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Corticosteroid-induced inhibition of the biosynthesis of human skin collagen
Abstract
The effects of hydrocortisone acetate, fluocinolone acetonide, fluclorolone acetonide, betamethasone-17-valerate, fluprednyliden-21-acetate and flumethasone pivalate on the biosynthesis of human skin collagen were studied in vitro. Skin specimens were incubated in a medium containing a test substance and radioactive proline, and the formation of radioactive hydroxyproline in nondialysable proteins was taken as an index of the rate of collagen biosynthesis.Hydroxyproline formation was inhibited by all the corticosteroids tested in concentrations of 30 μg/ml or higher. The effect on hydroxyproline formation was smallest with hydrocortisone acetate and most pronounced with betamethasone-17-valerate in all concentrations used. In general, the corticosteroid-induced inhibition of collagen biosynthesis was found to be dose-dependent. The differences in the degree to which collagen formation is inhibited by the different corticosteroids may have relevance to the extent of the local side-effects, such as atrophy of skin, reported to be produced by fluorinated corticosteroids.
... It is thought that the decreased productivity of hydroxyl-proline is the reason for such diminished structural stability of connective tissue. 17,18 Electrolyte abnormalities mainly involve the development of hypokalemia due to mineralocorticoid effects of prednisone, prednisolone and hydrocortisone. There exists the possibility, although rare, of occurrence of electrolyte abnormalities with topical (per rectum) use of corticosteroids. ...
Corticosteroids act as potent anti-inflammatory drugs and have been used in various sport settings for the treatment of both acute and chronic injuries. Basic physiology and mechanisms of action for gluco- and mineralocorticoids are discussed. Methods of administration, the action on the inflammatory response, and potential short and long-term side effects of corticosteroid use are also deliberated. More specifically, corticosteroid use in the treatment and management of sport-related injuries are discussed, elucidating on the ethical boundaries and anti-doping regulations pertaining to corticosteroid use in sports, as well as putting forth suggestions for the use of local steroid injections and their contraindications. In conclusion, it was found that, despite some controversy in the use of corticosteroid treatment in the sports environment, little empirical evidence exists that could conclusively rule for or against its use. It is however clear that, if clinically justified, there is a significant role for corticosteroid treatment in the realm of sports injury and notably with a relatively low risk profile, if administered correctly.
... It is thought that the decreased productivity of hydroxylproline is the reason for such diminished structural stability of connective tissue. 17,18 Electrolyte abnormalities mainly involve the development of hypokalemia due to mineralocorticoid effects of prednisone, prednisolone and hydrocortisone. There exists the possibility, although rare, of occurrence of electrolyte abnormalities with topical (per rectum) use of corticosteroids. ...
Corticosteroids act as potent anti-inflammatory drugs and have been used in various sport settings for the treatment of both acute and chronic injuries. Basic physiology and mechanisms of action for gluco- and mineralocorticoids are discussed. Methods of administration, the action on the inflammatory response, and potential short and long-term side effects of corticosteroid use are also deliberated. More specifically, corticosteroid use in the treatment and management of sport-related injuries are discussed, elucidating on the ethical boundaries and anti-doping regulations pertaining to corticosteroid use in sports, as well as putting
forth suggestions for the use of local steroid injections and their contraindications. In conclusion, it was found that, despite some controversy in the use of corticosteroid treatment in the sports environment, little empirical evidence exists that could conclusively rule for or against its use. It is however clear that, if clinically justified, there is a significant role for corticosteroid treatment in the realm of sports injury and notably with a relatively low risk profile, if administered correctly.
Keywords: Steroid hormones, athlete, sports, injury, inflammation.
... Subsequently, numerous studies have addressed the inhibition of collagen gene expression utilizing skin fibroblast cultures (see Ref 112). In general, the results indicate that fluorinated corticosteroids are more potent inhibitors of collagen biosynthesis than their non-fluorinated counterparts, and that the therapeutic efficacy of these compounds roughly parallels the inhibition of collagen synthesis [115,116]. The inhibition of collagen gene expression by steroids apparently occurs both on the transcriptional, translational and post-translational levels. ...
The extracellular connective tissue matrix of the skin is a complex aggregate of distinct collagenous and non-collagenous components. Optimal quantities and delicate interactions of these components are necessary to maintain normal physiologic properties of skin. This overview summarizes the progress made in understanding the normal biology and biochemistry of the extracellular matrix, and will highlight cutaneous diseases with underlying molecular defects in the structure and expression of extracellular matrix components.
In 1849, Thomas Addison discovered the link between chronic insufficiency of the adrenals and certain—unclear at the time—systemic features. This condition was later named Addison’s disease, but the mechanism of this deadly condition remained elusive. By 1894, other researchers concluded that the adrenal cortex produced a hormone, which they named cortin. During the 1930s, Edward Calvin Kendall, a researcher at Mayo Clinic, isolated six different substances produced by the adrenal cortex, which he coined as compounds A to F, according to the chronological order of their discovery. He distinguished compound E as a molecule with “…significant anti-rheumatic properties….” That molecule was named cortisone, and in September 1948, cortisone was the first glucocorticoid ever to be administered to a patient with rheumatoid arthritis [2], a condition that remained incurable.
