BCc1, the novel antineoplastic nanocomplex, showed potent anticancer effects in vitro and in vivo

Figures & data

Figure 1 (A) SEM image and (B) HRTEM image of BCc1.

Note: Scale of A is 500 nm and two yellow particles are 44 nm.
Abbreviations: SEM, scanning electron microscopy; HRTEM, high-resolution transmission electron microscopy.

Figure 2 Infrared spectrum analysis (A) and complexometric titration (B)

Table 1 IC₅₀ values in neoplastic and normal cells

	MEFs	MCF-7
24 hours	1,179±105	869±85
48 hours	705±50	493±75

Notes: Each IC₅₀ value (given in μg/mL) is the mean ± SEM of at least three experiments performed. The cells were incubated in the presence of different concentrations of BCc1 for 24 hours or 48 hours at 37°C. After this incubation period, cellular proliferation was determined by the MTT assay.

Abbreviations: IC₅₀, half maximal inhibitory concentration; MEFs, mouse embryonic fibroblasts; SEM, standard error of the mean.

Figure 3 BCc1 cell toxicity in malignant and normal cells after 24 hours (A) and 48 hours (B).

Note: Significant differences in the control cells have been specified by a single asterisk (*), meaning P-value ≤0.05.
Abbreviation: MEFs, mice embryonic fibroblasts.

Figure 4 The protection of MEF cells from H₂O₂-induced oxidative toxicity by BCc1.

Notes: Significant differences in the H₂O₂-treated cells have been specified by a single asterisk (*), meaning P-value ≤0.05. Significant differences in the control cells have been specified by a single hash (#), meaning P-value ≤0.05.
Abbreviation: MEF, mice embryonic fibroblast.

Table 2 BCc1 toxicity report

Group	Dose (mg/kg), IP injection	No of animals died after 24 hours	No of animals died after 48 hours	No of animals died after 72 hours	Total dead mice
1	2,000	6	0	0	6
2	1,000	6	0	0	6
3	500	6	0	0	6
4	300	6	0	0	6
5	200	6	0	0	6
6	100	2	0	0	2
7	80	1	0	0	1
8	60	0	0	0	0
9	50	0	0	0	0

Note: Intraperitoneal LD50 (mouse) – 109.17 mg/kg.

Abbreviation: IP, intraperitoneal.

Figure 5 Expression of cell markers CD44 and CD24 in MCF-7 cell line.

Note: The histogram outlined by a black line represents positive staining for CD44 and CD24, and color-filled histograms show negative control stained with matched isotype antibody.

Figure 6 Flow cytometry analysis in the control and test after annexin and PI staining.

Abbreviations: PI, propidium iodide; FITC, fluorescein isothiocyanate.

Figure 7 Flow cytometry analysis in the control and test for cell cycle analysis.

Figure 8 BCc1 tumor suppressing effect (A). Ratio of tumor size on the day 23 in relation to day 1 in the control and test groups (B).

Notes: The tumor size from each group at the end of the experiment (day 23) was divided over the tumor size from each group in the beginning of the treatment (day 1) to demonstrate the efficacy of the nanocomplex in controlling tumor growth by using mathematical logic.

Figure 9 Survival diagrams of the control and test groups.

Figure 10 The average amounts for Hb concentration (A), RBC count (B), HCT (C), and MCV (D).

Abbreviations: Hb, hemoglobin; RBC, red blood cell; HCT, hematocrit; MCV, mean cell volume.

Figure 11 Hematoxylin and eosin staining of (A) normal lung tissue, (B) clumps of metastatic tumor cells, (C) normal liver tissue, (D) clumps of metastatic cells, and (E) the megakaryocyte within the liver sinusoids.