Suction Blister Research Articles

Topically Applied Nitric Oxide Induces T-Lymphocyte Infiltration in Human Skin, but Minimal Inflammation

Nitric oxide (NO) plays an important role in the cutaneous response to UV radiation and in cutaneous inflammation. The presence of inducible NO synthase protein in a number of inflammatory dermatoses, coupled with the induction of an intense cutaneous inflammatory infiltrate following topical application of the NO donor-acidified nitrite (NO2(-)), has set the paradigm of NO being an inflammatory mediator in human skin. Using zeolite NO (Ze-NO), a chemically inert, pure NO donor, we have shown that NO per se produces little inflammation. Biologically, relevant doses of Ze-NO induce a dermal CD4-positive T-cell infiltrate and IFN-gamma secretion. In contrast acidified nitrite, releasing equal quantities of NO (measured by dermal microdialysis and cutaneous erythema), induces an intense epidermal infiltrate of macrophages with a similar dermal infiltrate of CD3-, CD4-, CD8-, and CD68-positive cells and neutrophils. Suction blisters were created in Ze-NO-treated and control skin. IFN-gamma, but not IL-4, was detected in Ze-NO-treated skin (mean control 0.1+/-0.07 pg mg(-1) protein, mean IFN-gamma 0.6+/-0.4 pg mg(-1) protein). We suggest that the potent inflammation induced by acidified NO2(-) is secondary to the release of additional mediators.

FLUORESCENCE DETECTION OF AN 8-METHOXYPSORALEN METABOLITE IN HUMAN INTERSTITIAL FLUID

— A fluorescent method has been used to study the suction blister fluid of human volunteers collected after 8-methoxypsoralen (8-MOP) oral intake. A fluorescent chromophore with spectral characteristics (Λmax= 390 nm, Λmax=470nm) distinct from 8-MOP has been detected. Our results suggest the existence of a metabolite form of 8-MOP within the patients's skin prior to any UV irradiation. This form might result in the opening of the4–5′ double bond of the 8-MOP molecule.

Using a microdermabrasion machine as a suction blister device

Using a microdermabrasion machine as a suction blister device

Autologous suction blister grafting for chronic leg ulcers

Non-healing leg ulcers represent a treatment problem. Investigate grafting of autologous suction blister roofs as treatment. Twenty-nine chronic, non-healing leg ulcers of various aetiologies in 18 inpatients were treated by autologous epidermal grafting using the roofs of suction blisters. 55% of ulcers completely healed 2 to 6 weeks after grafting. A 50-90% reduction in size was documented in 34% and no change was observed in 11% of ulcers. Twelve weeks after grafting, 89% of ulcers were healed completely. In most ulcers, we observed a stimulation of reepithelialization from the wound edge ('edge effect') and an accelerated formation of healthy granulation tissue. During a follow-up period of 12 months, 90% of the ulcers remained healed. Grafting of autologous suction blister roofs is an effective treatment option for non-healing leg ulcers. The advantages of the method are its lack of pain, low costs and immediate availability.

Punch Grafting versus Suction Blister Epidermal Grafting in the Treatment of Stable Lip Vitiligo

Among surgical methods, punch skin grafting (PSG) and suction blister epidermal grafting (SBEG) are simple, inexpensive, and effective treatment methods for resistant lip vitiligo cases, but there is a lack of comparison between these procedures for lip vitiligo. The objective was to compare the outcome following PSG and SBEG in stable lip vitiligo. Eighteen patients with stable lip vitiligo were randomly selected. Eight patients were included in PSG group and 10 patients in SBEG. All patients were kept on PUVA-SOL (psoralen+ultraviolet A therapy of solar origin) treatment after grafting. The results were evaluated after a follow-up period of 6 months. At the end of 6 months of grafting in PSG group, two patients had a repigmentation of 50% to 75%, one had 75% to 90%, and three had 90% to 100% in the recipient site in lip. In SBEG group, one patient had repigmentation of 50% to 75%, three had 75% to 90%, and two had 90% to 100%. The color match was statistically significant in PSG when compared to SBEG. In the recipient site, cobblestone appearance was the predominant complication in PSG whereas hyperpigmentation and thickening of grafts were common in SBEG. In the donor site, superficial scarring and hypopigmentation were the common findings in PSG group whereas hyperpigmentation was the main problem in SBEG group. Although both the procedures are effective in lip vitiligo, PSG gives a better color match than SBEG.