UV rays constitute an extremely important environmental factor known to operate adaptative mechanisms that maintain biological homeostasis in the skin, adrenal glands, and the brain. The skin is extremely vulnerable to UV rays. UV rays deform collagen, the main component of elastic fibers, decreasing its normal function, and ultimately reducing skin’s elasticity. We confirmed that psychological stress occurring during the early stages of UVB-irradiation degraded collagen function by inhibiting production rather than the decomposition of collagen, thereby promoting skin aging. UV irradiation for 0–2 weeks increased the level of a stress factor, corticosterone (CORT). High-performance liquid chromatography and western blot analysis confirmed that the increase was caused by enhanced CYP11B1/2 levels during steroid synthesis in the adrenal gland. Precursor levels decreased significantly during the two weeks of UV irradiation. Skin collagen and collagen fibers reduced drastically during this time. Furthermore, the administration of osilodrostat, a USFDA-approved drug that selectively inhibits CYP11B1/2, preserved skin collagen. The mechanism underlying the reduction of CORT by osilodrostat confirmed that the amount of skin collagen could be preserved with treatment. In addition, upon suppression of the CORT receptor, the amount of collagen was controlled, and skin aging was suppressed by the hypothalamic–pituitary–adrenal axis. Therefore, this study confirmed an inverse relationship between adrenal CYP11B1/2 levels and collagen during the initial stages of UV irradiation of the skin. The findings of this study may be useful for developing new detection mechanisms for aging, following their further verification.
The toxic reactions and side effects of glucocorticoids in humans are discussed.
The effects reviewed include metabolic (muscle and skin, carbohydrates and lipids), cardiovascular, cellular, immunologic, skeletal and growth, gastrointestinal, nervous system, ocular, and hypothalmic-pituitary-adrenal.
This article covers the normal anatomy and histology of the gastrointestinal tract (GIT), from the esophagus to the rectum, as well as pathologic responses to toxicants in the GIT. Because the GIT is a primary route of exposure to toxins and medications, the resulting lesions are important to understand and study. Many toxicopathologic lesions in the GIT, however, are nonspecific and follow a continuum, from necrosis and ulceration/inflammation to healing, fibrosis, or metaplasia/dysplasia. Nevertheless, these lesions are important in that they may lead either directly or indirectly to the death of the patient.
Millions of people suffer from tendon injuries in both occupational and athletic settings. However, the restoration of normal structure and function to injured tendons still remains as one of the greatest challenges in orthopaedics and sports medicine. In recent years, a remarkable advancement in tendon research field has been the discovery of tendon stem/ progenitor cells (TSCs). Unlike tenocytes, the predominant resident cell in tendons, TSCs have the ability to self-renew and multi-differentiate. Because of these distinct properties, TSCs may play a critical role in tendon physiology as well as pathology such as tendinopathy, which is a prevalent chronic tendon injury. Additionally, because TSCs are tendonspecific stem cells, they could potentially be used in tendon tissue engineering in vitro, and serve as a promising cell source for cell-based therapy to effectively repair or even regenerate injured tendons in clinical settings.
Glucocorticoids are generally used to relieve pain and/or inflammation in a wide variety of musculoskeletal disorders including osteoarthritis, inflammatory arthritis, tendinopathy and degenerative spine disease. Glucocorticoids reduce tendon derived cell proliferation in vitro and reduce extracellular matrix synthesis both in vitro and in vivo, in particular type I collagen synthesis. Glucocorticoids also appear to result in acute deleterious changes in healthy in vivo tendon including collagen necrosis, collagen disorganisation and inflammatory cell infiltration; while the overall effect of glucocorticoid administration on the mechanical properties of healthy in vivo tendon are generally negative. Overall the existing in vitro and in vivo evidence suggests that glucocorticoids should be used with caution in treating painful tendinopathy. Certainly a real need exists to follow up the long term clinical effects of glucocorticoid in treating tendinopathy, as there is currently a paucity of evidence in this area. However in this context while the short term benefits are clear, glucocorticoids remain a useful treatment option provided they are used in the right patients in sensible moderation.
Unklare Schulterbeschwerden mit schmerzhafter Bewegungseinschränkung des Glenohumeralgelenkes werden häufig unter dem Sammelbegriff Periarthropathia humeroscapularis (PHS) zusammengefasst. Dieser Begriff ist unpräzise, entspricht keinem eigenständigen Krankheitsbild und sollte daher möglichst vermieden werden.
Active patients of all ages often experience shoulder pain, and a physical exam and radiographs may help detect an injury or a degenerative cause. Though the problems vary, a trial of nonsurgical management is generally the first line of treatment, which can include nonsteroidal anti-inflammatory drugs, corticosteroid injections, physical therapy, and rest.
Collagen production by human skin fibroblast cultures was studied by incubation with [3H] proline and several glucocorticosteroids known to produce dermal atrophy in vivo. Collagen production was measured as formation of [3H]hydroxyproline or collagenase-digestible3H-polypeptides, and the values were corrected for changes in cell number in the same cultures. The steroids, in a wide concentration range, failed to elicit any consistent alterations in collagen production. Review of the literature dealing with corticosteroid-induced changes in collagen production by human skin fibroblasts indicate conflicting results even under apparently similar incubation conditions. Consequently, no unifying hypothesis for steroid-induced dermal atrophy can be developed presently based on the in vitro data.
Striae distensae are linear depressions in the skin that appear perpendicular to areas of increased skin tension. Striae are most commonly the result of changes in skin tension secondary to rapid fluctuations in weight and increases in muscle mass or height. They also appear frequently on the abdomen during pregnancy and in this context are referred to as striae gravidarum. Striae may also be the result of endogenous hypercortisolism like that seen in Cushing's syndrome or disease as well as exogenous hypercortisolism secondary to prolonged administration of systemic or topical corticosteroids. Regardless of the cause, clinically, striae appear initially as raised linear reddish/purple lesions that gradually progress to pale, wrinkled depressions in the skin. This chapter will address the pathophysiology and risks associated with the development of striae in the context of the above conditions as well as review preventive measures and known treatments. Finally, the chapter will conclude with a discussion on the future directions of the treatment of striae distensae and its subtypes.
