapy M. G roniowska, J. D abrowski, W. M aciejewski and M. Walski Department of Dermatology, Medical School, Warsaw
Key Words. Electron microscopy of collagen fibrils • Ruthenium red ■Atrophy induced by corticosteroids Abstract. A method used to assess the atrophogenic effect of steroids by electron microscopy after ruthenium red staining is described. In two patients more atrophy was observed after fluocinolone treatment than after hydrocortisone 17-butyrate treatment.
Hypotrophie effects of topical corticosteroids on connective tissue of the skin have recently been studied using light and electron microscopy [3,7,8], In this report the electron-microscopic appearance of dermal collagen in psoriatic patients treated with topical corticosteroids will be presented.
Patients with psoriasis who had no predisposition to atrophy of the skin after topical corticosteroid treatment were studied Hydrocortisone 17-butyrate (I.ocoid®) and fluocinolone acetonide (Synalar*) were used. In two patients with symmetrical skin lesions the effects of hydrocortisone 17-butyrate and fluocinolone acetonide applications were compared. Each patient was treated for 28-35 days. Punch biopsy specimens were obtained from the treated areas of the skin after the com pletion of the application. Control biopsies were taken from untreated skin. Tissue samples were fixed in 2% buffered glutaraldehyde cacodylate (pH 7.4), postfixed in 1.5% osmium tetroxide, dehydrated and embedded in Epon 812. Some ultra-thin sections were selected for measurements o f collagen fibrils (diameters). Measurements were restricted to the fibrils in true cross sections. Part of the tissue sample was processed using the ruthenium red method as described by Luft [4],
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/6/2018 12:18:22 AM
Material and Methods
148
G roniowska/D abrowski/M aciejewski/W alski
Results
On electron-microscopic examination of the skin treated with hydro cortisone 17-butyrate and fluocinolone acetonide, respectively, fibroblasts were characterized by a scarce, rough endoplasmic reticulum. Psoriatic lesions treated with fluocinolone acetonide only disclosed a few cells in the upper part of the dermis as compared with lesions treated with hydro cortisone 17-butyrate. In the subepithelial part of the dermis (fig. 1) collagen fibrils formed thin bundles separated by wide electron-translucent spaces. In the close vicinity of the bundles, randomly distributed collagen micro fibrils were observed. Agglomerations of intermingled microfibrils were also noted. In one case beneath the corium basement membrane, condensed collagen bundles and narrow interstitial spaces were observed. In the deeper parts of the dermis the bundles of collagen fibrils were generally thicker. The measurement of diameters of collagen fibrils disclosed statistically significant differences of the means of treated and untreated skin (table I). Areas of psoriatic skin treated with hydrocortisone 17-butyrate (fig. 2) and fluocinolone acetonide (fig. 3), respectively, revealed a clear difference of diameters of collagen fibrils, as compared with untreated skin (fig. 4). The mean diameter of collagen fibrils from areas treated with fluocinolone acetonide was significantly diminished as compared to the areas treated with hydrocortisone 17-butyrate. By means of the ruthenium red method the reaction product will form, as could be demonstrated, a close association with collagen fibrils. Ruthenium red-positive material formed transverse belts overlying the major periodic bands of collagen fibrils (fig. 5). Occasionally, individual fibrils were surTable I. Diameters of collagen fibrils
(1) HC17-B FCA (2) HC17-B FCA Control HC17-B
Number of measure ments
Mean
Range
SD
A
A
A
125 116 100 118 117
800 614 924 597 1,135
294-1,370 391-1,468 587-1,565 391-1,176 1,153-1,557
186 119 232 86 102
Hydrocortisone 17-butyrate; FCA
P
Electron-Microscopic Evaluation of Collagen Fibrils after Topical Corticosteroid Therapy M. G roniowska, J. D abrowski, W. M aciejewski and M. Walski Department of Dermatology, Medical School, Warsaw
Key Words. Electron microscopy of collagen fibrils • Ruthenium red ■Atrophy induced by corticosteroids Abstract. A method used to assess the atrophogenic effect of steroids by electron microscopy after ruthenium red staining is described. In two patients more atrophy was observed after fluocinolone treatment than after hydrocortisone 17-butyrate treatment.
Hypotrophie effects of topical corticosteroids on connective tissue of the skin have recently been studied using light and electron microscopy [3,7,8], In this report the electron-microscopic appearance of dermal collagen in psoriatic patients treated with topical corticosteroids will be presented.
Patients with psoriasis who had no predisposition to atrophy of the skin after topical corticosteroid treatment were studied Hydrocortisone 17-butyrate (I.ocoid®) and fluocinolone acetonide (Synalar*) were used. In two patients with symmetrical skin lesions the effects of hydrocortisone 17-butyrate and fluocinolone acetonide applications were compared. Each patient was treated for 28-35 days. Punch biopsy specimens were obtained from the treated areas of the skin after the com pletion of the application. Control biopsies were taken from untreated skin. Tissue samples were fixed in 2% buffered glutaraldehyde cacodylate (pH 7.4), postfixed in 1.5% osmium tetroxide, dehydrated and embedded in Epon 812. Some ultra-thin sections were selected for measurements o f collagen fibrils (diameters). Measurements were restricted to the fibrils in true cross sections. Part of the tissue sample was processed using the ruthenium red method as described by Luft [4],
Downloaded by: Univ. of California Santa Barbara 128.111.121.42 - 3/6/2018 12:18:22 AM
Material and Methods
148
G roniowska/D abrowski/M aciejewski/W alski
Results
On electron-microscopic examination of the skin treated with hydro cortisone 17-butyrate and fluocinolone acetonide, respectively, fibroblasts were characterized by a scarce, rough endoplasmic reticulum. Psoriatic lesions treated with fluocinolone acetonide only disclosed a few cells in the upper part of the dermis as compared with lesions treated with hydro cortisone 17-butyrate. In the subepithelial part of the dermis (fig. 1) collagen fibrils formed thin bundles separated by wide electron-translucent spaces. In the close vicinity of the bundles, randomly distributed collagen micro fibrils were observed. Agglomerations of intermingled microfibrils were also noted. In one case beneath the corium basement membrane, condensed collagen bundles and narrow interstitial spaces were observed. In the deeper parts of the dermis the bundles of collagen fibrils were generally thicker. The measurement of diameters of collagen fibrils disclosed statistically significant differences of the means of treated and untreated skin (table I). Areas of psoriatic skin treated with hydrocortisone 17-butyrate (fig. 2) and fluocinolone acetonide (fig. 3), respectively, revealed a clear difference of diameters of collagen fibrils, as compared with untreated skin (fig. 4). The mean diameter of collagen fibrils from areas treated with fluocinolone acetonide was significantly diminished as compared to the areas treated with hydrocortisone 17-butyrate. By means of the ruthenium red method the reaction product will form, as could be demonstrated, a close association with collagen fibrils. Ruthenium red-positive material formed transverse belts overlying the major periodic bands of collagen fibrils (fig. 5). Occasionally, individual fibrils were surTable I. Diameters of collagen fibrils
(1) HC17-B FCA (2) HC17-B FCA Control HC17-B
Number of measure ments
Mean
Range
SD
A
A
A
125 116 100 118 117
800 614 924 597 1,135
294-1,370 391-1,468 587-1,565 391-1,176 1,153-1,557
186 119 232 86 102
Hydrocortisone 17-butyrate; FCA
P
