Effect of PUVA therapy on melanocytes and keratinocytes in non-segmental vitiligo: histopathological, immuno-histochemical and ultrastructural study.

BACKGROUND AND AIMS: Psoralen ultraviolet A (PUVA) is an important modality in treating vitiligo. Its effect on melanocytes and keratinocytes is not sufficiently studied. In this work, we investigated 30 cases of non-segmental vitiligo regarding the changes of melanocytes and keratinocytes in both vitiliginous and nearby areas before and after PUVA therapy. METHODS: Three skin biopsies were obtained from each patient from the vitiliginous, marginal and perilesional areas before and after 12 months of PUVA. Biopsies were examined histologically using haematoxylin and eosin, Masson-Fontana stains and 3,4-dihydroxyphenylalanine (DOPA) reaction and histochemically using human melanoma black-45 (HMB-45) antibody while ultrastructural examination was performed on six patients. Control biopsies were taken from five healthy volunteers. RESULTS: In 10% of pretreated biopsies from the centre of vitiligo lesions, scanty melanocytes were detected histologically and ultrastructurally, while they did not stain with DOPA or HMB-45 antibody suggesting that these melanocytes were inactive. Moreover, degenerative changes were detected by electron microscopy in both melanocytes and keratinocytes in all areas. After PUVA therapy, obvious improvement of the histopathological changes occurred with significant increase in active melanocytes. The degeneration of melanocytes and keratinocytes was also reduced at the ultrastructural level. CONCLUSION: Vitiligo affects both melanocytes and keratinocytes causing degenerative changes. These changes were present in both the leucodermic and the apparently normal perilesional skin. PUVA increases the number of active epidermal melanocytes in the three tested areas and recovers the melanocyte and keratinocyte degeneration.

Patterns of repigmentation in two cases of hypopigmented type of vitiligo.

It has been observed that depigmentation in vitiligo passes through two stages; patches of light brown hypopigmentation which gradually changes into milky white patches. In this work, we studied two cases of hypopigmented vitiligo regarding the melanocytes and keratinocytes' changes before and after 7 months of psoralen plus ultraviolet A (PUVA) therapy. Skin biopsies were taken from the vitiliginous lesions before and after 7 months of PUVA therapy and were examined using haematoxylin and eosin and Masson Fontana stains, L-3,4-dihydroxyphenylalanine reaction, immuno-histochemical stains and ultrastructural examination. In the pretreated patients, the melanocytes present were inactive and degenerative changes were detected in both melanocytes and keratinocytes. After PUVA therapy, obvious histopathological improvement was detected. Clinically, the initial response to PUVA therapy was increased hypopigmentation indicating that degenerated cells in the vitiliginous patches might have continued the process of degeneration and did not recover. Meanwhile, the perifollicular and marginal pigmentation suggested that pigmentation occurred from those areas and not from activation of already degenerated melanocytes.

Preparation and clinical evaluation of nano-transferosomes for treatment of erectile dysfunction.

OBJECTIVE: The goal of the present study was to formulate topical nanocarriers of the low-cost vasodilator, papaverine hydrochloride (PH), as an alternative to the painful penile injections. The injections are used for both diagnosis and treatment of erectile dysfunction. Transdermal nano-transferosome (T), the ultraflexible nanoliposome, was used as a nanocarrier to enhance the penetration of the papaverine to the penis. METHODS: Different nano formulas were prepared and characterized for their encapsulation efficiency, particle size, zeta potential, and cumulative drug release. The formula acquired the best characteristics was incorporated into 2% (w/v) hydroxypropyl methylcellulose hydrogel base. The gel containing transferosomal papaverine hydrochloride (PH) and that containing free PH were clinically compared using color flow Doppler measurements. RESULTS: The results revealed that transferosome 3 (T3) had the highest entrapment efficiency approaching 72%, low particle size of 220 nm, and zeta potential of -33.4 mV. The formula released 73% of its initial drug content within 2 hours. The clinical evaluation showed the increase in the cavernous artery diameter from 0.53 mm to 0.78 mm and the increase in the peak systolic flow velocity from 5.95 cm/second to 12.2 cm/second, both of which were found to be significant at P<0.05. CONCLUSION: It is evident from the study that the transferosomes can be used as a carrier of papaverine hydrochloride for both diagnosis and treatment of the erectile dysfunction. This new strategy could be used successfully in the treatment of erectile dysfunction and in male impotency.

