Inhaled pyrazinamide proliposome for targeting alveolar macrophages

Figures & data

Figure 1.  The SEM images of (A) spray dried mannitol (bar = 1 µm), (B) porous mannitol (bar = 1 µm), (C–G) PZA- proliposome formulation I to V (bar = 3 µm) and (H) PZA (bar = 300 µm).

Table 1.  Aerosolization properties, encapsulation and dynamic light scattering size measurement after reconstitution of PZA-proliposome formulations (mean ± SD, n = 3).

Formulation	Content uniformity (%)	MMAD (µm)	FPF (%)	Emitted dose (%)	Encapsulation (%)	Size measurement (nm)
I	97.8 ± 4.8	4.26 ± 0.06	29.0 ± 4.1	97.8 ± 1.0	44.6 ± 0.5	216 ± 11
II	108.6 ± 0.6	4.26 ± 0.04	22.5 ± 6.9	94.5 ± 1.0	44.4 ± 1.1	359 ± 13
III	94.8 ± 0.4	4.26 ± 0.04	19.4 ± 5.1	94.4 ± 7.6	44.0 ± 1.9	516 ± 20
IV	94.3 ± 0.4	4.29 ± 0.03	25.8 ± 1.2	89.9 ± 4.1	37.3 ± 2.6	449 ± 15
V	93.4 ± 0.8	4.39 ± 0.21	23.7 ± 2.4	80.2 ± 7.3	26.7 ± 2.4	382 ± 12

Figure 2.  The X-ray diffraction patterns of (A) porous mannitol, (B–F) PZA-proliposome formulation I to V (containing PZA 10, 20, 40, 60 and 80% w/w, respectively) and (G) PZA.

Table 2.  Differential scanning calorimeter data of porous mannitol, PZA-proliposome formulations and PZA.

Material or formulation	Peak (°C)	Onset (°C)	End (°C)	Peak area (J/g)
Porous mannitol	163.5	151.8	166.5	266.5
I	161.9	149.3	165.4	249.9
II	162.3	150.3	166.0	234.3
III	157.8	145.4	162.9	221.3
IV	152.3	143.4	158.9	208.3
V	151.9	144.1	154.3	104.6
PZA	187.3	182.7	189.3	200.5

Figure 3.  Differential scanning calorimeter thermogram of (A) porous mannitol, (B–F) PZA-proliposome formulation I to V (containing PZA 10, 20, 40, 60 and 80% w/w, respectively) and (G) PZA.

Figure 4.  FT-IR spectra of (A) porous mannitol, (B–F) PZA-proliposome formulation I to V (containing PZA 10, 20, 40, 60 and 80% w/w, respectively) and (G) PZA.

Figure 5.  Viability of Calu-3 (A), A549 (B) and NR8383 (C) cell lines after exposed with different concentrations of PZA (•), PZA-proliposome formulation I (▴) and PZA-proliposome formulation V (▪) (mean ± SD, n ≥ 6).

Figure 6.  The level of inflammatory cytokine (IL-1β (A) and TNF-α (B)) and nitric oxide (C) produced from NR8383 cell lines after exposure with different concentrations of PZA (•), PZA-proliposome formulation I (▴), PZA-proliposome formulation V (▪) and LPS from E. coli (○) for 24 h (mean ± SD, n ≥ 6).

Figure 7.  Phagocytosis of NR8383 cells incubated with reconstituted PZA-proliposome formulation I stained with Lumidot^® 640 at (A–C) 2 and (D–F) 30 min in different mode; (A and D) bright field, (B and E) fluorescence and (C and F) overlay (bar = 50 µm).

Figure 8.  Serum biochemical parameters ((A) BUN, (B) Cr, (C) ALT and (D) AST) of control (□) and PZA-proliposome (▪) treated male Wistar rats in repeated dose toxicity study for 28 days.