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Preparation of temoporfin-loaded polyplex nanoparticles<br>As schematically presented in Fig. 1, preparations of<br>temoporfin-loaded polyplex nanoformulations were obtained<br>by multiple electrostatic interactions between cationic chitosan and anionic alginate with non-covalently encapsulated<br>temoporfin.<br>Briefly, 1 mL of temoporfin solution (1 mg/1 mL in methanol) was added dropwise above the surface of 7.5 mL of Algstock solution (pH ∼12) under magnetic stirring at room temperature. After that, 2.5 mL of chitosan (Chit or ChitOL, respectively) stock solution (pH ∼5) was added dropwise above<br>the surface of temoporfin-containing Alg solution under magnetic stirring. The particles were left stirring for 1 day. Prior to<br>analysis, samples were centrifuged (3 min/4000 rpm), and<br>1 mL of supernatant was removed and further analyzed.
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