Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution
Abstract Background ALM solution, a combination of adenosine, lidocaine and Mg2+, is an emerging small volume therapy that has been shown to prevent and correct coagulopathy and surgery-related inflammation in preclinical models, though its application in orthopaedic surgery is yet to be demonstrate...
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| Format: | Article |
| Language: | English |
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Wiley
2019-04-01
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| Series: | Journal of Experimental Orthopaedics |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40634-019-0185-5 |
| _version_ | 1827266683436269568 |
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| author | Andrew McCutchan Geoffrey P. Dobson Natalie Stewart Hayley L. Letson Andrea L. Grant Ivana-Aleksandra Jovanovic Kaushik Hazratwala Matthew Wilkinson Peter McEwen Jodie Morris |
| author_facet | Andrew McCutchan Geoffrey P. Dobson Natalie Stewart Hayley L. Letson Andrea L. Grant Ivana-Aleksandra Jovanovic Kaushik Hazratwala Matthew Wilkinson Peter McEwen Jodie Morris |
| author_sort | Andrew McCutchan |
| collection | DOAJ |
| description | Abstract Background ALM solution, a combination of adenosine, lidocaine and Mg2+, is an emerging small volume therapy that has been shown to prevent and correct coagulopathy and surgery-related inflammation in preclinical models, though its application in orthopaedic surgery is yet to be demonstrated. The effect of ALM solution on chondrocytes is unknown. The aim of this preliminary study was to investigate the effect of ALM solution on viability and inflammatory responses of chondrogenically-differentiated human bone marrow-derived mesenchymal stem cells (chondro-MSC), in vitro. Methods Chondro-MSC were exposed to media only, saline (0.9% NaCl or 1.3% NaCl) only, or saline containing ALM (1 mM adenosine, 3 mM lidocaine, 2.5 mM Mg2+) or tranexamic acid (TXA, 100 mg/ml) for 1 or 4 h. Responses to ALM solutions containing higher lidocaine concentrations were also compared. Chondrocyte viability was determined using WST-8 colorimetric assays and inflammatory cytokine (TNF-α, IL-1β, IL-8) and matrix metalloproteinases (MMP-3, MMP-12, MMP-13) concentrations using multiplex bead arrays. Results The viability of chondro-MSC was significantly greater after 1 h treatment with ALM compared to saline (96.2 ± 7.9 versus 75.6 ± 7.3%). Extension of exposure times to 4 h had no significant adverse effect on cell viability after treatment with ALM (1 h, 85.4 ± 5.6 v 4 h, 74.0 ± 15.2%). Cytotoxicity was evident following exposure to solutions containing lidocaine concentrations greater than 30 mM. There were no significant differences in viability (80 ± 5.4 v 57.3 ± 16.2%) or secretion of IL-8 (60 ± 20 v 160 ± 50 pg/ml), MMP-3 (0.95 ± 0.6 v 3.4 ± 1.6 ng/ml), and MMP-13 (4.2 ± 2.4 v 9.2 ± 4.3 ng/ml) in chondro-MSC exposed to saline, ALM or TXA. Conclusions Short-term, in vitro exposure to clinically-relevant concentrations of ALM solution had no adverse inflammatory or chondrotoxic effects on human chondro-MSC, with responses comparable to saline and TXA. These findings provide support for continued evaluation of ALM solution as a possible therapeutic to improve outcomes following orthopaedic procedures. |
| first_indexed | 2024-03-08T07:57:44Z |
| format | Article |
| id | doaj.art-6559b6eccfb14618946fec7c62c6262b |
| institution | Directory Open Access Journal |
| issn | 2197-1153 |
| language | English |
| last_indexed | 2025-03-22T04:21:45Z |
| publishDate | 2019-04-01 |
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| series | Journal of Experimental Orthopaedics |
