The in vitro effects of dexamethasone, insulin and triiodothyronine on degenerative human intervertebral disc cells under normoxic and hypoxic conditions

Bertolo, Alessandro; Ettinger, Ladina; Aebli, Niklaus; Haschtmann, Daniel; Baur, Martin; Berlemann, Ulrich; Ferguson, Stephen John; Stoyanov, Jivkov V (2011). The in vitro effects of dexamethasone, insulin and triiodothyronine on degenerative human intervertebral disc cells under normoxic and hypoxic conditions. European cells & materials eCM, 21, pp. 221-229. Aberystwyth (UK): University of Wales

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Degeneration of intervertebral discs (IVD) is one of the main causes of back pain and tissue engineering has been proposed as a treatment. Tissue engineering requires the use of highly expensive growth factors, which might, in addition, lack regulatory approval for human use. In an effort to find readily available differentiation factors, we tested three molecules – dexamethasone, triiodothyronine (T3) and insulin – on human IVD cells isolated after surgery, expanded in vitro and transferred into alginate beads. Triplicates containing 40 ng/ml dexamethasone, 10 nM T3 and 10 µg/ml insulin, together with a positive control (10 ng/mL transforming growth factor (TGF)-beta 1), were sampled weekly over six weeks and compared to a negative control. Furthermore, we compared the results to cultures with optimized chondrogenic media and under hypoxic condition (2% O2). Glycosaminoglycan (GAG) determination by Alcian Blue assay and histological staining showed dexamethasone to be more effective than T3 and insulin, but less than TGF-beta1. DNA quantification showed that only dexamethasone stimulated cell proliferation. qPCR demonstrated that TGF-beta1 and the optimized chondrogenic groups increased the expression of collagen type II, while aggrecan was stimulated in cultures containing dexamethasone. Hypoxia increased GAG accumulation, collagen type II and aggrecan expression, but had no effect on or even lowered cell number. In conclusion, dexamethasone is a valuable and cost-effective molecule for chondrogenic and viability induction of IVD cells under normoxic and hypoxic conditions, while insulin and T3 did not show significant differences.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute for Surgical Technology & Biomechanics ISTB

UniBE Contributor:

Ettinger, Ladina and Ferguson, Stephen John

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1473-2262

Publisher:

University of Wales

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 14:16

Last Modified:

16 Jul 2018 13:03

Web of Science ID:

000287766100003

Uncontrolled Keywords:

tervertebral disc degeneration, dexamethasone, insulin, triiodothyronine, cartilage, collagen, aggrecan, glycosaminoglycan

BORIS DOI:

10.7892/boris.4719

URI:

https://boris.unibe.ch/id/eprint/4719 (FactScience: 209335)

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