Evans blue as a selective dye marker for white-light diagnosis of non-muscle-invasive bladder cancer: an in vitro study.

Objective: To develop a diagnostic method relying on the preferential accumulation of a dye in non-muscle-invasive bladder cancer (NMIBC) that is visible in conjunction with white-light cystoscopy (WLC).

Materials And Methods: We investigated in detail the permeation of Evans blue in urothelial cell carcinoma (UCC) spheroids prepared from T24, J82 and RT-112 human cell lines and spheroids composed of normal human urothelial (NHU) cells. To gain more insight into the differential accumulation, all spheroids were investigated ultrastructurally using transmission electron microscopy (TEM).

Prevention of tumor cell reimplantation during transurethral resection: the in-vitro antiadhesive and cytotoxic properties of an irrigant containing polyethylene glycol 400.

A major challenge to urologists with respect to bladder cancer is the high rate of tumor recurrence after transurethral resection (TUR). Implantation of resected tumor cells on traumatized bladder urothelium is believed to be the main cause of tumor recurrence. The aim of this study was to find a safe irrigant fluid and modality that prevents reimplantation of malignant cells during TUR.

Differential accumulation of hypericin in spheroids composed of T-24 transitional cell carcinoma cells expressing different levels of E-cadherin.

Purpose: To obtain unambiguous evidence for the putative role of E-cadherin in the selective accumulation of hypericin after intravesical instillation in humans we investigated the accumulation of hypericin in spheroids from 3 clones of the human bladder carcinoma cell line T-24 that express different levels of E-cadherin, as determined by immunohistochemistry and reverse transcriptase-polymerase chain reaction.

Materials And Methods: Clones of T-24 cells transfected with the E-cadherin gene were analyzed for E-cadherin expression and 3 cell lines with different expression levels were selected. Spheroids of these cell lines were incubated with 10 microM hypericin in cell culture medium supplemented or not with fetal calf serum for 2 hours.

Influence of the glycosaminoglycan layer on the permeation of hypericin in rat bladders in vivo.

Objective: To investigate the influence of a glycosaminoglycan (GAG) layer on the specific location of hypericin in superficial urothelial carcinoma lesions of the bladder after intravesical instillation.

Materials And Methods: Fisher rat bladders were incubated with 15 or 30 microm hypericin for 2 h. To examine the influence of the GAG layer on the permeation of hypericin, bladders were pre-treated with chondroitinase ABC, n-dodecyl-beta-d-maltoside (DDM) or sodium dodecyl sulphate (SDS) to disrupt, or protamine to neutralise the GAG layer before incubating with hypericin.

In vitro accumulation and permeation of hypericin and lipophilic analogues in 2-D and 3-D cellular systems.

Previous studies have shown that hypericin is an excellent diagnostic tool for the fluorescence detection of carcinoma in situ in the human bladder. The present work was performed to get a better insight into the mechanism of cellular uptake of hypericin (HYP) using RT-112 human papillary TCC cells of the bladder. Using lipophilic hypericin acid amide derivatives like hypericin acid hexylamide (AM6), hypericin acid octylamide (AM8) and hypericin acid dodecylamide (AM12), the effect of increased lipophilicity on the binding to serum proteins was investigated, as well as the cellular accumulation and permeation, both in 2-D and 3-D cell conditions.

Enhancing the photodynamic effect of hypericin in human bladder transitional cell carcinoma spheroids by the use of the oxygen carrier, perfluorodecalin.

In the present study, we evaluated the possibility of enhancing the photodynamic effect of hypericin in transitional cell carcinoma (TCC) spheroids by the use of the oxygen carrier, perfluorodecalin. Following incubation with hypericin, RT-112 TCC spheroids were irradiated in the presence or absence of perfluorodecalin, at light doses of 7 J/cm(2) or 28 J/cm(2) , delivered at a fluence rate of 15 mW/cm(2) . The photodynamic therapy (PDT) efficacy was evaluated and apoptotic cells were visualized.

Synthesis, in vitro cellular uptake and photo-induced antiproliferative effects of lipophilic hypericin acid derivatives.

