The Effects of a Novel Curcumin Derivative Loaded Long-Circulating Solid Lipid Nanoparticle on the MHCC-97H Liver Cancer Cells and Pharmacokinetic Behavior

Figures & data

Figure 1 The structure of CU (A) and CU1 (B).

Table 1 The Factors and Levels of Orthogonal Design

Levels	Factors
	A (CU1, mg)	B (HSPC, mg)	C (DSPE-PEG2000, mg)
1	7.0	35.0	14.0
2	6.0	30.0	12.0
3	5.0	25.0	10.0

Table 2 The Design and Results of Orthogonal Table

No.	Levels			EE (%)	DL (%)	X
	A	B	C
1	2	1	2	97.52	5.32	96.23
2	3	1	3	94.90	5.49	94.01
3	1	3	2	91.85	5.83	91.60
4	2	3	3	93.75	5.15	92.56
5	1	1	1	92.70	8.41	95.40
6	2	2	1	92.15	4.71	90.57
7	1	2	3	87.63	8.54	90.88
8	3	3	1	88.11	4.91	87.07
9	3	2	2	89.29	4.34	87.49
K1/3	92.60	95.21	91.01
K2/3	93.12	89.65	91.77
K3/3	89.52	90.41	92.48
R	3.60	5.56	1.76

Abbreviations: EE, encapsulation efficiency of CU1-LSLN; DL, drug loading of CU1-LSLN; X, comprehensive score; R, the range analysis.

Table 3 Variance Analysis Results

No.	Sum of Squares	Free Degree	Mean Square	F value	P value
A	22.81	2	11.41	33.40	0.03
B	54.64	2	27.32	80.02	0.01
C	3.24	2	1.62	4.75	0.17
Errors	0.68	2	0.34

Figure 2 The particle size (A) and Zeta potential (B) of optimized CU1-LSLN.

Figure 3 XRD (A), DSC (B), and FTIR (C) patterns.

Figure 4 The results of stability test and in vitro release study. (A) The changes of particle size, PDI, DL of CU1-LSLN during the 180 days. (B) In vitro release profile of CU1 and CU1-LSLN.

Table 4 Drug Release Kinetic Parameters and Fitting Coefficients

	Zero-Order Equation		First-Order Equation		Higuchi’s Equation
	r^2	k0	r^2	k1	r^2	kh
CU1	0.7490	3.3640	0.9543	0.2374	0.9226	19.6434
CU1-LSLN	0.7355	0.8408	0.9922	0.0734	0.9180	9.3871

Abbreviations: r², fitting coefficient; K₀, K₁, and K_h are the rate constants for the zero-order, first-order, and Higuchi equations, respectively.

Figure 5 The uptake of CU1-LSLN by MHCC-97H cells. (A) Histogram showing uptake of CU, CU1 and CU1-LSLN by MHCC-97H cells using flow cytometry. (B) The mean fluorescence intensity of MHCC-97H cells after treatment with CU, CU1 and CU1-LSLN for 1 h and 3 h, respectively, *P < 0.05.

Figure 6 The effects of CU1-LSLN on MHCC-97H cells proliferation compared to CU and CU1. *P < 0.05 vs CU; ^#P < 0.05 vs CU1.

Figure 7 The effect of CU1-LSLN on apoptosis induction. The MHCC-97H cells (2×10⁵ cells/well) were treated with CU, CU1 and CU1-LSLN (5, 10, 20 μmol/L) then were analyzed by Annexin V-PI staining flow cytometry. *P < 0.05 vs CU; ^#P < 0.05 vs.CU1.

Figure 8 The effect of CU1-LSLN on cell migration and invasion in MHCC-97H cells compared to CU and CU1. (A) Representative images of migration cells stained with crystal violet. Columns, the mean migration rate (%) from three different experiments with three duplicates. (B) Representative images of invasive cells stained with crystal violet. Columns, the mean invasion rate (%) from three different experiments with three duplicates. *P < 0.05 vs CU; ^#P < 0.05 vs.CU1.

Figure 9 Expression of cells by Western blot (A), and quantitative analysis (B–F). * vs control P < 0.05; ** vs CU P < 0.05.

Table 5 The Main Pharmacokinetic Parameters After Given CU, CU1, or CU1-LSLN by Tail Vein Injection

Parameter	Unit	CU	CU1	CU1-LSLN
AUC(0-t)	mg/L*min	2.51	2.04	175.43
MRT(0-t)	min	142.89	49.14	632.31
t1/2z	min	160.86	40.62	1642.76
Tmax	min	10	10	15
Vz	L/kg	1268.93	426.65	96.57
CLz	L/min/kg	5.47	7.28	0.041
Cmax	mg/L	0.043	0.10	0.20

Abbreviations: AUC, area under the curve; MRT, mean residence time; t_1/2z, half-life; T_max, time of maximum concentration; V_z, apparent volume of distribution; CL_z, clearance rate; C_max, maximum concentration.

Figure 10 (A) Fluorescence images of organs at 1 h, 2 h, and 24 h after CU1-LSLN was injected into tail vein. (B) H&E staining of liver.