Ganoderic acid loaded nano-lipidic carriers improvise treatment of hepatocellular carcinoma

Figures & data

Table 1. MTT viability assay of GA solution and optimized GA-NLC.

Formulations	IC50 (µg/mL)
	24 h	48 h
GA solution	48.123	40.012
GA-NLC	40.121	32.612

IC₅₀: half maximal inhibitory concentration.

The GA-NLC shows significant (p<.05) cytotoxicity compared to GA solution after 24 and 48 h. Data are expressed as mean ± SD (n = 6).

Figure 1. Uptake of (A) free coumarin-6 (C-6) and (B) C-6 loaded GA-nano-lipid carrier, upon incubation at 1 µg/mL for 2 h. In all the images, (a) images under the green fluorescence channel; (b) corresponding differential interface contrast images of HepG2 cells. (c) Superimposition of figure (a) and figure (b). (d, e) In all, the images show horizontal line series analysis of fluorescence along the white line.

Figure 2. Macroscopical observation of DEN-induced HCC group rats. (A) DEN control group showed the expansion of hepatic nodules (white in color) and decolorization of tissue, (B) DEN control group treated with GA (25 mg/kg) illustrated the expansion of pre-cancerous nodules (white in color) and decolorization of tissue color, which was less as compared to DEN group, (C) DEN control group treated with GA (50 mg/kg) demonstrated the less pre-cancerous nodules (white in color) and decolorization of tissue color, (D) DEN control group treated with GA (100 mg/kg) illustrated the expansion of pre-cancerous nodules and decolorization of tissue color, which was less as compared to other group, and (E) DEN control group treated with GA-NLC (25 mg/kg) illustrated the decolorization of tissue color, which was less as compared to other group rats. Note: Normal control and normal control rats treated with GA (100 mg/kg) did not show any sign of precancerous cells and decolorization of skin (data not shown).

Table 2. The effect of GA-NLC on the number of rats, number of nodules and average number of nodules bearing rats.

S. no.	Groups	Number of rats with nodules/number of rats	Total number of nodules	Relative size (% of number size)
				≤1 mm	<3 mm >1 mm	≥3 mm
1	DEN control	9/9	202	101 (50)	54 (26.73)	47 (23.26)
2	DEN + GA (25 mg/kg)	10/11	184	94 (51.06)	46 (25)	44 (23.91)
3	DEN + GA (50 mg/kg)	7/10	112	54 (48.21)	35 (31.25)	23 (20.53)
4	DEN + GA (100 mg/kg)	4/9	45	22 (48.88)	14 (31.11)	9 (20)
5	DEN + GA-NLC	2/9	21	9 (42.85)	7 (33.33)	5 (23.80)

Group I and group II did not show any sign of hepatic nodules.

Figure 3. The histopathological evaluation of DEN-induced HCC group rats. (A) DEN control group showed the expansion of hepatic nodules (white in color) and decolorization of tissue, (B) DEN control group treated with GA (25 mg/kg) illustrated the expansion of pre-cancerous nodules (white in color) and decolorization of tissue color, which was less as compared to DEN group, (C) DEN control group treated with GA (50 mg/kg) demonstrated the less pre-cancerous nodules (white in color) and decolorization of tissue color, (D) DEN control group treated with GA (100 mg/kg) illustrated the expansion of pre-cancerous nodules and decolorization of tissue color, which was less as compared to other group, and (E) DEN control group treated with GA-NLC (25 mg/kg) illustrated the decolorization of tissue color, which was less as compared to other group rats. Note: Normal control and normal control rats treated with GA (100 mg/kg) did not show any sign of alteration in the histopathology (data not shown).

Table 3. The effect of GA-NLC on the histopathology character.

S. no.	Histopathology changes	Groups
		NC	NC + GA (50 mg/kg)	DEN	DEN + GA (25 mg/kg)	DEN + GA (100 mg/kg)	DEN + GA-NLC
1	Necrosis	–	–	+	+	+	+
2	Hydropic degeneration	–	–	+	+	+	–
3	HSCs focal proliferation	–	–	+	+	+	+
4	Bile cysts	–	–	+	–	–	–
5	Pseudo-nucleoli	–	–	+	+	+	–
6	Peliosis hepatis	–	–	+	+	–	–
7	Disorganized hepatic parenchyma	–	–	+	–	–	–
8	Apoptosis	–	–	+	+	–	–
9	Hepatocellular adenoma	–	–	+	+	+	+
10	Cell necrosis	–	–	+	+	–	–
11	Altered basophilic	–	–	+	+	+	+
12	Small dark cytoplasm	–	–	+	–	–	–
13	Enlargement of karyomegaly	–	–	+	+	–	–
14	Macro lipid droplets	–	–	+	+	+	+
15	Diffuse dysplasia	–	–	+	+	–	–
16	Hyperplastic foci	–	–	+	+	–	–