Abstract
Objective. Osteochondral allograft (OCA) transplantation has demonstrated good long-term outcomes in treatment of cartilage defects. Viability, a key factor in clinical success, decreases with peri-implantation storage at 4°C during pathogen testing, matching logistics, and transportation. Modern, physiologic storage conditions may improve viability and enhance outcomes. Design. Osteochondral specimens from total knee arthroplasty patients (6 males, 5 females, age 56.4 ± 2.2 years) were stored in media and incubated at normoxia (21% O2) at 22°C or 37°C, and hypoxia (2% O2) at 37°C. Histology, live-dead staining, and quantitative polymerase chain reaction (qPCR) was performed 24 hours after harvest and following 7 days of incubation. Tissue architecture, cell viability, and gene expression were analyzed. Results. No significant viability or gene expression deterioration of cartilage was observed 1-week postincubation at 37°C, with or without hypoxia. Baseline viable cell density (VCD) was 94.0% ± 2.7% at day 1. At day 7, VCD was 95.1% (37°C) with normoxic storage and 92.2% (37°C) with hypoxic storage (P ≥ 0.27). Day 7 VCD (22°C) incubation was significantly lower than both the baseline and 37°C storage values (65.6%; P < 0.01). COL1A1, COL1A2, and ACAN qPCR expression was unchanged from baseline (P < 0.05) for all storage conditions at day 7, while CD163 expression, indicative of inflammatory macrophages and monocytes, was significantly lower in the 37°C groups (P < 0.01). Conclusion. Physiologic storage at 37°C demonstrates improved chondrocyte viability and metabolism, and maintained collagen expression compared with storage at 22°C. These novel findings guide development of a method to optimize short-term fresh OCA storage, which may lead to improved clinical results.
Original language | English |
---|---|
Pages (from-to) | 280S-292S |
Journal | Cartilage |
Volume | 13 |
Issue number | 1_suppl |
Early online date | 28 Nov 2019 |
DOIs | |
Publication status | Published - Dec 2021 |
Keywords
- cartilage
- defect
- donor
- osteoarthritis
- osteochondral allograft
- storage
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In: Cartilage, Vol. 13, No. 1_suppl, 12.2021, p. 280S-292S.
Research output: Contribution to journal › Article › Academic › peer-review
TY - JOUR
T1 - Modernizing Storage Conditions for Fresh Osteochondral Allografts by Optimizing Viability at Physiologic Temperatures and Conditions
AU - Denbeigh, Janet M
AU - Hevesi, Mario
AU - Paggi, Carlo A
AU - Resch, Zachary T
AU - Bagheri, Leila
AU - Mara, Kristin
AU - Arani, Arvin
AU - Zhang, Chenghao
AU - Larson, A Noelle
AU - Saris, Daniel B F
AU - Krych, Aaron J
AU - van Wijnen, Andre J
N1 - Funding Information: Denbeigh Janet M. 1 * Hevesi Mario 1 * Paggi Carlo A. 1 Resch Zachary T. 3 Bagheri Leila 1 Mara Kristin 4 Arani Arvin 5 Zhang Chenghao 1 Larson A. Noelle 1 Saris Daniel B.F. 1 2 6 https://orcid.org/0000-0003-3248-8007 Krych Aaron J. 1 7 https://orcid.org/0000-0002-5882-709X van Wijnen Andre J. 1 1 Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA 2 Department of Orthopedics, University Medical Center Utrecht, Utrecht, Netherlands 3 Center for Regenerative Medicine, Mayo Clinic, Rochester, MN, USA 4 Department of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN, USA 5 Department of Radiology, Mayo Clinic, Rochester, MN, USA 6 Reconstructive Medicine, University of Twente, Enschede, Netherlands 7 Sports Medicine Center, Mayo Clinic, Rochester, MN, USA Aaron J. Krych MD, Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. Email: [email protected] Andre J. van Wijnen PhD, Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. Email: [email protected] * Janet M. Denbeigh and Mario Hevesi are co–first authors. 11 2019 1947603519888798 © The Author(s) 2019 2019 SAGE Publications Objective . Osteochondral allograft (OCA) transplantation has demonstrated good long-term outcomes in treatment of cartilage defects. Viability, a key factor in clinical success, decreases with peri-implantation storage at 4°C during pathogen testing, matching logistics, and transportation. Modern, physiologic storage conditions may improve viability and enhance outcomes. Design . Osteochondral specimens from total knee arthroplasty patients (6 males, 5 females, age 56.4 ± 2.2 years) were stored in media and incubated at normoxia (21% O 2 ) at 22°C or 37°C, and hypoxia (2% O 2 ) at 37°C. Histology, live-dead staining, and quantitative polymerase chain reaction (qPCR) was performed 24 hours after harvest and following 7 days of incubation. Tissue architecture, cell viability, and gene expression were analyzed. Results . No significant viability or gene expression deterioration of cartilage was observed 1-week postincubation at 37°C, with or without hypoxia. Baseline viable cell density (VCD) was 94.0% ± 2.7% at day 1. At day 7, VCD was 95.1% (37°C) with normoxic storage and 92.2% (37°C) with hypoxic storage ( P ≥ 0.27). Day 7 VCD (22°C) incubation was significantly lower than both the baseline and 37°C storage values (65.6%; P < 0.01). COL1A1, COL1A2, and ACAN qPCR expression was unchanged from baseline ( P < 0.05) for all storage conditions at day 7, while CD163 expression, indicative of inflammatory macrophages and monocytes, was significantly lower in the 37°C groups ( P < 0.01). Conclusion . Physiologic storage at 37°C demonstrates improved chondrocyte viability and metabolism, and maintained collagen expression compared with storage at 22°C. These novel findings guide development of a method to optimize short-term fresh OCA storage, which may lead to improved clinical