Summary Atrophic cutaneous side effects have recently been observed with increasing frequency during therapy with topical corticosteroids. An experimental model permitting histometric evaluation was developed to assess and, if possible, predict those effects. The activity of 5 widely used topical corticoids — Hydrocortisone, Flumethasone pivalate, Fluprednylidene acetate, Fluocinonide, and Fluocinolone acetonide — was compared in the guinea pig by this cutaneous hypotrophy test; in relation to untreated skin the substances tested exhibited characteristic and discernibly different hypotrophic effects on the subepidermal integument. The general relevance of these findings and the extent to which they may be correlated with the side effects observed in patients are discussed in detail.
Using the [17,16a-d]-2′-methyloxazoline derivatives of 3β-hydroxypregn-5-en-20-one (I) as an example, the α-hydroxylation
reaction has been studied with the use of rodosobenzene diacetate in a methanolic solution of NaOH as the α-hydroxylating
reagent. The reaction took place successfully through the stage of the formation and isolation of the corresponding [17,16a-d]-2′-methyloxazoline
derivative of 20, 20-dimethylpregn-15-ene-3β,21-diol (II), the dimethyl acetal protection of which was eliminated by acid
hydrolysis in methanol. The results of physicochemical investigations and biological trials of the [17,16a-d]-2′-methyloxazoline
derivatives of 21-hydroxy-21-acetoxy-20-ketosteroids obtained are given.
It has been observed that the beneficial anti-inflammatory effects of topical steroids in psoriasis are counteracted by increasing instability of the disease, with rebound phenomena associated with the cessation of these drugs. We report the occurrence of multi-layered fragmentation and disorganization of the basal laminae in active, untreated psoriatic lesions, resolving and uninvolved, inadvertently steroid-treated psoriatic skin, as well as in a variety of non-psoriatic dermatoses treated with fluorinated topical steroids for prolonged periods. These changes, which were associated with a moderate to severe loss of dermal collagen, were not found in untreated and treated psoriatic controls, with or without concomitant α1-antitrypsin deficiency, who had not received steroids, suggesting that they were probably the consequence of prolonged fluorinated steroid use. This conclusion is supported by the observation that the largest number of layers (10-15) of fragmented basal laminae was noted in the patients who had received fluorinated steroids for 6 years or more, while those on 4 years or less of fluorinated steroid therapy exhibited only three to seven layers of fragmented basal laminae. In psoriatic lesions, the fragmentation of the basal lamina was associated with the presence of basal keratinocyte herniations (BKH), the frequency of which has been shown to parallel clinical psoriatic activity. The persistence of these electron-microscopic markers of psoriatic activity (i.e. BKH) in psoriatic plaques treated with prolonged fluorinated steroids suggests that loss of integrity of the basement membrane, as indicated by the presence of multi-layered fragmentation of the basal lamina, may account for the instability of the psoriatic lesions treated with prolonged topical fluorinated steroids.
The effects of hydrocortisone and ascorbic acid on collagen and noncollagen protein synthesis, and on growth were examined in fibroblasts derived from normal human dermis. When the medium was supplemented with 0.28 mM ascorbic acid, the apparent rate of collagen production increased 2--3 fold over the culture cycle. Ascorbic acid also caused a small increase in the apparent rate of synthesis of noncollagen protein and an elevation in growth rate and maximum cell density. Growth was not required for the increase in collagen production since addition of ascorbate to confluent cultures induced a similar increase. Hydrocortisone (1.5 μM) blocked the ascorbate-related increase in collagen production during growth and in confluent cultures. The hormone simultaneously increased the apparent rate of noncollagen protein production and maximum cell density, suggesting that the effect on collagen synthesis was specific. Inhibition of collagen production by hydrocortisone was observed only in the presence of ascorbate, while the increase in growth and noncollagen protein production occurred in the presence and absence of the vitamin.
It is currently thought that most of the inhibitory and stimulatory actions of the glucocorticoid hormones are initiated by the binding of the steroid to specific glucocorticoid receptors, which are located in the cytoplasm, and by formation of the receptor-steroid complex. After a process called 'activation', the steroid receptor complex is translocated to the nucleus and binds to the chromatin, causing a modulation of genomic expression probably at the level of the formation of specific mRNAs. The translation products of these mRNAs then mediate the glucocorticoid response. From many aspects of the interactions of glucocorticoids with living cells the following will be discussed: the penetration of glucocorticoids into cells and the binding to the receptor, as well as effects of glucocorticoids on cell proliferation, the synthesis of matrix components and prostaglandins.
Confluent cultures of normal primary human skin fibroblasts were incubated with various glucocorticosteroids which are in current use clinically for the treatment of various skin disorders. For all steroids concentrations were found at which collagen hydroxyproline formation was inhibited, while total protein synthesis was little affected. The concentration effective for inhibition was highest for hydrocortisone and lowest for clobetasol-17-propionate. All other steroids (hydrocortisone-17-butyrate, triamcinolone acetonide and betamethasone-17-valerate) showed medium effectiveness. Fluorination as such was not a factor in the degree of inhibition. The inhibition observed was shown to be independent of concomitant specific effects on cell proliferation or cell turnover. The possible implications of these findings on the therapeutic effects in psoriasis and the frequently occurring atrophic side-effects of these steroids are discussed.Konfluente Zellkulturen normaler primrer menschlicher Hautfibroblasten wurden mit einer Anzahl Glucocorticosteroide inkubiert, die gegenwrtig in der Klinik zur Behandlung zahlreicher Hauterkrankungen verwendet werden. Bei allen diesen Steroiden gab es Konzentrationen, die die Synthese des Hydroxyprolins in Kollagen hemmten, whrend die Synthese des Gesamtproteins kaum beeinflut wurde. Die zur Hemmung erforderliche Konzentration war fr Hydrocortison am hchsten und fr Clobetasol-17-propionat am niedrigsten. Die Hemmung durch die anderen Steroide (Hydrocortison-17-butyrat, Triamcinolon-acetonid und Betamethason-17-valerat) erfolgte im Zwischenbereich. Die Hemmung wurde durch Fluorinierung an sich nicht beeinflut und wurde auch nicht durch spezifische Effekte auf die Zellvermehrung oder den Zell-turnover verursacht. Die mglichen Beziehungen wurden diskutiert zwischen diesen Ergebnissen und den therapeutischen Effekten der hier untersuchten Steroide bei Psoriasis, sowie die fters gefundenen atrophischen Seiteneffekten.
