Efficacy and safety of S-acyl glutathione 2% cream vs. placebo against UVB-induced erythema: a randomized, double-blinded clinical trial.

BACKGROUND: Reactive oxygen species have a major role in the UV-induced short- and long-term damage, thus the exogenous supplementation of antioxidant molecules may allow better skin protection. Despite glutathione has pivotal properties in the complex cytoplasmic antioxidant system, its supplementation is hampered by limited transmembrane absorption. Modification of glutathione pharmacokinetic properties via acetylation with long-chain polyunsaturated fatty acid may improve its passage through phospholipidic membranes. METHODS: This was a single center randomized double-blinded clinical trial, versus placebo, cross-sectional pairwise at time 0. The participants were 10 healthy volunteers with skin phototypes II or III and age ≥18 years interested in performing minimal erythema dose (MED) evaluation performed by photoallergology unit of Division of Dermatology at P. Palagi Hospital, in Florence. Each volunteer underwent UVB phototesting, treating four different areas with increasing UVB doses in four different conditions. One arm was treated as standard procedure (naïve arm), one applying linolenic-glutathione conjugate (Lin-GSH) cream 2% before irradiation (pre-Lin-GSH arm), one with placebo (placebo arm) and one applying Lin-GSH cream 2% (SoloSale Srl, Florence, Italy) after irradiation (post-Lin-GSH arm). The main endpoint was to evaluate efficacy of Lin-GSH cream 2% before UVB irradiation compared with placebo. A secondary endpoint was the evaluation of the same cream applied after irradiation compared to no treatment. Another secondary objective is the evaluation of safety in both conditions. Mean MED was evaluated at distinct conditions. Safety was evaluated reporting all grade 3-4 adverse events up to 30 days after treatment. All volunteers were treated in all four experimental conditions. The pre-Lin-GSH and placebo arms were applied in a double-blind condition for each volunteer. Neither the volunteer nor the investigator executing MED evaluation knew which arm was given Lin-GSH and which one placebo. RESULTS: Ten volunteers among 12 recruited for the study were correctly randomized and completed all study evaluations. Recruitment went from April 1st, 2016 to May 1st, 2016, up to adequate population number. Mean MED in pre-Lin-GSH arm was superior to mean MED in placebo arm (135±5.53 mJ/cm2 vs. 105±7.64 mJ/cm2, P=0.0003). No difference was observed among mean MED in post-Lin-GSH and naïve arms. No grade 3-4 adverse events were reported. CONCLUSIONS: Lin-GSH 2% cream seems a safe and effective in producing a significant increase in MED compared with placebo thanks to its antioxidant properties.

Photodynamic therapy with topical photosensitizers in mucosal and semimucosal areas: Review from a dermatologic perspective.

Photodynamic Therapy is a procedure based on the interaction between a Photosensitizer, a light source with a specific wavelength and oxygen. The aim of this review is to provide a brief and updated analysis of scientific reports on the use of PDT with topical PS in the management of oncological, infectious, and inflammatory disorders involving mucosal and semimucosal areas, with a specific focus on diseases of dermatologic interest.

ALA-PDT exerts beneficial effects on chronic venous ulcers by inducing changes in inflammatory microenvironment, especially through increased TGF-beta release: A pilot clinical and translational study.

A cohort of 19 patients affected by chronic venous ulcers was recruited from our centre. A 4-mm punch biopsy from wound bed was taken before application of ALA 20% gel and repeated one hour after the first PDT irradiation. We observed a significant and progressive reduction of wounds mean volumes right after three ALA-PDT sessions (once per week; 4479.9 +/- 345.5 mm3 vs 34599 +/- 190.3 mm3, p < .01). On immunofluorescence staining from biopsy specimens, we observed a change in all tested stains of post treatment specimens compared to pre-treatment ones. An increase of plasmacytoid dendritic cells (from 699 +/- 22 cells/0.018 mm2 to 1369 +/- 27 cells/0.018 mm2, p < .0001); MHC-II expression (260.39 +/- 99.7 Red, Green, Blue [RGB 0-255] to 370.2 +/- 162.6 RGB (0-255), p < .01), TNF-alpha positive mast cells expression (49 +/- 0.3 cells/0.018 mm2 to 69 +/- 0.4 cells/0.018 mm2, p < .001), TGF-beta expression (59.89 +/- 23.2 RGB (0-255)/cell vs 137.39 +/- 56.6 RGB (0-255)/cell, p < .01) and CD4+/CD25+ Treg cells (39 +/- 1 cells/0.018 mm2 vs 209 +/- 10 cells/0.018 mm2, p < .001) was observed. An increase of TGF-beta was correlated in a statistical significant manner with a reduction of wounds' mean volumes.

