Comparative In Vitro Study: Assessing Phytochemical, Antioxidant, Antimicrobial, and Anticancer Properties of Vaccinium macrocarpon Aiton and Vaccinium oxycoccos L. Fruit Extracts

Abstract

The phytochemical diversity and potential health benefits of V. oxycoccos and V. macrocarpon fruits call for further scientific inquiry. Our study aimed to determine the phytochemical composition of extracts from these fruits and assess their antioxidant, antibacterial, and anticancer properties in vitro. It was found that the ethanolic extracts of V. oxycoccos and V. macrocarpon fruits, which contained more lipophilic compounds, had 2-14 times lower antioxidant activity compared to the dry aqueous extracts of cranberry fruit, which contained more hydrophilic compounds. All tested cranberry fruit extracts (OE, OW, ME, and MW) significantly inhibited the growth of bacterial strains S. aureus, S. epidermidis, E. coli, and K. pneumoniae in vitro compared to the control. Cytotoxic activity against the human prostate carcinoma PPC-1 cell line, human renal carcinoma cell line (CaKi-1), and human foreskin fibroblasts (HF) was determined using an MTT assay. Furthermore, the effect of the cranberry fruit extract samples on cell migration activity, cancer spheroid growth, and viability was examined. The ethanolic extract from V. macrocarpon fruits (ME) showed higher selectivity in inhibiting the viability of prostate and renal cancer cell lines compared to fibroblasts. It also effectively hindered the migration of these cancer cell lines. Additionally, the V. macrocarpon fruit extract (ME) demonstrated potent cytotoxicity against PPC-1 and CaKi-1 spheroids, significantly reducing the size of PPC-1 spheroids compared to the control. These findings suggest that cranberry fruit extracts, particularly the ethanolic extract from V. macrocarpon fruits, have promising potential as natural remedies for bacterial infections and cancer therapy.

Keywords: anticancer activity; antimicrobial activity; antioxidant activity; cranberry; spheroids.

Figure 1

Determination of the reducing (A) and antiradical (B) activity of dried cranberry fruit extracts. OE—dry V. oxycoccos fruit extract obtained by extraction with 96% (v/v) ethanol; OW—dry V. oxycoccos fruit extract obtained by extraction with hot water; ME—dry V. macrocarpon fruit extract obtained by extraction with 96% (v/v) ethanol; MW—dry V. macrocarpon fruit extract obtained by extraction with hot water. Statistically significant differences between the antioxidant activity of cranberry extracts are indicated by different letters (p < 0.05).

Figure 2

Determination of the antimicrobial activity of dried cranberry fruit extracts. OE—dry V. oxycoccos fruit extract obtained by extraction with 96% (v/v) ethanol; OW—dry V. oxycoccos fruit extract obtained by extraction with hot water; ME—dry V. macrocarpon fruit extract obtained by extraction with 96% (v/v) ethanol; MW—dry V. macrocarpon fruit extract obtained by extraction with hot water. Statistically significant differences between the antibacterial activity of cranberry extracts are indicated by different letters (p < 0.05).

Figure 3

The effect of cranberry extracts on cell line viability using the MTT method. PPC-1—prostate carcinoma cell line; CaKi-1—kidney carcinoma cell line; HF—human skin fibroblast cell line. OE—dry V. oxycoccos fruit extract obtained by extraction with 96% (v/v) ethanol; OW—dry V. oxycoccos fruit extract obtained by extraction with hot water; ME—dry V. macrocarpon fruit extract obtained by extraction with 96% (v/v) ethanol; MW—dry V. macrocarpon fruit extract obtained by extraction with hot water. Statistically significant differences between the cytotoxic activity of cranberry extracts are indicated by different marks (*/^#/⁺/**/^##/⁺⁺/***/^###/⁺⁺⁺ p < 0.05).

Figure 4

Study of the effect of cranberry extracts on cell migration activity using the ‘wound healing’ method. (A) Quantitative calculations of the ‘wound’ area (%) of CaKi-1 cell line monolayer. (B) Photos of the ‘wound’ area in the CaKi-1 monolayer after 12 and 24 h of incubation with cranberry extracts and control. (C) Quantitative calculations of the ‘wound’ area (%) of PPC-1 cell line monolayer. (D) Photos of the ‘wound’ area in the PPC-1 monolayer after 24,48 and 72 h of incubation with cranberry extracts and control. PPC-1—prostate carcinoma cell line; CaKi-1—kidney carcinoma cell line. OE—dry V. oxycoccos fruit extract obtained by extraction with 96% (v/v) ethanol; OW—dry V. oxycoccos fruit extract obtained by extraction with hot water; ME—dry V. macrocarpon fruit extract obtained by extraction with 96% (v/v) ethanol; MW—dry V. macrocarpon fruit extract obtained by extraction with hot water. Statistically significant differences between the migration activity of cranberry extracts are indicated by marks (* p < 0.05).

Figure 5

The effect of cranberry extracts on spheroid diameter and viability using the magnetic 3D bioprinting method. (A) CaKi-1 spheroid size and viability of cells in spheroids at the end of the experiment. (B) PPC-1 spheroid size and viability of cells in spheroids at the end of the experiment. (C) Photos of CaKi-1 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment. (D) Photos of PPC-1 tumor spheroids at the beginning (Day 0), in the middle (Day 4), and at the end (Day 8) of the experiment. PPC-1—prostate carcinoma cell line; CaKi-1—kidney carcinoma cell line. OE—dry V. oxycoccos fruit extract obtained by extraction with 96% (v/v) ethanol; OW—dry V. oxycoccos fruit extract obtained by extraction with hot water; ME—dry V. macrocarpon fruit extract obtained by extraction with 96% (v/v) ethanol; MW—dry V. macrocarpon fruit extract obtained by extraction with hot water. Statistically significant differences between the spheroid diameter and viability of cranberry extracts are indicated by marks (* p < 0.05).