Chronic active liver diseases are characterized by the extensive substitution of liver parenchyma by connective tissue. An increased propagation of extracellular matrix components (collagens, structural glycoproteins, proteoglycans) finally leads to the dreaded complications of decompensated liver cirrhosis as well as to liver cell failure or oesophageal variceal bleeding. In Germany, about 14,000 individuals die annually from liver cirrhosis and its complications 1. The number of patients suffering from fibrotic liver diseases is estimated to exceed one million in this country. Needless to say that the disentanglement of basic pathogenetic mechanisms which lead to an increased deposition of connective tissue constituents is gaining high priority in basic science and practical gastroenterology.
The activity of hepatic collagen proline hydroxylase was examined in biopsy samples as a factor in collagen synthesis in 77 patients with alcoholic liver disease. The urinary excretion of peptide bound hydroxyproline was also measured in most of the patients, as an index of collagen degradation. The highest activities of collagen proline hydroxylase were found in the patients with alcoholic hepatitis. Enzyme activity was markedly increased in patients with non-specific changes on liver biopsy, whereas, patients with fatty infiltration had only mild elevations, and those with inactive cirrhosis had normal enzyme activity. Urinary hydroxyproline was elevated only in patients with alcoholic hepatitis and inactive cirrhosis. Follow-up determinations in 16 patients with alcoholic hepatitis, after 4 to 5 weeks, revealed a decrease in enzyme activity, but no change in urinary hydroxyproline. We conclude that among the types of alcohol-related liver diseases, alcoholic hepatitis is associated with the greatest turnover of hepatic collagen.
The goal of vital pulp therapy is to maintain pulp vitality and function. Fluocinolone acetonide is a potent topical glucocorticoid used in the treatment of skin disorders and oral lesions that could possibly be used to resolve inflammation and stimulate the healing process of inflamed dental pulp. The purpose of this study was to investigate the effects of fluocinolone acetonide (0.1-50 μmol/L) on cytotoxicity, cell proliferation, and fibronectin and type I collagen synthesis in human dental pulp cells (HDPCs).
HDPCs were prepared from freshly extracted human third molars. MTT assay was used to determine toxicity and cell proliferation. Western blot analysis was performed to detect fibronectin and type I collagen synthesis.
Low concentrations of fluocinolone acetonide were not only nontoxic but also significantly stimulated cell proliferation (P < .05). Fluocinolone acetonide significantly stimulated fibronectin and type I collagen synthesis (P < .05).
Low concentrations (0.1-10 μmol/L) of fluocinolone acetonide might have the potential to stimulate healing of inflamed dental pulp.
Experimental investigations on biological cell systems are in case of in vitro work often carried out with the help of cell suspensions, necessarily requiring a buffered, partly ‘inorganic’ medium. Such a procedure may of course involve a somewhat artifical situation, especially when living tissue regulation processes are the final goal of the description to be made. In this connection, one should realize that the dynamical basis of a purely phenomenological description of cell morphogenesis [1] as regulating its corresponding membrane transport does not conform to the same boundary conditions as those of cells incubated in a conditioning solution. However, in cell replication processes the code reading is always paralleled by a metabolic reaction catalyzed by the induced enzyme [2], so that an appreciable simplification can be reached by considering only a set of coupled metabolic reactions as driving forces of the biomembrane transport. Combining this with the idea that neurotransmitters may possess at least partially an inducer function and that their action can be either abolished or mimicked by drugs, the sections 2 and 3 of this paper have been outlined. The analytical method followed in these sections allows to reveal more specifically the action mechanism of some particular drugs, such as propranolol.
Hydrocortisone, betamethasone 17-valerate, clobetasone butyrate and clobetasol propionate were tested in vitro for effects on the proliferation and metabolism of six strains of fibroblasts from normal human skin.
The different cell strains gave similar results: hydrocortisone and betamethasone valerate slightly enhanced growth at all concentrations (0.0001-10 μg/ml), but high concentrations of clobetasone butyrate and clobetasol propionate significantly reduced proliferation.
Secretion of acid mucopolysaccharide was most inhibited by clobetasone butyrate and clobetasol propionate, then by betamethasone valerate, with hydrocortisone having the least effect. Clobetasol propionate and betamethasone valerate at 10 μg/ml both reduced collagen synthesis by about 50%; other protein synthesis was less affected.
Inhibition of dermal activity by cortisol in culture was partially reversed by two naturally occurring androgens, testosterone and dihydrotestosterone. ACTH and the androgen precursor dehydroepiandrosterone sulphate showed not such antagonistic effect. These results suggest that increased production of adrenal androgens during ACTH therapy may account for the relative absnece of 'skin-thinning' and 'steroid-bruising' which are common side-effects of corticosteroid therapy.