Currents: Searching the signaling subproteome for Alzheimer's biomarkers | Pseudo-internal standards for label-free quantitation | Top-down proteomics on a chromatographic timescale | Proteomics of suction blister fluid | Absolute quantitation with PSAQ | SILAC for top-down MS | HUPO BPP pilot-study results

Currents: Searching the signaling subproteome for Alzheimer's biomarkers | Pseudo-internal standards for label-free quantitation | Top-down proteomics on a chromatographic timescale | Proteomics of suction blister fluid | Absolute quantitation with PSAQ | SILAC for top-down MS | HUPO BPP pilot-study results

An action spectrum (290–320 nm) for TNFα protein in human skinin vivosuggests that basal-layer epidermal DNA is the chromophore

Terrestrial solar UVB radiation ( approximately 295-320 nm) readily induces cyclobutane pyrimidine dimers (CPDs) in human skin DNA that result in characteristic mutations associated with nonmelanoma skin cancer. The proinflammatory cytokine TNFalpha is important in mouse skin chemical carcinogenesis and is thought to also play a role in UVR-induced skin cancer by its immunomodulatory properties. There is some in vitro evidence that CPDs initiate the production of TNFalpha, and we tested this hypothesis by comparing the wavelength dependence (action spectrum) for TNFalpha protein induction in human skin in vivo with our earlier in vivo action spectra for CPD induction in four different epidermal layers of human skin. Normal volunteers (n = 35) were irradiated with physiologically relevant doses of monochromatic UVB (290-320 nm), and TNFalpha concentration was assessed, by high-sensitivity ELISA, in exudates from skin suction blisters raised 8 h after irradiation. An action spectrum, constructed from the slopes of the dose-response curves at the different wavelengths, showed maximal efficacy at 300 nm. An excellent match was observed for TNFalpha and the CPD action spectrum for cells in the lower basal epidermis. These data strongly suggest that UVB-induced photodamage to DNA in the epidermal basal layer is a major trigger for TNFalpha production. The TNFalpha may originate directly from the keratinocytes in this layer or inflammatory cells that are rapidly recruited into the upper dermis (e.g., neutrophils) as a consequence of DNA photodamage to basal-layer keratinocytes.

In vivo methods for the assessment of topical drug bioavailability.

This paper reviews some current methods for the in vivo assessment of local cutaneous bioavailability in humans after topical drug application. After an introduction discussing the importance of local drug bioavailability assessment and the limitations of model-based predictions, the focus turns to the relevance of experimental studies. The available techniques are then reviewed in detail, with particular emphasis on the tape stripping and microdialysis methodologies. Other less developed techniques, including the skin biopsy, suction blister, follicle removal and confocal Raman spectroscopy techniques are also described.

Surgical Pearl: Gloved finger as a transport platform for epidermal graft of suction blister

Epidermal grafting using the tops of suction blisters is one of the surgical repigmenting modalities in vitiligo. The procedure consists of raising blisters over the donor area through a suction cup. Once the bulla is formed, the roof is removed with a pair of curved iris scissors. This graft is then transferred to the dermabraded recipient donor vitiliginous area. Various techniques for transporting the epidermal graft have been used. These include the use of fine gauze, 1 Tang W.Y. De Han J. Lu N.Z. Chan L.Y. Lo K.K. Surgical pearl: fine gauze is a useful carrier for epidermal graft in the treatment of vitiligo by means of the suction blister method. J Am Acad Dermatol. 1999; 40: 247-249 Abstract Full Text Full Text PDF PubMed Scopus (9) Google Scholar microscopic slide, 2 Falabella R. Epidermal grafting: an original technique and its application in achromic and granulating areas. Arch Dermatol. 1971; 104: 592-600 Crossref PubMed Scopus (136) Google Scholar spatula, 3 Koga M. Epidermal grafting using the tops of suction blisters in the treatment of vitiligo. Arch Dermatol. 1988; 124: 1656-1658 Crossref PubMed Scopus (149) Google Scholar and acetate sheets. 4 Albert S. Shenoi S.D. Acetate sheets in the transfer of epidermal grafts in vitiligo. J Am Acad Dermatol. 2001; 44: 719-720 Abstract Full Text Full Text PDF PubMed Scopus (6) Google Scholar This step is crucial because of the flimsiness of the epidermal graft and its tendency to curl toward either surface (epidermal or dermal), which may result in graft failure if the wrong graft surface is applied. 1 Tang W.Y. De Han J. Lu N.Z. Chan L.Y. Lo K.K. Surgical pearl: fine gauze is a useful carrier for epidermal graft in the treatment of vitiligo by means of the suction blister method. J Am Acad Dermatol. 1999; 40: 247-249 Abstract Full Text Full Text PDF PubMed Scopus (9) Google Scholar An ideal transport material is one that provides a firm, flexible platform, and allows for easy manipulation especially for small grafts. The transport material or platform needs to be sterile, should avoid adherence of the graft to the material, and should prevent curling of the epidermal graft.

