Sclerotic Skin Research Articles

AB0229 Senescence associated β-galactosisdase expression in systemic sclerosis skin and fibroblasts

BackgroundSystemic sclerosis (SSc) is connective tissue disease characterized by fibrosis and vascular changes in the skin and internal organs with autoimmune background. However, the mechanism is still unknown. Recently, it...

Inhibition of RelA-Ser536 Phosphorylation by a Competing Peptide Reduces Mouse Liver Fibrosis Without Blocking the Innate Immune Response

Phosphorylation of the RelA subunit at serine 536 (RelA-P-Ser536) is important for hepatic myofibroblast survival and is mechanistically implicated in liver fibrosis. Here, we show that a cell-permeable competing peptide (P6) functions as a specific targeted inhibitor of RelA-P-Ser536 in vivo and exerts an antifibrogenic effect in two progressive liver disease models, but does not impair hepatic inflammation or innate immune responses after lipopolysaccharide challenge. Using kinase assays and western blotting, we confirm that P6 is a substrate for the inhibitory kappa B kinases (IKKs), IKKα and IKKβ, and, in human hepatic myofibroblasts, P6 prevents RelA-P-Ser536, but does not affect IKK activation of IκBα. We demonstrate that RelA-P-Ser536 is a feature of human lung and skin fibroblasts, but not lung epithelial cells, in vitro and is present in sclerotic skin and diseased lungs of patients suffering from idiopathic pulmonary fibrosis. Conclusion: RelA-P-Ser536 may be a core fibrogenic regulator of fibroblast phenotype. (Hepatology 2013)

Discoidin Domain Receptor 2–microRNA 196a–Mediated Negative Feedback against Excess Type I Collagen Expression Is Impaired in Scleroderma Dermal Fibroblasts

Systemic sclerosis (SSc) is characterized by excess collagen deposition in the skin, due to intrinsic transforming growth factor-β (TGF-β) activation. We tried to determine the expression and the role of discoidin domain receptor 2 (DDR2) in SSc. The expression of DDR2 mRNA and protein was significantly decreased in SSc dermal fibroblasts, which was recovered by knocking down TGF-β. The knockdown of DDR2 in normal fibroblasts induced microRNA-196a expression, which led to type I collagen downregulation, indicating that DDR2 itself has a negative effect on microRNA-196a expression and inducible effect on collagen expression. In SSc fibroblasts, however, the DDR2 knockdown did not affect TGF-β signaling and microRNA-196a expression. The microRNA-196a levels were significantly decreased in normal fibroblasts treated with TGF-β and in SSc fibroblasts. Taken together our data indicate that, in SSc fibroblasts, intrinsic TGF-β stimulation induces type I collagen expression, and also downregulates DDR2 expression. This probably acts as a negative feedback mechanism against excess collagen expression, as a decreased DDR2 expression is supposed to stimulate the microRNA-196a expression and further change the collagen expression. However, in SSc fibroblasts the microRNA-196a expression was downregulated by TGF-β signaling. DDR2-microRNA-196a pathway may be a previously unreported negative feedback system, and its impairment may be involved in the pathogenesis of SSc.

Bleomycin-induced Scleroderma in Nude Mice can be Reversed by Injection of Adipose Tissue: Evidence for a Novel Therapeutic Intervention in Systemic Sclerosis