The aim of the study was to observe pulpal collagen synthesis in response to trauma and to glucocorticoid medication. The material consisted of 290 rabbit pulps and 76 human premolar pulps. Collagen synthesis was determined by incubating whole pulps in a medium containing [14C]proline, and measuring the formation of [14C]hydroxyproline. The effect of glucocorticoids was studied in vitro using rabbit pulps. Hydrocortisone and dexamethasone inhibited collagen synthesis, whereas prednisolone had no marked effect. Hydrocortisone was found to inhibit the synthesis of [14C]hydroxyproline in neutral salt soluble and insoluble non-dialyzable collagen fractions. [14C]hydroxyproline in the dialyzable fraction was increased, suggesting that hydrocortisone increased collagen degradation. In the human material, premolar pulps were experimentally exposed and then medicated with capping agents. The contralateral teeth were exposed and capped with other capping materials, in some cases they were left as intact controls. The exposure led to an increase in the collagen synthesis as indicated by increased [14C]hydroxyproline formation and elevated protocollagen proline hydroxylase activity in the pulp. This enzyme activity was suppressed in pulps capped with a glucocorticoid paste. In addition, the collagen synthesis rate was lower in pulps treated with another glucocorticoid containing compound, when compared to pulps capped with a calcium hydroxide preparation.
The toxic reactions and side effects of glucocorticoids in humans are discussed.
The effects reviewed include metabolic (muscle and skin, carbohydrates and lipids), cardiovascular, cellular, immunologic, skeletal and growth, gastrointestinal, nervous system, ocular, and hypothalmic-pituitary-adrenal.
The effect of dexamethasone on skin collagen was studied in adult mice from strains with a high frequency (A/J) or low frequency (NIH Swiss Webster) of spontaneous or stress-induced facial malformations. Dexamethasone inhibited synthesis of collagen in both strains of mice. The proportion of collagen in the skin of dexamethasone-treated A/J but not Swiss Webster mice appeared to increase slightly due to a greater loss of noncollagenous proteins in the skin. These results indicate that dexamethasone causes degenerative effects in the skin by altering both the synthesis and degradation of proteins and that A/J mice are more affected by glucocorticoid treatment.
Specific pathogen free guinea pigs were treated with varying dosis of fluocortolone for 15 days. The treated animals showed the same per cent weight gain as the controls. The total hydroxyproline content of the skin and the hydroxyproline content in different collagen fractions was the same in treated and untreated animals. Thus fluocortolone seems to have no specific effect on the synthesis or the breakdown of collagen in the guinea pig skin. Also the physical development of the animals, kept under defined conditions, failed to show a catabolic effect of of fluorcortolone.
Topical corticosteroids produce atrophic changes in skin, including thinning of the epidermis and decrease in dermal ground substance. We observed that 12% ammonium lactate produced an increase in the thickness of epidermis and increased amounts of dermal glycosaminoglycans.
Our purpose was to determine whether 12% ammonium lactate could minimize cutaneous atrophy produced by a potent topical corticosteroid.
Clobetasol propionate, 12% ammonium lactate, and both agents were repetitively applied under occlusive patches as well as in open patches on the forearms of human volunteers for 3 to 4 weeks. Biopsy specimens were analyzed for thickness of the epidermis and dermal glycosaminoglycans by image analysis.
Twelve percent ammonium lactate produced a significant sparing of atrophy in both the epidermis and dermis without any influence on the bioavailability or antiinflammatory properties of the corticosteroid.
Twelve percent ammonium lactate may be useful in mitigating the adverse effects of corticosteroid on skin.
Low-dose corticosteroids (defined as less than or equal to 10 mg/d of prednisone or equivalent) are used increasingly for the management of rheumatoid arthritis. They are frequently substituted for nonsteroidal antiinflammatory drugs (NSAIDs), particularly in patients with gastrointestinal or other intolerance to NSAIDs, or as "bridge therapy" while patients await the benefits of delayed-acting, disease-modifying agents. Despite their clinical acceptance, published data concerning efficacy are meager. Adverse effects to low-dose corticosteroids are not so frequent nor so severe as those that occur with higher doses. Nevertheless, alterations in glucose metabolism, cutaneous atrophy, cataracts, and glaucoma are common. Osteoporosis, steroid-myopathy, a steroid-withdrawal syndrome, and dysfunction of the hypothalamic-pituitary-adrenal axis appear in some patients. Osteonecrosis, gastrointestinal, cardiovascular, infectious, or neurological complications probably do not occur. Fetal wastage, prematurity, or congenital malformations have not been proven with this dosage.
Glucocorticoids selectively decrease procollagen synthesis in animal and human skin fibroblasts. beta-Actin content and beta-actin mRNA are not affected by glucocorticoid treatment of chick skin fibroblasts. The inhibitory effect of glucocorticoids on procollagen synthesis is associated with a decrease in total cellular type I procollagen mRNAs in chick skin fibroblasts. These effects of dexamethasone are receptor mediated as determined by pretreatment with the glucocorticoid antagonists progesterone and RU-486 and with the agonist beta-dihydrocortisol. Dexamethasone has a small but significant inhibitory effect on cell growth of chick skin fibroblasts. The ability of this corticosteroid to decrease the steady-state levels of type I procollagen mRNAs in nuclei, cytoplasm, and polysomes varies. The largest decrease of type I procollagen mRNAs is observed in the nuclear and cytoplasmic subcellular fractions 24 h after dexamethasone treatment. Type I procollagen hnRNAs are also decreased as determined by Northern blot analysis of total nuclear RNA. The synthesis of total cellular type I procollagen mRNAs is reversibly decreased by dexamethasone treatment. In addition the synthesis of total nuclear type I procollagen mRNA sequences is decreased at 2, 4, and 24 h following the addition of radioactive nucleoside and dexamethasone to cell cultures. Although the synthesis of pro alpha 1(I) and pro alpha 2(I) mRNAs is decreased in dexamethasone-treated chick skin fibroblasts, the degradation of the total cellular procollagen mRNAs is not altered while the degradation of total cellular RNA is stabilized. These data indicate that the dexamethasone-mediated decrease of procollagen synthesis in embryonic chick skin fibroblasts results from the regulation of procollagen gene expression.