Indole 3-acetic acid-photodynamic therapy in the treatment of multiple actinic keratoses: A proof of concept pilot study.

BACKGROUND: Actinic Keratoses (AK) are considered a form of in situ Squamous Cell Carcinoma (SCC) arising on chronically photoexposed skin. PDT with ALA or MAL is an effective treatment for multiple AK due its high Overall Response Ratio (ORR) but is burdened by important drawbacks: time-consumption, pain and high costs. Indole-3 acetic acid (IAA) is a newly described photosensitizer with proven clinical efficacy on seborrheic dermatitis and acne vulgaris. The aim of the study was to assess efficacy, safety and tolerability of a cycle of IAA-PDT at 0.015% in liposomal gel in the treatment of multiple AK of face or scalp. METHODS: We treated 12 patients affected by multiple AK on face/scalp with 4 weekly applications. The product was applied for 15min under occlusion; medicated area was then irradiated for 15min with a LED light at 520nm wavelength for a total fluency of 9J/cm2. RESULTS: No grade 3/4 SAE have been reported; all 12 patients successfully completed the cycle. CR at 3 months follow-up was 25%, with an ORR of 50%. Patients reported almost no pain during irradiations, with mean overall Visual Analogic Scale (VAS) score of 0.3±0.7. IAA-PDT led to a significant improve on Disease Life Quality Index (DLQI) mean score, but no variations on Actinic Keratosis Quality of Life (AKQoL) mean score. CONCLUSIONS: Although larger studies are needed, this is a first "proof of concept" of IAA- PDT as a possible treatment for multiple AK on face/scalp.

Early activation of fibroblasts during PDT treatment in leg ulcers.

BACKGROUND: This pilot study was aimed to assess the variations of some microscopical parameters in skin ulcers, caused by chronic venous insufficiency of the lower extremities (chronic leg ulcers), in 15 patients refractory to previous conventional treatments during photodynamic therapy (PDT). METHODS: Samples of control, wounded and PDT treated skin were taken and analyzed by immunohistochemistry. RESULTS: The cellular infiltrate, as well as the thickness of epidermis, vascularization, mast cell and fibroblast numbers, were increased in chronic wounds as compared to healthy skin. After completion of PDT, fibroblasts appeared further increased in number. Mast cells, closely clustered with fibroblasts, also showed an increase in their numbers, degranulation index and expression of basic fibroblast growth factor. CONCLUSIONS: The present findings support a primary role of fibroblasts in the wound healing process upon PDT treatment, given their early and intense reaction to injury. Mast cells seem to play an accessory yet important role, on the basis of their number and degranulation index variations and expression of basic FGF. In addition, the clustering of mast cells with fibroblasts around blood vessels suggest that these cells may stimulate angiogenesis and, in parallel, fibroblasts to secrete extracellular matrix during PDT therapy.

Photodynamic therapy for chronic venous ulcers.

Some laboratory findings and a few case reports support the treatment of chronic venous ulcers with photodynamic therapy. We treated 19 patients with refractory chronic venous ulcers using photodynamic therapy. The ulcers healed in 15 cases (78.9%) after an average of 6.8 photodynamic therapy sessions (range 6.0-8.0). In the remaining 4 cases, the ulcers showed marked improvement after 10 photodynamic therapy sessions. Photodynamic therapy seems to represent a good alternative therapeutic choice for refractory chronic venous ulcers.

Depigmentation patterns of nonsegmental vitiligo: a prospective study of macromorphologic changes in lesions.

