Effect of supplemented and topically applied antioxidant substances on human tissue

Systemic and topical application of antioxidant substances for the medical treatment and prophylaxis of many diseases as well as additional protection of the skin against the destructive action of free radicals and other reactive species has become very popular during the past years. Stimulated by the positive results of a fruit and vegetable diet in supporting medical treatment and in cosmetics, artificial and extracted antioxidant substances have been broadly applied. Surprisingly, not only positive but also strong negative results have been obtained by different authors. According to study reports artificial and extracted antioxidant substances support different kinds of medical therapies, if they are applied in mixtures of different compounds at low concentration levels. In the case of the application of high concentration of some single compounds, side effects were often observed. Regarding skin treatment by systemically applied antioxidant substances for cosmetic purposes, positive cosmetic effects as well as no effects, but almost no side effects, apart from a number of allergic reactions, were reported. One reason for this seems to be the lower concentration of systemically applied antioxidant substances in comparison with a medical application. Topical application of antioxidant substances is closely related to cosmetic treatment for skin protection and anti-aging. Positive results were also obtained in this case. The present review is an attempt to classify and summarize the published literature concerning the efficiency of action of systemic and topical applications of antioxidant substances, such as carotenoids and vitamins, on human organism and especially on the skin. The available literature on this topic is very extensive and the results are often contradictory. Nevertheless, there are some clear tendencies concerning systemic and topical application of antioxidant substances in medicine and cosmetics, and we summarize them in the present paper.     

醋酸曲安奈德鼻喷剂长期应用对鼻粘膜影响动物实验

目的观察醋酸曲安奈德鼻喷剂长期应用对鼻粘膜和鼻纤毛有元损害作用.方法杂种狗,高剂量组4只;低剂量组4只;对照组4只,鼻腔给药0.64%醋酸曲安奈德鼻喷剂组每日一次,每次2个鼻孔各喷药1次(880ug);0.14%醋酸曲安奈德鼻喷剂每日一次,每次2个鼻孔各喷药1次(220ug,);0.9%生理盐水组每日一次,每次2个鼻孔各喷100ul.用药前后观察鼻腔粘膜,血常规,用药后,光镜和电镜观察鼻粘膜.结果用药后,鼻粘膜三组间无差异,血常规无差异,光镜电镜未见鼻粘膜病损.结论醋酸曲安奈德鼻喷剂长期应用对鼻粘膜无毒害作用.     

Glutathione transferases in human nasal mucosa

Glutathione transferase (GST) was investigated with 1-chloro-2,4-dinitrobenzene as substrate in tissues speciments of human nasal mucosa. The average ±(SD) of GST activity in the cytosol was 76.8 ±21 nmol/min/mg with a range of 47–113. Using affinity chromatography and isoelectric focusing, the isozymes of GST from human nasal mucosa have been purified and characterized. On the criteria of isoelectric point, substrate specificities, apparent subunit molecular weight, sensitivity to characteristic inibitors and immunological properties the major GST purified (about 85% of total activity) can be identified as class pi GST. Although a limited amount of class alpha GST was expressed by human nasal mucosa, no class mu isoenzymes was noted. In addition, we have also identified a GST subunit that cannot be related to any of three major classes of human GST.     

Eosinophil chemotactic activity of topical PAF on the human nasal mucosa

Summary Ten patients with strictly seasonal allergic rhinitis were challenged with 132 g PAF-acether in each nasal cavity outside their relevant pollen season. Cells from the nasal mucosa were collected by nasal lavage prior to and every second hour up to 8 h after the PAF challenge. At the same times the volume of methacholine-induced secretory responsiveness was measured. A brush specimen was harvested from the nasal mucosa prior to and 8 h after the PAF challenge.PAF led to an increase in the number of eosinophils from an initial 6.1 to 64.4 per glass 2 h later. The number of eosinophils in the nasal lavage fluid then decreased to its initial baseline value. By 8 h after PAF challenge the number of eosinophils collected with the brush was still elevated as compared to the initial brush sample (3.1 vs 24.1). PAF did not produce any change in methacholine-induced secretory responsiveness at any time.It appears that PAF possesses eosinophil chemotactic properties in the human nasal airway without altering the nasal secretory responsiveness. This confirms previous findings that the induction of nasal responsiveness is a more complex phenomenon than just the recruitment of eosinophils into the airway mucosa.     