The use of Botulinum toxin-a injection for facial wrinkles: a histological and immunohistochemical evaluation.

Botulinum toxin (BTX)-A has been used for years in the reduction of facial wrinkles; however, histological and immunohistochemical changes after its use were not previously investigated. To evaluate histological and immunohistochemical changes after BTX-A injection for facial wrinkles, sixteen volunteers, with wrinkles on the upper third of the face, were subjected to single injection of BTX-A. Skin biopsy specimens were obtained from peri-orbital wrinkle site (crow's feet area) before and after 3 months of BTX-A injection. Using histological and immunohistochemical evaluation coupled with computerized morphometric analysis, measurement of epidermal thickness, wrinkle depth and width as well as quantitative evaluation of collagen types I and III and elastin was performed for skin biopsies. After BTX-A injections, there were significant increase in wrinkle width and granular layer thickness (P < 0.001), while the other histometrical measures as well as the immunohistochemical expression of collagen types I and III and elastin showed no significant difference (P > 0.05). However, collagen fibers showed better organization and orientation after BTX-A injection. The histological changes observed after BTX-A injection for facial wrinkles may help in better understanding of its mechanism of action.

Effect of topical tretinoin, chemical peeling and dermabrasion on p53 expression in facial skin.

The tumour suppressor protein p53 is a phosphoprotein that is activated by DNA damage. It is involved in the decision whether the cells should stop replication and proceed to repair their DNA, or to die by apoptosis. In the present study, we evaluate the effect of some treatment modalities on the expression of p53 in facial skin. Biopsy specimens were obtained from the facial skin of 20 patients before and after treatment using topical tretinoin (11 cases), TCA chemical peeling (5 cases) and dermabrasion (4 cases). Biopsy specimens were also obtained from 12 control subjects representing the same age groups of the patients. Topical tretinoin therapy was found to induce a significant decrease in the expression of p53 up to 6 months of therapy followed by a significant increase after 10 months of therapy. On the contrary, superficial TCA peeling did not induce any statistically significant change in the expression of p53. On the other hand dermabrasion was found to induce a significant decrease in the level of expression of p53 in biopsies obtained after complete re-epithelialization followed by a significant increase. These changes in the expression of p53 may play a role in mediating the effects of such treatment modalities on the epidermis, as well as prevention of actinic neoplasia by adjusting any disturbance in the proliferation/apoptosis balance observed in photoaged facial skin.

Expression of IFN-γ, IL-4, and IL-17 in cutaneous schistosomal granuloma.

OBJECTIVES: Cutaneous schistosomal granuloma (CSG) is a rare dermatological disease, the clinical and histopathological features of which are well defined. Although a panoramic picture of its immunopathogenesis in humans is not yet available, it is believed to be induced by T helper 1 (Th1), Th2, or Th17 cytokines in animals. This study evaluated the expression of different types of Th cytokines, including Th1 cytokine interferon-γ (IFN-γ), Th2 cytokine interleukin-4 (IL-4), and Th17 cytokine IL-17, in human CSG. METHODS: This study included nine patients with CSG. Dermatological examinations were conducted in all subjects. Skin biopsy specimens were stained with hematoxylin and eosin (H&E). Immunohistochemical examination was performed using three monoclonal anti-human antibodies against IFN-γ, IL-4, and IL-17 to evaluate Th1, Th2, and Th17 cytokines, respectively. RESULTS: The most common site of CSG manifestation was the paraumbilical area, which was affected in 66.7% of patients. All lesional skin biopsy specimens revealed multiple dermal granulomas surrounding schistosomal eggs. Positive immunoreactivity for IFN-γ, IL-4, and IL-17 was present in dermal inflammatory infiltrate in 88.9, 11.1, and 88.9% of subjects, respectively. There were statistically significant negative correlations between the duration of disease and both IFN-γ and IL-17 (P ≤ 0.05), and a statistically significant positive correlation between IFN-γ and IL-17 (P ≤ 0.05). CONCLUSIONS: This study suggests that CSG is formed by the action of both Th1 (IFN-γ) and Th17 (IL-17) cytokines, which have been shown to be directed against the schistosomal egg to induce a cell-mediated immune response.

Effect of laser resurfacing on p53 expression in photoaged facial skin.