Subcutaneously implanted cylindrical hollow viscose cellulose sponges were used to study the effect of locally applied epidermal growth factor (EGF) on methylprednisolone-induced inhibition of granulation tissue formation in rats. In in vivo studies the sponges were treated immediately after implantation with a single injection of 2 mg (approximately 1.7 x 10(-3) M) of depot methylprednisolone or with its carrier solution only. Thereafter the implants were injected daily with 5 micrograms of EGF or with its carrier solution 0.1% albumin for 7 days. Methylprednisolone treatment decreased the accumulation of nucleic acids, collagen, and glycosaminoglycans in the developing granulation tissue. After daily injections of EGF the concentrations of these tissue components returned close to the control values. In cultures of rat granulation tissue fibroblasts, 10(-4) M and 10(-3)M methylprednisolone decreased collagen synthesis by 41 and 81% from the control level, respectively. In the presence of methylprednisolone EGF treatment could not increase collagen synthesis of fibroblasts. Methylprednisolone treatment resulted in a dose-dependent reduction in pro alpha 1(I) collagen mRNA levels, which was partially inhibited by low EGF concentrations (1 and 10 ng/ml). In the presence of methylprednisolone all concentrations of EGF increased fibronectin mRNA levels in a dose-dependent manner. It is concluded that EGF treatment can prevent the inhibitory effect of methylprednisolone on wound healing by stimulating fibroblast proliferation but does not stimulate collagen synthesis per cell.
We tested the ability of all-trans-retinoic acid to prevent corticosteroid-induced skin atrophy without lessening the anti-inflammatory effect of the steroids. Histologic study and skin-fold thickness in hairless mice treated topically with various steroids, followed by topical all-trans-retinoic acid, were used to measure prevention of atrophy. By both assessments, all-trans-retinoic acid prevented atrophy. Noninterference with the anti-inflammatory property of steroids was tested in a phorbol ester-induced mouse ear edema model and by histologic assessment of croton oil-induced inflammation of mouse dermis. We found that all-trans-retinoic acid did not interfere with steroid suppression of either edema or dermal inflammation. Thus all-trans-retinoic acid was effective in preventing steroid-induced atrophy without affecting the steroid's anti-inflammatory property.
Dexamethasone disodium phosphate was found to inhibit in vitro fibrillogenesis in a buffered collagen solution that otherwise formed in vivo like fibrils. A nonlinear relationship was observed between the steroid salt concentration and the kinetic parameter half transition time. Full fibril inhibition occurred at dexamethasone phosphate concentrations above 15 mM. At lower concentrations, sample buffers that also contained inorganic phosphate were not different from the control in activation energy of 224.3 +/- 29.3 kJ/mol (53.6 +/- 7.0 kcal/mol). The idea is advanced that the soluble steroid salt associates directly to the collagen and prevents the formation of lateral, hydrophobic interactions between adjacent collagen aggregates.
Several glucocorticoids as a cream formulation were applied to the recto-anus of the croton-oil-induced hemorrhoid rat. Among the steroids tested, i.e. diflucortolone valerate (DFV), prednisolone (PS), hydrocortisone caproate (HC), and hydrocortisone (H), DFV was found to suppress inflammation most effectively. The effect of DFV was not affected by combination with lidocaine. In this model, the analgesic effect of lidocaine was apparently prolonged by an increase of the threshold for pain by the anti-inflammatory effect of DFV. This additive effect is regarded as a merit of the combination in Neriproct. Therapeutic effects of Neriproct and several anti-hemorrhoid drugs were also examined by using a hemorrhoid model with abrasive irritation compared to those obtained by the croton-oil model. In both models, efficacy of Neriproct was superior to that of the other drugs such as Scheriproct, Proctosedyl, Posterisan forte, Borraginol N, Posterisan and Borraza G. Microscopic observation showed that destruction of the mucus epithelium, necrosis of the mucus layer, infiltration of inflammatory cells and vasodilatation in the croton-oil model were also suppressed markedly by Neriproct application. No difference was observed in the efficacy between the cream and suppository formulation of Neriproct. Suppression of wound healing was found with a dosage of DFV lower than those of PS, HC and H. However, the efficacy ratio of the wound-healing suppression and anti-inflammation of DFV was the largest among the steroids tested.
Osteoporosis is a common complication of corticosteroid therapy and it is associated with both decreased bone formation and increased bone resorption. We have measured radiocalcium absorption and the fasting urinary calcium/creatinine and hydroxyproline/creatinine ratios in 30 postmenopausal women receiving prednisolone therapy and compared the patients with normal spine radiographs (N = 14) with those whose spine radiographs showed osteoporosis (N = 16). The osteoporotic cases had lower radiocalcium absorption (p less than 0.001), higher fasting urinary calcium (p less than 0.05), and higher fasting urinary hydroxyproline excretion (p less than 0.001). As calcium absorption has a positive effect on calcium balance and urinary calcium a negative effect, the difference between these two variables was calculated in each case. This derived variable (radiocalcium absorption--fasting urinary calcium/creatinine) disclosed a greater difference between the osteoporotic and normal groups (p less than 0.0001) than either variable alone.
Topical corticosteroids are utilized in the treatment of a wide variety of skin diseases, primarily those involving an inflammatory component. Recent investigations have revealed that one of the effects of long-term usage of steroids is the depletion of skin mast cells. This led to the treatment of patients with urticaria pigmentosa with topical high potency corticosteroids for 6 weeks. At the end of treatment there was a marked reduction in tissue histamine and an absence of mast cells as well as a disappearance of pruritus and Darier's sign. Treated areas remained clinically improved for at least 9-12 months. Observations that corticosteroids profoundly affect mast cells in vivo provides a rationale to devise new treatment regimens for mast-cell-related diseases.