| spelling | doaj.art-6559b6eccfb14618946fec7c62c6262b2024-04-28T07:43:36ZengWileyJournal of Experimental Orthopaedics2197-11532019-04-01611910.1186/s40634-019-0185-5Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solutionAndrew McCutchan0Geoffrey P. Dobson1Natalie Stewart2Hayley L. Letson3Andrea L. Grant4Ivana-Aleksandra Jovanovic5Kaushik Hazratwala6Matthew Wilkinson7Peter McEwen8Jodie Morris9Department of Haematology and Bone Marrow Transplantation, Townsville HospitalHeart, Trauma and Sepsis Research Laboratory, College of Medicine and Dentistry, James Cook UniversityDepartment of Haematology and Bone Marrow Transplantation, Townsville HospitalHeart, Trauma and Sepsis Research Laboratory, College of Medicine and Dentistry, James Cook UniversityThe Orthopaedic Research Institute of QueenslandThe Orthopaedic Research Institute of QueenslandHeart, Trauma and Sepsis Research Laboratory, College of Medicine and Dentistry, James Cook UniversityHeart, Trauma and Sepsis Research Laboratory, College of Medicine and Dentistry, James Cook UniversityHeart, Trauma and Sepsis Research Laboratory, College of Medicine and Dentistry, James Cook UniversityHeart, Trauma and Sepsis Research Laboratory, College of Medicine and Dentistry, James Cook UniversityAbstract Background ALM solution, a combination of adenosine, lidocaine and Mg2+, is an emerging small volume therapy that has been shown to prevent and correct coagulopathy and surgery-related inflammation in preclinical models, though its application in orthopaedic surgery is yet to be demonstrated. The effect of ALM solution on chondrocytes is unknown. The aim of this preliminary study was to investigate the effect of ALM solution on viability and inflammatory responses of chondrogenically-differentiated human bone marrow-derived mesenchymal stem cells (chondro-MSC), in vitro. Methods Chondro-MSC were exposed to media only, saline (0.9% NaCl or 1.3% NaCl) only, or saline containing ALM (1 mM adenosine, 3 mM lidocaine, 2.5 mM Mg2+) or tranexamic acid (TXA, 100 mg/ml) for 1 or 4 h. Responses to ALM solutions containing higher lidocaine concentrations were also compared. Chondrocyte viability was determined using WST-8 colorimetric assays and inflammatory cytokine (TNF-α, IL-1β, IL-8) and matrix metalloproteinases (MMP-3, MMP-12, MMP-13) concentrations using multiplex bead arrays. Results The viability of chondro-MSC was significantly greater after 1 h treatment with ALM compared to saline (96.2 ± 7.9 versus 75.6 ± 7.3%). Extension of exposure times to 4 h had no significant adverse effect on cell viability after treatment with ALM (1 h, 85.4 ± 5.6 v 4 h, 74.0 ± 15.2%). Cytotoxicity was evident following exposure to solutions containing lidocaine concentrations greater than 30 mM. There were no significant differences in viability (80 ± 5.4 v 57.3 ± 16.2%) or secretion of IL-8 (60 ± 20 v 160 ± 50 pg/ml), MMP-3 (0.95 ± 0.6 v 3.4 ± 1.6 ng/ml), and MMP-13 (4.2 ± 2.4 v 9.2 ± 4.3 ng/ml) in chondro-MSC exposed to saline, ALM or TXA. Conclusions Short-term, in vitro exposure to clinically-relevant concentrations of ALM solution had no adverse inflammatory or chondrotoxic effects on human chondro-MSC, with responses comparable to saline and TXA. These findings provide support for continued evaluation of ALM solution as a possible therapeutic to improve outcomes following orthopaedic procedures.http://link.springer.com/article/10.1186/s40634-019-0185-5Chondro-MSCChondrocyteALMTranexamic acidViabilityInflammation |
| spellingShingle | Andrew McCutchan Geoffrey P. Dobson Natalie Stewart Hayley L. Letson Andrea L. Grant Ivana-Aleksandra Jovanovic Kaushik Hazratwala Matthew Wilkinson Peter McEwen Jodie Morris Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution Journal of Experimental Orthopaedics Chondro-MSC Chondrocyte ALM Tranexamic acid Viability Inflammation |
| title | Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution |
| title_full | Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution |
| title_fullStr | Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution |
| title_full_unstemmed | Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution |
| title_short | Absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically-differentiated human mesenchymal stem cells to adenosine, lidocaine and Mg2+ solution |
| title_sort | absence of cytotoxic and inflammatory effects following in vitro exposure of chondrogenically differentiated human mesenchymal stem cells to adenosine lidocaine and mg2 solution |
| topic | Chondro-MSC Chondrocyte ALM Tranexamic acid Viability Inflammation |
| url | http://link.springer.com/article/10.1186/s40634-019-0185-5 |
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