Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
BMC Musculoskeletal Disorders
Published in: BMC Musculoskeletal Disorders 1/2016

Open Access 01-12-2016 | Research article

Surgical removal and controlled trypsinization of the outer annulus fibrosus improves the bioactivity of the nucleus pulposus in a disc bioreactor culture

Authors: Pei Li, Rongmao Shi, Daosen Chen, Yibo Gan, Yuan Xu, Lei Song, Songtao Li, Qiang Zhou

Published in: BMC Musculoskeletal Disorders | Issue 1/2016

Login to get access

Abstract

Background

The maintenance of nucleus pulposus (NP) viability in vitro is difficult. The annulus fibrosus (AF) pathway reflects one nutrient transport channel and may have an important effect on NP viability in disc organ cultures. The present study describes a feasible disc pre-treatment involving the AF and investigates its efficacy in improving NP bioactivity in an in vitro disc bioreactor culture.

Methods

Rabbit discs that were randomly assigned to the experimental group (EG) were pretreated via the surgical removal and controlled trypsinization of the outer AF. The discs in the control group (CG) did not receive any special treatment. All discs were organ-cultured in a self-developed bioreactor. Solute transport into the central NP was measured using a methylene blue solution. On days 7 and 14, histological properties, cell viability, cell membrane damage, gene expression and matrix composition within the NP in these two groups were compared with each other and with the corresponding parameters of fresh NP samples. Additionally, the structures of the outer AF and the cartilage endplate (CEP) following pre-treatment were also assessed.

Results

The outer AF in the EG became disorganized, but no specific changes occurred in the CEP or the inner AF following pre-treatment. The discs in the EG exhibited increased penetration of methylene blue into the central NP. On days 7 and 14, the NP bioactivity in the EG was improved compared with that of the CG in terms of cell viability, cell membrane damage, gene expression profile and matrix synthesis. Moreover, cell viability and matrix synthesis parameters in the EG were more similar to those of fresh samples than they were to the same parameters in the CG on day 14.

