Preparation and ocular pharmacokinetics of hyaluronan acid-modified mucoadhesive liposomes

Figures & data

Table 1. Effect of the phospholipid/cholestenone ratio on physicochemical properties of DOX-loaded liposomes.

Sample codes	Phospholipid/ cholestenone ratio	Particle size (nm)	PDI	EE (%)
S1	3:1	517.3 ± 90.1	0.622 ± 0.05	8.15 ± 1.6
S2	4:1	204.1 ± 8.2	0.329 ± 0.04	58.7 ± 2.3
S3	5:1	134.6 ± 5.5	0.194 ± 0.02	84.9 ± 1.9
S4	6:1	137.1 ± 7.4	0.225 ± 0.04	86.5 ± 2.4

Data are the mean of three determinations ± SE. PDI, polydispersity index; EE, entrapment efficiency (%).

Figure 1. Zeta potential of blank liposomes (A), SA-added liposomes (B), and HA-modified liposomes (C).

Figure 2. TEM morphology of DOX-loaded liposome.

Figure 3. In vitro release profile of DOX solution and liposomes in PBS solution (pH = 7.4) at 37 °C (n = 3, mean ± SD).

Figure 4. Cytotoxicity of HA-modified liposomes in human epithilium cells.

Figure 5. CLSM of DOX uptakes by human corneal epithelia cell after incubated alternately with 40 PPM (A1–A3) free DOX group, (B1–B3) non-modified liposome group, and (C1–C3) HA-modified liposome group after (A1, B1, and C1) 0.5 h, (A2, B2, and C2) 3 h, and (A3, B3, and C3) 6 h. (Up) blue filter; (middle) red filter; (down) overlay of both (up and middle) sections.

Figure 6. In vitro transcorneal permeation of DOX liposome preparation and solution (X ± SD, n = 3).

Figure 7. Kinetics of DOX disappearance from tear fluid following topical administration of the liposome and the control solution (X ± SD, n = 4).

Figure 8. Concentration–time profles of DOX in aqueous humor after instillation of 0.5 mg/mL DOX liposome preparation and DOX solution in rabbits (ng/mL, X ± SD, n = 4).

Table 2. Pharmacokinetic parameters of DOX in the aqueous humor of rabbits (n = 4).

Parameters	HA-LP	NM-LP	Free DOX
Tmax (min)	48.75 ± 12.43	27.5 ± 14.36	33.75 ± 6.49
Cmax (ng·mL^−1)	525.46 ± 204.47	409.95 ± 17.54	387.58 ± 201.18
t1/2 (min)	389.70 ± 61.07	182.79 ± 81.86	114.96 ± 42.10
AUC (ng mL^−1min^−1)	61 155 ± 29 366.76	56 217.05 ± 392.87	36 462.6 ± 14 925.04
Cl/F	4.39 ± 4.52	9.65 ± 1.46	10.60 ± 14.09
MRT (min)	527.11 ± 604.89	211.45 ± 52.04	152.73 ± 3.72

HA-LP indicates HA-modified liposome group; NM-LP, non-modified liposome group; DOX, doxorubicin; AUC, area under the curve; Cl/F, clearance rate; MRT, mean retention time.

Figure 9. CLSM of DOX absorption by corneal tissues after rabbits’ eyes in vivo treated alternately with free DOX (A1–A3), (B1–B3) non-modified liposome, and (C1–C3) HA-modified liposome after (A1, B1, and C1) 0.5 h, (A2, B2, and C2) 2 h, and (A3, B3, and C3) 3 h. (Up) blue filter; (middle) red filter; (down) overlay of both (up and middle) sections.