title: The combined effects of limited nutrition and high-frequency loading on intervertebral discs with endplates.
abstract: METHODS
  Whole ovine caudal intervertebral discs were cultured under simulated-physiologic or high-frequency loading and either sufficient or limited nutrition for 7 days.
  
  
  OBJECTIVE
  To study the effect of high-frequency loading under sufficient or limited glucose conditions and to investigate the additive effects of load and nutrition on cell survival, gene expression, and cell activity after 7 days of culture.
  
  
  BACKGROUND
  Limited nutrition and certain mechanical stimuli are generally believed to be etiologic factors for disc degeneration. Although these effects and their interactions have been demonstrated in cell culture, no investigations have been reported in entire discs.
  
  
  METHODS
  Discs were maintained in a whole organ culture bioreactor system under simulated-physiologic (0.2 Hz) or high-frequency (10 Hz) loading, in media with either limited (2 g/L) or sufficient (4.5 g/L) glucose concentration. After 7 days, cell viability, relative gene expression, newly synthesized chondroitin sulfate content, glycosaminoglycan synthesis rate, and disc morphology were assessed after culture and compared with fresh tissue.
  
  
  RESULTS
  Culture under either limited glucose or high-frequency loading conditions led to a significant drop in cell viability. Combined treatment with limited glucose and high-frequency loading resulted in an additive increase in cell death in both the anulus fibrosus and nucleus pulposus and in an increase in MMP13 gene expression.
  
  
  CONCLUSIONS
  Supporting in vivo studies and cell culture experiments, high-frequency loading simulating vibration conditions shows detrimental effects on intervertebral disc cells in whole organ culture. The effect on cell viability was exacerbated by limited nutrition culture. However, neither frequency nor limited glucose affected cell metabolism, measured by glycosaminoglycan synthesis rate. Longer culture periods may be required to detect changes at the extracellular matrix level.
authors.affiliation: AO Research Institute, Davos, Switzerland.
authors.forename: Svenja
authors.initials: S
authors.lastname: Illien-Jünger
authors.forename: Benjamin
authors.initials: B
authors.lastname: Gantenbein-Ritter
authors.forename: Sibylle
authors.initials: S
authors.lastname: Grad
authors.forename: Patrick
authors.initials: P
authors.lastname: Lezuo
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Ferguson
authors.forename: Mauro
authors.initials: M
authors.lastname: Alini
authors.forename: Keita
authors.initials: K
authors.lastname: Ito
mesh_terms: D000818:Animals; D019149:Bioreactors; D002470:Cell Survival; D002809:Chondroitin Sulfates / Q000096:biosynthesis; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000172:deficiency / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000096:biosynthesis; D007403:Intervertebral Disc / Q000378:metabolism* / Q000473:pathology*; D055959:Intervertebral Disc Degeneration / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D053509:Matrix Metalloproteinase 13 / Q000235:genetics; D009752:Nutritional Status; D009924:Organ Culture Techniques; D012756:Sheep; D013997:Time Factors; D014732:Vibration
chemical_list: D006025:Glycosaminoglycans; D002809:Chondroitin Sulfates; D053509:Matrix Metalloproteinase 13; D005947:Glucose