Mesenchymal stem cells (MSCs) and acellular dermal matrices (ADMs) represent innovative and effective strategies in the setting of wound healing. The aim of our study was to assess the efficacy of a combination therapy based on the use of both MSCs and ADMs in vitro. Adipose-derived MSCs (ADSC) were plated on 8-mm punch-biopsies of a commercially available ADM (Integra®) and maintained in culture for 3 weeks. Then, conventional histology with hematoxylin-eosin staining, environmental scanning electron microscopy and confocal-laser scanning microscopy were used to obtain imaging of ADSC-seeded ADMs. Collagen autofluorescence was used to quantify collagen production by mean fluorescence intensity (MFI), expressed in terms of positive pixels/field, obtained through ImageJ software processing of threedimensional projections from confocal scanning images. ADSCs were efficiently seeded on Integra® and were perfectly incorporated in the pores of the scaffold. ADSCs were also demonstrated to actively proliferate in ADMs. Even more importantly, collagen autofluorescence was efficiently used as a surrogate marker of ECM production. In fact, based on MFI measurements, collagen production turned out to be significantly higher when ADSCs were seeded on ADM rather than in control conditions. In conclusion, our work supports the use of combined therapies based on MSCs and collagenic ADMs. In fact, not only ADSCs can be efficiently seeded on ADMs, but ADMs also seem to enhance their regenerative properties therefore making it a promising therapeutic option for chronic wounds.

Mesenchymal stem cells (MSCs) have recently been shown to have not only regenerative capabilities but also immunomodulating properties. For this reason, they are currently under investigation in clinical trials for the treatment of several autoimmune systemic disorders. Psoriasis is a systemic immune-mediated disease for which MSCs could have therapeutic potential. We analysed the existing literature with regard to MSC-based strategies for the treatment of psoriasis, using the MEDLINE, Embase, Scopus and Cochrane Library electronic databases from inception to the date of study. A number of studies confirm the involvement of MSCs in psoriasis pathogenesis and therefore designate MSCs as an important potential therapeutic tool in this setting. Preclinical data are mostly based on imiquimod-induced murine models of psoriasis, and confirm the anti-inflammatory and immunomodulatory action of MSCs in the setting of psoriasis. Six patients affected by psoriasis were described in four clinical studies. Despite significant differences in terms of therapeutic protocols and clinical outcomes, the MSC-based regimens were efficacious in 100% of the cases. Despite more data still being needed, MSCs could be a promising therapy for psoriasis.

Adipose-derived stem cells (ADSCs) are a particular subset of mesenchymal stem cells present in the stromal-vascular fraction of the adipose tissue. These cells are considered very attractive for their relative abundance and accessibility in the human body. Keratinocytes and fibroblasts cells play important roles in the skin-wound healing process and are the cell types activated by trauma. The aim of our study was focused to evaluate ADSCs ability to build a dermal matrix to be potentially used as a dermal substitute in the field of wound healing. ADSCs were defined as Cluster Differentiation (CD) markers CD90, CD73 and CD105 positive cells and were capable of chondrogenic, osteoblastic and adipogenic differentiation using established protocols. We evaluated several ADSCs abilities in comparison with fibroblasts in order to secrete crucial components of the extracellular matrix such as collagen I and fibronectin. Moreover the culture medium conditioned by ADSCs enhanced the rate of wound closure of fibroblasts and keratinocytes on an in vitro skin wound healing model. Ki67 immunostaining evaluated keratinocyte migrating capacity and colonization on this ADSC-induced matrix demonstrating its ability to promote re-epithelization. Furthermore in the angiogenic 3D assay on Matrigel, ADSCs grown in EGM-2 medium started to form microvessels within 24 hours. This study suggests that ADSCs support vascular network formation in vitro and demonstrates that the ADSCs trophic activity may play an important role in skin wound healing by affecting keratinocytes migration and dermal fibroblasts contribution to generate extracellular matrix.

Sigma receptors are highly expressed in various cancer cells and are involved in the modulation of cellular proliferation and cell death. Several classes of sigma-1 and sigma-2 ligands developed in our laboratory were assayed for the capability to inhibit cell proliferation in two melanoma cell lines SK-MEL-2 and SK-MEL-28. Cells were treated with increasing concentrations (from 20 to 100μM) of the compounds and monitored at 24 and 48 hours. In order to identify the best candidate for potential application to melanoma therapy we have selected a novel sigma-2 receptor agonist (BS148) based on its cytotoxic effect. BS148 was able to significantly induce cell death in SK-MEL-2 cells. Particularly about 40% positive cells were detected by TUNEL assay in SK-MEL-2 cells at 72 hours while almost no cell death occurred in healthy human melanocytes and SK-MEL-28 cells. Immunofluorescence analysis, carried out against human mitochondrial protein and cleaved caspase 3, was aimed to evaluate apoptosis in SK-MEL-2 cells. Our data revealed that most of SK-MEL-2 cells stained positive for cleaved caspase 3 expression after 48 hours of exposure, besides showing altered cell morphology and mitochondria distribution. The activation of apoptotic effector caspase 3 was also confirmed by Western blot analysis. According to our findings, BS148 owns a remarkable selective toxicity for SK-MEL-2 cells and further investigations might provide the basis for representing a promising compound for the treatment of human metastatic melanoma.

