title: No Effects of Hyperosmolar Culture Medium on Tissue Regeneration by Human Degenerated Nucleus Pulposus Cells Despite Upregulation Extracellular Matrix Genes.
abstract: An in vitro study using human degenerated nucleus pulposus cells.
  
  
  
  To determine the effect of osmolality and different osmolytes on the regeneration by human nucleus pulposus cells through gene expression and extracellular matrix production.
  
  
  
  Intervertebral disc (IVD) degeneration is a major problem in developed countries. Regeneration of the IVD can prevent pain and costs due to diminished work absence and health care, and improve quality of life. The osmotic value of a disc decreases during degeneration due to loss of proteoglycans and might increase degeneration. It is known that gene expression of matrix genes of nucleus pulposus (NP) cells increases when cultured in hyperosmotic medium. Thus, increasing the osmolality of the disc might be beneficial for disc regeneration.
  
  
  
  In the current study, isolated degenerated human NP cells were used in regeneration culture with medium of different osmolalities, adjusted with different osmolytes. NaCl, urea and sucrose. The cells were cultured for 28 days and expression of matrix genes and production of glycosaminoglycans and collagen II were measured.
  
  
  
  Gene expression for both collagen II and aggrecan increased with increasing osmolality using NaCl or sucrose, but not urea. Protein production however, was not affected by increasing osmolality and was decreased when using urea and sucrose. Expression of genes for Col1A1, MMP13, and MMP14 decreased with increasing osmolality, whereas expression of LOXL2 and LOXL3 increased. Transient expression of TonEBP was found 6 hours after the start of culture, but not at later time points.
  
  
  
  Although expression of matrix genes is upregulated, hyperosmolality does not enhance matrix production by nucleus pulposus cells. Raising osmolality can potentially increase matrix production, but in itself is not sufficient to accomplish regeneration in the current in vitro culture system.
  
  
  
  N /A.
authors: Department of Orthopedics, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: Anita
authors: A
authors: Krouwels
authors: Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands.
authors: Jelena
authors: J
authors: Popov-Celeketic
authors: Department of Orthopedics, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: Saskia G M
authors: SGM
authors: Plomp
authors: Department of Orthopedics, University Medical Center Utrecht, Utrecht, The Netherlands.|Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors: Wouter J A
authors: WJA
authors: Dhert
authors: Department of Orthopedics, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: F Cumhur
authors: FC
authors: Öner
authors: Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands.
authors: Ruud A
authors: RA
authors: Bank
authors: Department of Orthopedics, University Medical Center Utrecht, Utrecht, The Netherlands.
authors: Laura B
authors: LB
authors: Creemers
mesh_terms: D000328:Adult; D000368:Aged; D053674:Aggrecans / Q000096:biosynthesis / Q000235:genetics; D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006025:Glycosaminoglycans / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D055959:Intervertebral Disc Degeneration / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D000070614:Nucleus Pulposus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011509:Proteoglycans / Q000096:biosynthesis / Q000235:genetics; D012038:Regeneration / Q000187:drug effects / Q000502:physiology*; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology*
chemical_list: D053674:Aggrecans; D003470:Culture Media; D006025:Glycosaminoglycans; D011509:Proteoglycans; D000594:Amino Acid Oxidoreductases; C119091:LOXL2 protein, human