Indication-Specific Effect of a Phytotherapeutic Remedy on Human Fetal Osteoblastic Cells: An in vitro Analysis

Abstract

Background Impaired fracture healing is a recurring interdisciplinary medical challenge. Alternative treatment concepts, apart from conventional medicine are popular, but scientific evidence on their effects is still lacking. Plant-derived substances are widely assumed to support bone homeostasis. To clarify the effects on bone healing mechanisms, a commercially available, homeopathic-spagyric remedy, containing inter alia two herbal substances with assumed osteogenic potential, equisetum arvense and bellis perennis, was analyzed. Methods Human fetal osteoblastic cells (hFOB1.19) were incubated with the test substance, in serial dilutions from 10 to 0,00001 %. Cell viability has been evaluated through ATP level (CTG assay) and MTT tetrazolium reduction. Cell proliferation was analyzed by BrdU incorporation and cell migration by wound healing assay (WHA) via image analysis. Additionally, determination of the expression of key genes via real-time PCR and proteins via Proteome Array for inflammation, cell proliferation and angiogenesis were performed. Results An incubation of hFOB 1.19 with the test substance for 24/72 hours showed no reduction in cell number, viability or proliferation. Cell migration was unimpaired. The test substance induced inflammatory genes and growth factors along with genes of osseous regeneration. (ALP, Col1, IL-1α, IL-6, IL-8, IL-10, Osteocalcin, Osteonectin, RUMX2, TGF, VEGFA). Increased protein expression was found in multiple cytokines, chemokines and acute phase proteins. Conclusion The test substance did not impair cell vitality parameters (MTT, CTG, BrdU and WHA). A tendency to activate growth factors, bone regeneration genes and proteins was shown for osteoblasts, indicating a possible positive effect on osteogenic processes.