BACKGROUND: Efforts have been recently made to investigate simple, objective, accurate, and reproducible methods of clinical/noninvasive assessment of nonsegmental vitiligo. However, studies have mostly considered quantitative or semiquantitative parameters, almost neglecting the purely qualitative appearance of vitiligo lesions at a given moment and over time. OBJECTIVE: The objective of this study was to investigate the dynamics of macromorphologic alterations taking place within vitiligo patches. STUDY DESIGN: This was a prospective study of a vitiligo cohort. PATIENT POPULATION: Consecutive patients affected by nonsegmental vitiligo. METHODS: Enrolled patients affected by nonsegmental vitiligo underwent a dermatology visit once monthly for 12 months. Vitiligo lesions were photographed at each visit under both room light and Wood's light, and analyzed via a morphometric, digitalized software capable of detecting quantitative changes of white areas. Pictures depicting changing patches were evaluated in order to assess clinical morphology. RESULTS: Ninety patients were included for the final analysis and 360 lesions were evaluated, 102 of which (28.9 %) showed changes of white areas. Subjective evaluation highlighted two distinct depigmentation patterns, which were present either alone or in combination: (1) a sharply defined band of intermediate color between the depigmented center and the surrounding normal skin, which was defined as 'marginal hypopigmentation;' and (2) pinpoint hypopigmented/depigmented macules centered by a follicle, which we named 'perifollicular depigmentation.' On the other hand, only one repigmentation pattern was detected, the already known 'perifollicular repigmentation.' CONCLUSION: The depigmentation process in vitiligo seems to follow only two specific patterns. This preliminary study represents, in our opinion, a valuable background for future research aiming to investigate the dynamics of vitiligo pathogenesis or assess depigmentation/repigmentation patterns for monitoring treatment response.

Treatment of anogenital condylomata acuminata with topical photodynamic therapy: report of 14 cases and review.

Photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is an emerging technique for the treatment of genital human papillomavirus (HPV)-induced benign and premalignant lesions. We report herein a case series of anogenital CA successfully treated with ALA-PDT and review the literature available to date on this topic. Thirteen out of 14 cases of anogenital condylomata acuminata were successfully treated with topical PDT. We conclude that PDT can be considered a highly effective and safe treatment option for anogenital condylomata acuminata. Also, considering the available literature, the best results are likely to be achieved with a 16-20% gel formulation of 5-ALA and a red light dose of 100-150J/cm(2).

Application of optical coherence tomography in non-invasive characterization of skin vascular lesions.

BACKGROUND: Optical coherence tomography (OCT) is a new non-invasive approach for real-time in vivo tissue characterization. A promising use of OCT can be the assessment of the architecture of lesions with some degree of inhomogeneities, such as vascular lesions. Knowledge of the size and depth of the vascular structures can be useful for the diagnosis and for choosing the best treatment. OBJECTIVE: The purpose of this study was to investigate a series of vascular lesions by means of OCT in order to obtain new insights into the non-invasive, pre-operative analysis of these lesions. METHODS: Seven vascular lesions were included in the study. Histopathological diagnosis showed two haemangiomas and one haemolymphangioma; the remaining four cases were classified as haemangiomas on the basis of their clinical appearance. RESULTS: In all lesions, OCT analysis was able to visualize different areas of the lesion from the horny layer to the dermis showing a clear image of the vascular proliferation. Specifically, oval to roundish signal-poor areas sharply demarcated by a surrounding signal-rich layer were observed in good correlation with histopathology. CONCLUSION: The analysis of vascular lesions by OCT provides a new insight into non-invasive diagnosis and can be helpful in the selection of the most appropriate treatment.

Topical photodynamic therapy for primary cutaneous B-cell lymphoma: a pilot study.

Photodynamic therapy (PDT) is a treatment based on the accumulation of a photosensitizer in the target cells and their selective destruction by irradiation with visible light. In the past 10 years, several patients with cutaneous T cell lymphoma have been successfully treated. The use of PDT in cutaneous B cell lymphoma has not been reported to date. We report the successful PDT treatment of 3 patients with early primary cutaneous B cell lymphoma.

Vascular endothelial growth factor receptor-2 and low affinity VEGF binding sites on human glomerular endothelial cells: Biological effects and advanced glycosilation end products modulation.

