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No Effects of Hyperosmolar Culture Medium on Tissue Regeneration by Human Degenerated Nucleus Pulposus Cells Despite Upregulation Extracellular Matrix Genes
- Source :
- SPINE, 43(5), 307-315. LIPPINCOTT WILLIAMS & WILKINS
- Publication Year :
- 2018
- Publisher :
- Ovid Technologies (Wolters Kluwer Health), 2018.
-
Abstract
- Study Design. An in vitro study using human degenerated nucleus pulposus cells.Objective. To determine the effect of osmolality and different osmolytes on the regeneration by human nucleus pulposus cells through gene expression and extracellular matrix production.Summary of Background Data. 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.Methods. 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.Results. 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.Conclusion. 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.
- Subjects :
- Male
0301 basic medicine
osmolyte
PROTEIN EXPRESSION
INTERVERTEBRAL DISC CELLS
OSMOLARITY
Intervertebral Disc Degeneration
Extracellular matrix
DNA
0302 clinical medicine
MAMMALIAN-CELLS
Gene expression
Orthopedics and Sports Medicine
Aggrecans
Cells, Cultured
Glycosaminoglycans
GAG
collagen II
nucleus pulposus
CHONDROCYTES
hyperosmotic
sucrose
Middle Aged
musculoskeletal system
Extracellular Matrix
Up-Regulation
Cell biology
medicine.anatomical_structure
Biochemistry
Osmolyte
sodium chloride
Female
Proteoglycans
Amino Acid Oxidoreductases
aggrecan
MESSENGER-RNA
Adult
musculoskeletal diseases
urea
METABOLISM
Biology
03 medical and health sciences
Downregulation and upregulation
CARTILAGE
medicine
Humans
Regeneration
osmolality
GLYCOSAMINOGLYCAN PRODUCTION
Aggrecan
Aged
030203 arthritis & rheumatology
Cartilage
Regeneration (biology)
COLLAGEN
Culture Media
030104 developmental biology
intervertebral disc
Neurology (clinical)
Nucleus
Subjects
Details
- ISSN :
- 15281159 and 03622436
- Volume :
- 43
- Database :
- OpenAIRE
- Journal :
- Spine
- Accession number :
- edsair.doi.dedup.....31efaa00cdeea0a0c03708d7724b6c12