The studies presented here establish that steroids have an effect on at least two of the cellular elements of the dermis: an inhibitory effect on the synthetic ability of fibroblasts which results in the depletion of ground substance and consequent reorganization of the fibrous elements, and a toxic effect on mast cells as evidenced by mast cell disappearance. This latter effect provides a reversible in vivo system to study correlations such as those described, as well as to further study the biology of the mast cell. Furthermore, aggressive treatment with topical steroids of mast cell-dependent disorders may have a role in appropriately severe cutaneous disease states.
The effect of topical betamethasone-17-valerate on collagen biosynthesis in skin was studied in patients with psoriasis. The skin specimens, taken from both lesion and normal-looking skin, were incubated in a medium containing 14C-proline and the rate of 14C-hydroxyproline formation was taken as an index of collagen biosynthesis. 10-day treatment decreased the rate of collagen formation in the lesion, whereas in normal-looking skin, no effect was detectable. The findings may have relevance to the local side-effects, such as atrophy of the skin, produced by fluorinated corticosteroids.Copyright © 1971 S. Karger AG, Basel
Five male patients with atrophic striae in the groins are reported. All patients received Mycolog cream* topically. None received corticosteroids orally or parenterally. No corticotropin was given. Current theories of the mechanism of the production of striae are discussed. We suggest that the initial change is nonunion of groups of connective tissue fibrils. This is most clearly demonstrated for elastic tissue.
The possibility is raised that local inflammation and topically applied corticosteroids may influence the production of striae. The serious consequences of striae are mentioned. Caution is advised in the use of corticosteroids in the treatment of intertrigo.
A methyl Cellosolve-toluene phosphor solvent system is described for the differential determination of absolute levels of tritium and C14, as well as for the determination of H3/C14 dpm ratios in water-soluble biological samples by liquid scintillation counting techniques. A plot of H3/C14 cpm vs. H3/C14 dpm determined using the above system is linear over wider range than are two other counting systems developed for the counting of water-soluble samples. H3/C14 cpm ratios counted with the methyl Cellosolve-toluene system are relatively stable in the presence of HCl and NaCl contamination, and thus this solvent system can be used in the initial determinations of H3/C14 ratios of evaporated HCl hydrolyzates and other partially purified biological materials. Activities in dilute solutions of C14- and tritium-labeled proteins can also be differentially determined using this solvent system.
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(Second of Three Parts) Intracellular Biosynthesis of Collagen Work from several laboratories has demonstrated that the biosynthesis of collagen occurs by a series of sequential steps: assembly on ribosomal complexes of a polypeptide precursor of collagen that we have called "protocollagen"; hydroxylation of appropriate Pro and Lys residues in protocollagen to Hypro and Hylys; and substitution of some of the Hylys residues with Gal or glucosylgalactose in O-glycosidic linkage before the molecule is extruded (Fig. 2). Our knowledge of the biosynthesis of collagen began with the experiments of Stetten and Schoenheimer, who in the 1940's prepared some of the first . . .
: Previously reported changes in dermal chemistry associated with cortisol administration are explained on the basis of: (1) a hormonally potentiated release of a cellular protease into the extracellular compartment of the connective tissue; (2) the limited proteolytic activation of collagenase zymogen by this protease; and (3) the digestion of insoluble collagen by the activated dermal collagenase. The dermal chemical responses of rats to cortisol were inhibited by 'Dilantin' and by age, as was the ability of the steroid to effect the release of cellular protease into the ground substance.
The effect of hydrocortisone acetate, fluocinolone acetonide, fluclorolone acetonide, betamethasone-17-valerate and fluprednyliden-21-acetate on collagen biosynthesis was studied both in vivo and in vitro. In experiments in vivo, test substances and [14C]proline were injected on the chorioallantoic membrane of 11-day-old chick embryos. In experiments in vivo, chick embryo tibiae were incubated in a medium containing the corticosteroids to be tested and [14C]proline. In both types of experiments, the formation of [14C]hydroxyproline was taken as a measure of the rate of collagen biosynthesis. In addition, the effect of these corticosteroids on the activity of protocollagen proline hydroxylase was tested.[14C]hydroxyproline formation in vivo was inhibited by all the corticosteroids tested, and no clear difference in the degree of inhibition could be observed between the various corticosteroids. in vitro, betamethasone-17-valerate inhibited hydroxyproline formation more than the other substances tested. The concentration of corticosteroids required to inhibit collagen formation was considerably higher in vivo than in vitro. The corticosteroids tested did not affect the activity of protocollagen proline hydroxylase in the bones. In all these experiments, total protein synthesis, measured as incorporation of total 14C-radioactivity into the bones, was inhibited by corticosteroids to the same extent as hydroxyproline formation. The results seem to suggest that corticosteroids inhibit collagen biosynthesis by inhibiting the formation of polypeptide precursors of collagen.
Biosynthesis of collagen was studied by incubating slices of human skin in a medium containing [14C]proline. The presence of hydroxy[14C]proline in nondialysable protein of the skin was taken as a measure of collagen formation. The synthesis of radioactive hydroxyproline was studied under various experimental conditions, and an incubation medium (20 mM N-2-hydroxyethylpiperazine-N′-2-ethanesulphonic acid buffer (pH 7.4), so mM glucose, 0.05 mg/ml ampicillin, I μC [14C]proline, all in phosphate-free Krebs-Ringer solution) for an optimal rate of collagen synthesis is presented. The separate components of the incubation medium had no inhibitory effect on the hydroxylation of [14C]proline when partially purified preparations of protocollagen proline hydroxylase were incubated with radioactive protocollagen substrate.When skin biopsy specimens from living human subjects of various ages were used for incubation, younger subjects demonstrated a relatively high rate of collagen synthesis and a decrease with advancing age was observed.