Objective: Systemic sclerosis is an autoimmune disease characterised by uncontrolled fibrosis and vascular insufficiency of the skin and internal organs, without efficient treatments. Subcutaneous adipose transplants have been shown to exert trophic effects on surrounding tissue. We aimed to determine the effects of human adipose tissue in a murine model of sclerotic skin. Methods: Scleroderma was induced in 48 nude mice by daily subcutaneous injection of bleomycin during four weeks. Immediately after the final bleomycin injection, human subcutaneous adipose tissue was implanted into the subcutaneous space of mice using either the Coleman method or the micro-injection method. Epidermal and dermal thicknesses were assessed on skin biopsy specimens six weeks after fat implantation. Capillary density was assessed using immunohistochemistry with an endothelial-specific marker (CD31). Results: Bleomycin increased dermal thickness (p<0.01), collagen network, and density of elastic fibers. Adipose tissue implantation reduced dermal thickness by 10% (p<0.01) with the Coleman method and by 14% (p<0.001) with the micro-injection method. Delivery of adipose tissue by the micro-injection method led to significantly greater neovascularization than the Coleman method (p<0.001). Conclusion: Implantation of human adipose tissue resulted in significant improvement of both fibrosis and peripheral vascular sufficiency in this mouse model of sclerotic skin. Use of a micro-injection method was associated with superior outcome. We postulate that this positive effect is likely due to the activity of the stromal vascular fraction found within adipose tissue. Thus, injection of autologous adipose tissue may be a promising novel therapy for patients with hand or perioral functional impairment.

Interleukin‐13–producing CD8+ T cells mediate dermal fibrosis in patients with systemic sclerosis

Fibrosis is a major contributor to morbidity and mortality in systemic sclerosis (SSc). T cells are the predominant inflammatory infiltrate in affected tissue and are thought to produce cytokines that drive the synthesis of extracellular matrix (ECM) proteins by fibroblasts, resulting in excessive fibrosis. We have previously shown that aberrant interleukin-13 (IL-13) production by peripheral blood effector CD8+ T cells from SSc patients correlates with the extent of skin fibrosis. The present study was undertaken to investigate the role of IL-13 production by CD8+ T cells in dermal fibrosis, an early and specific manifestation of SSc. ECM protein production by normal dermal fibroblasts cocultured with SSc CD8+ T cell supernatants was determined by quantitative polymerase chain reaction and Western blotting. Skin-homing receptor expression and IL-13 production by CD8+ T cells in the peripheral blood of SSc patients were measured by flow cytometry. IL-13+ and CD8+ cells in sclerotic skin were identified by immunohistochemistry. IL-13-producing circulating CD8+ T cells from patients with SSc expressed skin-homing receptors and induced a profibrotic phenotype in normal dermal fibroblasts, which was inhibited by an anti-IL-13 antibody. High numbers of CD8+ T cells and IL-13+ cells were found in the skin lesions of SSc patients, particularly during the early inflammatory phase of the disease. These findings show that IL-13-producing CD8+ T cells are directly involved in modulating dermal fibrosis in SSc. The demonstration that CD8+ T cells homing to the skin early in the course of SSc are associated with accumulation of IL-13 is an important mechanistic contribution to the understanding of the pathogenesis of dermal fibrosis in SSc and may represent a potential target for therapeutic intervention.

Risk Factors to Develop Sclerotic Chronic GvHD After Allogeneic HSCT: Preliminary Results of a Multicentre Retrospective study From the Société Française De Greffe De Moelle Et De Thérapie Cellulaire (SFGM-TC)