Ocular anti-inflammatory and systemic immunosuppressive effects of topically applied fluorometholone.

Anti-inflammatory and systemic immunosuppressive effects of topically applied steroidal and nonsteroidal anti-inflammatory agents on experimental immunogenic uveitiis in rabbits were studied. The onset of the immunogenic uveitis induced by a single intravitreous injection of bovine serum albumin (BSA) was significantly correlated with the appearance of anti-BSA antibody in the serum. A significant correlation was also found between the maximum serum level of antibody and the maximum severity of uveitis. Suspensions of fluorometholone, dexamethasone or indomethacin were applied topically twice a day for a month to eyes injected with BSA. Ocular inflammation was suppressed by fluorometholone and dexamethasone, fluorometholone having the greater anti-inflammatory activity. The circulating antibody titer was also suppressed by both these steroids, but conversely fluorometholone had the lower systemic immunosuppressive activity. Indomethacin did not suppress the uveitis or circulating antibody production. The results of these investigations indicate that the anti-inflammatory effect of the steroid fluorometholone surpasses the immunosuppressive effect, unlike usual anti-inflammatory steroids.     

Xenobiotic-metabolizing enzymes in human respiratory nasal mucosa

Study of oxidative and non-oxidative xenobiotic-metabolizing enzymes was undertaken in microsomal and cytosolic fractions of two human livers, 10 individual and several pooled samples of human respiratory nasal mucosa obtained by surgical operation of male and female patients affected by hypertrophy of the inferior turbinates. The purity of nasal microsomes was checked by electron microscopy and marker enzyme assay. The pooled samples of respiratory nasal epithelium contained, relative to liver, a low amount of cytochrome P450 (about 25 pmol/mg protein) and associated biotransformation activities, and a low level of other components of the mixed-function oxidase system such as cytochrome b5, NADH and NADPH-cytochrome c reductase however the NADH-cytochrome b5 reductase activity was comparable to that of liver. The P450-dependent monooxygenase activities such as ethoxycoumarin O-deethylase, ethoxyresorufin O-deethylase and the dimethylnitrosamine N-demethylase were found in nearly all nasal microsomal specimens. The aniline hydroxylase and the aminopyrine or hexamethylphosphoramide N-demethylases were detected only in the pooled nasal samples. With regard to the non-oxidative enzymes, the activities of glutathione S-transferase, DT-diaphorase, epoxide hydrolase, UDP-glucuronyl-transferase, carbonyl reductase, benzaldehyde and propionaldehyde dehydrogenases, were investigated both in the individual and pooled nasal tissues and livers. These activities were similar in nasal and liver tissue, except for UDP-glucuronyltransferase which was not detected in nasal mucosa. The present findings demonstrate that the respiratory section of human nose contains a wide array of oxidative and non-oxidative enzymes, which could play a crucial role in the bioactivation or detoxication in situ of inhaled xenobiotics.     

Permeability of porcine nasal mucosa correlated with human nasal absorption.

The Ussing chamber diffusion system was used as a model to study the apparent permeability across porcine nasal mucosa of eight drugs and molecules with different physicochemical characteristics, namely insulin, lidocaine, nicotine, PEG 4000, propranolol, sumatriptan, melagatran and an amino diether. A weak correlation was found between the apparent permeability coefficients and the corresponding literature data on the fraction absorbed after nasal administration in humans. In the case of passively transported drugs, a closer correlation was found than for the substances where other mechanisms such as carrier-mediated transport or possible efflux were involved. Factors influencing the correlation between in vitro and in vivo data are discussed and the importance of electrophysiological control of the viability status of the excised mucosa is emphasised. Although caution has to be exercised in view of the limitations of the in vitro system, it seems to be a useful tool when evaluating different factors influencing permeability of nasal mucosa.     