Transdermal Penetration of UV Filters

A penetration study of 2-ethylhexyl-4-methoxycinnamate (EHMC), 4-methyl benzylidenecamphor (MBC), butyl methoxydibenzoylmethane (BMBM), 2-ethylhexyl-2,4,5-trimethoxycinnamate (EHTMC) and di(2-ethylhexyl)-2,4,5-trimethoxybenzalmalonate (TMB) through baby mouse skin (Mus musculus Linn.) was carried out using a vertical Franz diffusion cell. At 4.4 mg/cm² coverage of UV filter on the skin, 2.98 ± 0.38, 1.15 ± 0.14 and 0.80 ± 0.28% of the applied EHMC, MBC and BMBM were detected in the receptor fluid at 24 h after application. Penetrations of UV filter in an ethanolic solution and lotion forms were comparable. EHTMC and TMB showed insignificant penetration across the baby mouse skins. Baby mouse skins kept at 4, –20 and –80°C gave similar EHMC penetration results. Penetrations of EHMC, BMBM, EHTMC and TMB across human epidermis were carried out upon 5 volunteers using the suction blister technique. The results also confirmed the significant penetrations of EHMC and BMBM and the insignificant penetrations of EHTMC and TMB.

Suction blister fluid as potential body fluid for biomarker proteins

Early diagnosis is important for effective disease management. Measurement of biomarkers present at the local level of the skin could be advantageous in facilitating the diagnostic process. The analysis of the proteome of suction blister fluid, representative for the interstitial fluid of the skin, is therefore a desirable first step in the search for potential biomarkers involved in biological pathways of particular diseases. Here, we describe a global analysis of the suction blister fluid proteome as potential body fluid for biomarker proteins. The suction blister fluid proteome was compared with a serum proteome analyzed using identical protocols. By using stringent criteria allowing less than 1% false positive identifications, we were able to detect, using identical experimental conditions and amount of starting material, 401 proteins in suction blister fluid and 240 proteins in serum. As a major result of our analysis we construct a prejudiced list of 34 proteins, relatively highly and uniquely detected in suction blister fluid as compared to serum, with established and putative characteristics as biomarkers. We conclude that suction blister fluid might potentially serve as a good alternative biomarker body fluid for diseases that involve the skin.

Autologous Cultured Melanocytes in Vitiligo Treatment

BACKGROUND Surgical treatment of vitiligo is indicated when lesions are localized in poorly responding areas. OBJECTIVES The objectives were: (1) to establish the melanocyte culture obtained from the epidermis of vitiligo patients for future treatment; (2) to estimate the influence of selected factors on the formation of suction blisters and the results of culture; and (3) to compare the results of treatment of vitiliginous macules localized in the dorsum of the hands and lower limbs by transplantation of cultured autologous melanocytes plus psoralen and ultraviolet A (PUVA) therapy (CMP), suction blister transplantation plus PUVA therapy (SBP), cryotherapy plus PUVA-therapy (CP), and only PUVA therapy (OP). METHODS Forty patients were qualified for the study. The roofs of the suction blisters were used as a melanocyte source for culture establishment or were directly transplanted. RESULTS The CMP procedure was successfully performed on only 10 of 20 patients because of the difficulties in cell culture establishment. The SBP method was carried out on all 20 patients. A total lack of effectiveness was found in CP and OP methods. CONCLUSIONS The effectiveness of culture depends on time of suction blister forming, phototype, and previous PUVA therapy. This study demonstrated the advantage of the SBP over the CMP method.