AbstractAbstract 1947 IntroductionSclerotic cGvHD is one of the most severe complications after allogeneic hematopoietic stem cell transplantation (HSCT). The sclerotic skin manifestations of cGvHD result from inflammation and fibrosis of the dermis, subcutaneous tissue, or muscular fascia, leading to functional disability. Risk factors associated with sclerotic cGvHD remain not well studied. TBI, PBSC, female recipient, and prior severe skin acute GvHD were identified as factors associated with an increased risk of sclerotic cGvHD in a large previous retrospective study from Seattle (n=986 patients) which was presented at the ASH 2011 meeting. ObjectiveIn the aim to validate the previous risk factors in France, we have conducted this retrospective multicentric analysis. We present here the preliminary results from four centers, seven are expected in the next months. Between 2005 and 2010, 437 consecutive patients who received systemic therapy for chronic GvHD after a first allogeneic HSCT were included. Chronic GvHD was diagnosed by the NIH consensus criteria. Sclerotic GvHD was defined by manifestations of cutaneous sclerosis, fasciitis or joint contracture in the medical record. Analyses to determine risk factors included as variable patient and donor age, kind of donor, HLA matching, diagnosis, stem cell source, patient and donor gender, intensity of conditioning regimen, use of total body irradiation (TBI) or antithymocyte globulin (ATG) in the conditioning regimen, and prior acute GvHD. ResultsAll patients had hematological malignancies, the median age is 45 (17–65), 227 patients (52%) had HLA-matched related donors, 125 (29%) had HLA-matched unrelated donors, 48 (11%) had HLA- 9/10 unrelated donors and 37 (8%) had one or two cord blood unit (HLA-4/6 or 5/6 matched). The median time from allogeneic HSCT to chronic GvHD was 6.5 (1.8–41.5) months; 69 patients (15.7%) presented sclerotic features at initial diagnosis or after the onset of chronic GvHD.The cumulative incidence of sclerotic cGvHD was 12% (95%CI, 11–13.5) at 5 years after onset of chronic GvHD (Figure). From the 69 patients who experienced sclerotic cGvHD, 49 patients (71%) had HLA-matched related donors, 19 patients (27.5%) had HLA-matched unrelated donors, and 1 patient (1.5%) had HLA-mismatched unrelated donor. In univariate analysis, we found more sclerotic cGvHD in patients who had multiple myeloma (p=0.018); this effect seems to be associated with the prophylaxis for GvHD with CsA alone. There was no effect of TBI or intensity of conditioning. There was more sclerotic GvHD in female recipient (p=0.032) and for patients who had an unrelated donor (p=0.0028).In multivariate analysis (Table), factors associated with an increased risk of sclerotic cGvHD were female recipient, patients with unrelated donor, and patients with previous aGvHD. Cord blood as stem cell source was associated with a lower risk of sclerotic GvHD. Prophylaxis of GvHD with CsA alone showed a trend to a higher risk. At the last follow-up, of the 69 patients, 51 (74%) were alive and 10 (14.5%) died from cGvHD. ConclusionIn this intermediate analysis, we found that female recipient, unrelated donor, previous aGvHD are risk factors for developing sclerotic cGvHD. Moreover, we found that cord blood as stem cell source is a protector factor. Study of GvHD prophylaxis, ABO matching, type of HLA-mismatching, acute skin GvHD is ongoing on the whole cohort (n= 1587 patients from 7 centers) and results will be communicated later.After the final analysis on the whole cohort we would like to establish a score risk of sclerotic cGvHD in the goal to conduct a prospective study in France with a prophylactic treatment for high risk patients.TableMultivariate analysisHRpStem cells source and conditioning intensityMAC + Bone marrow1.0RIC + Bone marrow2.03 (0.39–10.6).4MAC + Cord blood0.0.00001RIC + Cord blood0.68 (0.02–18.01).82MAC + PBSC0.32 (0.081–1.28).11RIC + PBSC1.15 (0.5–2.5).73HLA and DonorMatched related1.0Matched unrelated0.04 (0.011–0.181).00001Mismatched unrelated0.62 (0.13–2.7).53Patient ageNSDonor ageNSSexmatchingMale to male1.0Female to female2.7 (1.2–6.06).016Female to male1.8 (0.78–4.33).16Male to female1.44 (0.67–3.13).34Disease diagnosisNSATG in conditioningNSTBI in conditioning1.35 (0.69–2.64).38Prior acute GvHD1.8 (0.9–3.36).064NS= not statistically significant. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Bilateral stenosis of carotid siphon in Hutchinson-Gilford progeria syndrome