Gold nanoparticles (AuNPs) represent an effective choice for topical drug delivery systems thanks to their small size, general non-toxicity, ease of functionalization and high surface to volume ratio. Even if systemic, methotrexate still plays an important role in psoriasis treatment: its topical use shows insufficient percutaneus penetration owing to limited passive diffusion, high molecular weight and dissociation at physiological pH. The aim of our study was to design a new drug delivery nanocarrier for Methotrexate and to improve its solubility, stability and biodistribution. AuNPs were on purpose prepared with a hydrophilic stabilizing layer, in order to improve the colloidal stability in water. Water-soluble gold nanoparticles functionalized by sodium 3-mercapto-1-propansulfonate (Au-3MPS) were prepared and loaded with methotrexate (MTX). The loading efficiency of MTX on Au-3MPS was assessed in the range 70-80%, with a fast release (80% in one hour). The release was studied up to 24h reaching the value of 95%. The Au-3MPS@MTX conjugate was fully characterized by spectroscopic techniques (UV-vis, FTIR) and DLS. Preliminary toxicity tests in the presence of keratinocytes monolayers allowed to assess that the used Au-3MPS are not toxic. The conjugate was then topically used on C57BL/6 mouse normal skin in order to trace the absorption behavior. STEM images clearly revealed the distribution of gold nanoparticles inside the cells. In vitro studies showed that Methotrexate conjugated with Au-3MPS is much more efficient than Methotrexate alone. Moreover, DL50, based on MTT analysis, is 20 folds reduced at 48 h, by the presence of nanoparticles conjugation. UV-vis spectra for in vivo tracing of the conjugate on bare mouse skin after 24h of application, show increased delivery of Methotrexate in the epidermis and dermis using Au-3MPS@MTX conjugate, compared to MTX alone. Moreover we observed absence of the Au-3MPS in the dermis and in the epidermis, suggesting that these layers of the skin do not retain the nanoparticles. Based on our data, we found that the novel Au-3MPS@MTX conjugate is an effective non-toxic carrier for the satisfactory percutaneous absorption of Methotrexate and could help in possible topical treatment of psoriasis.

Efficacy of synthesized bioconjugate of hydrophilic gold nanoparticles (Au 3MPS) loaded MTX was evaluated by topical applicaction on imiquimod-induced psoriasis like mice model by comparing the epidermal thickness and cell proliferation after treatment with nanoparticle alone versus the bioconjugate. Using 50 mg of 5% Imiquimod on shaved back of 8 weeks C57BL/6 mice model for 6 days resulted psoriasis like inflammation. Mice were divided to 4 groups, topically treated with bio-conjugate or nanoparticle alone for 24 or 48 hours and on day 7 or 8 erythema and scaling were scored from 0 to 4. Mice were euthanized and biopsy to evaluate epidermal thickness by H&E-stain and CD3, CD8 &Ki-67 by immunohistochemical stain. The epidermal thickness was significantly reduced in group treated by the conjugate than by nanoparticle alone. It was 0.322μm ±SEM 0.038 vs 0.707μm ±SEM 0.097 and p<0.001. We noticed reduction in the immunopositive Ki-67 cells with significant p<0.001, plus the decrease in CD3 in MTX -Au3MPS treated group compared to the Au 3MPS treated mice after 24h &48 h in epidermis respectively P<0.05, P<0.001 and in dermis 24h &48 h respectively P<0.001, P<0.05. The decrease in CD8 lymphocyte count after 24h &48 h in epidermis was significant P<0.001, P<0.01 and in dermis 24h &48 h respectively P<0.05, P<0.001. After systematic literature review, this is the first study assessing those parameters in psoriasis model under the cutaneous effect of bio-conjugate of MTX and gold nanoparticles. In addition, the data may contribute to clarify the possibility of enhancing the topical delivery of methotrexate by the aid of gold nanoparticle delivery.

