Influence of diurnal hyper-osmotic loading on the metabolic activity and gene expression of a whole-organ intervertebral disc model

Increasing evidence implicates intervertebral disc (IVD) degeneration as a major contributor to low back pain. In addition to a series of pathogenic processes, degenerated IVDs become vascularized in contrast to healthy IVDs. In this context, angiopoietin (Ang) plays a crucial role and is involved in cytokine recruitment, and anabolic and catabolic reactions within the extracellular matrix (ECM). Over the last decade, a progenitor cell population has been described in the nucleus pulposus (NP) of the IVD to be positive for the Tie2 marker (also known as Ang-1 receptor). In this study, we investigated the influence of Ang-1 and Ang-2 on human NP cell (Tie2+, Tie2- or mixed) populations isolated from trauma patients during 7 days in normoxia (21% O2) or hypoxia (≤ 5% O2). At the end of the process, the proliferation and metabolic activity of the NP cells were analyzed. Additionally, the relative gene expression of NP-related markers was evaluated. NP cells showed a higher proliferation depending on the Ang treatment. Moreover, the study revealed higher NP cell metabolism when cultured in hypoxia. Additionally, the relative gene expression followed, with an increase linked to the oxygen level and Ang concentration. Our study comparing different NP cell populations may be the start of new approaches for the treatment of IVD degeneration.

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Influence of diurnal hyperosmotic loading on the metabolism and matrix gene expression of a whole-organ intervertebral disc model

Journal of Orthopaedic Research® ◽

10.1002/jor.20243 ◽

2006 ◽

Vol 24 (10) ◽

pp. 1957-1966 ◽

Cited By ~ 42

Author(s):

Daniel Haschtmann ◽

Jivko V. Stoyanov ◽

Stephen J. Ferguson

Keyword(s):

Gene Expression ◽

Intervertebral Disc ◽

Matrix Gene ◽

Disc Model

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DNA Demethylation Regulates Anabolic and Catabolic Gene Expression in Intervertebral Disc Cells

Global Spine Journal ◽

10.1055/s-0034-1376587 ◽

2014 ◽

Vol 4 (1_suppl) ◽

pp. s-0034-1376587-s-0034-1376587

Author(s):

N. Chutkan ◽

R. Sangani ◽

H. Zhou ◽

S. Fulzele

Keyword(s):

Gene Expression ◽

Intervertebral Disc ◽

Dna Demethylation ◽

Catabolic Gene ◽

Disc Cells

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A Hyaluronan and Platelet-Rich Plasma Hydrogel for Mesenchymal Stem Cell Delivery in the Intervertebral Disc: An Organ Culture Study

International Journal of Molecular Sciences ◽

10.3390/ijms22062963 ◽

2021 ◽

Vol 22 (6) ◽

pp. 2963

Author(s):

Fabrizio Russo ◽

Luca Ambrosio ◽

Marianna Peroglio ◽

Wei Guo ◽

Sebastian Wangler ◽

...

Keyword(s):