results. cartilage osteochondral allograft storage defect osteoarthritis donor Foundation for the National Institutes of Health https://doi.org/10.13039/100000009 NIH R01 AR049069 edited-state corrected-proof Supplementary material for this article is available on the Cartilage website at https://journals.sagepub.com/home/CAR . Author Contributions Janet M. Denbeigh: Conception and design, analysis and interpretation of the data, drafting of the article, critical revision of the article for important intellectual content, final approval of the article, statistical expertise, administrative, technical, or logistic support, collection and assembly of data. Mario Hevesi: Conception and design, analysis and interpretation of the data, drafting of the article, critical revision of the article for important intellectual content, final approval of the article, statistical expertise, administrative, technical, or logistic support, collection and assembly of data. Carlo A. Paggi: Critical revision of the article for important intellectual content, final approval of the article, collection and assembly of data. Zachary T. Resch: Critical revision of the article for important intellectual content, final approval of the article, provision of study materials or patients, administrative, technical, or logistic support. Leila Bagheri: Collection and assembly of data, critical revision of the article for important intellectual content, final approval of the article. Kristin Mara: Analysis and interpretation of the data, critical revision of the article for important intellectual content, final approval of the article, statistical expertise. Arvin Forghanian-Arani: Critical revision of the article for important intellectual content, final approval of the article, collection and assembly of data. Chenghao Zhang: Critical revision of the article for important intellectual content, final approval of the article, collection and assembly of data. Daniel B.F. Saris: Conception and design, critical revision of the article for important intellectual content, final approval of the article. Aaron J. Krych: Conception and design, analysis and interpretation of the data, critical revision of the article for important intellectual content, final approval of the article, provision of study materials or patients, obtaining of funding. Andre van Wijnen: Conception and design, analysis and interpretation of the data, critical revision of the article for important intellectual content, final approval of the article, provision of study materials or patients, obtaining of funding, administrative, technical, or logistic support. Acknowledgments and Funding We would like to acknowledge Catherine Gray for assistance with histology, and the teams at the Biomaterials and Histomorphometry Laboratory (Rochester) and Biorepository Cores (Arizona) for processing histology tissues. We also thank Michella H. Hagmeijer, Koen Dijkstra, and Roeland Huitsing for assistance with histological scoring, as well as the members of our research group for stimulating discussions. We are also indebted to the kind support of the Mayo Clinic Bone Bank and the significant efforts of Susan Puffer and Renae Boyum. The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was in part supported by NIH R01 AR049069 (to AJvW) for general lab supplies and investigator effort. We also thank William and Karen Eby for their generous support of our program in joint preservation. Declaration of Conflicting Interests The author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: M. Hevesi is a paid consultant for Moximed.D. B.F. Saris is a paid consultant for Cartiheal, Ivy Sports, and Smith & Nephew; is on the editorial or governing board, of Cartilage; and receives research support from Ivy Sports and Smith & Nephew. A.J. Krych receives research support from Aesculap/B. Braun, Arthrex, Inc., Arthritis Foundation, Ceterix, and Histogenics; receives IP royalties from Arthrex, Inc.; is a paid consultant from Arthrex, Inc., JRF Ortho, and Vericel; is on the editorial or governing board of the American Journal of Sports Medicine; and is a board or committee member of the International Cartilage Repair Society, International Society of Arthroscopy, Knee Surgery, and Orthopaedic Sports Medicine, Minnesota Orthopedic Society, and Musculoskeletal Transplantation Foundation. The other authors have no disclosures to make. Ethical Approval All aspects of this study were performed following institutional review board approval (IRB 13-005619). Informed Consent Written informed consent was obtained from all subjects before the study. Trial Registration Not applicable. ORCID iDs Aaron J. Krych https://orcid.org/0000-0003-3248-8007 Andre J. van Wijnen https://orcid.org/0000-0002-5882-709X Funding Information: We would like to acknowledge Catherine Gray for assistance with histology, and the teams at the Biomaterials and Histomorphometry Laboratory (Rochester) and Biorepository Cores (Arizona) for processing histology tissues. We also thank Michella H. Hagmeijer, Koen Dijkstra, and Roeland Huitsing for assistance with histological scoring, as well as the members of our research group for stimulating discussions. We are also indebted to the kind support of the Mayo Clinic Bone Bank and the significant efforts of Susan Puffer and Renae Boyum. The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was in part supported by NIH R01 AR049069 (to AJvW) for general lab supplies and investigator effort. We also thank William and Karen Eby for their generous support of our program in joint preservation. Publisher Copyright: © The Author(s) 2019.
