Enhancement of Pancreatic Cancer Therapy Efficacy by Type-1 Matrix Metalloproteinase-Functionalized Nanoparticles for the Selective Delivery of Gemcitabine and Erlotinib

Figures & data

Figure 1 Schematic diagram and TEM image of M-M GEM/ERL NPs. M-M GEM/ERL NPs was synthesized by conjugating MT1-MMP to the surface of GEM/ERL NPs and exhibited uniform spherical morphology.

Table 1 Characterization of NPs

Formulation	GEM/ERL NPs	GEM NPs	ERL NPs	M-M GEM/ERL NPs	Blank M-M NPs
Particle size (nm)	123.51 ± 4.66	120.93 ± 5.01	125.4 ± 4.37	167.82 ± 5.34*	165.43 ± 5.27
Polydispersity index	0.16 ± 0.03	0.15 ± 0.02	0.14 ± 0.02	0.20 ± 0.03	0.19 ± 0.03
Zeta potential (mV)	14.21 ± 1.92	13.17 ± 2.06	15.32 ± 1.89	20.13 ± 2.24*	19.85 ± 2.17
GEM DL (%)	5.15 ± 0.48	5.29 ± 0.43	/	4.97 ± 0.52	/
ERL DL (%)	5.37 ± 0.61	/	5.21 ± 0.55	4.88 ± 0.42	/
GEM EE (%)	81.31 ± 2.98	80.93 ± 2.37	/	83.14 ± 3.32	/
ERL EE (%)	79.64 ± 2.81	/	81.13 ± 2.59	80.35 ± 3.57	/

Notes: Data are presented as Mean ± SD, n = 10; *P < 0.05 by Student’s t-test between M-M GEM/ERL NPs and GEM/ERL NPs.

Figure 2 Plasma stability of NPs characterized by the changes of size (A), GEM EE (B), and ERL EE (C). During the 72 h of stability study, the particle sizes, GEM and ERL EEs of nanocarriers exhibited negligible changes. Data are presented as mean ± standard deviation (n=10).

Figure 3 In vitro GEM (A) and ERL (B) release profiles. M-M GEM/ERL NPs showed slower release than GEM/ERL NPs. The drug release behaviors of single drug encapsulated GEM NPs and ERL NPs were similar with dual drugs loaded GEM/ERL NPs. Data are presented as mean ± standard deviation (n=10).

Figure 4 Cellular uptake efficiency of M-M GEM/ERL NPs and GEM/ERL NPs: cell images (A) and flow cytometry (B). *P < 0.05 compared with GEM/ERL NPs. MT1-MMP decorated M-M GEM/ERL NPs exhibited higher uptake ability (67.65 ± 2.87%) than GEM/ERL NPs (43.17 ± 2.37%). Data are presented as mean ± standard deviation (n=10).

Figure 5 In vitro cell viabilities of NPs evaluated on two kinds of pancreatic tumor cell lines: PANC-1 (A) and AsPC-1 cells (B). M-M GEM/ERL NPs illustrated the most remarkable tumor cell inhibition ability on both cell lines. GEM/ERL NPs exhibited significantly higher cell inhibition efficiency than free GEM/ERL. Single drug-loaded GEM NPs and ERL NPs showed higher cell viability than their double drugs GEM/ERL NPs counterparts. Data are presented as mean ± standard deviation (n=10). *P < 0.05.

Table 2 Pharmacokinetics Data

Drug	Formulation	GEM/ERL NPs	M-M GEM/ERL NPs	Free GEM/ERL
GEM	T1/2 (h)	8.31 ± 0.92*	11.51 ± 1.23*	1.29 ± 0.34
	Cmax (L/kg/h)	23.94 ± 2.13	25.17 ± 2.66	20.68 ± 1.95
	AUC (mg/L h)	86.74 ± 3.33*	106.92 ± 4.64*	39.45 ± 2.37
	Vd (L/kg)	3.31 ± 0.51	4.12 ± 0.44	1.21 ± 0.36
	CL (L/h)	0.20 ± 0.03	0.34 ± 0.05	0.65 ± 0.04
ERL	T1/2 (h)	9.63 ± 0.86*	12.41 ± 1.53*	1.91 ± 0.78
	Cmax (L/kg/h)	17.65 ± 1.83	19.32 ± 1.49	18.32 ± 1.73
	AUC (mg/L h)	76.95 ± 4.68*	113.65 ± 7.14*	29.53 ± 3.16*
	Vd (L/kg)	3.81 ± 0.41	4.98 ± 0.47	1.39 ± 0.33
	CL (L/h)	0.27 ± 0.05	0.28 ± 0.04	0.50 ± 0.06

Notes: Data are presented as Mean ± SD, n = 10; *P < 0.05 by Student’s t-test between M-M GEM/ERL NPs and Free GEM/ERL, or GEM/ERL NPs and Free GEM/ERL.

Table 3 BUN, AST, and ALT Measurement

Formulation	BUN (mg/dl)	AST (U/L)	ALT (U/L)
GEM/ERL NPs	29.51 ± 1.88	121.6 ± 8.63	75.42 ± 7.32
GEM NPs	31.17 ± 2.15	126.5 ± 9.14	73.13 ± 6.56
ERL NPs	28.76 ± 1.65	130.4 ± 11.16	70.39 ± 8.64
M-M GEM/ERL NPs	27.65 ± 2.77	128.3 ± 7.61	76.34 ± 10.63
Blank M-M NPs	26.88 ± 2.01	120.7 ± 6.59	79.85 ± 9.35
Free GEM/ERL	42.74 ± 2.35 *	136.9 ± 17.32	80.23 ± 12.31

Notes: Data are presented as Mean ± SD, n = 10; *P < 0.05 by Student’s t-test between Free GEM/ERL and M-M GEM/ERL NPs.

Figure 6 In vivo tumor growth curves (A) and tumor inhibition efficiencies (B). M-M GEM/ERL NPs group showed the most efficient tumor growth inhibition, and GEM/ERL NPs also suppressed tumor growth effectively compared to GEM NPs, ERL NPs and free GEM/ERL. Data are presented as mean ± standard deviation (n=10). *P < 0.05.