Gene Expression ◽

Intervertebral Disc ◽

Organ Culture ◽

Platelet Rich Plasma ◽

Cell Activity ◽

Human Mesenchymal Stem Cells ◽

Cell Phenotype ◽

Cytokeratin 19 ◽

Stem Cell Delivery ◽

Normal Metabolism

The purpose of the present pilot study was to evaluate the effect of a hydrogel composed of hyaluronic acid (HA) and platelet-rich plasma (PRP) as a carrier for human mesenchymal stem cells (hMSCs) for intervertebral disc (IVD) regeneration using a disc organ culture model. HA was mixed with batroxobin (BTX) and PRP to form a hydrogel encapsulating 1 × 106 or 2 × 106 hMSCs. Bovine IVDs were nucleotomized and filled with hMSCs suspended in ~200 μL of the PRP/HA/BTX hydrogel. IVDs collected at day 0 and nucleotomized IVDs with no hMSCs and/or hydrogel alone were used as controls. hMSCs encapsulated in the hydrogel were also cultured in well plates to evaluate the effect of the IVD environment on hMSCs. After 1 week, tissue structure, scaffold integration, hMSC viability and gene expression of matrix and nucleus pulposus (NP) cell markers were assessed. Histological analysis showed a better preservation of the viability of the IVD tissue adjacent to the gel in the presence of hMSCs (~70%) compared to the hydrogel without hMSCs. Furthermore, disc morphology was maintained, and the hydrogel showed signs of integration with the surrounding tissues. At the gene expression level, the hydrogel loaded with hMSCs preserved the normal metabolism of the tissue. The IVD environment promoted hMSC differentiation towards a NP cell phenotype by increasing cytokeratin-19 (KRT19) gene expression. This study demonstrated that the hydrogel composed of HA/PRP/BTX represents a valid carrier for hMSCs being able to maintain a good cell viability while stimulating cell activity and NP marker expression.

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An organ culture system to model early degenerative changes of the intervertebral disc II: profiling global gene expression changes

Arthritis Research & Therapy ◽

10.1186/ar4301 ◽

2013 ◽

Vol 15 (5) ◽

pp. R121 ◽

Cited By ~ 25

Author(s):

Dessislava Z Markova ◽

Christopher K Kepler ◽

Sankar Addya ◽

Hallie B Murray ◽

Alexander R Vaccaro ◽

...

Keyword(s):

Gene Expression ◽

Intervertebral Disc ◽

Organ Culture ◽

Culture System ◽

Global Gene Expression ◽

Degenerative Changes ◽

Organ Culture System

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Cell shape and gene expression in human intervertebral disc cells: in vitro tissue engineering studies

Biotechnic & Histochemistry ◽

10.1080/10520290310001593793 ◽

2003 ◽

Vol 78 (2) ◽

pp. 109-117 ◽

Cited By ~ 32

Author(s):

He Gruber ◽

Ja Ingram ◽

K Leslie ◽

Hj Norton ◽

En Hanley Jr

Keyword(s):

Gene Expression ◽

Tissue Engineering ◽

Intervertebral Disc ◽

Cell Shape ◽

Engineering Studies ◽

Disc Cells

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Biotribological Tests of Osteochondral Grafts after Treatment with Pro-Inflammatory Cytokines

Cartilage ◽

10.1177/1947603521994900 ◽

2021 ◽

pp. 194760352199490

Author(s):

Christoph Bauer ◽

Hakan Göçerler ◽

Eugenia Niculescu-Morzsa ◽

Vivek Jeyakumar ◽

Christoph Stotter ◽

...

Keyword(s):

Gene Expression ◽

Inflammatory Cytokines ◽

Metabolic Activity ◽

Wear Debris ◽

Cartilage Tissue ◽

Test Fluid ◽

Osteochondral Grafts ◽

Extracellular Matrix Components ◽

Proteoglycan Content ◽

Pro Inflammatory Cytokines

ObjectiveDuring osteoarthritis progression, cartilage degrades in a manner that influences its biomechanical and biotribological properties, while chondrocytes reduce the synthesis of extracellular matrix components and become apoptotic. This study investigates the effects of inflammation on cartilage under biomechanical stress using biotribological tests.MethodsBovine osteochondral grafts from five animals were punched out from the medial condyle and treated with or without pro-inflammatory cytokines (interleukin-1β [IL-1β], tumor necrosis factor-α [TNF-α], IL-6) for 2 weeks. After incubation, biotribological tests were performed for 2 hours (alternating 10 minutes test and pause respectively at 39°C, 180 N, 1 Hz, and 2 mm stroke). Before and after testing, the cartilage surface was imaged with a 3-dimensional microscope. During testing, the coefficient of friction (COF) was measured, while gene expression analysis and investigation of metabolic activity of chondrocytes were carried out after testing. Histological sections of the tissue and wear debris from the test fluid were also analyzed.ResultsAfter biotribological tests, surface cracks were found in both treated and untreated osteochondral grafts. In treated grafts, the COF increased, and the proteoglycan content in the cartilage tissue decreased, leading to structural changes. Chondrocytes from treated grafts showed increased expression of genes encoding for degradative enzymes, while cartilage-specific gene expression and metabolic activity exhibited no significant differences between treated and untreated groups. No measurable difference in the wear debris in the test fluid was found.ConclusionsTreatment of osteochondral grafts with cytokines results in a significantly increased COF, while also leading to significant changes in cartilage proteoglycan content and cartilage matrix compression during biotribological tests.