PY - 2021/12
Y1 - 2021/12
N2 - Objective. Osteochondral allograft (OCA) transplantation has demonstrated good long-term outcomes in treatment of cartilage defects. Viability, a key factor in clinical success, decreases with peri-implantation storage at 4°C during pathogen testing, matching logistics, and transportation. Modern, physiologic storage conditions may improve viability and enhance outcomes. Design. Osteochondral specimens from total knee arthroplasty patients (6 males, 5 females, age 56.4 ± 2.2 years) were stored in media and incubated at normoxia (21% O2) at 22°C or 37°C, and hypoxia (2% O2) at 37°C. Histology, live-dead staining, and quantitative polymerase chain reaction (qPCR) was performed 24 hours after harvest and following 7 days of incubation. Tissue architecture, cell viability, and gene expression were analyzed. Results. No significant viability or gene expression deterioration of cartilage was observed 1-week postincubation at 37°C, with or without hypoxia. Baseline viable cell density (VCD) was 94.0% ± 2.7% at day 1. At day 7, VCD was 95.1% (37°C) with normoxic storage and 92.2% (37°C) with hypoxic storage (P ≥ 0.27). Day 7 VCD (22°C) incubation was significantly lower than both the baseline and 37°C storage values (65.6%; P < 0.01). COL1A1, COL1A2, and ACAN qPCR expression was unchanged from baseline (P < 0.05) for all storage conditions at day 7, while CD163 expression, indicative of inflammatory macrophages and monocytes, was significantly lower in the 37°C groups (P < 0.01). Conclusion. Physiologic storage at 37°C demonstrates improved chondrocyte viability and metabolism, and maintained collagen expression compared with storage at 22°C. These novel findings guide development of a method to optimize short-term fresh OCA storage, which may lead to improved clinical results.
AB - Objective. Osteochondral allograft (OCA) transplantation has demonstrated good long-term outcomes in treatment of cartilage defects. Viability, a key factor in clinical success, decreases with peri-implantation storage at 4°C during pathogen testing, matching logistics, and transportation. Modern, physiologic storage conditions may improve viability and enhance outcomes. Design. Osteochondral specimens from total knee arthroplasty patients (6 males, 5 females, age 56.4 ± 2.2 years) were stored in media and incubated at normoxia (21% O2) at 22°C or 37°C, and hypoxia (2% O2) at 37°C. Histology, live-dead staining, and quantitative polymerase chain reaction (qPCR) was performed 24 hours after harvest and following 7 days of incubation. Tissue architecture, cell viability, and gene expression were analyzed. Results. No significant viability or gene expression deterioration of cartilage was observed 1-week postincubation at 37°C, with or without hypoxia. Baseline viable cell density (VCD) was 94.0% ± 2.7% at day 1. At day 7, VCD was 95.1% (37°C) with normoxic storage and 92.2% (37°C) with hypoxic storage (P ≥ 0.27). Day 7 VCD (22°C) incubation was significantly lower than both the baseline and 37°C storage values (65.6%; P < 0.01). COL1A1, COL1A2, and ACAN qPCR expression was unchanged from baseline (P < 0.05) for all storage conditions at day 7, while CD163 expression, indicative of inflammatory macrophages and monocytes, was significantly lower in the 37°C groups (P < 0.01). Conclusion. Physiologic storage at 37°C demonstrates improved chondrocyte viability and metabolism, and maintained collagen expression compared with storage at 22°C. These novel findings guide development of a method to optimize short-term fresh OCA storage, which may lead to improved clinical results.
KW - cartilage
KW - defect
KW - donor
KW - osteoarthritis
KW - osteochondral allograft
KW - storage
UR - http://www.scopus.com/inward/record.url?scp=85076209344&partnerID=8YFLogxK
U2 - 10.1177/1947603519888798
DO - 10.1177/1947603519888798
M3 - Article
C2 - 31777278
SN - 1947-6035
VL - 13
SP - 280S-292S
JO - Cartilage
JF - Cartilage
IS - 1_suppl
ER -