A Degenerative/Proinflammatory Intervertebral Disc Organ Culture: An Ex Vivo Model for Anti-inflammatory Drug and Cell Therapy

2016 ◽  
Vol 22 (1) ◽  
pp. 8-19 ◽  
Author(s):  
Graciosa Q. Teixeira ◽  
Antje Boldt ◽  
Ines Nagl ◽  
Catarina Leite Pereira ◽  
Karin Benz ◽  
...  
Keyword(s):  
Cell Therapy ◽  
Intervertebral Disc ◽  
Organ Culture ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Inflammatory Drug ◽  
Anti Inflammatory

Anti-Inflammatory Effects of Novel Standardized Platelet Rich Plasma Releasates on Knee Osteoarthritic Chondrocytes and Cartilage in vitro

2019 ◽  
Vol 20 (11) ◽  
pp. 920-933 ◽  
Author(s):  
Lucía Gato-Calvo ◽  
Tamara Hermida-Gómez ◽  
Cristina R. Romero ◽  
Elena F. Burguera ◽  
Francisco J. Blanco
Keyword(s):  
Gene Expression ◽  
Ex Vivo ◽  
Platelet Rich Plasma ◽  
Cartilage Explants ◽  
Ex Vivo Model ◽  
Chondrocyte Viability ◽  
Platelet Concentration ◽  
The Absolute ◽  
Anti Inflammatory

Background: Platelet Rich Plasma (PRP) has recently emerged as a potential treatment for osteoarthritis (OA), but composition heterogeneity hampers comparison among studies, with the result that definite conclusions on its efficacy have not been reached. Objective: 1) To develop a novel methodology to prepare a series of standardized PRP releasates (PRP-Rs) with known absolute platelet concentrations, and 2) To evaluate the influence of this standardization parameter on the anti-inflammatory properties of these PRP-Rs in an in vitro and an ex vivo model of OA. Methods: A series of PRPs was prepared using the absolute platelet concentration as the standardization parameter. Doses of platelets ranged from 0% (platelet poor plasma, PPP) to 1.5·105 platelets/µl. PRPs were then activated with CaCl2 to obtain releasates (PRP-R). Chondrocytes were stimulated with 10% of each PRP-R in serum-free culture medium for 72 h to assess proliferation and viability. Cells were co-stimulated with interleukin (IL)-1β (5 ng/ml) and 10% of each PRP-R for 48 h to determine the effects on gene expression, secretion and intra-cellular content of common markers associated with inflammation, catabolism and oxidative stress in OA. OA cartilage explants were co-stimulated with IL-1β (5 ng/ml) and 10% of either PRP-R with 0.75·105 platelets/µl or PRP-R with 1.5·105 platelets/µl for 21 days to assess matrix inflammatory degradation. Results: Chondrocyte viability was not affected, and proliferation was dose-dependently increased. The gene expression of all pro-inflammatory mediators was significantly and dose-independently reduced, except for that of IL-1β and IL-8. Immunoblotting corroborated this effect for inducible NO synthase (NOS2). Secreted matrix metalloproteinase-13 (MMP-13) was reduced to almost basal levels by the PRP-R from PPP. Increasing platelet dosage led to progressive loss to this anti-catabolic ability. Safranin O and toluidine blue stains supported the beneficial effect of low platelet dosage on cartilage matrix preservation. Conclusion: We have developed a methodology to prepare PRP releasates using the absolute platelet concentration as the standardization parameter. Using this approach, the composition of the resulting PRP derived product is independent of the donor initial basal platelet count, thereby allowing the evaluation of its effects objectively and reproducibly. In our OA models, PRP-Rs showed antiinflammatory, anti-oxidant and anti-catabolic properties. Platelet enrichment could favor chondrocyte proliferation but is not necessary for the above effects and could even be counter-productive.

scholarly journals Resveratrol displays anti-inflammatory properties in an ex vivo model of immune mediated inflammatory arthritis

2018 ◽  
Vol 2 (1) ◽  
Author(s):  
S. Lomholt ◽  
A. Mellemkjaer ◽  
M. B. Iversen ◽  
S. B. Pedersen ◽  
T. W. Kragstrup
Keyword(s):  
Inflammatory Arthritis ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Immune Mediated ◽  
Anti Inflammatory

scholarly journals Unravelling the role of Mesenchymal Stem/Stromal Cells Secretome in intervertebral disc degeneration in a pro-inflammatory/degenerative ex vivo model

2020 ◽  
Author(s):  
JR Ferreira ◽  
GQ Teixeira ◽  
E Neto ◽  
C Ribeiro-Machado ◽  
AM Silva ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Disc Degeneration ◽  
Stromal Cells ◽  
Intervertebral Disc Degeneration ◽  
Ex Vivo ◽  
Ex Vivo Model

Abstract The authors have withdrawn this preprint due to author disagreement.

