Safety and efficacy of human juvenile chondrocyte-derived cell sheets for osteochondral defect treatment

Makoto Kondo; Sumako Kameishi; Kyungsook Kim; Nicolas F Metzler; Travis G Maak; Douglas T Hutchinson; Angela A Wang; Miki Maehara; Masato Sato; David W Grainger; Teruo Okano

doi:10.1038/s41536-021-00173-9

Safety and efficacy of human juvenile chondrocyte-derived cell sheets for osteochondral defect treatment

NPJ Regen Med. 2021 Oct 15;6(1):65. doi: 10.1038/s41536-021-00173-9.

Authors

Makoto Kondo¹, Sumako Kameishi², Kyungsook Kim², Nicolas F Metzler^{2

3}, Travis G Maak⁴, Douglas T Hutchinson^{4

5}, Angela A Wang^{4

5}, Miki Maehara⁶, Masato Sato⁶, David W Grainger^{2

3}, Teruo Okano^{7

8}

Affiliations

¹ Cell Sheet Tissue Engineering Center (CSTEC), Department of Pharmaceutics and Pharmaceutical Chemistry, Health Sciences, University of Utah, 30 South 2000 East, Salt Lake City, UT, 84112, USA. makoto.kondo@utah.edu.
² Cell Sheet Tissue Engineering Center (CSTEC), Department of Pharmaceutics and Pharmaceutical Chemistry, Health Sciences, University of Utah, 30 South 2000 East, Salt Lake City, UT, 84112, USA.
³ Department of Biomedical Engineering, University of Utah, 36 S. Wasatch Drive SMBB 3100, Salt Lake City, UT, 84112, USA.
⁴ Department of Orthopaedic Surgery, University of Utah Orthopaedic Center, University of Utah, 590 Wakara Way, Salt Lake City, UT, 84108, USA.
⁵ Pediatric Orthopaedics Surgery, Primary Children's Hospital Orthopedics, 100 North Mario Capecchi Dr. Suite 4550, Salt Lake City, UT, 84113, USA.
⁶ Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, 259-1193, Japan.
⁷ Cell Sheet Tissue Engineering Center (CSTEC), Department of Pharmaceutics and Pharmaceutical Chemistry, Health Sciences, University of Utah, 30 South 2000 East, Salt Lake City, UT, 84112, USA. teruo.okano@utah.edu.
⁸ Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan. teruo.okano@utah.edu.

Abstract

Knee cartilage does not regenerate spontaneously after injury, and a gold standard regenerative treatment algorithm has not been established. This study demonstrates preclinical safety and efficacy of scaffold-free, human juvenile cartilage-derived-chondrocyte (JCC) sheets produced from routine surgical discards using thermo-responsive cultureware. JCCs exhibit stable and high growth potential in vitro over passage 10, supporting possibilities for scale-up to mass production for commercialization. JCC sheets contain highly viable, densely packed cells, show no anchorage-independent cell growth, express mesenchymal surface markers, and lack MHC II expression. In nude rat focal osteochondral defect models, stable neocartilage formation was observed at 4 weeks by JCC sheet transplantation without abnormal tissue growth over 24 weeks in contrast to the nontreatment group showing no spontaneous cartilage repair. Regenerated cartilage was safranin-O positive, contained type II collagen, aggrecan, and human vimentin, and lacked type I collagen, indicating that the hyaline-like neocartilage formed originates from transplanted JCC sheets rather than host-derived cells. This study demonstrates the safety of JCC sheets and stable hyaline cartilage formation with engineered JCC sheets utilizing a sustainable tissue supply. Cost-benefit and scaling issues for sheet fabrication and use support feasibility of this JCC sheet strategy in clinical cartilage repair.