Effect of topically applied flurbiprofen on ultraviolet-induced erythema

The effect of flurbiprofen, a nonsteroidal antiinflammatory agent, on ultraviolet B-induced erythema was studied in normal volunteers. The effect of various concentrations as well as the effect of multiple applications were evaluated at 4, 8, and 24 hours after irradiation with three MEDs of ultraviolet B. Repeated applications of flurbiprofen during the four- and eight-hour periods following ultraviolet B exposure did not increase the blanching response obtained following a single treatment. A concentration dependent effect of flurbiprofen on blanching was observed with suppression of erythema increasing with increasing concentration of flurbiprofen up to 3%. Further increase in concentration up to 5% offered no added advantage. Significant differences in blanching were also observed at different postirradiation time periods. No cutaneous or systemic complications were reported during the entire study.     

Risk assessment of topically applied products

The human risk of harmful substances in semisolid topical dosage forms applied topically to normal skin and broken skin, respectively, was assessed. Bisphenol A diglycidyl ether (BADGE) and three derivatives of BADGE previously quantified in aqueous cream and the UV filters 3-BC and 4-MBC were used as model compounds. Tolerable daily intake (TDI) values have been established for BADGE and derivatives. Endocrine disruption was chosen as endpoint for 3-BC and 4-MBC. Skin permeation of the model compounds was investigated in vitro using pig skin membranes. Tape stripping was applied to simulate broken skin associated with various skin disorders. BADGE and derivatives had a tendency to permeate pig skin membranes in vitro with higher fluxes in the tape stripped membranes compared to the non-treated membranes. Data from the in vitro skin permeation study and from the literature were used as input parameters for estimating the risk. The immediate human risk of BADGE and derivatives in topical dosage forms was found to be low. However, local treatment of broken skin may lead to higher exposure of BADGE and derivatives compared to application to normal skin. 3-BC permeated skin at higher flux than 4-MBC. Both UV filters are endocrine disrupting compounds with 3-BC being the more potent. UV filters in sunscreen are often present in high concentrations, which potentially may lead to high systemic exposure dosages. Thus, the risk associated with use of 3-BC and 4-MBC containing sunscreen with regards to endocrine disrupting effects was found to be high and more data is urgently needed in order to fully assess the human risk of 3-BC and 4-MBC in commercial sunscreen.     

Analgesic effect of topically applied pranoprofen-gel

The analgesic effect of topically applied pranoprofen-gel (1% and 3%) was investigated in comparison with indomethacin-gel in experimental animals. Applied topically, 1% and 3% pranoprofen-gel inhibited the inflammatory pain induced by Randall and Selitto's method and the pain response (abnormal gait) of concanavalin A-induced arthritis in rats dose-dependently. Furthermore, in antigen (methylated bovine serum albumin)-induced arthritis in rats, pranoprofen-gel had a concentration and application-dependent therapeutic effect on knee joint swelling and the pain response. Pranoprofen-gel had a stronger analgesic effect than indomethacin-gel in these experimental models. Both drugs inhibited the flexor reflexes of the hind limb induced by injecting bradykinin (BK) in combination with arachidonic acid into the common iliac artery of the spinal rat, but failed to do so with BK combined with prostaglandin E2 (PGE2). Moreover, pranoprofen-gel inhibited the BK-induced increase in the firing rate of the saphenous nerve of the spinal cat. These results show that pranoprofen-gel, applied topically, permeates well from the skin to the nociceptor site, relieving the hyperalgesia caused by PGs-induced sensitization of pain receptors by inhibiting their production. As a topical anti-inflammatory and analgesic agent, pranoprofen-gel is at least as effective as indomethacin-gel, so that it should have good clinical potential.     

