The combined effects of limited nutrition and high-frequency loading on intervertebral discs with endplates.
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.
Illien-Jünger
Svenja
S
AO Research Institute, Davos, Switzerland.
Gantenbein-Ritter
Benjamin
B
Grad
Sibylle
S
Lezuo
Patrick
P
Ferguson
Stephen J
SJ
Alini
Mauro
M
Ito
Keita
K
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
D006025:Glycosaminoglycans; D002809:Chondroitin Sulfates; D053509:Matrix Metalloproteinase 13; D005947:Glucose