Vascular Endothelial Growth Factor (VEGF), binding to its receptor in endothelial cells, seems to modulate the increased blood flow in the early phase of diabetic renal disease. The aim of the study was to evaluate, in a diabetic milieu, the expression, biological function and modulation of VEGF binding sites in human glomerular endothelial cells (GENC). We demonstrated the presence of VEGF binding sites with high (VEGFR-2) and low (heparan sulfate proteoglycans, HSPG) affinity. VEGF165 and VEGF121 working through VEGFR-2 stimulated nitric oxide (NO) production at low doses (0.1-1 nM), whereas only VEGF165 at high doses (10-100 nM) increased thymidine incorporation. 1 nM VEGF165 and VEGF121 induced in GENC a significant peak of inducible NO synthase (iNOS) production and, at a lower level, of endothelial NOS (eNOS). The copresence of VEGF165 with aminoguanidine (iNOS inhibitor) determined an increase of eNOS and a significant increase in thymidine incorporation. Advanced glycation end products (AGEs) working through specific receptors (RAGE) up-regulated the expression of VEGFR-2, decreased the expression of HSPG sites and reduced GENC growth. These results identify in GENC VEGFR-2 as a mediator of iNOS and eNOS release under control of VEGF, whereas HSPG binding sites seem to mediate the weak growth effect. The presence of AGEs, up-regulating the VEGFR-2 and decreasing HSPG sites might participate to the block of glomerular angiogenesis addressing the VEGF effects on glomerular permeability.

Possible histopathologic correlates of dermoscopic features in pigmented melanocytic lesions identified by means of optical coherence tomography.

Optical coherence tomography (OCT) is a novel non-invasive imaging technique for in vivo histologic characterization of tissues. Besides pure morphology, additional functional parameters of the tissue investigated can be evaluated at the same time, such as the refractive index and the scattering coefficient. The purpose of our study is to correlate in vivo the dermoscopic structures that first appear in the melanocytic pigmented lesion (pigment network and brown globules) using this new method, with the histopathologic correlates, in order to improve their characterization and achieve easier recognition and inter-observer reproducibility. We concentrated in particular on the pigment network and the brown globules, as these are dermoscopic parameters of great diagnostic importance in melanocytic lesions. Moreover, as these parameters are the histopathologic equivalents of structures located at the level of the dermo-epidermal junction, they enable a correct evaluation to be made using OCT, that at present has only a few millimetres penetration power. The results of our trial, performed using the histopathological preparation as an evaluation gold standard, show that in selected cases OCT allows an in vivo correlation to be made between surface dermoscopic parameters and histopathologic correlates, in particular the pigment network and brown globules. The resolution is not high enough to reveal the morphology of the single cells, but it is possible to evaluate the architecture of a lesion.

Monochromatic excimer light (308 nm) in patch-stage IA mycosis fungoides.

Recently, numerous studies have been reported concerning the treatment of early-stage mycosis fungoides (MF) with narrowband (311-nm) UVB, claiming a beneficial response. We have used for the first time a 308-nm monochromatic excimer light, a new kind of xenon-chloride lamp, in the treatment of patch stage IA MF. We treated 7 patch lesions in 4 patients with unequivocal clinicopathologic diagnosis of MF. All lesions achieved clinical and histologic complete remission. The number of weekly sessions varied from 4 to 11 (mean 6.5; median 5.5). The total UVB 308-nm irradiation dose ranged from 5 to 9.3 J/cm(2) (mean 7.1 J/cm(2); median 7 J/cm(2)). All lesions remained in stable complete remission after a follow-up of 3 to 28 months. No remarkable side effects were reported. Our preliminary results suggest that monochromatic excimer light phototherapy is a possibly very useful treatment modality in patch stage IA MF.

AT1 and AT2 receptors in human glomerular endothelial cells at different passages.

In human adult kidney angiotensin II (AngII) effects are mediated by the AT1 receptor, while the functions of AT2 receptors are mostly unknown. Since AngII regulates endothelial cell growth by AT1 and AT2 receptors, we analysed their functional aspects at different passages in human glomerular endothelial cells (GENC). Semiquantitative reverse transcription-polymerase chain reaction revealed the presence of AT1 and AT2 receptors between 2p and 15p cell passages with different levels of expression. In fact, binding studies of different families of displacement curves using AngII, DUP753 (AT1 antagonist), and PD123177 (AT2 antagonist) showed the presence of AT1a and AT2 receptors at 4p-9p while in GENC 2p only the presence of AT2. In terms of mitogenic activity, AngII was unable to stimulate GENC 2p growth. On the contrary, in GENC 4p-9p and 15p a significant thymidine incorporation was observed. This stimulatory effect seemed to be induced also by the concomitant release of PDGF-BB AT1a mediated. In conclusion, AT1a and AT2 receptors are represented in GENC with a different ratio depending upon the cell passage. AngII regulates the mitogenic effect through AT1a receptors (in later cell passages 4p-15p) involving the release of PDGF-BB, while AT2 (in early cell passage 2p) showed a predominant negative growth control.