An ex vivo model of human corneal rim perfusion organ culture

2021 ◽  
pp. 108891
Author(s):  
Michael Peng ◽  
Tyler J. Margetts ◽  
Chenna Kesavulu Sugali ◽  
Naga Pradeep Rayana ◽  
Jiannong Dai ◽  
...  
Keyword(s):  
Organ Culture ◽  
Ex Vivo ◽  
Ex Vivo Model

Glucan particles as suitable carriers for the natural anti-inflammatory compounds curcumin and diplacone – Evaluation in an ex vivo model

2020 ◽  
Vol 582 ◽  
pp. 119318
Author(s):  
Dominik Rotrekl ◽  
Bert Devriendt ◽  
Eric Cox ◽  
Lenka Kavanová ◽  
Martin Faldyna ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Ex Vivo Model ◽  
Anti Inflammatory

Development of a Large Animal Long-Term Intervertebral Disc Organ Culture Model That Includes the Bony Vertebrae for Ex Vivo Studies

2016 ◽  
Vol 22 (7) ◽  
pp. 636-643 ◽  
Author(s):  
Michael Grant ◽  
Laura M. Epure ◽  
Omar Salem ◽  
Nizar AlGarni ◽  
Ovidiu Ciobanu ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Organ Culture ◽  
Ex Vivo ◽  
Large Animal ◽  
Culture Model

Human Ocular Perfusion Organ Culture: A Versatile Ex Vivo Model for Glaucoma Research

2000 ◽  
Vol 9 (6) ◽  
pp. 468-479 ◽  
Author(s):  
Iok-Hou Pang ◽  
Mitchell D. McCartney ◽  
Thomas H. Steely ◽  
Abbot F. Clark
Keyword(s):  
Organ Culture ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Ocular Perfusion

scholarly journals Unravelling the role of Mesenchymal Stem/Stromal Cells Secretome in intervertebral disc degeneration in a pro-inflammatory/degenerative ex vivo model

2020 ◽  
Author(s):  
JR Ferreira ◽  
GQ Teixeira ◽  
E Neto ◽  
C Ribeiro-Machado ◽  
AM Silva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Intervertebral Disc ◽  
Stromal Cells ◽  
Ex Vivo ◽  
Low Oxygen ◽  
Ex Vivo Model ◽  
A Cell ◽  
New Perspective ◽  
Regulated Gene Expression ◽  
Ivd Degeneration

Abstract Background: Mesenchymal stem/stromal cells (MSCs) have been increasingly used in clinical trials for intervertebral disc (IVD) degeneration. Here, we aimed to evaluate the potential of a cell-free approach to degenerated IVD, testing if MSCs secretome can stimulate a regenerative response by modulating the IVD inflammatory cascade. Methods: Human bone marrow-derived MSCs were pre-conditioned with IL-1β (10 ng/mL) and low oxygen (6% O2). The secretome of MSCs (MSCsec) was collected after 48h. Bovine IVD tissue explants cultured in pro-inflammatory/degenerative conditions (needle puncture + IL-1β) were treated with MSCsec or co-cultured with MSCs. Results: MSCsec obtained upon IL-1β-pre-conditioning, as well as MSCs co-culture, down-regulated gene expression of pro-inflammatory cytokines, bIL-6 and bIL-8 after 48h, in IVD. IVD matrix degrading enzymes, bMMP1 and bMMP3, were downregulated and upregulated, respectively, in the presence of MSCsec, but not MSCs. After 14 days, MSCsec-treated IVDs revealed increased aggrecan content at the protein level, contrarily to MSCs/IVD co-cultures. Interestingly, IL-1β-preconditioning only, but not IL-1β-IVD, increased gene expression of hADAMTS5 and hTIMP-1in MSCs. Additionally, conditioned medium from MSCsec-treated IVDs did not promote angiogenesis or neurogenesis. In MSCsec-treated IVD, an increase in MCP-3 and GCP-2 was observed, while SDF-1α, TNF-α, IGF-1, Eotaxin 3, FGF-9, MIP-1δ, IFN-γ, IL-5, TNF-β, IL-4, TGF-β1, IL-16, IGFBP-3 and IGFBP-4 were decreased, compared with MSCs/IVD co-cultures. Conclusions: MSCsec obtained upon IL1β-preconditioning, present an immunomodulatory role in degenerated IVD, as well as MSCs. Nevertheless, MSCsec but not MSCs, potentiate aggrecan deposition in IVD in pro-inflammatory/degenerative conditions. This finding can open new perspective on the use of MSCsec as a cell-based/cell-free approach to LBP.

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