Hutchinson-Gilford progeria syndrome (HGPS) is a rare premature aging disease, caused by a de novo mutation of lamin-A gene, LMNA G608G. Accumulation of abnormal lamin-A (progerin) compromises nuclear membrane integrity and results in the accelerated senescence. Affected patients show a typical feature of birdlike face, alopecia, sclerotic skin, loss of subcutaneous fat, and short stature with advancing years. Neonatal scleroderma is the first presentation, although early diagnosis is challenging. The leading cause of death is cardio-/cerebro-vascular accidents associated with atherosclerosis. However, not all findings may recapitulate the aging process. We herein report a 9-year-old Japanese male with HGPS who developed cerebral infarction. The genetic study of peripheral blood-derived DNA determined a heterozygous c.1824C>T mutation, p.G608G. Telomere length of lymphocytes was normal. Bilateral stenosis of carotid siphons was prominent, while systemic arteriosclerosis was unremarkable assessed by the ankle-brachial index, carotid ultrasound imaging and funduscopic study. HGPS patients have marked loss and functional defects in vascular smooth muscle cells, leading to the vulnerability to circulatory stress. Symmetrical stenosis of siphons might occur as a distinctive cerebral vasculopathy of HGPS, rather than simple vascular senescence. Peripheral blood study on LMNA G608G and telomere length could screen progerias in infancy for early therapeutic intervention.

“Assessment of chronic sclerodermoid Graft‐versus‐Host Disease patients, using 20MHz high‐frequency ultrasonography and cutometer methods”

The development of an adverse graft-versus-host disease (GvHD) is a major complication of stem cell transplantations, which are widely used to cure increasing number of hematologic malignancies. Patients with chronic GvHD are at risk of joint contractures secondary to sclerodermatous skin changes. Several clinical scores or serologic markers have been used to assess skin sclerosis in scleroderma patients. Evaluation of sclerotic skin changes using biometric tools remains to be challenging. The purpose of this study was to illustrate and exemplify ultrasound measurement and measurement of skin elasticity of five chronic sclerodermoid GvHD patients. There is still a substantial lack of studies using objective and non-invasive methods helpful in assessment of patients with skin involvement of GvHD. Although ultrasound is not the ideal method, it is worth emphasizing that it is still useful, non-invasive, and repeatable device in monitoring patients suffering from GvHD. It should also be added, that it seems to be advisable to repeat USG examination at an interval of 3 months after the treatment. In addition, skin echogenicity may be a more sensitive parameter than skin thickness in assessment of cGvHD patients.

Type 1 IFN‐induced protein MxA and plasmacytoid dendritic cells in lesions of morphea

Morphea is an autoimmune sclerotic skin disease of unknown pathogenesis. As type 1 interferons (IFN) have been implicated in the pathogenesis of systemic sclerosis, we proposed that type 1 IFN promote localized inflammation and fibrosis in morphea. To investigate the expression of the type 1 IFN-inducible protein myxovirus A (MxA) and the presence of plasmacytoid dendritic cells (pDC) in lesions of morphea, lesional skin of 10 patients with morphea was examined by immunohistochemistry for the presence of the type 1 IFN-inducible protein, myxovirus A (MxA), and the pDC markers, CD123 and BDCA-2, and was compared with lesional skin of cutaneous lupus erythematosus, lichen planus and keloid. Lesional and non-lesional morphea skin was compared. MxA was expressed in the epidermis as well as the reticular dermis and subcutis in morphea. pDCs were abundant around vessels and between fibrous bundles. Non-lesional biopsies demonstrated little or no expression of MxA and pDC markers. Keloid showed minimal expression of MxA and pDC markers. We demonstrate the expression of type 1 IFN-related protein MxA and plasmacytoid DCs in lesional but not in non-lesional biopsies of morphea. These findings suggest a potential role for type 1 interferons in the pathogenesis of morphea.

