Evaluation of Preclinical Assays to Investigate an Anthroposophic Pharmaceutical Process Applied to Mistletoe (Viscum album L.) Extracts

Abstract

Extracts from European mistletoe (Viscum album L.) developed in anthroposophic medicine are based on specific pharmaceutical procedures to enhance remedy efficacy. One such anthroposophic pharmaceutical process was evaluated regarding effects on cancer cell toxicity in vitro and on colchicine tumor formation in Lepidium sativum. Anthroposophically processed Viscum album extract (APVAE) was produced by mixing winter and summer mistletoe extracts in the edge of a high-speed rotating disk and was compared with manually mixed Viscum album extract (VAE). The antiproliferative effect of VAE/APVAE was determined in five cell lines (NCI-H460, DU-145, HCC1143, MV3, and PA-TU-8902) by WST-1 assay in vitro; no difference was found between VAE and APVAE in any cell line tested (P > 0.14). Incidence of colchicine tumor formation was assessed by measurement of the root/shoot-ratio of seedlings of Lepidium sativum treated with colchicine as well as VAE, APVAE, or water. Colchicine tumor formation decreased after application of VAE (-5.4% compared to water, P < 0.001) and was even stronger by APVAE (-8.8% compared to water, P < 0.001). The high-speed mistletoe extract mixing process investigated thus did not influence toxicity against cancer cells but seemed to sustain morphostasis and to enhance resistance against external noxious influences leading to phenomenological malformations.

Figure 1

Schematic representation of the turning disc of the apparatus that has been used since autumn of 1996 at Hiscia Institute (Arlesheim, Switzerland) to produce anthroposophically processed Viscum album extract (APVAE). The winter mistletoe extract flows into the center of a 1 m diameter titanium disc rotating at 10,000 rpm and then spreads out horizontally. At the upturned disc edge (“mixing zone”), it combines with summer mistletoe extract dropping vertically from a height of 1 meter. Summer mistletoe extract enters the rotating disk not only at one position as schematically outlined above but also at 12 positions, equally spaced along the edge of the disk.

Figure 2

Colchicine (17 μg/mL) induces shortening and thickening of the shoots when applied to Lepidium sativum.

Figure 3

ED50 [μg/mL] of Viscum album extract (VAE) Mali and anthroposophically processed Viscum album extract (APVAE) Mali in five different cell lines. Mean ± SE based on 8 independent experiments each. No significant differences (P < 0.05) were observed for any pairwise comparison between VAE and APVAE.

Figure 4

Root/shoot-elongation-ratio (mean ± SE) of colchicine-treated Lepidium sativum, with either water, Viscum album extract (VAE) Mali or Pini [2 mg/mL], or anthroposophically processed Viscum album extract (APVAE) Mali or Pini [2 mg/mL] added. (a) Average over all colchicine concentrations used. Parameters with different letters (a, b, and c) are statistically different (P < 0.001, LSD test); parameters with identical letters are statistically indistinguishable (P > 0.53, LSD test). (b) The same data as in (a) but differentiated according to the colchicine concentration used (17, 18, or 20 μg/mL). Data from VAE Mali and VAE Pini as well as APVAE Mali and APVAE Pini were pooled. Parameters with different letters (a, b, c,…) are statistically different (P < 0.05, LSD test); parameters with identical letters are statistically indistinguishable (P > 0.05, LSD test).