154
10. Add the suspended microspheres to the aqueous solution of
cellulose.
11. Leave the mixture under stirring at room temperature for
24 h.
12. Centrifuge the emulsion (see Note 9).
Forming of the third layer by chitosan deposition (CS) (see
Note 10):
13. Dissolve 0.05 g of chitosan in 25 mL aqueous acidic solution
(see Note 5).
14. Add and disperse 0.4 g of modified spheres in 50 mL of basic
aqueous solution (see Note 6).
15. Add the dispersion to the aqueous solution of chitosan.
16. Leave the mixture under continuous stirring for 24 h at room
temperature.
17. Centrifuge the emulsion (see Note 9).
Aiming at cross-linking the three layers are deposited on vehicle’s surface:
18. Disperse 0.4 g of coated PMMA@HPC@CS@CH spheres in
2 mL water.
19. Add aqua solution of EDC (10  mg, 0.06  mmol) and NHS
(5 mg, 0.04 mmol) in the resulting emulsion.
20. Leave the mixture for 30 min under vigorous stirring.
21. Centrifuge the final mixture (see Note 9) and purify after suspended in water.
MMA Core
MMA@HPC
CS
HPC
MMA
HPC
CHIT
CHIT
EDC/NHS
DAUNORUBICIN
CS
cro cros ss lin link k
Fig. 1 Synthetic scheme of the present study
Aikaterini-Foteini Metaxa et al.