Epidermal repair results from activation of follicular and epidermal progenitor keratinocytes mediated by a growth factor cascade

Reepithelialization of human suction blister wounds was examined in five normal human volunteers over a period of 14 days postwounding to understand the control of keratinocyte migration, proliferation, and differentiation in acute wound healing in a controlled model. The hypothesis that morphological changes and progenitor activation result from altered cytokines and growth factor expression [in particular interleukin-1 beta (IL-1beta), interleukin-6 (IL-6), transforming growth factor alpha (TGF-alpha), TGF-beta 1, and keratinocyte growth factor] was tested using semiquantitative immunohistochemistry combined with reverse transcriptase-polymerase chain reaction of samples from the blister roof, edge, and base. Parallel changes in keratin expression were examined using a wide range of well-established antibodies to multiple keratins and in situ hybridization for keratin 16 (K16), a marker of the hyperproliferative (mucoregenerative) phenotype. Longitudinal morphological, semiquantitative cytokine and growth factor expression, and histometric histone and cytokeratin profiles suggest three phases to reepithelialization: phase 1, or the acute activation phase, early in the first 24 hours postwounding is characterized by epidermal expression of IL-1beta and IL-6, and dermal expression of TGF-beta1, as basal, upper outer root sheath, and putative interfollicular transit amplifying keratinocytes become committed to mitosis; phase 2, or the early activation phase, late in the second 24 hours postwounding, characterized by epidermal expression of TGF-alpha and IL-6 with concurrent suprabasal K16 expression and migration with continued proliferation, and dermal expression of keratinocyte growth factor and IL-6; and phase 3 or restitution over the following 2 weeks, characterized by the return of normal homeostasis, including bulge activation as evidenced by K19 expression.

Successful treatment of scleroderma-related cutaneous ulcer with suction blister grafting

We presented a case of systemic sclerosis (SSc) in a 53year-old Japanese woman with a refractory toe ulcer, which was successfully treated with suction-blister grafting. A 53year-old Japanese woman with scleroderma presented with the ischemic lesions on the toe tips in November 2001. Over the preceding 20 years, she had suVered from sclerodactylia, arthralgia and Raynaud’s phenomina during cold weather. For more than 5 years, she had repeatedly several episodes of pitting ulcers on the Wnger and toe tips, which were treated with intravenous alprostadil injections and topical antibiotics. In October 2001, a painful ulcer Wrst appeared on her right third toe and worsened rapidly. She was introduced to our hospital. Physical examination revealed diVuse cutaneous sclerosis over the hands, arms, legs, and face. Results of laboratory studies showed the following abnormal values: anti-nuclear antibody 320£ (homogeneous), anti-SS-A antibody 190 IU dLi1, hemoglobin 9.9 g dLi1 and CRP 0.7 mg dLi1. The diagnosis of a diVuse type of systemic sclerosis (SSc) was made on the basis of typical clinical presentations. She was admitted to our hospital and started on intravenous alprostadil therapy once a day. Despite additional conservative treatments including systemic antibiotics and topical prostandin, the ulcer had expanded in size and it had remained extremely painful. Over the following 2 months, the ulcer extended to the entire middle toe with necrotic distal phalanx (Fig. 1a, b). After control of the infection with prolonged courses of intraveneous antibiotics, she underwent surgical therapy. The necrotic bone was removed at the PIP joint, and the ulcer was thoroughly debrided and cleansed under local anesthesia. Subsequently, artiWcial bilayer dermis (Pelnac, Gunze, Osaka, Japan) was applied to the ulcer bed. Twentyeight days later, the ulcer was cleaned accurately (Fig. 1c) and a suction blister was applied as described below. The donor area, the inner thigh, was disinfected with popidone iodine. The proximal end of a 20 ml disposal syringe was placed on the donor skin and suction was applied with the same size syringe connected through a three-way tap (Fig. 1d). It was locked and the negative pressure in the syringe was retained carefully. Vesicles started to form after about 1 h of continuous suction, and they enlarged and coalesced to form a complete blister after 2 h (Fig. 1e). After the roof of the bulla was removed by cutting the periphery of the blister with a small scissors, the epidermal graft was lifted gently with intertulle gauze, which was precut same size as the graft. The graft stuck to the intertulle gauze was placed on the cleaned ulcer (Fig. 1f) and Wxed spokewise with steristrip tape, and thereafter a pressure dressing was applied. The dressing gauze was removed after 7 days. There was complete improvement of the graft after another 7 days (Fig. 1g, h). SSc is a connective tissue disease characterized by cutaneous and visceral Wbrosis, as well as vascular disease involving arterioles, and medium arteries of peripheral circulation [1]. Skin ulcers are clinical problems in SSc that are diYcult to resolve. Although various medicines-like vasodilators, hemorheologic agents, and endothelin receptor antagonist (Bosentan) have been reported to be beneWcial in treating acral ulcers and ischemia, the most eVective therapy remains elusive [2]. Surgical procedures such as microvascular revascularization, digital arterial reconstruction, and peripheral sympathetectomy are used as a last resort, as they are invasive Y. Ichiki (&) · Y. Kitajima Department of Dermatology, School of Medicine, Gifu University, 1-1 Yanagido, Gifu 501-1194, Japan e-mail: yoichiki@gifu-u.ac.jp