The effectiveness of topically applied capsaicin

To undertake a quantitative overview of trials of topical capsaicin for the treatment of diabetic neuropathy, osteoarthritis, post-herpetic neuralgia, and psoriasis.A systematic search of the literature using both computerized and manual methods for identifying clinical trials of capsaicin. The trials identified were abstracted for response data, which then were analysed using established meta-analytic methods for both fixed and random effects modelling.The odds ratio of the response rate of subjects receiving topical capsaicin relative to that of subjects on placebo was used as the main outcome measure. The difference in the response rate was used as the response variable under the random effects model. When dropouts were mentioned and unambiguous assignment could not be made, the analysis was made on the basis of intention to treat.Capsaicin cream give more pain relief to patients with diabetic neuropathy than placebo did. The odds ratio (OR) and corresponding 95% confidence interval (95% CI) in favour of capsaicin cream were OR=2.74 (95% CI=1.73, 4.32). Using a random effect model the rate difference (RD) in favour of capsaicin cream was RD=0.25 (95% CI=0.15, 0.35).Capsaicin cream was also better than placebo in providing pain relief in osteoarthritis: OR=4.36 (95% CI=2.77, 6.88) and RD=0.29 (95% CI=0.20, 0.37) and in psoriasis: OR=2.80 (95% CI=1.69, 4.62) and RD=0.35 (95% CI=0.14, 0.56). There was, however, evidence of heterogeneity in the individual RDs in psoriasis, and complete blinding was difficulty because of the initial discomfort associated with topical capsaicin.In post-herpetic neuralgia the results were even less convincing.Topical capsaicin appears to be effective in the management of a variety of painful clinical conditions affecting the skin. However, totally blind trials are difficult to conduct with this substance. Future trials will need to address this problem more rigorously if a definitive answer about the effectiveness of capsaicin is to be obtained.     

Anti-inflammatory effect of topically applied pranoprofen-gel

The anti-inflammatory activity and mode of action of topically applied pranoprofen-gel were investigated in comparison with indomethacin-gel in experimental animals. Applied topically, 0.3 approximately 3% pranoprofen-gel inhibited carrageenin-induced paw edema, fracture-induced paw edema, carrageenin-induced increase in vascular permeability and adjuvant arthritis in rats and ultraviolet ray-induced erythema in guinea pigs dose-dependently, with a potency slightly greater than that of indomethacin-gel. In addition, pranoprofen-gel inhibited dose-dependently the formation of granuloma caused by a cotton pellet implantation, without affecting the weight of the thymus or adrenals and without inducing gastrointestinal lesions. Moreover, applied topically to carrageenin-treated rats, pranoprofen-gel inhibited dose-dependently, and more potently than indomethacin-gel, the production of a prostaglandin E2 (PGE2)-like substance in both the exudate of the carrageenin air pouch and the inflamed synovial membrane. Both drugs inhibited the potentiation of bradykinin-induced vascular permeability in rabbit skin by arachidonic acid, but not by PGE2. Pranoprofen was only one third as potent as indomethacin in inhibiting the production of PGE2 from the phagocytosing of killed bacteria by rat peritoneal leucocytes in vitro. These results show that pranoprofen-gel, applied topically, permeates well from the skin to the deep inflammatory site, relieving potently and long lastingly the inflammation by inhibiting PG production. As a topical anti-inflammatory agent, pranoprofen-gel is at least as effective as indomethacin-gel, so it may have good clinical use.     