Hypericin, a naturally occurring hydroxylated phenanthroperylene dione, is used as a powerful photosensitizer for photodynamic therapy as well as a diagnostic tool for the fluorescence detection of flat neoplastic lesions in the bladder of patients. Both applications are based on the tumouritropic characteristics of the compound. To get more insight into some of the physicochemical properties of hypericin affecting its tumouritropic characteristics, we set out to synthesize a series of more lipophilic hypericins.

Permeation of hypericin in spheroids composed of different grade transitional cell carcinoma cell lines and normal human urothelial cells.

Purpose: We investigated the importance of E-cadherin expression on the selective accumulation of hypericin in superficial bladder cancer after intravesical instillation.

Materials And Methods: Spheroids obtained from a panel of 3 transitional cell carcinoma cell lines, namely J-82, RT-4 (American Type Culture Collection, Manassas, Virginia) and RT-112 (German Collection of Micro-organisms and Cell Cultures, Braunschweig, Germany), and normal human urothelial (NHU) cells were incubated with hypericin. Accumulation was examined with fluorescence microscopy.

Enhancing the photodynamic effect of hypericin in tumour spheroids by fractionated light delivery in combination with hyperoxygenation.

The aim of this study was to explore the hypothesis of oxygen depletion during light irradiation as a possible explanation for the incomplete response seen after hypericin-mediated photodynamic therapy (PDT) under specific conditions. To investigate this, we performed PDT experiments using transitional cell carcinoma spheroids with fractionated light irradiation and hyperoxygenation. After 2-h incubation with 3 different hypericin concentrations, spheroids were irradiated either continuously or with fractionated light delivery.

Stability of different formulations and ion pairs of hypericin.

Hypericin, solubilized in an instillation fluid consisting of an aqueous buffer supplemented with 1% plasma proteins, is currently used as a clinical diagnostic tool for the detection of superficial TCC (transitional cell carcinoma) tumors. However, the development of a sterile and stable hypericin stock formulation, excluding the presence of plasma constituents, would be an important factor in a more general clinical application of the method. Therefore, we investigated the stability of several heat sterilized hypericin formulations and ion pairs.

In vivo accumulation of different hypericin ion pairs in the urothelium of the rat bladder.

Objective: To optimise the diagnostic and phototherapeutic efficacy of hypericin in superficial bladder cancer, by developing a bladder instillation fluid that does not depend on the presence of plasma proteins for an appropriate and reliable urothelial uptake of hypericin.

Materials And Methods: Sodium hypericinate (in distilled water, in sodium phosphate buffer, or in polyethylene glycol) and several other hypericinate salts (potassium, lysine, TRIS or hexylamine) were instilled with no plasma constituents into the rat bladder. The accumulation of hypericin was assessed with fluorescence microscopy.

Accumulation and photocytotoxicity of hypericin and analogs in two- and three-dimensional cultures of transitional cell carcinoma cells.

The aim of this study was to investigate the in vitro cellular accumulation, distribution and photocytotoxic effect of hypericin in two-dimensional (2-D) and three-dimensional (3-D) cultured RT-112 transitional cell carcinoma cells of the bladder. In addition, two iodinated derivatives of hypericin were incorporated to investigate whether these analogs, with their increased lipophilicity and heavy-atom effect, display a different biological behavior and optimized photodynamic effect. The results indicate that hypericin and mono-iodohypericin behave similarly in terms of cellular accumulation, spheroidal distribution and photocytotoxic effect.

Photodynamic therapy of transitional cell carcinoma multicellular tumor spheroids with hypericin.

The objective of the present study was to evaluate the sensitivity of the bladder transitional cell carcinoma (TCC) to hypericin PDT in a 3-D system using multicellular tumor spheroids. The photodynamic response in RT-112 human bladder TCC spheroids was also compared to 2-D cultured monolayer cells. Following a 2-4 h incubation with 8-30 microM hypericin, spheroids or monolayer cells were irradiated at the light dose of 12 J/cm2, delivered at a fluence rate of 10-100 mW/cm2.