Cytotoxic and chemosensitizing effects of glycoalkaloidic extract on 2D and 3D models using RT4 and patient derived xenografts bladder cancer cells.

Glycoalkaloids have been widely demonstrated as potential anticancer agents. However, the chemosensitizing effect of these compounds with traditional chemotherapeutic agents has not been explored yet. In a quest for novel effective therapies to treat bladder cancer (BC), we evaluated the chemosensitizing potential of glycoalkaloidic extract (GE) with cisplatin (cDDP) in RT4 and PDX cells using 2D and 3D cell culture models. Additionally, we also investigated the underlying molecular mechanism behind this effect in RT4 cells. Herein, we observed that PDX cells were highly resistant to cisplatin when compared to RT4 cells. IC50 values showed at least 2.16-folds and 1.4-folds higher in 3D cultures when compared to 2D monolayers in RT4 cells and PDX cells, respectively. GE + cDDP inhibited colony formation (40%) and migration (28.38%) and induced apoptosis (57%) in RT4 cells. Combination therapy induced apoptosis by down-regulating the expression of Bcl-2 (p < 0.001), Bcl-xL (p < 0.001) and survivin (p < 0.01), and activating the caspase cascade in RT4 cells. Moreover, decreased expression of MMP-2 and 9 (p < 0.01) were observed with combination therapy, implying its effect on cell invasion/migration. Furthermore, we used 3D bioprinting to grow RT4 spheroids using sodium alginate-gelatin as a bioink and evaluated the effect of GE + cDDP on this system. Cell viability assay showed the chemosensitizing effect of GE with cDDP on bio-printed spheroids. In summary, we showed the cytotoxicity effect of GE on BC cells and also demonstrated that GE could sensitize BC cells to chemotherapy.

FDG-PET may predict the effectiveness of hyperbaric oxygen therapy in a patient with bisphosphonate-related osteonecrosis of the jaw: report of a case.

We report an advanced case of bisphosphonate-related osteonecrosis of the jaw (BRONJ) in an osteoporotic patient treated with oral risedronate sodium for 2 years. An 80-year-old woman presented to our hospital complaining of pain, swelling and pus discharge in the lower alveolar ridge. Fluorine-18 labeled fluorodeoxyglucose positron emission tomography (FDG-PET) and bone scintigraphy showed definite uptake in the mandible. Under clinical diagnosis of BRONJ, we applied systematic treatments including antibiotic therapy, irrigation, cessation of bisphosphonate, hyperbaric oxygen (HBO) therapy, and debridement of necrotic bone. After pre-operative 20 sessions of HBO therapy, her clinical symptoms disappeared. SUVmax of FDG-PET decreased definitely from 4.5 to 2.5, although magnetic resonance image and bone scintigraphy did not show remarkable changes. After minor surgery with debridement of necrotic bone, she received another ten sessions of HBO therapy. After the treatment, her clinical course was excellent. In conclusion, this report demonstrates FDG-PET may predict the effect of HBO therapy in BRONJ.

Analgesic, anti-inflammatory, and anti-oxidant activities of Phlogacanthus thyrsiflorus leaves.

BACKGROUND: Our present study was carried out to explore the potential role of the methanol extract from the leavesof Phlogocanthus thyrsiflorus (PT) Nees. in central and peripheral analgesic activities using hot plate and acetic acid-induced writhing methods. We also tested the antiinflammatory effects and anti-oxidant activity using carrageenan-induced paw edema and the DPPH method, respectively. METHODS: Methanol extracts of PT leaves were prepared using 500 g powder in 1.8 L methanol by percolation method, followed by evaporation in a rotary evaporator under controlled temperature and pressure. The crude methanol extract was dried by freeze drier and preserved at 4 °C. RESULTS: Oral administration of PT significantly (p < 0.05)increased the reaction time at 55.73% (250 mg/kg) and 72.81% (500 mg/kg) inhibition (p < 0.05) in the hot plate test at 3 h. PT significantly (p < 0.05) inhibited 42.17% (250 mg/kg) and 56.63% (500 mg/kg) acetic acid-induced writhing. PT leaves (250 and 500 mg/kg) also significantly (p < 0.05) inhibited paw edema 6 h after carrageenan injection. Furthermore, this plant showed significant (p < 0.05) free radical-scavenging activity at a dose range of 25­800 µg/mL. CONCLUSIONS: Based on the findings, we can conclude that PT leaf possesses analgesic, anti-inflammatory, and antioxidant activities. Preliminary phytochemical study of PT leaves revealed the presence of flavonoids, tannins and triterpens in methanol extract which could be correlated with its observed biological activities.

Evaluation of changes in the tumor microenvironment after sorafenib therapy by sequential histology and 18F-fluoromisonidazole hypoxia imaging in renal cell carcinoma.

The mechanistic dissociation of 'tumor starvation' versus 'vascular normalization' following anti-angiogenic therapy is a subject of intense controversy in the field of experimental research. In addition, accurately evaluating changes of the tumor microenvironment after anti-angiogenic therapy is important for optimizing treatment strategy. Sorafenib has considerable anti-angiogenic effects that lead to tumor starvation and induce tumor hypoxia in the highly vascularized renal cell carcinoma (RCC) xenografts. 18F-fluoromisonidazole (18F­FMISO) is a proven hypoxia imaging probe. Thus, to clarify early changes in the tumor microenvironment following anti-angiogenic therapy and whether 18F-FMISO imaging can detect those changes, we evaluated early changes in the tumor microenvironment after sorafenib treatment in an RCC xenograft by sequential histological analysis and 18F-FMISO autoradiography (ARG). A human RCC xenograft (A498) was established in nude mice, for histological studies and ARG, and further assigned to the control and sorafenib-treated groups (80 mg/kg, per os). Mice were sacrificed on Days 1, 2, 3 and 7 in the histological study, and on Days 3 and 7 in ARG after sorafenib treatment. Tumor volume was measured every day. 18F-FMISO and pimonidazole were injected intravenously 4 and 2 h before sacrifice, respectively. Tumor sections were stained with hematoxylin and eosin and immunohistochemically with pimonidazole and CD31. Intratumoral 18F-FMISO distribution was quantified in ARG. Tumor volume did not significantly change on Day 7 after sorafenib treatment. In the histological study, hypoxic fraction significantly increased on Day 2, mean vessel density significantly decreased on Day 1 and necrosis area significantly increased on Day 2 after sorafenib treatment. Intratumoral 18F-FMISO distribution significantly increased on Days 3 (10.2-fold, p<0.01) and 7 (4.1-fold, p<0.01) after sorafenib treatment. The sequential histological evaluation of the tumor microenvironment clarified tumor starvation in A498 xenografts treated with sorafenib. 18F-FMISO hypoxia imaging confirmed the tumor starvation. 18F-FMISO PET may contribute to determine an optimum treatment protocol after anti-angiogenic therapy.