硝酸铈对豚鼠变应性鼻炎鼻黏膜的影响

目的探讨1%硝酸铈溶液对豚鼠变应性鼻炎鼻黏膜的影响。方法健康豚鼠39只随机分成正常对照组,变应性鼻炎组：用2,4-二异氰酸甲苯酯（2,4-Toluene-Dissocya-nate,TDI）作致敏原,经鼻腔滴入建立变应性鼻炎模型。治疗期间变应性鼻炎组（n=30）动物随机分成3组：阳性对照组（n=10）、药物治疗对照组（n=10）、硝酸铈治疗组（n=10）,正常对照组（n=9）设为阴性对照组,治疗15d。观察症状体征、鼻分泌物涂片、鼻黏膜组胺含量和光镜和电镜下鼻黏膜组织形态学的变化。结果阳性对照组出现鼻痒、喷嚏、流清涕,鼻分泌物嗜酸性粒细胞增多,鼻黏膜组胺含量增多,嗜酸性粒细胞浸润,鼻黏膜细胞超微结构改变。与阳性对照组相比,硝酸铈治疗组上述变化明显减轻。结论 1%硝酸铈溶液能有效控制豚鼠变应性鼻炎的症状体征,使鼻黏膜嗜酸性粒细胞减少、组胺含量减低。     

Enhancing the nail permeability of topically applied drugs

The topical therapy of nail diseases, especially of onychomycosis, and to a smaller extent, of nail psoriasis, is desirable to avoid the side effects associated with their systemic therapy, to increase patient compliance and reduce the cost of treatment. Systemic therapy is however the mainstay of treatment due to the poor permeability of the nail plate to topically applied drugs. For effective topical therapy, ungual drug permeation must be enhanced. This can be achieved by disrupting the nail plate using physical techniques or chemical agents. Alternatively, drug permeation into the intact nail plate may be encouraged, for example, by iontophoresis or by formulating the drug within a vehicle which enables high drug partition out of the vehicle and into the nail plate. The physical techniques (manual and electrical nail abrasion, acid etching, ablation by lasers, microporation, application of low-frequency ultrasound and electric currents) and chemicals (thiols, sulphites, hydrogen peroxide, urea, water, enzymes) that have shown ungual enhancer activity are discussed in this review. Optimal drug formulation, while crucial to ungual drug delivery, is only briefly reviewed due to the limited literature.     

Enhancing the nail permeability of topically applied drugs

The topical therapy of nail diseases, especially of onychomycosis, and to a smaller extent, of nail psoriasis, is desirable to avoid the side effects associated with their systemic therapy, to increase patient compliance and reduce the cost of treatment. Systemic therapy is however the mainstay of treatment due to the poor permeability of the nail plate to topically applied drugs. For effective topical therapy, ungual drug permeation must be enhanced. This can be achieved by disrupting the nail plate using physical techniques or chemical agents. Alternatively, drug permeation into the intact nail plate may be encouraged, for example, by iontophoresis or by formulating the drug within a vehicle which enables high drug partition out of the vehicle and into the nail plate. The physical techniques (manual and electrical nail abrasion, acid etching, ablation by lasers, microporation, application of low-frequency ultrasound and electric currents) and chemicals (thiols, sulphites, hydrogen peroxide, urea, water, enzymes) that have shown ungual enhancer activity are discussed in this review. Optimal drug formulation, while crucial to ungual drug delivery, is only briefly reviewed due to the limited literature.     

Vascular responses of the rat to bradykinin

The effects of oestrogens and of sympathetic blockade on the vascular response of rats to bradykinin were studied. No difference in response was found between males and oestrous or dioestrous females. The previous administration of an oestrogen did not affect the response. An augmentation of the depressor response to bradykinin was seen in both sexes after the intravenous administration of either dihydroergotamine or hexamethonium, but not after atropine. Pithing reduced the response to bradykinin. The differences between these results and those previously obtained with oxytocin are discussed.     

