Statistička analiza nanokapsuliranja niskomolekularnog heparina

The objective of this study was to use Box-Behnken design (BBD) to investigate the influence of formulation variables on the properties of heparin-loaded poly(lactic-co-glycolic acid) (PLGA)-Eudragit-RLPO (E-RLPO) nanoparticles (NP) in terms of mean diameter (as size) and drug encapsulation efficiency. The NPs were prepared by the double emulsion solvent evaporation method. The independent variables were: X1 olymer mass ratio (PLGA:E-RLPO) in the oil phase, X2 concentration of polyvinyl alcohol (PVA) as emulsion stabilizer, and X3 volume of the external aqueous phase (W2). Particle size (analyzed by dynamic light scattering) and encapsulation efficiency (EE, estimated by spectrophotometry) were the investigated responses. The polynomial equation obtained from regression analysis of the reduced model (p = 0.0002, F = 25.7952 and R2 = 0.96) provided an excellent fit. The optimal size for the NP was found to be 134.2 ± 16.5 nm with formulation variables of 48.2:61.8, 0.321 (%, m/V) and 263 mL for X1, X2 and X3, respectively. Probably, due to electrostatic interaction between the negatively charged drug and the positively charged E-RLPO, the percent EE of heparin was between 74.4 ± 6.5 % (lowest value) and 92.1 ± 5.3 % (highest value). The data suggest that BBD is a useful tool in rational design of heparin-loaded NPs.Box-Behnkenovo dizajniranje (BBD) primijenjeno je za praćenje utjecaja formulacijskih varijabli na svojstva nanočestica (NP) s heparinom. Za izradu nanočestica korišten je kopolimer mliječne i glikolne kiseline (PLGA) i Eudragit-RLPO (E-RLPO). Nanočestice su pripravljene metodom dvostruke evaporacije otapala iz emulzije. Nezavisne varijable bile su: X1 omjer masa polimera (PLGA : E-RLPO) u uljnoj fazi, X2 koncentracija polivinil alkohola (PVA) kao stabilizatora emulzije i X3 volumen vanjske vodene faze (W2). Zavisne varijable bile su veličina čestica (analizirana pomoću dinamičkog rasapa svjetlosti) i učinkovitost inkapsuliranja (EE) (praćena spektrofotometrijski). Polinomska jednadžba dobivena regresijskom analizom reduciranog modela odlično je odgovarala (p = 0,0002, F = 25,7952 i R2 = 0,96). Optimalna veličina nanočestica bila je 134,2 ± 16,5 nm s formulacijskim varijablama 48,2:61,8, 0,321 (%, m/V) i 263 mL for X1, X2 odnosno X3. Vjerojatno je zbog elektrostatskih interakcija između negativno nabijene ljekovite tvari i pozitivno nabijenog E-RLPO učinkovitost inkapsuliranja heparina varirala od 74,4 ± 6,5 % (najniža vrijednost) do 92,1 ± 5,3 % (najviša vrijednost). Rezultati sugeriraju da je BBD vrlo korisno u racionalnom dizajniranju nanočestica s heparinom