Altered decorin expression of systemic sclerosis by UVA1 (340–400 nm) phototherapy: Immunohistochemical analysis of 3 cases

BACKGROUND: Ultraviolet A1 (340–400 nm, UVA1) phototherapy is highly effective in sclerotic lesions of systemic sclerosis (SSc). Histological evaluation of skin specimens obtained before and after UVA1 phototherapy revealed loosening of collagen bundles and the appearance of small collagen fibers. We have previously shown that UVA1 irradiation induced collagenase in vitro study by using SSc fibroblasts. The increased levels of mRNA and protein of decorin in SSc fibroblasts were reported. In this study, we focus on the lesional expression of small dermatan sulfate proteoglycan, decorin that has a role of binding to collagen and fibrillogenesis. CASE PRESENTATION: We employed immunohistochemical analysis of decorin before and after UVA1 phototherapy. The skin specimens from three patients who were effectively treated with UVA1 phototherapy were analysed. Monoclonal antibody 6B6 as the specific reactivity to decorin was used. The increased decorin was focally accumulated in the newly synthesized collagen fibers in the sclerotic lesion of SSc. After UVA1 phototherapy, decorin was decreased in upper to middle dermis, although decorin was slightly increased in papillary dermis. CONCLUSIONS: These results suggest that decreased and normalized levels of accumulated decorin may relate to the efficacy of sclerotic lesions in UVA1 phototherapy

An Ex Vivo Model Employing Keloid-Derived Cell–Seeded Collagen Sponges for Therapy Development

The most distinctive feature of keloid is the extreme deposition of extracellular matrix, including collagens and proteoglycans (PGs). The focus of this study was the PG versican, which presumably defines keloid volume because of its ability to retain large amounts of water through its component glycosaminoglycans (GAGs). The excessive deposition of versican in keloids was examined by immunohistochemical analysis and by upregulation of the versican gene in these lesions by real-time PCR. The latter showed that mesenchymal cells derived from keloid lesion (KL) cells continue to exhibit above-normal versican production in culture. To establish a model of GAG deposition in keloids, collagen sponges seeded with KL cells (KL-SPos) were implanted in the subcutaneous space of nude mice. After 1 month, the KL-SPos were significantly heavier than the fibroblast (Fb)-seeded sponges (Fb-SPos). This ex vivo model was subsequently used to examine an inhibitory ability of IL-1β that was identified to reduce versican in vitro. IL-1β or chondroitinase ABC, when injected directly, successfully reduced the weight of the KL-SPos. Thus, on the basis of the change in weight of the seeded sponges, this ex vivo model can be used to test therapies aimed at reducing or inhibiting keloid formation and to study the pathogenesis of this aberrant response

Chemical toxicity of indocyanine green damages retinal pigment epithelium. Invest Ophthalmol Vis Sci.

PURPOSE. To investigate the chemical toxicity of indocyanine green (ICG). METHODS. Surface active and precipitating effects of ICG were quantitatively analyzed by determining bovine serum albumin dissolved or precipitated in the presence or absence of salt solutions. The effects of precipitation on serum and cytotoxicity were evaluated by measuring the viability of retinal pigment epithelium (RPE) in vitro. RESULTS. ICG functioned as a surfactant without salts, but with nearly physiological concentrations of balanced salts, it functioned as a unique precipitating factor. This rendered the soluble molecules in serum that are indispensable in the culture of RPE cells insoluble during a 12-hour exposure, resulting in poor cell survival in vitro. Cytotoxicity in serum-free medium was also shown during brief exposures. CONCLUSIONS. Commonly used dosages of ICG directly applied into the vitreous cavity, which not only contact the retina but also invade the space between the retina and RPE through a macular hole, may be sufficient to induce retinal disorders after the damaging chemical property of ICG has disturbed the microenvironment. (Invest Ophthalmol Vis Sci. 2005;46: 2531-2539) DOI:10.1167/iovs.04-1521 B efore 1990, there was no treatment available for macular holes. Kelly and Wendel 1 first reported that vitreous surgery improved visual acuity in some eyes with macular holes, and since then an effective internal limiting membrane (ILM) peeling technique has been used to improve outcomes. 2 However, it is generally very difficult to perform efficient and complete removal of the ILM, because the ILM is thin and transparent. After indocyanine green (ICG) was introduced for selective staining of the ILM, 3 ICG-facilitated ILM peeling appeared to be beneficial, because it facilitated functional and anatomic success. 4 -10 Although some reports have shown no negative effects and/or excellent anatomic results and visual retinal function after ICG-assisted ILM peeling for macular hole surgery, 18,21 Unusual atrophic changes in the retinal pigment epithelium (RPE) at the site of the macular hole and undesirable postoperative visual acuity were reported, despite successful anatomic closure of the macular hole. 22 After macular hole surgery with ICG-guided ILM peeling, hypertrophic and atrophic RPE changes were apparent. 26 ICG injected into the vitreous cavity showed b-and a-wave amplitude, and latency abnormalities, suggesting impairment of retinal function. 27 ICG caused cytotoxicity in cultured RPE cells in a dose-and time-dependent manner, and necrotic cell death occurred. 29 ICG injected into the subretinal space caused destructive degeneration of photoreceptors and RPE, 32 Although all these data obviously indicate toxic effects of ICG on retinal function, we hardly consider these results to be caused by ICG alone. ICG-induced cytotoxicity in cultured RPE was reduced with the removal of sodium from the solvent. 33 Toxic effects of ICG on cultured RPE cells have been related to osmolarity of the solvent. 34 Factors such as sodium or osmolarity modulate the degree of toxicity, implying that some chemical mechanisms may be involved in the effects. The dye ICG is an amphipathic molecule and has both hydrophilic (two anions and one ammonium cation) and hydrophobic (mainly aromatic series) properties, suggesting that ICG is a surface active compound and may have unknown chemical and toxic properties that affect the retina. In the present study, we demonstrated unique chemical properties of ICG-acting as a precipitating factor in the presence of salts, but as a surface active compound in the absence of salts. Our data support the proposition that the clinically used dosage of ICG in buffered salt conditions directly applied into the vitreous cavity and subretinal space through a macular hole may be sufficient to induce retinal disorder. From th

The pulp tissue regeneration induced by MMP-3 in the irreversible pulpitis model.

<p>A. H&E staining of pulp tissues from dogs with mild pulpitis at 14 and 28 days after MMP-3 or saline treatment, as indicated. The arrows show the imaginary amputated site, with the indicated areas magnified in C. Scale bar, 200 µm. B. H&E staining of pulp tissues from dogs with severe pulpitis at 14 days after MMP-3 or saline treatment, as indicated. The arrows indicate the imaginary amputated site. Scale bar, 200 µm. C. Immunohistochemical analysis for BS-1-lectin, TuJ1, and GAP43 and Masson’s trichrome staining of pulp tissues from dogs with mild pulpitis at 14 or 28 days after MMP-3 treatment, as indicated. The representative staining of the cells is indicated by arrowheads. Scale bar, 50 µm.</p

Hyaluronan (HA), SHAP and versican localization in the mild pulpitis model.

<p>Immunofluorescence-stained pulp tissues were prepared 3 days after treatment. A. H&E-stained pulp tissues from dogs with mild pulpitis 3 days after treatment with MMP-3, MMP-3 plus NNGH, NNGH alone or saline as indicated, with the indicated area magnified in B. The arrows indicate the imaginary amputated site. Scale bar, 200 µm. B. Pulpitis tissues stained with biotinylated HABP (HA), anti-SHAP (SHAP) and anti-versican as indicated. Scale bar, 20 µm.</p