Irinotecan and 5-fluorouracil-co-loaded, hyaluronic acid-modified layer-by-layer nanoparticles for targeted gastric carcinoma therapy

Figures & data

Figure 1 Scheme graph and TEM images of HA-CH-IRN/5-FU NPs and other NPs.

Notes: (A) Scheme graph of the structure of HA–CH–IRN/5-FU NPs. (B) TEM image of HA–CH–IRN/5-FU NPs. (C) TEM images of HA–CH–IRN/5-FU NPs (1), CH–IRN/5-FU NPs (2), IRN NPs (3), 5-FU NPs (4) and HA–CH NPs (5) (yellow scale bars represent 200 nm).
Abbreviations: HA, hyaluronic acid; CH, chitosan; IRN, irinotecan; 5-FU, 5-fluorouracil; NP, nanoparticle; PLGA, poly(d,l-lactide-co-glycolide); TEM, transmission electron microscopy.

Table 1 Characterization of NPs

Formulations	HA–CH–IRN/5-FU NPs	CH–IRN/5-FU NPs	IRN NPs	5-FU NPs	HA–CH NPs
Particle size (nm)	153.8±5.6	127.3±4.8	91.4±3.6	86.9±3.9	155.1±5.2
Size distribution (PDI)	0.187±0.021	0.164±0.018	0.118±0.012	0.147±0.016	0.184±0.025
Zeta potential (mV)	−13.7±1.9	+17.5±2.6	−16.9±2.2	+25.8±3.7	−15.2±2.4
EE of IRN (%)	94.5±2.3	93.6±2.1	92.8±2.8	N/A	N/A
EE of 5-FU (%)	91.4±3.1	92.3±2.9	N/A	93.4±3.2	N/A

Abbreviations: NPs, nanoparticles; HA, hyaluronic acid; CH, chitosan; IRN, irinotecan; 5-FU, 5-fluorouracil; PDI, polydispersity index; EE, entrapment efficiency; N/A, not applicable.

Figure 2 Changes in size in the presence of serum.

Note: Data are presented as mean ± SD (n=3).
Abbreviations: HA, hyaluronic acid; CH, chitosan; IRN, irinotecan; 5-FU, 5-fluorouracil; NPs, nanoparticles.

Figure 3 In vitro release of IRN (A) and 5-FU (B) from NPs.

Note: Data are presented as mean ± SD (n=3).
Abbreviations: IRN, irinotecan; 5-FU, 5-fluorouracil; NPs, nanoparticles; HA, hyaluronic acid; CH, chitosan.

Figure 4 Cellular uptake efficiency of the NPs.

Note: Data are presented as mean ± SD (n=3).
Abbreviations: NPs, nanoparticles; HA, hyaluronic acid; CH, chitosan; IRN, irinotecan; 5-FU, 5-fluorouracil.

Figure 5 In vitro cytotoxicity of IRN and/or 5-FU-contained solutions or NPs investigated in MGC803 cells.

Note: Data are presented as mean ± SD (n=3).
Abbreviations: IRN, irinotecan; 5-FU, 5-fluorouracil; NPs, nanoparticles; HA, hyaluronic acid; CH, chitosan.

Table 2 Synergistic effect evaluation and selection of the ratio of drugs by CI calculation

Formulations	IRN/5-FU ratio (w/w)	IC50 IRN (μM)	IC50 5-FU (μM)	CI50
IRN NPs	N/A	0.261	N/A	N/A
5-FU NPs	N/A	N/A	0.123	N/A
CH–IRN/5-FU NPs	4/1	0.242	0.061	1.423
CH–IRN/5-FU NPs	2/1	0.112	0.056	0.884
CH–IRN/5-FU NPs	1/1	0.096	0.096	1.148
HA–CH–IRN/5-FU NPs	2/1	0.066	0.033	0.521
IRN solution	N/A	1.961	N/A	N/A
5-FU solution	N/A	N/A	1.162	N/A
IRN/5-FU solution	2/1	0.837	0.419	0.787

Abbreviations: CI, combination index; IRN, irinotecan; 5-FU, 5-fluorouracil; IC₅₀, median inhibitory concentration; NPs, nanoparticles; N/A, not applicable; CH, chitosan; HA, hyaluronic acid.

Figure 6 In vivo tissue distribution of IRN (A) and 5-FU (B) investigated in GC-bearing mice model.

Note: Data are presented as mean ± SD (n=8).
Abbreviations: IRN, irinotecan; 5-FU, 5-fluorouracil; GC, gastric cancer; HA, hyaluronic acid; CH, chitosan; NPs, nanoparticles.

Figure 7 In vivo antitumor effect and system toxicity of the NPs evaluated in terms of tumor volume (A) and body weight changes (B). Note: Data are presented as mean ± SD (n=8).

Abbreviations: NPs, nanoparticles; HA, hyaluronic acid; CH, chitosan; IRN, irinotecan; 5-FU, 5-fluorouracil.