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Effect of hyperlipidaemia to accelerate intervertebral disc degeneration in the injured rat caudal disc model

Journal of Orthopaedic Science ◽

10.1016/j.jos.2018.08.006 ◽

2019 ◽

Vol 24 (1) ◽

pp. 42-49 ◽

Cited By ~ 1

Author(s):

Yuedong Zhang ◽

Meng Si ◽

Chunpu Li ◽

Yi Liu ◽

Yingguang Han ◽

...

Keyword(s):

Intervertebral Disc ◽

Disc Degeneration ◽

Intervertebral Disc Degeneration ◽

Disc Model

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Immune deposit and vasculopathy in metabolic-active lung tissues of patients with pulmonary tuberculosis

10.1101/2021.02.09.430558 ◽

2021 ◽

Author(s):

Hui Ma ◽

Lin Wang ◽

Zilu Wen ◽

Xinchun Chen ◽

Haiying Liu ◽

...

Keyword(s):

Gene Expression ◽

Systemic Lupus Erythematosus ◽

Inflammatory Response ◽

Pulmonary Tuberculosis ◽

Autoimmune Diseases ◽

Metabolic Activity ◽

Lupus Erythematosus ◽

Immune Complexes ◽

Systemic Lupus ◽

Dsdna Antibodies

ABSTRACTMetabolic activity in pulmonary lesion is associated with disease severity and relapse risk in tuberculosis. However, the nature of the metabolic activity associated with tuberculosis in humans remains unclear. Previous works indicate that tuberculosis bears resemblance transcriptionally with systemic lupus erythematosus in peripheral blood, except that the plasma cell component was absent in tuberculosis. Here we reported that the missing transcriptional component was present within the metabolic active tissues in the lung of patients with sputum culture-negative tuberculosis, within which increased levels of circulating immune complexes and anti-dsDNA antibodies were found relative to nearby non-metabolic active tissues. Histological examination revealed specific vascular deposition of immune complexes, neutrophil extracellular traps, and vascular necrosis in the metabolic-active tissue. Thus, tuberculosis-initiated metabolic activity was associated with hyperactive antibody responses and vascular pathology, and shared features with systemic lupus erythematosus and other autoimmune diseases. We discussed these observations in the context of earlier literatures demonstrating that similar effects could be induced in humans and animal models by complete freund’s adjuvant, the most potent antibody response inducer ever reported. Our small case series, if verified in a larger size study, might help inform host-directed therapies to alleviate disease progression and augment treatment efficacy.IMPORTANCEIn patients with pulmonary tuberculosis, lung tissues were destroyed by a hyperactive inflammatory response towards M. tuberculosis. The mechanisms underlying the inflammatory response are still poorly understood. Using 18F-FDG avidity as a surrogate marker of inflammation, we have identified that hyper-inflamed tissues possessed features associated with systemic lupus erythematosus: gene expression signatures of plasma cell and immunoglobulins and increased levels of anti-dsDNA antibodies, immune deposits, and vasculopathy. This observation might suggest an explanation to why patients with tuberculosis share more gene expression signatures with autoimmune diseases than infectious diseases and why they are more likely to develop autoimmune diseases. Defining the inflammatory responses at the lesion could help inform host-directed therapies to intervene disease progression or even accelerate cure.

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