UVA irradiation following treatment with topical 8-methoxypsoralen improves bleomycin-induced scleroderma in a mouse model, by reducing the collagen content and collagen gene expression levels in the skin

Recent studies have demonstrated that systemic or topical PUVA therapy, i.e., ultraviolet A (UVA) irradiation following treatment with 8-methoxypsoralen (8-MOP), is effective against the sclerotic skin lesions in systemic sclerosis. However, the mechanisms still remain unknown. To clarify the mechanisms of this therapy, we created a mouse model of bleomycin (BLM) injection-induced scleroderma and evaluated the effects of PUVA on the fibrotic lesions of scleroderma in this mouse model. BLM was injected subcutaneously once a day into the mice for 24 days. During the injection period, one group of mice was irradiated with UVA following local application of 8-MOP. Control groups were also set up, which were injected with phosphate-buffered saline, instead of BLM. Skin tissue samples examined histopathologically changes, measured of the content of hydroxyproline, and checked for the expression of genes encoding type I collagen, type III collagen, and transforming growth factor-β1 (TGF-β1). The mouse models of scleroderma was found to show an increase in the density of the collagen fibers and thickening of the dermis and increased expressions of type I collagen, type III collagen, and TGF-β1. However, the combination of BLM treatment and topical PUVA treatment mice appeared reduced the dermal thickness and hydroxyproline content, down-regulation of expressions of the type I and type III collagen genes was observed while the expression of the TGF-β1 gene remained unchanged. These results suggest that the effectiveness of topical PUVA therapy is attributable to the down-regulation of the expressions of the collagen genes by this treatment. The results additionally suggest that is not mediated by down-regulated expression of the TGF-β1.

Phase II Pilot Study of Imatinib Mesylate for the Treatment of Severe Sclerotic Skin Chronic Graft Versus Host Disease (ScGVHD)

Phase II Pilot Study of Imatinib Mesylate for the Treatment of Severe Sclerotic Skin Chronic Graft Versus Host Disease (ScGVHD)

Graft-Versus-Host Disease (GVHD) After Double-Unit Cord Blood Transplantation (DCBT) Is Associated with Unique Clinical Features Including a Higher Incidence of Grade III-IV Acute GVHD in Children

