PMID- 24999113 OWN - NLM STAT- MEDLINE DCOM- 20150512 LR - 20211021 IS - 1522-9653 (Electronic) IS - 1063-4584 (Print) IS - 1063-4584 (Linking) VI - 22 IP - 9 DP - 2014 Sep TI - A high-throughput model of post-traumatic osteoarthritis using engineered cartilage tissue analogs. PG - 1282-90 LID - S1063-4584(14)01155-8 [pii] LID - 10.1016/j.joca.2014.06.032 [doi] AB - OBJECTIVE: A number of in vitro models of post-traumatic osteoarthritis (PTOA) have been developed to study the effect of mechanical overload on the processes that regulate cartilage degeneration. While such frameworks are critical for the identification therapeutic targets, existing technologies are limited in their throughput capacity. Here, we validate a test platform for high-throughput mechanical injury incorporating engineered cartilage. METHOD: We utilized a high-throughput mechanical testing platform to apply injurious compression to engineered cartilage and determined their strain and strain rate dependent responses to injury. Next, we validated this response by applying the same injury conditions to cartilage explants. Finally, we conducted a pilot screen of putative PTOA therapeutic compounds. RESULTS: Engineered cartilage response to injury was strain dependent, with a 2-fold increase in glycosaminoglycan (GAG) loss at 75% compared to 50% strain. Extensive cell death was observed adjacent to fissures, with membrane rupture corroborated by marked increases in lactate dehydrogenase (LDH) release. Testing of established PTOA therapeutics showed that pan-caspase inhibitor [Z-VAD-FMK (ZVF)] was effective at reducing cell death, while the amphiphilic polymer [Poloxamer 188 (P188)] and the free-radical scavenger [N-Acetyl-L-cysteine (NAC)] reduced GAG loss as compared to injury alone. CONCLUSIONS: The injury response in this engineered cartilage model replicated key features of the response of cartilage explants, validating this system for application of physiologically relevant injurious compression. This study establishes a novel tool for the discovery of mechanisms governing cartilage injury, as well as a screening platform for the identification of new molecules for the treatment of PTOA. CI - Copyright (c) 2014 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved. FAU - Mohanraj, B AU - Mohanraj B AD - McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: mbhavana@mail.med.upenn.edu. FAU - Meloni, G R AU - Meloni GR AD - McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: gmeloni@mail.med.upenn.edu. FAU - Mauck, R L AU - Mauck RL AD - McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA 19104, USA; Collaborative Research Partner Acute Cartilage Injury Program of AO Foundation, Davos, Switzerland; Translational Musculoskeletal Research Center, Philadelphia Veterans Administration Medical Center, Philadelphia, PA 19104, USA. Electronic address: lemauck@mail.med.upenn.edu. FAU - Dodge, G R AU - Dodge GR AD - McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA; Collaborative Research Partner Acute Cartilage Injury Program of AO Foundation, Davos, Switzerland; Translational Musculoskeletal Research Center, Philadelphia Veterans Administration Medical Center, Philadelphia, PA 19104, USA. Electronic address: gdodge@mail.med.upenn.edu. LA - eng GR - I01 RX001213/RX/RRD VA/United States GR - P30 AR050950/AR/NIAMS NIH HHS/United States PT - Evaluation Study PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20140704 PL - England TA - Osteoarthritis Cartilage JT - Osteoarthritis and cartilage JID - 9305697 RN - 0 (Amino Acid Chloromethyl Ketones) RN - 0 (Caspase Inhibitors) RN - 0 (Glycosaminoglycans) RN - 0 (benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone) RN - 106392-12-5 (Poloxamer) RN - WYQ7N0BPYC (Acetylcysteine) SB - IM MH - Acetylcysteine/pharmacology MH - Amino Acid Chloromethyl Ketones/pharmacology MH - Animals MH - Cartilage, Articular/drug effects/*injuries/pathology/physiopathology MH - Caspase Inhibitors/pharmacology MH - Cattle MH - Cell Death/drug effects MH - Disease Models, Animal MH - Drug Evaluation, Preclinical/methods MH - Glycosaminoglycans/metabolism MH - High-Throughput Screening Assays/methods MH - Materials Testing/methods MH - Osteoarthritis/*etiology MH - Pilot Projects MH - Poloxamer/pharmacology MH - Stress, Mechanical MH - Tissue Engineering/*methods PMC - PMC4313617 MID - NIHMS618719 OTO - NOTNLM OT - Cartilage tissue analog (CTA) OT - High-throughput screening OT - Impact loading OT - Injurious compression OT - Tissue engineering COIS- Conflicts of Interest All authors have no conflicts of interest to report. EDAT- 2014/07/08 06:00 MHDA- 2015/05/13 06:00 PMCR- 2015/09/01 CRDT- 2014/07/08 06:00 PHST- 2014/02/08 00:00 [received] PHST- 2014/05/31 00:00 [revised] PHST- 2014/06/25 00:00 [accepted] PHST- 2014/07/08 06:00 [entrez] PHST- 2014/07/08 06:00 [pubmed] PHST- 2015/05/13 06:00 [medline] PHST- 2015/09/01 00:00 [pmc-release] AID - S1063-4584(14)01155-8 [pii] AID - 10.1016/j.joca.2014.06.032 [doi] PST - ppublish SO - Osteoarthritis Cartilage. 2014 Sep;22(9):1282-90. doi: 10.1016/j.joca.2014.06.032. Epub 2014 Jul 4.