The action of cortisone on the metabolism of collagen was studied by injecting 14C-proline into rats and determining the effect of cortisone on the specific activity and total activity of 14C-hydroxyproline in the urine and in the skin collagen fractions. In the first series of experiments the administration of cortisone was begun 10 days before the administration of 14C-proline. Both the amount of hydroxyproline excreted and the specific activity of urine 14C-hydroxyproline Were lower in the cortisonetreated rats than in the controls. The total activity of urine 14C-hydroxyproline was therefore still more reduced, and during scme of the urine collection periods it decreased to almost half the value of the controls. The mean values for the specific activity and total activity of 14C-hydroxyproline in the soluble collagen fraction of the skin were likewise lower in the cortisone-treated rats than in the controls. In the second series of experiments the 14C-proline was injected 30 days before the start of cortisone administration. A small decrease occurred in the amount of hydroxyproline excreted in the urine in the cortisone-treated rats compared with the controls, but the total activity of urine 14C-hydroxyproline remained unchanged. The total activity of 14C-hydroxyproline in the skin collagen fractions was likewise not changed by the cortisone treatment. The results of the present study suggest that pharmacological doses of cortisone have an antianabolic action on the formation of soluble collagen, but no effect on the catabolism of the insoluble collagen fibres.
Previously published procedures for the assay of radioactively labeled hydroxyproline have been modified so that the assay is simpler, more reproducible, and less affected by the presence of other C14- and H3-labeled compounds. Although the present procedure measures total radioactive hydroxyproline rather than the specific radioactivity of hydroxyproline, it is more readily applicable to samples from a variety of sources.
Several modifications of a specific chemical assay for hydroxyproline in urine are presented which make the procedure more rapid without reducing its specificity or accuracy. Also, a new method for measuring the recoveries in individual samples with hydroxyproline-14C is presented. Results obtained with the new procedure in two laboratories over a number of years are reviewed.
Twelve weanling male rats each received 15μc of l-proline-14C in 3 equal and consecutive daily i.p. injections. Fourteen days after the third isotope injection, 4 of the animals were sacrificed (0-day control). Four of the remaining rats each received 9 daily injections of 10 mg cortisone acetate/kg body wt. and then were sacrificed with the other animals (9-day control) 24 hr after the final steroid injection.Salt-soluble skin collagen represented a significantly lower fraction of the total skin collagen in the cortisone-treated rats than in the 9-day control animals. The sp. act. of hydroxyproline from salt-soluble skin collagen, mature skin collagen, and total femur collagen from the cortisone-treated rats were each significantly greater than those of the 9-day control animals.The mean urinary hydroxyproline excretion of the steroid-treated rats was significantly less after 4 days of treatment until termination of the experiment. Slightly greater urinary hydroxyproline specific activities of the cortisone-treated rats reflected the higher sp. act. of the collagen hydroxyproline pool of the steroid-treated rats. No significant differences were observed between the 24-hr urinary hydroxyproline radio-activities of control and cortisone-treated rats.The above observations, in conjunction with additional data given in this report and the findings of earlier experiments, were interpreted as providing further evidence in agreement with anti-anabolic changes in collagen metabolism (decreased collagen synthesis) resulting from the administration of pharmacologie quantities of cortisone to rats which would normally undergo rapid body growth. Although the data from the various experiments indicate that the steroid had an anticatabolic effect on femur collagen (decreased collagen degradation), urinary hydroxyproline excretions and radioactivities were inconclusive for the demonstration of an anticatabolic effect of the hormone on the total body pool of collagen.
The Journal of Investigative Dermatology publishes basic and clinical research in cutaneous biology and skin disease.
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This was prompted by the well known retardation in the development of fibrous tissue after treatment with hormones of the adrenal cortex. It seemed justifiable to investigate whether the synthesis of the polypeptide precursors of the insoluble collagen was affected or whether the defect in the fibre formation was present in other constituents. Some histological description was thought relevant.
EXPERIMENTAL Treatment of the Animals. — Twelve adult guinea pigs (360—410 gm.) were divided into three groups (two males and two females in each) which were kept in separate cages. They were fed ad libitum with our standard diet. Hydrocortisone acetate was injected intramuscularly as a crystalline aqueous suspension. The animals in one group received 10 mg. and in the other 25 mg. daily for 22 days. The third group served as control. The animals remained healthy, except one, which developed a suppurating lymphadenitis in the neck after 18 days and
HYPERACTIVITY of the adrenal cortex induced by treatment of rats with ACTH results in a general suppression of growth. In the skin, the epidermis becomes thinner, hair stops growing and the dermal connective tissue assumes a more compact arrangement (Baker, Ingle, Li and Evans, 1948). These effects appear to be mediated by C-11 oxygenated steroid which are secreted by the adrenal glands. If one of these compounds, cortisone, or an extract of the adrenal cortex is applied directly to the skin, similar changes are induced locally in the epidermis and hair follicles (Baker and Whitaker, 1948). In these early short-term experiments consistent changes in the connective tissue were not observed but, subsequently, it was demonstrated that adrenocortical extract inhibits growth of granulation tissue by local action (Baker and Whitaker, in press).
This report deals with the cutaneous modifications elicited by prolonged local treatment of non-traumatized skin with adrenocortical compounds. The experiments had two general purposes: (1) to observe the changes occurring in the epidermal cells and fibro-elastic connective tissue of the dermis and (2) to find out if a refractory state will develop toward the local action of adrenocortical steroids in rats treated for long periods of time.