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Influence of dendrimers and iontophoresis in protoporphyrin IX penetration into skin tumors

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Author(s):
Laura Freire Cardoso Pimenta
Total Authors: 1
Document type: Master's Dissertation
Press: Ribeirão Preto.
Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto
Defense date:
Examining board members:
Renata Fonseca Vianna Lopez; Maria Bernadete Riemma Pierre; Antonio Claudio Tedesco
Advisor: Renata Fonseca Vianna Lopez
Abstract

Photodynamic therapy (PDT) associated with topical administration of photosensitizer agents is a promising therapy for topical treatment of skin cancer. Protoporphyrin IX (PpIX) is a photosensitizer commonly used in PDT; however, due to its high lipophilicity it aggregates in aqueous medium, which decreases its photodynamic activity and hinders its penetration through the skin. In this way, nanoparticles have been designed to improve the distribution of PpIX in the skin and enhance its tumor cell penetration. The polyamidoamine dendrimers (PAMAM) represent a new generation of nanosystems that has aroused great interest in recent years. They are hyberbranched polymers capable to form complexes with PpIX (PpIX-PAMAM), increasing PpIX aqueous solubility. The application of a low intensity electrical current, known as iontophoresis, may influence the nanoparticles skin penetration, directing them to the tumor cells. Therefore, the aim of this study was to evaluate the influence of iontophoresis and PpIX-PAMAM G4-OH complexes in PpIX subcellular localization and penetration into skin tumors. The complexes were prepared and characterized as a function of particle size and zeta potential. The subcellular localization of PpIX from the complexes was investigated in squamous cell carcinoma. The influence of PpIX-PAMAM on the generation of singlet oxygen after irradiation was also evaluated. Finally, the penetration of PpIX from the PpIX-PAMAM complexes was evaluated in vivo in healthy skin and in tumors induced in BalbC nude mice with and without application of iontophoresis. The average size of PpIX-PAMAM nanoparticles dispersed in aqueous medium was approximately 220 nm. When evaluated as a function of time, this size was increased only 5% after 24 h and remained constant for 7 days. The zeta potential of the dispersions was 10 mV at pH 7 and 30 mV at pH 5.5, allowing the contribution of electromigration during iontophoresis. In studies in culture tumor cells it was observed that complexation with PAMAM increased 30 times the localization of PpIX in the mitochondria compared to free PpIX. Furthermore, the amount of singlet oxygen generated when PpIX-PAMAM was irradiated was similar to that generated by the irradiation of the non-aggregated free PpIX, 4.6 x 10-3 and 4.3 x 10-3, respectively, suggesting that PAMAM did not modify the photodynamic activity of PpIX. In vivo experiments on healthy skin have shown that PpIX from the PpIX-PAMAM was homogeneously distributed throughout the skin, whereas free PpIX fluorescence was visualized only in some restricted areas of the skin surface. Iontophoresis facilitated PpIX diffusion to deep layers of the skin. Finally, the treatment of skin tumors have shown that the topical administration of the PpIX-PAMAM for only 30 min, passively or by iontophoresis, allowed the penetration of PpIX into the tumors located below the skin. Therefore, the PpIX complexation with PAMAM is a promising nanoparticle delivery system for the topical treatment of skin tumors by PDT. (AU)

FAPESP's process: 11/14940-0 - Influence of dendrimers and iontophoresis on the tumor penetration of Protoporphyrin IX in the treatment of skin tumors by photodynamic therapy
Grantee:Laura Freire Cardoso Pimenta
Support type: Scholarships in Brazil - Master