Acetaminophen, bupivacaine, Duramorph, and Toradol: A comparison of chondrocyte viability and gene expression changes in osteoarthritic human chondrocytes

Materials on the basis of cycloolefin copolymers (COC) are suitable for subchondral defect repairs. The objective of this study was to evaluate the influence of surface modification of COC and COC/LLDPE blends on the viability and gene expression of chondrocytes. Human chondrocytes were incubated on the surface of the studied materials. Half of the materials were plasmatically modified with a subsequent type II collagen application. The gene expression of matrix metalloproteinases (MMP-1,-3,-13), pro-inflammatory cytokines (IL-1, TNF-alpha) and apoptotic molecules (BAX, Bcl-2) was evaluated using quantitative Taq-Man PCR after 48 h incubation. Chondrocyte viability was evaluated by the MTT test after 2, 4 and 8 days of incubation. The synthesis of MMPs was measured by ELISA assay in cell culture medium after 48 h of incubation. Chondrocytes incubated on plasmatically modified in contrast to unmodified materials demonstrated significantly increased gene expression of IL-1 (p<0.05), MMP-1 and MMP-3 (p<0.05 for both comparisons) as well as MMP-13 (p<0.001). Increased gene expression was confirmed by significantly increased production of active forms of particular MMPs into the cell culture medium. Unlike surface unmodified polymers, the modified materials showed timedependent reduction of chondrocyte viability. The gene expression of TNF-α and apoptotic molecules by chondrocytes was not significantly changed by different materials. Cycloolefin copolymers and their blends may represent suitable materials for tissue engineering, however, their surface modification followed by collagen type II application may, at least under in vitro conditions, reduce the viability of chondrocytes and induce their pro-destructive behavior. The potential benefit or disadvantage of surface modifications of materials for osteochondral defect repairs needs to be further elucidated.

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Acetaminophen, Bupivacaine, Duramorph, and Toradol: A comparison of Gene Expression changes in Osteoarthritic Human Chondrocytes

Orthopaedic Journal of Sports Medicine ◽

10.1177/2325967120s00390 ◽

2020 ◽

Vol 8 (7_suppl6) ◽

pp. 2325967120S0039

Author(s):

Philip Sakalian ◽

Christopher Cooke

Keyword(s):

Gene Expression ◽

Cell Death ◽

Cross Sections ◽

Caspase 3 ◽

Alginate Bead ◽

Culture Method ◽

Culture Technique ◽

Human Chondrocytes ◽

Chondrocyte Viability ◽

Cellular Apoptosis

Objectives: To determine if exposure to Duramorph results in a lower level of chondrotoxicity than Acetaminophen, Bupivacaine, or Toradol. Methods: Distal femur and proximal tibia cross sections were obtained during total knee arthroplasty and a 3D Chondrocyte culture technique was performed. Digested cartilage was then treated via an alginate bead culture method to ensure lower rates of differentiation. Chondrocyte cells were exposed to the study chemicals and gene expression and chondrocyte viability were measured by RT-PCR and flow cytometry, respectively. Results: Exposure to Bupivacaine leads to cellular apoptosis with a % PI+ (dead) cells of 30.45, significantly greater than the other test groups P<0.001. Exposure to Toradol, duramorph and acetaminophen did not result in a significant increase in cell death. Bupivicaine also lead to an increase in Caspase 3 gene expression (p<0.001) when compared to all other groups. Acetophinophen also lead to an increased Caspase 3 gene expression when compared to control. Neither Toradol nor Duramorph led to an increase in caspase 3 gene expression. Conclusion: Our data show Duramorph and Toradol were not cytotoxic to human chondrocytes and may be better alternatives to the frequently used and more cytotoxic Bupivacaine. Acetominophen did not result in increased cell death however did show increased caspase 3 gene expression and caution should be considered.

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Faculty Opinions recommendation of LEF1-mediated MMP13 gene expression is repressed by SIRT1 in human chondrocytes.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.727488527.793556145 ◽

2019 ◽

Author(s):

David Young

Keyword(s):

Gene Expression ◽

Human Chondrocytes

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

Current Pharmaceutical Biotechnology ◽

10.2174/1389201020666190619111118 ◽

2019 ◽

Vol 20 (11) ◽

pp. 920-933 ◽

Cited By ~ 3

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.

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Phenotypic characteristics and gene expression profiles of human chondrocytes in tissuegene C

Osteoarthritis and Cartilage ◽

10.1016/j.joca.2021.02.288 ◽

2021 ◽

Vol 29 ◽

pp. S211-S212

Author(s):

I. Uzieliene ◽

E. Bagdonas ◽

J. Denkovskij ◽

E. Bernotiene ◽

H.-J. Yoon ◽

...

Keyword(s):

Gene Expression ◽

Expression Profiles ◽

Gene Expression Profiles ◽

Human Chondrocytes ◽

Phenotypic Characteristics

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Cytotoxic responses of human chondrocytes to bupivacaine, levobupivacaine, and ropivacaine

Asian Biomedicine ◽

10.1515/abm-2019-0017 ◽

2019 ◽

Vol 12 (4) ◽

pp. 169-178

Author(s):

Pudkrong Kaewpichit ◽

Somrat Charuluxananan ◽

Monpichar Srisa-Art ◽

Sarocha Sisawat ◽

Vitavat Aksornkitti ◽

...

Keyword(s):

Colorimetric Assay ◽

Control Cell ◽

Human Chondrocytes ◽

No Production ◽

Chondrocyte Viability ◽

Chondrocyte Death ◽

Phosphate Buffered Saline ◽

Cytotoxic Responses ◽

Dependent Pathway

AbstractBackgroundIntra-articular injections of local anesthetics are used commonly in articular surgery. However, chondrocyte viability and metabolism may be adversely affected by various anesthetics.ObjectivesTo assess the chondrotoxic effects of bupivacaine, levobupivacaine, and ropivacaine on human chondrocytes and elucidate possible mechanisms of chondrocyte death.MethodsCultured human chondrocytes (CHON-001) were exposed to 0.25% or 0.5% of bupivacaine, levobupivacaine, and ropivacaine in vitro. Cell viability was determined by flow cytometry after 15, 30, 60, and 120 min of exposure. Chondrocyte reactive oxygen species (ROS) production was measured every 10 min for up to 1 h using 2ʹ,7ʹ-dichlorodihydrofluorescein staining. Chondrocyte production of glycosaminoglycan was measured by capillary electrophoresis. NO production was measured using a colorimetric assay kit.ResultsWe found a significant increase in chondrotoxicity dependent on exposure time and concentration of the anesthetic. At 60 min, chondrocyte viability was significantly (P < 0.05) decreased when exposed to 0.5% levobupivacaine (32.5%), or 0.25% or 0.5% bupivacaine (34.3% or 46.5%, respectively) compared with exposure to phosphate-buffered saline (PBS) vehicle as a control. Cell death at 120 min was mainly necrosis. There was no difference in viability after treatment with either concentration (0.25% or 0.5%) of ropivacaine at any time compared with exposure to PBS. We found increased production of NO, while ROS decreased after exposure to any of the anesthetics tested.ConclusionsRopivacaine may be safer than bupivacaine or levobupivacaine as an intra-articular analgesic. Chondrotoxicity of anesthetics in vitro may be mediated via a reactive nitrogen species-dependent pathway.

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