Suction blister grafting with CO2 laser resurfacing of the graft recipient site for vitiligo

Suction blister grafting with CO₂ laser resurfacing of the graft recipient site for vitiligo

Autologous Cultured Melanocytes in Vitiligo Treatment

Surgical treatment of vitiligo is indicated when lesions are localized in poorly responding areas. The objectives were: (1) to establish the melanocyte culture obtained from the epidermis of vitiligo patients for future treatment; (2) to estimate the influence of selected factors on the formation of suction blisters and the results of culture; and (3) to compare the results of treatment of vitiliginous macules localized in the dorsum of the hands and lower limbs by transplantation of cultured autologous melanocytes plus psoralen and ultraviolet A (PUVA) therapy (CMP), suction blister transplantation plus PUVA therapy (SBP), cryotherapy plus PUVA-therapy (CP), and only PUVA therapy (OP). Forty patients were qualified for the study. The roofs of the suction blisters were used as a melanocyte source for culture establishment or were directly transplanted. The CMP procedure was successfully performed on only 10 of 20 patients because of the difficulties in cell culture establishment. The SBP method was carried out on all 20 patients. A total lack of effectiveness was found in CP and OP methods. The effectiveness of culture depends on time of suction blister forming, phototype, and previous PUVA therapy. This study demonstrated the advantage of the SBP over the CMP method.

Epithelial grafting for vitiligo requires ultraviolet A phototherapy to increase success rate

Phototherapeutic techniques were introduced into medical practice by the ancient Egyptians. It is considered a cornerstone in the management of resistant vitiligo; yet, failures are very well known. Recently, the introduction of surgical techniques provided a major development in the management of vitiligo and replaced other conventional unsuccessful therapies. The aim of this work is to find out if phototherapy, which failed to resolve the vitiligo problem in patients, is still required in the treatment strategy after epithelial grafting of the same cases. Twenty-five vitiligo patients, nonresponding to classic phototherapy, were treated surgically. Ultrathin Thiersch grafts and suction blister grafts were used. Phototherapy using ultraviolet A (UVA) bulbs in combination with psoralen or khellin was used postoperatively immediately after take of grafts onto recipient sites. In spite of reactivation of depigmentary effects at grafted areas, phototherapy acted as a stimulator for melanocytic proliferation and function and as an immunosuppressant, halting the melanocytic destructive process. The application of UVA phototherapy resulted in successful treatment in the patients receiving it. The success of epithelial grafting in patients with vitiligo can be increased by UVA phototherapy.

