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Effects of initial boost with TGF-beta 1 and grade of intervertebral disc degeneration on 3D culture of human annulus fibrosus cells

Hegewald, Aldemar Andres ; Cluzel, Jessie ; Krüger, Jan Philipp ; Endres, Michaela ; Kaps, Christian ; Thomé, Claudius

In: Journal of Orthopaedic Surgery and Research, 9 (2014), Nr. 73. pp. 1-8. ISSN 1749-799X

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Download (1MB) | Lizenz: Creative Commons LizenzvertragEffects of initial boost with TGF-beta 1 and grade of intervertebral disc degeneration on 3D culture of human annulus fibrosus cells by Hegewald, Aldemar Andres ; Cluzel, Jessie ; Krüger, Jan Philipp ; Endres, Michaela ; Kaps, Christian ; Thomé, Claudius underlies the terms of Creative Commons Attribution 3.0 Germany

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Abstract

Background: Three-dimensional (3D) culture in porous biomaterials as well as stimulation with growth factors are known to be supportive for intervertebral disc cell differentiation and tissue formation. Unless sophisticated releasing systems are used, however, effective concentrations of growth factors are maintained only for a very limited amount of time in in vivo applications. Therefore, we investigated, if an initial boost with transforming growth factor-beta 1 (TGF-beta 1) is capable to induce a lasting effect of superior cartilaginous differentiation in slightly and severely degenerated human annulus fibrosus (AF) cells. Methods: Human AF tissue was harvested during surgical treatment of six adult patients with lumbar spinal diseases. Grading of disc degeneration was performed with magnet resonance imaging. AF cells were isolated and expanded in monolayer culture and rearranged three-dimensionally in a porous biomaterial consisting of stepwise absorbable poly-glycolic acid and poly-(lactic-co-glycolic) acid and a supportive fine net of non-absorbable polyvinylidene fluoride. An initial boost of TGF-beta 1 or TGF-beta 1 and hyaluronan was applied and compared with controls. Matrix formation was assessed at days 7 and 21 by (1) histological staining of the typical extracellular matrix molecules proteoglycan and type I and type II collagens and by (2) real-time gene expression analysis of aggrecan, decorin, biglycan, type I, II, III, and X collagens as well as of catabolic matrix metalloproteinases MMP-2 and MMP-13. Results: An initial boost with TGF-beta 1 or TGF-beta 1 and hyaluronan did not enhance the expression of characteristic AF matrix molecules in our 3D culture system. AF cells showed high viability in the progressively degrading biomaterial. Stratification by grade of intervertebral disc degeneration showed that AF cells from both, slightly degenerated, or severely degenerated tissue are capable of significant up-regulations of characteristic matrix molecules in 3D culture. AF cells from severely degenerated tissue, however, displayed significantly lower up-regulations in some matrix molecules such as aggrecan. Conclusions: We failed to show a supportive effect of an initial boost with TGF-beta 1 in our 3D culture system. This underlines the need for further investigations on growth factor releasing systems.

Item Type: Article
Journal or Publication Title: Journal of Orthopaedic Surgery and Research
Volume: 9
Number: 73
Publisher: BioMed Central
Place of Publication: London
Date Deposited: 03 Feb 2016 10:38
Date: 2014
ISSN: 1749-799X
Page Range: pp. 1-8
Faculties / Institutes: Medizinische Fakultät Mannheim > Neurochirurgische Klinik
Subjects: 610 Medical sciences Medicine
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