title: Milk thistle extract and silymarin inhibit lipopolysaccharide induced lamellar separation of hoof explants in vitro.
abstract: The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS) in this process remains unclear. Phytogenic substances, like milk thistle (MT) and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT and silymarin are capable of inhibiting LPS-induced effects in an in vitro/ex vivo model. In preliminary tests, LPS neutralization efficiency of these phytogenics was determined in an in vitro neutralization assay. Furthermore, tissue explants gained from hooves of slaughter horses were tested for lamellar separation after incubation with different concentrations of LPS. By combined incubation of explants with LPS and either Polymyxin B (PMB; positive control), MT or silymarin, the influence of these substances on LPS-induced effects was assessed. In the in vitro neutralization assay, MT and silymarin reduced LPS concentrations by 64% and 75%, respectively, in comparison PMB reduced 98% of the LPS concentration. In hoof explants, LPS led to a concentration dependent separation. Accordantly, separation force was significantly decreased by 10 µg/mL LPS. PMB, MT and silymarin could significantly improve tissue integrity of explants incubated with 10 µg/mL LPS. This study showed that LPS had a negative influence on the structure of hoof explants in vitro. MT and silymarin reduced endotoxin activity and inhibited LPS-induced effects on the lamellar tissue. Hence, MT and silymarin might be used to support the prevention of laminitis and should be further evaluated for this application.
authors.affiliation: Biomin Research Center, Technopark 1, Tulln 3430, Austria. nicole.reisinger@biomin.net.
authors.forename: Nicole
authors.initials: N
authors.lastname: Reisinger
authors.affiliation: Biomin Research Center, Technopark 1, Tulln 3430, Austria. simone.schaumberger@biomin.net.
authors.forename: Simone
authors.initials: S
authors.lastname: Schaumberger
authors.affiliation: Center for Analytical Chemistry, Department for Agrobiotechnology (IFA Tulln), University of Natural Resources and Life Sciences, Vienna, Tulln (BOKU), Konrad Lorenz Str. 20, Tulln 3430, Austria. veronika.nagl@boku.ac.at.
authors.forename: Veronika
authors.initials: V
authors.lastname: Nagl
authors.affiliation: Biomin Research Center, Technopark 1, Tulln 3430, Austria. sabine.hessenberger@biomin.net.
authors.forename: Sabine
authors.initials: S
authors.lastname: Hessenberger
authors.affiliation: Biomin Research Center, Technopark 1, Tulln 3430, Austria. gerd.schatzmayr@biomin.net.
authors.forename: Gerd
authors.initials: G
authors.lastname: Schatzmayr
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D004731:Endotoxins / Q000633:toxicity; D006724:Hoof and Claw / Q000187:drug effects* / Q000473:pathology; D066298:In Vitro Techniques; D008070:Lipopolysaccharides / Q000633:toxicity; D020944:Silybum marianum / Q000737:chemistry*; D010936:Plant Extracts / Q000494:pharmacology*; D012838:Silymarin / Q000494:pharmacology*
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D004731:Endotoxins; D008070:Lipopolysaccharides; D010936:Plant Extracts; D012838:Silymarin