Abstract 3044While the GVHD incidence after unrelated donor CBT is lower than expected for the degree of human leukocyte antigen (HLA)-mismatch, GVHD can be a serious complication and at our center has been the second most common cause of transplant-related mortality after DCBT. However, relatively little is known about DCBT GVHD manifestations, treatment response, and risk factors. Therefore, we evaluated 108 DCBT recipients (median 37 years, range 0.9–69) transplanted for hematologic malignancies. The majority had acute leukemia and high-risk disease. Patients received either myeloablative (n = 81) or non-myeloablative (n = 27) conditioning and 4–6/6 HLA-matched grafts. GVHD prophylaxis consisted of a calcineurin-inhibitor with mycophenolate mofetil, and no patient received anti-thymocyte globulin (ATG). With a median follow-up of 28 months (range 9–64), the cumulative incidences of day 180 grade II-IV and III-IV acute GVHD (aGVHD) were 52% (95%CI :42–62) and 24% (95%CI :15–32), respectively. The median onset was 40 days (range 14–161); the gut was most commonly affected (43/54, 80%) followed by skin (35/54, 65%). Twenty-five patients with mainly grade II gut aGVHD were treated with budesonide alone, 26 patients with predominantly grade III-IV aGVHD received systemic corticosteroids, and complete or partial treatment response was achieved in over 80% by day 56 of therapy. However, 41 patients had active GVHD after day 100 with the majority (25/41, 61%) having aGVHD (persistent, recurrent or late onset), particularly of the gut. Overlap syndrome and classical chronic GVHD were uncommon. Only 1 patient had oral ulceration, and no patient had moderate or severe ocular or sclerotic skin involvement, joint, or pulmonary GVHD manifestations. Univariate analysis of the association between patient/ graft characteristics and grade III-IV aGVHD showed the only significant factor associated with a higher severe aGVHD incidence was age 0–15 years (Figure). Diagnosis, patient ancestry, cytomegalovirus seropositivity, conditioning intensity, and infused cell doses/kg (total graft and engrafting-unit nucleated cell, CD34+ and CD3+) were not significant. A higher engrafting unit-recipient HLA-match of 8–9/10 was associated with a lower incidence of severe aGVHD, and a better unit-unit HLA-match of 6–10/10 was associated with a higher incidence of severe aGVHD, although these differences were not significant (p = 0.128 and 0.266, respectively). To further investigate these findings multivariate Cox regression analysis was performed (Table). Younger age was independently associated with a higher incidence of severe aGVHD (p = 0.042) whereas better engrafting unit-recipient match at 8–9/10 HLA-alleles was protective (p = 0.053). There was a trend toward better unit-unit HLA-match being associated with a higher incidence of grade III-IV aGVHD, but, surprisingly, total infused TNC/kg had no relationship. The 2-year PFS of 72% (95%CI :51–94) in children was higher than the 56% (95%CI :45–66) in adults despite their greater incidence of severe aGVHD. Nine patients (all adult) have died of GVHD including 5 patients initially treated with systemic corticosteroids and 4 with budesonide. We conclude that aGVHD after DCBT is common in the absence of ATG, predominantly affects the gut, and has a high rate of treatment response. Furthermore, GVHD after day 100 frequently has acute features. While the GVHD incidence does not preclude a high rate of survival, improved prophylaxis and treatment are needed. Notably, in contrast to single-unit CBT and adult hematopoietic stem cell transplantation, children receiving DCBT are at a higher risk for severe disease. A possible approach to reduce aGVHD in pediatric DCBT recipients with adequate CB units doses would be to prioritize high resolution HLA-match. Moreover, our data does not currently support an upper limit of infused TNC/kg in DCBT recipients. Further investigation of the biology underlying these unique observations (including the role of specific cellular subsets) should be a major priority. [Display omitted] [Display omitted] Disclosures:Off Label Use: Mycophenolate Mofetil as GvHD prophylaxis.

Cracking the cement overcoat

In this issue of Blood , Martires et al report on the risk factors for sclerotic skin manifestations in 206 patients with established chronic graft-versus-host disease (cGVHD). 1 Sclerotic skin changes are one of the most dreaded manifestations of cGVHD, encasing the patients in unmovable skin. Patients become progressively immobilized in skin like layers of cement, making normal, daily living impossible (see figure).

Sclerotic-type chronic GVHD of the skin: clinical risk factors, laboratory markers, and burden of disease

AbstractChronic GVHD is one of the most severe complications of allogeneic HSCT. The sclerotic skin manifestations of cGVHD (ScGVHD) result from inflammation and fibrosis of the dermis, subcutaneous tissue, or fascia, leading to significant functional disability. Risk factors and clinical markers associated with ScGVHD remain largely unexamined. By using a single-visit, cross-sectional design, we evaluated 206 patients with cGVHD at the National Institutes of Health. Most patients manifested severe (ie, 63% National Institutes of Health score “severe”), refractory disease (median treatments = 4). ScGVHD was detected in 109 (52.9%) patients. ScGVHD was associated with greater platelet count (P < .001) and C3 (P < .001), and decreased forced vital capacity (P = .013). Total body irradiation (TBI) was associated with development of ScGVHD (P = .002). TBI administered in reduced-intensity conditioning was most strongly associated with ScGVHD (14/15 patients, P < .0001). Patients with ScGVHD had significant impairments of joint range of motion and grip strength (P < .001). Greater body surface area involvement was associated with poorer survival (P = .015). We conclude that TBI, particularly in reduced-intensity regimens, may be an important risk factor for ScGVHD. Widespread skin involvement is associated with significant functional impairment, distressing symptoms, and diminished survival. This trial is registered at http://www.clinicaltrials.gov as NCT00331968.