Recovery of rat nasal mucosa from the effects of aminopeptidase inhibitors

Aminopeptidase inhibitors may be useful for improving the systemic bioavailability of peptide drugs administered nasally or by other routes. Preferably, their effects would be rapidly reversible. The recovery of peptide hydrolytic activity after exposure of the rat nasal cavity to various aminopeptidase inhibitors was evaluated. Leucine enkephalin (0.1 mM) was used as a model peptide which is predominantly metabolized by aminopeptidases nasally. All experiments involved in situ perfusion of the rat nasal cavity with leucine enkephalin and the inhibitor for 90 min, followed by a washout with saline, and finally a second experimental phase of perfusion with leucine enkephalin but no inhibitor. Boroleucine (0.1 microM) was a potent inhibitor of leucine enkephalin metabolism, and, after its removal, the leucine enkephalin metabolism rate returned to control levels. Much higher concentrations of bestatin (0.1 mM) and puromycin (1 mM) did not inhibit leucine enkephalin metabolism as much as did boroleucine. Furthermore, when these inhibitors were washed out, the rates of leucine enkephalin disappearance rebounded to higher than control levels. With bestatin this could have been partially due to membrane disruption, as evidenced by significantly increased protein concentrations in the perfusates. However, protein release was much lower than that caused by sodium glycocholate, a nasal membrane permeation enhancer. In considering the use of peptidase inhibitors as pharmaceutical adjuvants for peptide drugs, the aminoboronic acid derivatives, including boroleucine, have the advantages of efficacy at very low concentrations and reversibility of effects.     

The effects of water-soluble cyclodextrins on the histological integrity of the rat nasal mucosa

The aim of this study was to investigate the effect of highly water-soluble cyclodextrins (CDs) on the histological integrity of the nasal mucosa. In order to evaluate their effects, the in vivo single and repeated nasal exposure studies were performed using male Wistar rats. The rat nasal cavity was excised after an application of various CD solutions at different times. The morphological appearances of the rat nasal mucosae were analyzed with the light microscopic and the scanning electron microscopic studies. By utilizing 5-min exposure of each CD solution to the nasal mucosa, no tissue damage was visible for 1.5% w/v beta-CD and 5 and 20% w/v hydroxypropyl beta-CD (HP beta-CD), and the effects were quite similar to controls. However, using 20% w/v randomly methylated beta-CD (RM beta-CD) showed severe damage on the integrity of nasal mucosa. The severity was similar to 1% w/v polyoxyethylene-9-lauryl ether or l% w/v sodium deoxycholate. Meanwhile, 30 or 60 min exposure to 10% w/v HP beta-CD or RM beta-CD resulted in no obvious mucosal damage. In addition, in vivo repeated dosing of RM beta-CD did not show any toxicity up to 20% w/v. These results suggest that at least, less than 10% w/v CD solutions do not induce gross tissue damage and can keep the histological integrity of the nasal mucosa.     

Toxicological investigations of the effects carboxymethylcellulose on ciliary beat frequency of human nasal epithelial cells in primary suspension culture and in vivo on rabbit nasal mucosa

The objective of this study was to investigate the safety of a mucoadhesive carboxymethylcellulose (CMC) formulation for intranasal administration of apomorphine. The effect of different concentrations of CMC on ciliary beat frequency (CBF) was studied using a human nasal epithelial suspension cell culture system. The CBF was determined by computerized microscope photometry. The in vivo rabbit nasal mucosal tolerance of the mucoadhesive polymer was investigated using light microscopy. Twice daily, six rabbits received CMC powder in one nostril and CMC/apomorphine powder in the alternate nostril for 4 weeks. Two control rabbits received air puffs in one nostril and nothing in the alternate nostril. The rabbits were subsequently sacrificed and the stained nasal sections examined microscopically. CMC showed both concentration- and time-dependent inhibitory effects on the CBF. Only mild-to-moderate cilio-inhibition was recorded with the different concentrations of the polymer. CMC (both with and without apomorphine) caused mild-to-moderate inflammation after 4 weeks. Necrosis, squamous metaplasia or ciliary degeneration was not observed. Based on: (1) the mild-to-moderate cilio-inhibition induced by different concentrations of CMC; and (2) the mild-to-moderate nasal mucosal inflammation caused by CMC with and without apomorphine, we conclude that this polymer can be considered as a safe carrier for short-term intranasal administration. However, further investigations are required for its use in the treatment of chronic diseases such as with apomorphine in Parkinson's disease.