Structural Health Monitoring (SHM) is an important requirement to handle military aircraft safety. In contrast with civil aircraft, missions, configurations and environment are changing frequently and therefore also the load spectra, which lead to various life consumptions. Event and fatigue monitoring, remaining life assessment and damage detection monitoring are the basic functions for economic and safe in-service operations of a flying weapon system.

Malignant melanoma is particularly resistant to conventional chemotherapy and radiotherapy. For this reason in the past years a huge variety of new compounds has been developed with potential chemotherapeutic activity which needs to be tested in vitro and in vivo. We investigated the in vitro action of three new experimental antifolate substances (MR7, MR21 and MR36) with a critical target for thymidylate synthase (TS), an essential enzyme for DNA synthesis. The response of two melanoma cell lines (SK-MEL-2 derived from malignant melanoma metastasis and SK-MEL-28 derived from primary malignant melanoma) was examined after treatment with these substances. The antifolate agents induced apoptosis in SK-MEL-2 and SK-MEL-28 cells as confirmed by the TUNEL technique and Comet Assay. Western-blot analysis showed a down-regulation of Bcl-2 protein level and PARP cleavage, otherwise p53 and Bax expressions were not modulated. Moreover, these antifolate-induced apoptosis was accompanied by both pro-caspase-9 and -8 activations. These results were supported by the use of the pan-caspases inhibitor Z-VAD-FMK that almost completely decreased the amount of apoptosis in both the melanoma cell lines treated with antifolate. In conclusion our results show that TS inhibitors are able to induce apoptosis through a caspase-mediated pathway, but without the involvement of the p53/Bax signalling.

BACKGROUND:It is generally accepted that sunlight may contribute to the development of melanoma.OBJECTIVES:To analyse gene expression of melanocytes obtained from clinically unaffected skin of patients with melanoma and healthy controls before and after exposure to ultraviolet B radiation.METHODS:Using GeneChip array technology, the gene expression of melanocytes obtained from the two donor groups was profiled, in order to identify transcriptional differences affecting susceptibility to melanoma.RESULTS:The data collected did not show any difference between the expression profiles of melanocytes purified from normal donors and from patients with melanoma that was able to give a statistically significant class separation. However, by means of unsupervised clustering our data could be divided into two main classes. The first class included the transcriptome profiles of melanocytes obtained from skin samples of patients with a vertical growth phase (VGP) melanoma, while the second class included the transcriptome profiles of melanocytes obtained from skin samples of patients with a radial growth phase (RGP) melanoma.CONCLUSIONS:These data suggest that melanocytes in patients with VGP and RGP melanomas show significant differences in gene expression profiles, which allow us to classify patients with melanoma also from clinically unaffected skin.

Strategies for the suppression of plate vibration are investigated by considering the equivalent impedance of power-minimising vibration controllers. The total power transmitted to a plate by both a primary and secondary point forces is used as a function to be minimised. If the system is linear, then the total power has a known minimum value that is associated with an optimal solution for the secondary force. This minimum power, applied to infinite and finite plates, has been compared to the power reduction that can be achieved with passive vibration treatments. The ratio of the optimal secondary force and the resultant velocity at the secondary force location is termed the equivalent impedance of the active control system, and if only a single primary source is present, this equivalent impedance is entirely reactive but generally unrealisable.

Strategies for the suppression of plate vibration are Investigated by considering approximations to the equivalent impedance of power-minimizing vibration controllers. The total power transmitted to a plate by both a primary and a secondary point force is minimized and the equivalent impedance presented by the secondary source to the plate is considered.A novel device for active vibration control, based on an inertial actuator with displacement sensor and local PID controller and an outer velocity feedback control loop, is used to control the vibrating flexible plate. The impedance presented to the plate by this actuator is compared with the equivalent impedance of the optimal active control system. A frequency-domain formulation is used to analyse the stability and performance of an active vibration suppression system using this modified inertial actuator.The results of an experimental study of active vibration suppression on a flexible plate using the modified inertial actuator are then described. Theory and experiments agree well, demonstrating the effectiveness of the modified inertial actuator.

The design of inertial actuators with local force feedback control and their use in active vibration isolation systems is considered. Unlike reactive actuators, inertial actuators do not need to react off a base structure and can therefore be directly installed on a vibrating structure. In order to guarantee good stability margins in the active isolation controller, however, the actuator resonance must have a low natural frequency and it must be well damped.The behaviour of an inertial actuator with different local force feedback control schemes is first analysed, and it is shown that a phase-lag controller has a good stability margin and can effectively damp the actuator resonance using relatively low gains, compared with a direct force feedback or integrated force feedback controller.A frequency-domain formulation is then used to analyse the stability and performance of an active isolation system using an inertial actuator with local force feedback control and an outer velocity feedback control loop. The plant response, from force actuator input to sensor output, is derived in terms of the mechanical mobilities of the equipment structure being isolated and the vibrating base structure, and the mechanical impedance of the intervening mount.An experimental study of active vibration isolation using an inertial actuator with local feedback control was then carried out. Theory and experiments agree well, demonstrating the effectiveness of the phase-lag controller. However, the need to have an inertial actuator with a low resonance frequency leads to problems with static deflections. (C) 2003 Elsevier Ltd. All rights reserved.