Localization of Sclerotic-type Chronic Graft-vs-Host Disease to Sites of Skin Injury

The mechanisms responsible for the variable manifestations of chronic cutaneous graft-vs-host disease (cGVHD) are poorly understood. Localization of sclerotic-type chronic graft-vs-host disease to sites of skin injury (isomorphic and isotopic responses), a recognized phenomenon in morphea, suggests a potential common pathway between cGVHD and other sclerotic skin conditions. Four cases of sclerotic-type cGVHD developed at the site of disparate skin injuries (ionizing radiotherapy, repeated needle sticks, central catheter site, and varicella-zoster virus infection). We review the spectrum of previously reported cases of sclerotic and nonsclerotic cGVHD relating to external forces on the skin. Localization of sclerotic-type cGVHD may occur after many types of skin injury, including UV and ionizing radiotherapy, needle sticks, viral infection, and pressure or friction. Recognition of this phenomenon may be helpful for the early diagnosis of sclerotic disease. Recent insights into the immunological consequences of minor skin injury may provide important clues to the underlying pathogenesis of cGVHD-mediated skin disease.

Skin Disinfection by Plasma-Tissue Interaction: Comparison of the Effectivity of Tissue-Tolerable Plasma and a Standard Antiseptic

Wound healing disorders frequently occur due to biofilm formation on wound surfaces requiring conscientious wound hygiene. Often, the application of conventional liquid antiseptics is not sufficient and sustainable as (1) the borders and the surrounding of chronic wounds frequently consist of sclerotic skin, impeding an effectual penetration of these products, and (2) the hair follicles representing the reservoir for bacterial recolonization of skin surfaces are not affected. Recently, it has been reported that tissue-tolerable plasma (TTP), which is used at a temperature range between 35 and 45°C, likewise has disinfecting properties. In the present study, the effectivity of TTP and a standard liquid antiseptic was compared in vitro on porcine skin. The results revealed that TTP was able to reduce the bacterial load by 94%, although the application of the liquid antiseptic remained superior as it reduced the bacteria by almost 99%. For in vivo application, however, TTP offers several advantages. On the one hand, TTP enables the treatment of sclerotic skin as well, and on the other hand, a sustainable disinfection can be realized as, obviously, also the follicular reservoir is affected by TTP.

Effects of different doses of UVA1 irradiation on the expressions of CD34 and M30 in skin homogenate from a mouse model of scleroderma

Objective To observe the expression changes of CD34 and M30 in skin homogenate from a mouse model of scleroderma after irradiation with different doses of UVA1, and to investigate the effect of UVA1 phototherapy on vascular endothelial cell function in scleroderma. Methods The experimental mouse models of scleroderma were established by the injection with bleomycin and randomly divided into model control group (n = 10), UVA1 irradiation group (n = 30) and unirradiated group (n = 10). The UVA1 irradiation group was further equally divided into 3 groups, HD-UVA1 group irradiated with UVA1 at 100 J/cm2, MD-UVA1group with UVA1 at 60 J/cm2, and LD-UVA1 group with UVA1 at 20 J/cm2; phototherapy was performed thrice weekly for 10 weeks followed by the sacrifice of mice. The mice in model control group were killed immediately after the establishment of models, and the mice in unirradiated group received no irradiation after the establishment of models and were maintained till the killing of mice in UVA1 irradiation groups. Skin specimens were obtained from the bleomycin-induced scleroderma lesions of mice and separated into two parts, one was subjected to histopathological examination, and the other one was used to prepare skin homogenate for the detection of CD34 and M30 content with ELISA assay. Results After 30 sessions of treatment with UVA1,the softening and thinning of sclerotic skin were seen by the naked eye, with the most obvious changes in HDUVA1 group; pathological examination revealed a reduction in dermal thickness and the presence of hair follicular structures in subcutaneous fat tissue with no obvious proliferation of collagen in these mice. Compared with the mice in model control group and unirradiated group, there was an increase in CD34 and decrease in M30 content in skin homogenate from UVA 1-irradiated mice, with the most marked changes in mice irradiated with UVA1 at 100 J/cm2. The concentration of CD34 and M30 in skin homogenate from unirradiated group and model control group was significantly different from that in HD-UVA1 group (22.25 ± 8.91 μg/L and 31.97 ±17.97 μg/L vs. 72.39 ± 13.04 μg/L, 162.41 ± 58.00 U/L and 195.71 ± 71.09 U/L vs. 38.06 ± 19.89 U/L, all P ＜ 0.01 ). Additionally, significant differences were observed between the three UVA1 groups in the concentration of CD34 and M30 (F = 21.23, 15.32, respectively, both P ＜ 0.01 ). Conclusions UVA1 phototherapy could up-regulate the expression of CD34 but down-regulate that of M30 in skin homogenate from the mouse model of scleroderma, and the effect is correlated with the intensity and cumulative dose of irradiation. Key words: Ultraviolet therapy; Scleroderma, systemic; Disease models, animal; Antigens, CD34; M30