BACKGROUND: p53 overexpression has been reported in photoaged skin. Meanwhile, p53 gene mutations have been implicated as an important factor in the pathogenesis of ultraviolet (UV) light-induced skin cancer. OBJECTIVE: The objective was to evaluate the effect of laser resurfacing on the epidermal thickness and expression of p53 in photoaged skin. METHODS: Specimens were obtained from the facial skin of 10 patients before and after 3 months and 1 year of treatment using CO(2) (five cases) and erbium (Er):YAG (five cases) lasers. Specimens were also obtained from six age-matched controls. These biopsies were used for routine histopathology, histometry, and p53 immunoperoxidase staining. RESULTS: Both CO(2) and Er:YAG lasers were found to induce a significant decrease in p53 expression in biopsies obtained after 3 months (p=.0004 and .002, respectively) followed by gradual increase (p=.01 in both groups). A significant increase (p<.01) in epidermal thickness was also observed after 1 year of resurfacing. This increase, however, is inversely correlated with the level of p53 expression in such patients. CONCLUSION: The decrease in epidermal p53 expression after CO(2) and Er:YAG lasers may account for some of the benefits of resurfacing on the epidermis, as well as prevention of actinic neoplasia by adjusting any disturbance in the proliferation/apoptosis balance observed in photoaged facial skin.

Trichloroacetic acid peeling versus dermabrasion: a histometric, immunohistochemical, and ultrastructural comparison.

BACKGROUND: Trichloroacetic acid (TCA) chemical peel and dermabrasion are beneficial methods for treatment of photoaged skin. OBJECTIVE: In this study, we evaluated the changes induced by these therapies on various structures of facial skin of nine dark-skinned patients (Fitzpatrick types IV-V; TCA, five patients; dermabrasion, four patients) demonstrating different degrees of photodamage. METHODS: Routine histopathology coupled with histometric computer-assisted image analysis was used to assess epidermal changes. Alcian blue stain was used to evaluate changes in glycosaminoglycans. Immunoperoxidase techniques with antibodies against types I and III collagen and elastin were used to evaluate quantitatively changes in collagen and elastic fibers, and their ultrastructure was examined by transmission electron microscopy. RESULTS: Similar histologic, immunohistochemical, as well as ultrastructural changes were observed in the two groups, including epidermal and dermal rejuvenation with new collagen deposition and normalization of the elastic tissue. However, these changes were more prominent in patients treated with dermabrasion than those treated with TCA. CONCLUSION: The results of this study suggest that beneficial effects of such modalities on facial skin were accomplished primarily by increasing the amounts of collagen I and collagen III and improving the morphologic appearance of collagen and elastic fibers.

Effect of topical tretinoin on photoaged facial skin: a histometric, immunohistochemical and ultrastructural study.

BACKGROUND: Topical tretinoin is a recognized treatment for photoageing. AIM: To evaluate the microscopic changes induced by topical tretinoin used to treat mild to moderate photodamage in dark-skinned patients aged 30 to 50 years. PATIENTS AND METHODS: Biopsy specimens were obtained from the facial skin of 11 patients before and after treatment with topical tretinoin. Routine histopathology coupled with histometric computer-assisted image analysis was used to assess epidermal changes. Alcian blue stain was used to measure changes in glycosaminoglycans (GAGs). Immunoperoxidase technique for type I and III collagens and elastin, as well as transmission electron microscopy, were used to measure changes in collagen and elastic fibres. RESULTS: Epidermal hyperplasia occurs following tretinoin application, which is reversible with continued therapy. GAGs decreased (p < 0.05) after 6 months of tretinoin application but with no significant change thereafter. Quantitatively, there was an insignificant decrease of type I (p = 0.7) and III (p = 0.3) collagens during the first 6 months of tretinoin usage. However, biopsies taken after 10 months revealed a statistically significant increase in collagen I from a mean of 75.2% +/- 9.6 before treatment to 94.2% +/- 4.1 after treatment (p = 0.05). Similarly, the amount of type III collagen increased from a mean of 74.6% +/- 9.96 to 90.6% +/- 2.1 after 10 months of treatment (p = 0.05). On the other hand, the amount of elastin significantly (p = 0.02) decreased from a mean of 54.5% +/- 3.68 before treatment to 43.4% +/- 4.42 after 6 months of tretinoin application but with no significant change thereafter. Such changes were associated ultrastructurally with new collagen deposition and improvement of the quality of elastic fibres. CONCLUSION: Topical tretinoin benefits facial skin, mainly by increasing collagen I and III and also by improving the morphological appearance of collagen and elastic fibres.