A frequency-domain formulation is used to analyze the stability and performance of an active vibration isolation system which uses feedback control. The active mount is modelled as a single-axis force actuator in parallel with a passive spring and damper. The feedback sensor measures either the absolute velocity of the equipment to be isolated at one end of the mount, or the integral of the transmitted force through the mount. The plant response, from force actuator input to sensor output, is derived for these two cases in terms of the mechanical mobilities of the two structures connected by the active mount.The limits of the phase of the plant response are derived for the two feedback strategies and these are used to explain the stability and performance of several specific examples of active isolation systems. It is shown that, in the absence of actuator and sensor dynamics, the integrated force feedback system is unconditionally stable. The stability of the absolute velocity feedback system is, however, threatened if the vibrating base structure becomes very mobile, with a small effective mass, at the same frequency as the equipment structure becomes very stiff.By quantifying the conditions under which velocity feedback systems can become unstable, these conditions can be avoided. If the stability of an absolute velocity feedback system can be assured, it is shown to be more effective at controlling resonances caused by equipment dynamics than integrated force feedback. (C) 2003 Elsevier Ltd. All rights reserved.

Journal of the European Academy of Dermatology and VenereologyVolume 17, Issue 3 p. 370-370 Another case of dyshidrosiform pemphigoid A Patrizi, Corresponding Author A Patrizi Department of Clinical and Experimental Medicine, Division of Dermatology, University of the Study of Bologna, Bologna Italy,Corresponding author, Via Massarenti 1, 40138 Bologna, Italy, tel. +39 51 6364848/6363475; fax +39 51 347847; E-mail: [email protected]===Search for more papers by this authorL Rizzoli, L Rizzoli Department of Clinical and Experimental Medicine, Division of Dermatology, University of the Study of Bologna, Bologna Italy,Search for more papers by this authorL Benassi, L Benassi Department of Dermatology, University of the Study of Modena and Reggio Emilia, , Italy.Search for more papers by this authorI Neri, I Neri Department of Clinical and Experimental Medicine, Division of Dermatology, University of the Study of Bologna, Bologna Italy,Search for more papers by this author A Patrizi, Corresponding Author A Patrizi Department of Clinical and Experimental Medicine, Division of Dermatology, University of the Study of Bologna, Bologna Italy,Corresponding author, Via Massarenti 1, 40138 Bologna, Italy, tel. +39 51 6364848/6363475; fax +39 51 347847; E-mail: [email protected]===Search for more papers by this authorL Rizzoli, L Rizzoli Department of Clinical and Experimental Medicine, Division of Dermatology, University of the Study of Bologna, Bologna Italy,Search for more papers by this authorL Benassi, L Benassi Department of Dermatology, University of the Study of Modena and Reggio Emilia, , Italy.Search for more papers by this authorI Neri, I Neri Department of Clinical and Experimental Medicine, Division of Dermatology, University of the Study of Bologna, Bologna Italy,Search for more papers by this author First published: 16 April 2003 https://doi.org/10.1046/j.1468-3083.2003.00792_17.xCitations: 10Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL References 1 Levine N, Freilich A, Barland P. Localized pemphigoid simulating dyshidrosiform dermatitis. Arch Dermatol 1979; 115: 320– 321. 2 Gammon WR, Fine JD, Forbes M, Briggman RA. Immunofluorescence on split skin for the detection and differentation of basement zone autoantibodies. J Am Acad Dermatol 1992; 27: 79– 87. 3 Descamps V, Flageul B, Vignon-Pennamen D et al. Dyshidrosiform pemphigoid: report of three cases. J Am Acad Dermatol 1992; 26: 651– 652. 4 Scola F, Telang GH, Swartz C. Dyshidrosiform pemphigoid. J Am Acad Dermatol 1995; 32: 516– 517. 5 Atakan N, Tuzun J, Karaduman A. Dyshidrosiform pemphigoid induced by nickel in the diet. Contact Dermatitis 1993; 29: 159– 160. 6 Beylot-Barry M, Doutre MS, Beylot C. Dyshidrotic pemphigoid. Ann Dermatol Venereol 1995; 122: 81– 83. Citing Literature Volume17, Issue3May 2003Pages 370-370 ReferencesRelatedInformation