Differential Contribution of Dermal Resident and Bone Marrow–Derived Cells to Collagen Production during Wound Healing and Fibrogenesis in Mice

Recent studies show that bone marrow (BM)-derived cells migrating into a dermal wound promote healing by producing collagen type I. However, their contribution to the repair process has not been fully verified yet. It is also unclear whether BM-derived cells participate in dermal fibrogenesis. We have addressed these issues using transgenic mice that harbor tissue-specific enhancer/promoter sequences of α2(I) collagen gene linked to either enhanced green fluorescent protein (COL/EGFP) or the luciferase (COL/LUC) reporter gene. Following dermal excision or subcutaneous bleomycin administration, a large number of EGFP-positive collagen-producing cells appeared in the dermis of COL/EGFP reporter mice. When wild-type mice were transplanted with BM cells from transgenic COL/EGFP animals and subjected to dermal excision, no EGFP-positive BM-derived collagen-producing cells were detected throughout the repair process. Luciferase assays of dermal tissues from COL/LUC recipient mice also excluded collagen production by BM-derived cells during dermal excision healing. In contrast, a limited but significant number of CD45-positive collagen-producing cells migrated from BM following bleomycin injection. These results indicate that resident cells in the skin are the major source of de novo collagen deposition in both physiological and pathological conditions, whereas BM-derived cells participate, in part, in collagen production during dermal fibrogenesis.

Case of diabetic scleredema: Diagnostic value of magnetic resonance imaging

We report the case of a patient with a 20-year history of diabetes mellitus type 2 who developed sclerotic skin lesions on his neck and upper back. Physical and histological findings were compatible with diabetic scleredema. T(2)-weighted magnetic resonance imaging (MRI) revealed diffuse thickening of the dermis and subcutaneous tissue, with hyperintense signals. The MRI findings in the dermis corresponded with the accumulation of collagen tissue with deposition of glycosaminoglycans.

A Case of Paclitaxel Induced Scleroderma in a Patient with Ovarian Cancer

Paclitaxel takes effects as an antineoplastic agent by inter-fering with microtubules and then blocking cell growth. It has been used to treat patients with lung, ovarian, breast, head and neck cancer, and advanced forms of Kaposi's sarcoma. Along with its reputation as an effective chemo-therapeutic drug, paclitaxel has numerous adverse effects. Among them, cutaneous adverse effects of paclitaxel in-clude pruritis, bullous fixed eruption, onycholysis, and transient erythrodysesthesia. Only several cases of scleroderma-like lesions have been reported throughout the world, and in Korea, only one case of paclitaxel induced scleroderma has been reported in 2006. We report a case of paclitaxel induced scleroderma in an 83-year old woman with ovarian cancer. After administration of paclitaxel and cisplatin, the patient presented with edema in both bra-chial areas, Raynaud's phenomenon, and sclerotic skin lesions in both extremities.