title: The biomedical potential of genetically modified flax seeds overexpressing the glucosyltransferase gene.
abstract: BACKGROUND
  Flax (Linum usitatissimum) is a potential source of many bioactive components that can be found in its oil and fibers, but also in the seedcake, which is rich in antioxidants. To increase the levels of medically beneficial compounds, a genetically modified flax type (named GT) with an elevated level of phenylopropanoids and their glycoside derivatives was generated. In this study, we investigated the influence of GT seedcake extract preparations on human fibroblast proliferation and migration, and looked at the effect on a human skin model. Moreover, we verified its activity against bacteria of clinical relevance.
  
  
  METHODS
  The GT flax used in this study is characterized by overexpression of the glucosyltransferase gene derived from Solanum sogarandinum. Five GT seedcake preparations were generated. Their composition was assessed using ultra pressure liquid chromatography and confirmed using the UPLC-QTOF method. For the in vitro evaluation, the influence of the GT seedcake preparations on normal human dermal fibroblast proliferation was assessed using the MTT test and the wound scratch assay. A human skin model was used to evaluate the potential for skin irritation. To assess the antimicrobial properties of GT preparations, the percentage of inhibition of bacterial growth was calculated.
  
  
  RESULTS
  The GT seedcake extract had elevated levels of phenylopropanoid compounds in comparison to the control, non-transformed plants. Significant increases in the content of ferulic acid, p-coumaric acid and caffeic acid, and their glucoside derivatives, kaempferol, quercitin and secoisolariciresinol diglucoside (SDG) were observed in the seeds of the modified plants. The GT seedcake preparations were shown to promote the proliferation of normal human dermal fibroblasts and the migration of fibroblasts in the wound scratch assay. The superior effect of GT seedcake extract on fibroblast migration was observed after a 24-hour treatment. The skin irritation test indicated that GT seedcake preparations have no harmful effect on human skin. Moreover, GT seedcake preparations exhibited inhibitory properties toward two bacterial strains: Staphylococcus aureus and Escherichia coli.
  
  
  CONCLUSIONS
  We suggest that preparations derived from the new GT flax are an effective source of phenylopropanoids and that their glycoside derivatives and might be promising natural products with both healing and bacteriostatic effects. This flax-derived product is a good candidate for application in the repair and regeneration of human skin and might also be an alternative to antibiotic therapy for infected wounds.
authors.affiliation: Len Pharma, ul, Zamkowa 17, 82-200 Malbork, Poland. czemplik@ibmb.uni.wroc.pl
authors.forename: Magdalena
authors.initials: M
authors.lastname: Czemplik
authors.forename: Anna
authors.initials: A
authors.lastname: Kulma
authors.forename: Karolina
authors.initials: K
authors.lastname: Bazela
authors.forename: Jan
authors.initials: J
authors.lastname: Szopa
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003879:Dermatologic Agents / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D019597:Flax / Q000235:genetics / Q000378:metabolism*; D005964:Glucosyltransferases / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology; D030821:Plants, Genetically Modified / Q000378:metabolism*; D020005:Propanols / Q000378:metabolism / Q000494:pharmacology*; D012639:Seeds / Q000378:metabolism*; D012867:Skin / Q000187:drug effects / Q000382:microbiology; D032322:Solanum / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects; D014945:Wound Healing / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D003879:Dermatologic Agents; D010936:Plant Extracts; D020005:Propanols; C439395:1-phenylpropanol; D005964:Glucosyltransferases