A novel albumin wrapped nanosuspension of meloxicam to improve inflammation-targeting effects

Figures & data

Figure 1 Appearance of MX-BSA-NS before lyophilization (A), after lyophilization (B), and after redispersion (C).

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure 2 Particle size and distribution of MX-BSA-NS before lyophilization (A) and after lyophilization (B).

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure 3 Transmission electron microscopy morphology of MX-BSA-NS.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure 4 FTIR spectrum of BSA, MX-BSA-NS, and MX mixture (MX, mannitol).

Abbreviations: BSA, bovine serum albumin; FTIR, Fourier transform infrared; MX, meloxicam; NS, nanosuspension.

Figure 5 X-ray powder diffraction patterns of (A) raw MX, (B) dried MX nanosuspensions, (C) BSA, (D) mannitol, and (E) physical mixture.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam.

Figure 6 In vitro release profile of MX from different vehicles at pH 7.4.

Note: The data are shown as mean ± standard deviation (SD).

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Table 1 Stability of MX-BSA-NS after lyophilization under 4°C

Batches	Time (months)	Appearance	Content (%)	PS (nm)	PI	Zeta potential (mV)
20160826	0	Yellow bulk powder	99.7	90.43	0.155	−8.7
	1	Yellow bulk powder	100.2	91.37	0.136	−8.8
	2	Yellow bulk powder	99.5	91.98	0.147	−8.5
	3	Yellow bulk powder	98.9	92.29	0.158	−9.0
	6	Yellow bulk powder	98.6	92.47	0.154	−8.7
20160827	0	Yellow bulk powder	99.2	89.75	0.147	−8.9
	1	Yellow bulk powder	98.9	90.12	0.156	−8.8
	2	Yellow bulk powder	98.7	90.55	0.149	−8.6
	3	Yellow bulk powder	99.2	91.18	0.153	−8.3
	6	Yellow bulk powder	98.3	90.82	0.166	−8.5
20160828	0	Yellow bulk powder	99.4	90.91	0.139	−8.5
	1	Yellow bulk powder	99.1	90.49	0.148	−9.2
	2	Yellow bulk powder	98.9	91.38	0.154	−8.8
	3	Yellow bulk powder	99.1	92.74	0.145	−8.4
	6	Yellow bulk powder	98.2	91.89	0.143	−8.7

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension; PI, polydispersity index; PS, particle size.

Figure 7 Cell viability of RAW264.7 murine macrophage cells assessed by MTT assay after exposure to different concentrations of MX (0–120 μg mL⁻¹) in different vehicles for 24 h.

Note: The data are shown as mean±SD of three independent experiments.

Abbreviations: BSA, bovine serum albumin; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide; MX, meloxicam; NS, nanosuspension.

Table 2 Main pharmacokinetic parameters of MX solution and MX-BSA-NS following intravenous injection in rabbits (mean±SD, n=6)

Preparation	t1/2 (h)	AUC0–∞ (ng L^−1 h)	Cl (L h^−1 kg^−1)	MRT (h)
MX solution	4.299±0.682	12,351.267±3,038.82	0.06±0.015	5.683±0.666
MX-BSA-NS	11.6±2.16	30,730.213±3,741.436	0.023±0.003	14.215±3.144

Note: t_1/2, half-life.

Abbreviations: AUC_0–∞, area under the concentration–time curve; Cl, clearance; MRT, mean residence time; BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure 8 Mean blood concentration–time curve of MX in rabbits after intravenous administration of MX solution and MX-BSA-NS.

Note: Each value represents the mean±SD, n=6.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure 9 Mean concentration of MX in rats’ paws after intravenous administration of MX solution and MX-BSA-NS.

Note: *P<0.05 compared to MX solution group.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure 10 Effects of meloxicam (at an equivalent dose of 25 mg kg⁻¹) on carrageenan-induced paw edema in rats.

Notes: Data are the average values of six experiments (±SD). *P<0.05 compared to control group.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; NS, nanosuspension.

Figure S1 The collapsed (A) and smooth and compact (B) appearances of freeze-dried products containing different cryoprotectants.

Figure S2 Appearance of MX nanosuspension without BSA coating before (A) and after (B) 24 h of placement.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam.

Table S1 Summary of the experiments performed

Experiment	Content of Tween-80 (w/v) (%)	MX:BSA	Content of MX (mg mL^−1)	Particle size (nm)	Particle size after 24 h (nm)	Stability time (at 4°C)
1	0.1	1:5	5	255.5	592.3	ND
2	0.2	1:5	5	179.4	445.6	ND
3	0.3	1:5	5	78.42	78.61	>15 days
4	0.4	1:5	5	71.5	187.9	ND
5	0.3	1:1	5	90.12	Precipitation appeared	<1 day
6	0.3	1:3	5	87.82	114.37	3 days
7	0.3	1:10	5	108.17	139.25	3 days
8	0.3	1:7	5	85.81	90.49	7 days
9	0.3	1:5	2.5	92.35	ND	>15 days
10	0.3	1:5	7.5	59.21	ND	<3 days

Note: Stability time: the time that precipitation did not appear.

Abbreviations: BSA, bovine serum albumin; MX, meloxicam; ND, not determined.

Table S2 Solubility of MX in different media (37°C±0.5°C)

Medium	Solubility (mg mL^−1)
Distilled water	0.0079
0.1 mol L^−1 HCl	0.0013
0.01 mol L^−1 HCl	0.0019
pH 4.0 PBS	0.0034
pH 5.0 PBS	0.0055
pH 5.8 PBS	0.0183
pH 6.5 PBS	0.1630
pH 7.4 PBS	0.6403
pH 8.5 PBS	1.5502
10^−4 mol L^−1 NaOH	2.0568
10^−3 mol L^−1 NaOH	3.1594
0.01 mol L^−1 NaOH	3.6188
0.1 mol L^−1 NaOH	32.4790
0.4 mol L^−1 NaOH	5.4864

Abbreviation: MX, meloxicam.

Table S3 The appearance of formulations containing different cryoprotectants

Cryoprotectants	3% (w/v)	5% (w/v)	7% (w/v)	10% (w/v)
None	+	+	+	+
Glucose	−	−	−	−
Sucrose	−	−	−	−
Lactose	−	−	−	+
Trehalose	−	−	−	−
Mannitol	+	+	+	+
Maltose	−	−	−	−

Notes: +, smooth and compact; −, collapse.

Table S4 Reconstitution of various cryoprotectants on the character of the freeze-dried product

Cryoprotectants	3% (w/v)	5% (w/v)	7% (w/v)	10% (w/v)
None	+	+	+	+
Glucose	−	−	−	−
Sucrose	−	−	−	−
Lactose	−	−	−	−
Trehalose	+	−	−	−
Mannitol	+	++	++	+
Maltose	−	−	−	−

Notes: ++, translucent; +, opalescent; −, milky or sedimentary.