Tight Control of Matrix Metalloproteinase-1 Activity in Human Skin¶

Chronic ultraviolet irradiation leads to photoaging in human skin, which is associated with degradation of connective tissue. This is partly due to the fibroblast collagenase (matrix metalloproteinase 1 [MMP-1]). Using complementary DNA array technique we demonstrate that after UV irradiation, MMP-1, MMP-3 and the tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) are time-dependently induced on the messenger RNA level in dermal fibroblasts in vitro and in vivo in human buttock skin. This increase in gene expression is paralleled by an increase of latent and active MMP-1 protein after low-dose UV-A exposure in vitro. In vivo the concentration of latent MMP-1 in suction blister fluids peaks 24 h after irradiation with 2 minimal erythema doses of solar simulated radiation. However, only a small proportion of MMP-1 in vitro (5.5 ± 1.5%) and in vivo is active, whereas the majority of MMP-1 remains in its inactive proform. Interestingly, in suction blister fluid the concentration and duration of TIMP-1 expression exceeds that of MMP-1. Taken together, these data indicate that MMP-1 activity is tightly regulated transcriptionally and posttranscriptionally. Furthermore, the pronounced individual differences in all targets investigated provide a possible explanation for the different susceptibility of individuals to UV exposure and, thus, to the clinical features of photodamage.

Tannic Acid Induces in vitro Acantholysis of Keratinocytes via IL-1α and TNF-α

The mechanism of acantholysis in pemphigus vulgaris (PV) is an intriguing argument since several chemical mediators are implicated. We previously reported a central role for IL-1alpha and TNF- alpha, both able to regulate complement activation and plasminogen activators. Very little is known about what triggers the disease (drugs, viruses or food). In this study, we evaluate the molecular role of tannins in acantholysis. By HPLC chromatography we measured tannic acid (TA) and gallic acid (GA) in blister fluid of 4 groups of patients divided according to their dietary habits, including a regular diet, a diet rich in tannins, a diet free of tannins, and a group of pemphigus patients. Blister fluid was obtained from patients using a suction blister apparatus. We show that people with a diet rich in tannins have increased tannin metabolites (TA and GA) in the skin in respect to controls (tannin-rich diet: GA = 194.52+/-2.39 nmol/ml; TA = 348.28+/-1.4 nmol/ml versus tannin-Mediterranean diet: GA = 15.28+/-1.63 nmol/ml; TA = 22.81+/-1.68 nmol/ml). PV patients showed similar values to the Mediterranean diet population (PV patients: GA = 95.8+/-1.97 nmol/ml; TA = 199.09+/-4.15 nmol/ml versus Mediterranean diet: GA = 83.53+/-2.35 nmol/ml; TA = 195.1+/-2.50 nmol/ml). In an in vitro acantholysis system using TA and PV-IgG we show that TA 0.1 mM in NHEK culture is able to induce acantholysis. This effect was able to amplify the acantholytic action of PV-IgG in vitro. A blocking study using anti IL-1 alpha and anti TNF-alpha antibodies showed a reduction in TA-induced acantholysis. Taken together, these results suggest that a diet rich in tannins could be a trigger in genetically predisposed patients. If these data are confirmed, a complementary diet poor in tannins may be useful in patients affected by PV.

Quantification of Human β-Defensin-2 and -3 in Body Fluids: Application for Studies of Innate Immunity

Human beta-defensins (hBDs) are epithelial cell-derived antimicrobial and immunoregulatory cationic peptides. Our objective was to establish an analytical tool to quantify inducible hBD-2 and -3 in body fluids. We developed sandwich ELISAs using commercially available capture and detection antibodies and determined optimal assay conditions (with 250 mmol/L CaCl(2)) to overcome masking by endogenous components of body fluids. We used recombinant hBD as calibrators and for recovery testing. hBD-2 and -3 detection limits were approximately 75 ng/L and approximately 3 microg/L, respectively. Mean (SD range) values in saliva samples from healthy donors (n = 60) were 9.5 (1.2-21) microg/L for hBD-2 and 326 (50-931) microg/L for hBD-3. We did not detect hBD-3 in suction blister fluid (BF; n = 10) or bronchoalveolar lavage (BAL; n = 5) from healthy participants. We detected low hBD-2 peptide concentrations in BF and BAL, 0.16 (0.03-0.32) and 0.04 (0-0.049) microg/g total protein, respectively. We observed no correlation of hBD-2 in BF and saliva or BAL and saliva from the same person. In vaginal swabs from healthy women (n = 2), mean hBD-2 and -3 concentrations were 3.42 and 103 microg/g total protein, respectively. Cervicovaginal lavage from the same women contained mean concentrations of 1.46 and 55.5 microg/g total protein. These ELISA assays can measure inducible hBD peptide concentrations in body fluids by overcoming masking effects of anionic molecules. This approach may therefore be applicable for quantifying these peptides in health and disease.