Evaluation of Anti-Neoplastic Effects of a New Hemostatic Agent Ankaferd Blood Stopper on Myeloma Cell Line and Plasmocytoma Development in Balb/c Mice: Results of the First in Vitro and in Vivo Study

Abstract

Ankaferd Blood Stopper (ABS), a unique traditional herbal mixture, has been used topically to stop bleeding for centuries in Anatolia. As well as ABS has been used as a blood-stopping agent, it may also have a considerable therapeutic benefit, because of its anti-infective, anti-neoplastic, and wound healing properties. The aim of this study is to investigate the anti-neoplastic effects of the ABS on myeloma cell line, in vitro and on the plasmocytoma development in Balb/c mice by intraperitoneal injection of pristane, in vivo. We therefore sought to evaluate the efficacy of ABS on MM cells and to study the modulation of cell-death pathways. The cytotoxicity of ABS against the MM cell lines (RPMI-8226, and ARH-77) was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-dye reduction assay. Responses to ABS by RPMI-8226 and ARH-77 cell lines were dose dependent but not time dependent. The IC50 values for RPMI 8226 and ARH 77 myeloma cell lines in 24h were 12,84 μL/mL and 13,86 μL/mL, respectively. Various cell-death characteristics such as caspase-3, Bcl-2, Bax were studied in response to ABS, but we couldn’t demonstrate specific features of apoptotic cell death, in vitro. We have also investigated the effect of the ABS on the pristane (2.6.10.14-tetramethylpentadecane)-induced plasmacytoma (PCT) development on six-week-old BALB/c mice. Three groups of mice were treated with intraperitoneal ABS (1 mg/kg, 0.5mg/kg, and 0,1mg/kg) per-week for eight weeks after pristane-induced PCT development. The study was stopped at twelfth week, the remaining mice were autopsied, and peritoneal tissues were examined histologically for PCTs. A database of different groups’ mice was analyzed using Kaplan-Meier and Cox regression statistics based on variables. Kaplan-Meier analysis revealed a difference of the survival of pristane-induced alone between the groups of pristane-induced plus ABS 1 mg/kg, 0.5mg/kg, and 0.1mg/kg. (Log-rank, p=0.016; p<0.001 and p<0.001; respectively). The present results indicate that direct anti-tumor effect of ABS on pristane-induced PCT and significantly increased survival. This hypothesis needs to now be further investigated in clinical trials. Disclosures No relevant conflicts of interest to declare.

Qualitative/Chemical Analyses of Ankaferd Hemostat and Its Antioxidant Content in Synthetic Gastric Fluids

“…Similarly, ABS can induce apoptosis, regulates cellular proliferation [ 21 , 67 – 69 ], and decreases tumor vascularization [ 52 ]. Experimental antineoplastic activities of ABS have been shown in rats and cancer cell lines [ 20 – 22 ]. The interrelationships between ABS, its antioxidant content, and tocotrienol should be investigated in future experimental neoplastic studies.…”

“…For instance, a recent clinical study by Patiroglu et al has demonstrated that oral topical ABS use at the beginning of chemotherapy could provide less oral mucositis when compared to the control group in the pediatric patients with cancer [ 19 ]. Moreover, experimental antineoplastic activities of ABS have been shown in rats and cancer cell lines [ 20 – 22 ].…”

Proteomic and transcriptomic analyses to explain the pleiotropic effects of Ankaferd blood stopper

“…ABS has demonstrated cytotoxicity against human erythrocytes at some concentrations, as well as against tumoral cells in hematologic malignancies such as multiple myeloma, chronic myelogenous leukemia, and lymphoma. 35 , 37 , 44 Possible related ingredients of Ankaferd are CREBZF, PIAS-2, HNF-4a, ME-1, P18INK4C, and Midkine and addressed below in alphabetical order.…”

“…35 It has also demonstrated toxicity toward multiple myeloma and plasmacytoma cells both in vitro and in vivo, while decreasing M protein production. 36 , 37 Other solid malignancies such as sarcoma and colon cancer are also targets of the anti-neoplastic action of Ankaferd.…”

The effect of Ankaferd blood stopper on liver damage in experimental obstructive jaundice

“…regulate cellular proliferation and suppress tumor angiogenesis. Experimental antineoplastic activities of ABS have been demonstrated in rats and cancer cell lines[24][25][26][27][28][29]. Since the desirable feature of an anticancer drug is that it can kill cancer cells and does not damage normal human cells, natural products are widely preferred in the production of anticancer drugs.…”