Conclusions

Using this disc pre-treatment, i.e., the surgical removal and controlled trypsinization of the outer AF, NP bioactivity was better maintained for up to 14 days in an in vitro disc bioreactor culture.
Literature
1.
Powell M, Szypryt P, Wilson M, Symonds E, Worthington B. Prevalence of lumbar disc degeneration observed by magnetic resonance in symptomless women. Lancet. 1986;328(8520):1366–7.CrossRef
2.
Smith LJ, Nerurkar NL, Choi KS, Harfe BD, Elliott DM. Degeneration and regeneration of the intervertebral disc: lessons from development. Dis Model Mech. 2011;4(1):31–41.CrossRefPubMed
3.
Kim JH, Studer RK, Sowa GA, Vo NV, Kang JD. Activated macrophage-like THP-1 cells modulate anulus fibrosus cell production of inflammatory mediators in response to cytokines. Spine (Phila Pa 1976). 2008;33(21):2253–9.CrossRef
4.
Collin EC, Grad S, Zeugolis DI, Vinatier CS, Clouet JR, Guicheux JJ, Weiss P, Alini M, Pandit AS. An injectable vehicle for nucleus pulposus cell-based therapy. Biomaterials. 2011;32(11):2862–70.CrossRefPubMed
5.
Woods BI, Sowa G, Vo N, Kang JD. A change in strategy: the use of regenerative medicine and tissue engineering to augment the course of intervertebral disc degeneration. Oper Tech Orthop. 2010;20(2):144–53.CrossRef
6.
Martin JT, Milby AH, Chiaro JA, Kim DH, Hebela NM, Smith LJ, Elliott DM, Mauck RL. Translation of an engineered nanofibrous disc-like angle-ply structure for intervertebral disc replacement in a small animal model. Acta Biomater. 2014;10(6):2473–81.CrossRefPubMedPubMedCentral
7.
8.
9.
OGATA K, WHITESIDE LA. Nutritional pathways of the intervertebral disc: an experimental study using hydrogen washout technique. Spine. 1981;6(3):211–6.CrossRefPubMed
10.
Roberts S, Menage J, Urban J. Biochemical and structural properties of the cartilage end-plate and its relation to the intervertebral disc. Spine. 1989;14(2):166–74.CrossRefPubMed
11.
Gantenbein B, Grünhagen T, Lee CR, van Donkelaar CC, Alini M, Ito K. An in vitro organ culturing system for intervertebral disc explants with vertebral endplates: a feasibility study with ovine caudal discs. Spine. 2006;31(23):2665–73.CrossRefPubMed
12.
Haglund L, Moir J, Beckman L, Mulligan KR, Jim B, Ouellet JA, Roughley P, Steffen T. Development of a bioreactor for axially loaded intervertebral disc organ culture. Tissue Eng Part C Methods. 2011;17(10):1011–9.CrossRefPubMed
13.
Hassler O. The human intervertebral disc: a micro-angiographical study on its vascular supply at various ages. Acta Orthop. 1969;40(6):765–72.CrossRef
14.
Nachemson A, Lewin T, Maroudas A, Freeman M. In vitro diffusion of dye through the end-plates and the annulus fibrosus of human lumbar inter-vertebral discs. Acta Orthop. 1970;41(6):589–607.CrossRef
15.
Huang C-YC, Yuan T-Y, Jackson AR, Hazbun L, Fraker C, Gu WY. Effects of low glucose concentrations on oxygen consumption rates of intervertebral disc cells. Spine. 2007;32(19):2063.CrossRefPubMedPubMedCentral
16.
Mokhbi Soukane D, Shirazi-Adl A, Urban J. Computation of coupled diffusion of oxygen, glucose and lactic acid in an intervertebral disc. J Biomech. 2007;40(12):2645–54.CrossRefPubMed
17.
Ejeskär A, Holm S. Oxygen tension measurements in the intervertebral disc: a methodological and experimental study. Ups J Med Sci. 1979;84(1):83–93.CrossRefPubMed
18.
Lim T-H, Ramakrishnan PS, Kurriger GL, Martin JA, Stevens JW, Kim J, Mendoza SA. Rat spinal motion segment in organ culture: a cell viability study. Spine. 2006;31(12):1291–7.CrossRefPubMed
19.
Farndale RW, Sayers CA, Barrett AJ. A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res. 1982;9(4):247–8.CrossRefPubMed
20.
Sun W, Zhang K, Liu G, Ding W, Zhao C, Xie Y, Yuan J, Sun X, Li H, Liu C, et al. Sox9 gene transfer enhanced regenerative effect of bone marrow mesenchymal stem cells on the degenerated intervertebral disc in a rabbit model. PLoS One. 2014;9(4):e93570.CrossRefPubMedPubMedCentral
21.
Gawri R, Mwale F, Ouellet J, Roughley PJ, Steffen T, Antoniou J, Haglund L. Development of an organ culture system for long-term survival of the intact human intervertebral disc. Spine. 2011;36(22):1835–42.CrossRefPubMed
22.
Jim B, Steffen T, Moir J, Roughley P, Haglund L: Development of an intact intervertebral disc organ culture system in which degeneration can be induced as a prelude to studying repair potential. Eur Spine J. 2011;20(8):1244–54.CrossRefPubMedPubMedCentral
23.
Haschtmann D, Stoyanov JV, Ettinger L, Nolte LP, Ferguson SJ. Establishment of a novel intervertebral disc/endplate culture model: analysis of an ex vivo in vitro whole-organ rabbit culture system. Spine (Phila Pa 1976). 2006;31(25):2918–25.CrossRef
24.
Cs-Szabo G, Ragasa-San Juan D, Turumella V, Masuda K, Thonar EJA, An HS. Changes in mRNA and protein levels of proteoglycans of the anulus fibrosus and nucleus pulposus during intervertebral disc degeneration. Spine. 2002;27(20):2212–9.CrossRefPubMed
25.
Rinkler C, Heuer F, Pedro MT, Mauer UM, Ignatius A, Neidlinger-Wilke C. Influence of low glucose supply on the regulation of gene expression by nucleus pulposus cells and their responsiveness to mechanical loading: Laboratory investigation. J Neurosurg Spine. 2010;13(4):535–42.CrossRefPubMed
26.
Lee CR, Iatridis JC, Poveda L, Alini M. In vitro organ culture of the bovine intervertebral disc: effects of vertebral endplate and potential for mechanobiology studies. Spine. 2006;31(5):515–22.CrossRefPubMed
27.
Gorth DJ, Lothstein KE, Chiaro JA, Farrell MJ, Dodge GR, Elliott DM, Malhotra NR, Mauck RL, Smith LJ. Hypoxic regulation of functional extracellular matrix elaboration by nucleus pulposus cells in long-term agarose culture. J Orthop Res. 2015;33(5):747–54.CrossRefPubMedPubMedCentral
28.
Horner HA, Urban JP. 2001 Volvo award winner in basic science studies: effect of nutrient supply on the viability of cells from the nucleus pulposus of the intervertebral disc. Spine (Phila Pa 1976). 2001;26(23):2543–9.CrossRef
29.
Smith LJ, Byers S, Costi JJ, Fazzalari NL. Elastic fibers enhance the mechanical integrity of the human lumbar anulus fibrosus in the radial direction. Ann Biomed Eng. 2008;36(2):214–23.CrossRefPubMed
30.
Smith LJ, Fazzalari NL. The elastic fibre network of the human lumbar anulus fibrosus: architecture, mechanical function and potential role in the progression of intervertebral disc degeneration. Eur Spine J. 2009;18(4):439–48.CrossRefPubMedPubMedCentral
Metadata
Title
Surgical removal and controlled trypsinization of the outer annulus fibrosus improves the bioactivity of the nucleus pulposus in a disc bioreactor culture
Authors
Pei Li
Rongmao Shi
Daosen Chen
Yibo Gan
Yuan Xu
Lei Song
Songtao Li
Qiang Zhou
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Musculoskeletal Disorders / Issue 1/2016
Electronic ISSN: 1471-2474
DOI
https://doi.org/10.1186/s12891-016-0990-2

Other articles of this Issue 1/2016

BMC Musculoskeletal Disorders 1/2016 Go to the issue

Research article

Identification in GRMD dog muscle of critical miRNAs involved in pathophysiology and effects associated with MuStem cell transplantation

Research article

The influence of calcitonin gene-related peptide on markers of bone metabolism in MG-63 osteoblast-like cells co-cultured with THP-1 macrophage-like cells under virtually osteolytic conditions

Research article

Macrophage migration inhibitory factor enhances lipopolysaccharide-induced fibroblast proliferation by inducing toll-like receptor 4

Research article

Prevalence of osteoporosis in the Italian population and main risk factors: results of BoneTour Campaign

Research article

Non-ossifying fibroma: natural history with an emphasis on a stage-related growth, fracture risk and the need for follow-up

Research article

The frequency of and risk factors for osteoporosis in Korean patients with rheumatoid arthritis