Anticoagulant effect of Feijoa sellowiana extracts generated by different biotechnological techniques

Abstract

Blood clotting has become one of the most dangerous side effects associated with Corona virus, as well as the high level of cholesterol and triglycerides in the blood. Therefore, it has become necessary to use medicinal plants that are biologically safe and containing anti-clotting compound. Feijoa sellowiana represents a prolific source diverse compounds that may have thrombolytic activity. Therefore, the main research point is the production and scaling up of a target contents that have anticoagulants by using biotechnological techniques; calli production, and bioreactors and assessed their activity through in-vivo study. Murashige and Skoog (MS) medium enriched with varying concentrations of benzyl adenine (BA) and naphthalene acetic acid (NAA) was used to cultivate calli and cell suspension cultures from F. sellowiana seeds. Bioreactors were employed to boost active constituent's production. Moreover, the bioreactor physical factors such as effect of controlled or uncontrolled pH medium were investigated. The leaves of the main plant were extracted by ethanol 70% and polar and non-polar extracts were also prepared. The ethanol extract of calli and cells resulting from bioreactors were also prepared. All prepared extracts were subjected to chemical analysis by HPLC, in-vitro antioxidant assays, in-vivo anticoagulant activity and histopathological examination. Calli and cell suspension cultures were produced by using MS medium fortified with 1 mg/L BA+ 0.1 mg/L NAA. It was found that culturing of cell cultures in a bioreactor with uncontrolled pH and aeration at the value of 0.5 L/min gave the maximum and economical fresh and dry weights of the plants. After evaluation of all extracts; it was found that the calli ethanol extract for each plant was the highest value of total phenolic and total flavonoid contents either quantitatively or qualitatively. All extracts of Feijoa had antioxidant activity. The IC₅₀ of the DPPH of Feijoa calli extract was 13.45 μg/mL, it was also confirmed by FRAP and ABTs values. Feijoa calli extract decreased platelet aggregation by suppression of thrombin, extended aPTT, PT, bleeding and clotting times. It was safer than warfarin medication. From these findings the authors can conclude that Feijoa had highly anticoagulant activity and the calli production achieved the goal of the enhancement of the phenolic constituent and thus their activity.

Keywords: Anticoagulant; Bioreactor; Feijoa sellowiana; Natural products; Plant biotechnology- cell suspension culture.

Graphical abstract

Fig. 1

Calli culture production fromFeijoa sellowianaleaf explants.

Fig. 2

Cell number x 10⁵and P.C.V. ofFeijoa sellowianaduring 24 days of cultivation.

Fig. 3

Effect ofFeijoa sellowianaon Plasma Coagulation Factors Measurement in male rats Data are presented as the mean ± S.E. of (n = 8) for each group. Statistical analysis was carried out by one-way analysis of variance followed by Dunnett multiple comparisons test. ^a Statistically significant from control group. ^b Statistically significant from warfarin group at P < 0.05. ^c Statistically significant from callus group at P < 0.05. ^d Statistically significant from bioreactor group at P < 0.05.

Fig. 4

Effect ofFeijoa sellowianaon TXB 2 and COX 1 in male rats Data are presented as the mean ± S.E. of (n = 8) for each group. Statistical analysis was carried out by one-way analysis of variance followed by Dunnett multiple comparisons test. ^a Statistically significant from control group. ^b Statistically significant from warfarin group at P < 0.05. ^c Statistically significant from callus group at P < 0.05.

Fig. 5

EffectofFeijoa sellowiana on Plasma Coagulation Factors Measurement in female rats. Data are presented as the mean ± S.E. of (n = 8) for each group. Statistical analysis was carried out by one-way analysis of variance followed by Dunnett multiple comparisons test. ^a Statistically significant from control group. ^b Statistically significant from warfarin group at P < 0.05. ^c Statistically significant from callus group at P < 0.05. ^d Statistically significant from bioreactor group at P < 0.05.

Fig. 6

Effect of Feijoa sellowiana on TXB 2 and COX 1 in female rats. Data are presented as the mean ± S.E. of (n = 8) for each group. Statistical analysis was carried out by one-way analysis of variance followed by Dunnett multiple comparisons test. ^a Statistically significant from control group. ^b Statistically significant from warfarin group at P < 0.05. ^c Statistically significant from callus group at P < 0.05.

Fig. 7

Mechanism of action ofFeijoa sellowianaextractsthrough in vitroassessment as antioxidant andin vivoinvestigation as anticoagulant. Created withBioRender.com(Agreement number:BS24PE8WFC).

Fig. 8

A photomicrograph of (a) spleen of the control group, (b) spleen of the warfarin group, (c) spleen of the Callus F. sellowiana group (d) bioreactor F. sellowiana group, (e) F. sellowiana group, (f) Non polar F. sellowiana group, (g) polar F. sellowiana group (H&E X 200).