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Methods & Approach

Two cancer prostate cellular models, namely human prostate adenocarcinoma LNCaP C4-2 (PSMA-positive, low metastatic potential, high radiosensitivity), human prostate carcinoma DU 145 (PSMA-negative, moderate metastatic potential, high radioresistance) and two normal prostate cell models, namely human primary prostate epithelial cells and normal epithelial prostate cells RWPE-1 are used for in vitro experiments. For in vivo experiments BALB/c and BALB/c nude with LNCaP C4-2 and DU145 xenografts are used. A wide spectrum of methods is utilized (i.g. physico-chemical methods, methods of cellular and mollecular biology, immunological and biochemical methods).The project involves different methodological approaches:

Synthesis and physicochemical characterization of anti-PSMA-functionalized NaA nanozeolites labeled with the alpha-emitter 225Ra

Synthesis of NaA nanozeolites, modification of their surface with silane-polyethylene glycol (PEG) molecules and immobilization of 223Ra.

Physicochemical characterization of NaA nanozeolites and anti-PSMA-functionalized NaA nanozeolites unlabeled and labeled with the alpha-emitter 225Ra.

Analysis of in vitro toxicity of unlabeled anti-PSMA-functionalized NaA nanozeolitesand unlabeled NaA nanozeolites as carriers in vitro.

The effects of unlabeled anti-PSMA-functionalized NaA nanozeolites and unlabeled NaA nanozeolites on cellular cytotoxicity in vitro.

The effects of unlabeled anti-PSMA-functionalized NaA nanozeolites and unlabeled NaA nanozeolites on the NF-κB signaling pathway in vitro.

The effects of unlabeled anti-PSMA-functionalized NaA nanozeolites and unlabeled NaA nanozeolites on the mRNA expression of inflammatory cytokines and stress-related proteins in vitro.

The effects of unlabeled anti-PSMA-functionalized NaA nanozeolites and unlabeled NaA nanozeolites on secretion of pro-inflammatory cytokines and oxidative stressin vitro.

Analysis of in vitro efficacy of anti-PSMA-functionalized NaA nanozeolites labeled withthe alpha-emitter 223Ra

Stability studies of 223Ra labeled anti-PSMA-functionalized NaA nanozeolites and 223Ra labeled NaA nanozeolites.

Binding affinity studies of anti-PSMA-functionalized NaA nanozeolites to PSMA receptors on human prostate cells.

The effects of 223Ra labeled anti-PSMA-functionalized NaA nanozeolites on invasion and migration of human prostate cells.

The effects of 223Ra labeled anti-PSMA-functionalized NaA nanozeolites and 223Ra labeled NaA nanozeolites on induction of double strand breaks (DSBs).

Preclinical pharmacokinetics and toxicity studies of NaA nanozeolite functionalized withanti-PSMA antibodies and labeled with barium-133 and radium-223 radioisotopes in normal mice.

Analysis of pharmacokinetics of anti-PSMA-functionalized NaA nanozeolites labeled with the radioizotopic marker 133Ba in BALB/c mice.

Acute toxicity study of anti-PSMA-functionalized NaA nanozeolites labeled with the alphaemitter 223Ra in normal Balb/c mice.

Analysis of localization in vivo and biodistribution ex vivo of NaA nanozeolite functionalized with anti-PSMA antibodies and labeled with radium-223 and barium-133 radioizotope and NaA nanozeolite labeled with barium radioizotope as emitter of beta particles in murine model of human prostate cancer.

Analysis of in vivo localization of anti-PSMA-functionalized NaA nanozeolites labeled with the alpha-emitter 223Ra in BALB/c nude mice with xenograft prostate tumors.

Analysis of biodistribution of NaA nanozeolites in BALB/c nude mice with xenograft prostate tumors.