A Gómez-Aristizábal1, A Sharma2, M A Bakooshli3, M Kapoor2, P M Gilbert4, S Viswanathan5, R Gandhi6. 1. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada; The Arthritis Program, Toronto Western Hospital, Toronto, ON, Canada. 2. The Arthritis Program, Toronto Western Hospital, Toronto, ON, Canada; Division of Genetics and Development, Krembil Research Institute, Toronto, ON, Canada. 3. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, ON, Canada. 4. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, ON, Canada; Department of Biochemistry, University of Toronto, ON, Canada. 5. Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada; The Arthritis Program, Toronto Western Hospital, Toronto, ON, Canada; Division of Genetics and Development, Krembil Research Institute, Toronto, ON, Canada; Cell Therapy Program, University Health Network, Toronto, Canada. Electronic address: sowmya.viswanathan@uhnresearch.ca. 6. The Arthritis Program, Toronto Western Hospital, Toronto, ON, Canada; Division of Orthopaedic Surgery, Toronto Western Hospital, University of Toronto, Toronto, ON, Canada. Electronic address: rajiv.gandhi@uhn.ca.
Abstract
OBJECTIVE: Although, mesenchymal stromal cells (MSCs) are being clinically investigated for their use in osteoarthritis (OA), it is unclear whether their postulated therapeutic properties are equally effective in the early- and late-stages of OA. In this study we investigated MSC cytokine secretion post-exposure to synovial fluid (SF), obtained from early- vs late-stage knee OA patients to justify a potential patient stratification strategy to maximize MSC-mediated treatment effects. METHOD: Subjects were recruited and categorized into early- [Kellgren-Lawrence (KL) grade I/II, n = 12] and late-stage (KL-III/IV, n = 12) knee OA groups. SF samples were obtained, and their proteome was tested using multiplex assays, after 3-days culture, with and without MSCs. SFs cultured without MSCs were used as a baseline to identify MSC-secreted factors into SFs cultured with MSCs. Linear mixed-effect models and non-parametric tests were used to identify alterations in the MSC secretome during exposure to OA SF (3-days). MSCs cultured for 3-days in 0.5% fetal bovine serum (FBS)-supplemented medium were used to compare SF results with culture medium. RESULTS: Following exposure to OA SF, the MSC secretome contained proteins that are involved in tissue repair, angiogenesis, chemotaxis, matrix remodeling and the clotting process. However, chemokine (C-X-C motif) ligand-8 (CXCL8; chemoattractant), interleukin-6 (IL6) and chemokine (C-C motif) ligand 2 (CCL2) were elevated in the MSC-secretome in response to early- vs late-stage OA SF. CONCLUSION: Early- vs late-stage OA SF samples elicit a differential MSC secretome response, arguing for stratification of OA patients to maximize MSC-mediated therapeutic effects.
OBJECTIVE: Although, mesenchymal stromal cells (MSCs) are being clinically investigated for their use in osteoarthritis (OA), it is unclear whether their postulated therapeutic properties are equally effective in the early- and late-stages of OA. In this study we investigated MSC cytokine secretion post-exposure to synovial fluid (SF), obtained from early- vs late-stage knee OA patients to justify a potential patient stratification strategy to maximize MSC-mediated treatment effects. METHOD: Subjects were recruited and categorized into early- [Kellgren-Lawrence (KL) grade I/II, n = 12] and late-stage (KL-III/IV, n = 12) knee OA groups. SF samples were obtained, and their proteome was tested using multiplex assays, after 3-days culture, with and without MSCs. SFs cultured without MSCs were used as a baseline to identify MSC-secreted factors into SFs cultured with MSCs. Linear mixed-effect models and non-parametric tests were used to identify alterations in the MSC secretome during exposure to OA SF (3-days). MSCs cultured for 3-days in 0.5% fetal bovine serum (FBS)-supplemented medium were used to compare SF results with culture medium. RESULTS: Following exposure to OA SF, the MSC secretome contained proteins that are involved in tissue repair, angiogenesis, chemotaxis, matrix remodeling and the clotting process. However, chemokine (C-X-C motif) ligand-8 (CXCL8; chemoattractant), interleukin-6 (IL6) and chemokine (C-C motif) ligand 2 (CCL2) were elevated in the MSC-secretome in response to early- vs late-stage OA SF. CONCLUSION: Early- vs late-stage OA SF samples elicit a differential MSC secretome response, arguing for stratification of OA patients to maximize MSC-mediated therapeutic effects.
Authors: Quan Li; Guangyan Qi; Dylan Lutter; Warren Beard; Camila R S Souza; Margaret A Highland; Wei Wu; Ping Li; Yuanyuan Zhang; Anthony Atala; Xiuzhi Sun Journal: Biomolecules Date: 2022-09-17