Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration
(1) Background: Inexplicable low back and neck pain frequently results from spinal disc degeneration with an imbalanced intervertebral disc (IVD) cell homeostasis. We hypothesize that introducing MSC expressing a sustained cartilage-anabolic factor in the IVD may stimulate the mucoid materials secre...
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MDPI AG
2022-02-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/10/3/555 |
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| author | Long-Yi Chan Cheng-Chung Chang Po-Liang Lai Tomoji Maeda Horng-Chaung Hsu Chin-Yu Lin Shu-Jui Kuo |
| author_facet | Long-Yi Chan Cheng-Chung Chang Po-Liang Lai Tomoji Maeda Horng-Chaung Hsu Chin-Yu Lin Shu-Jui Kuo |
| author_sort | Long-Yi Chan |
| collection | DOAJ |
| description | (1) Background: Inexplicable low back and neck pain frequently results from spinal disc degeneration with an imbalanced intervertebral disc (IVD) cell homeostasis. We hypothesize that introducing MSC expressing a sustained cartilage-anabolic factor in the IVD may stimulate the mucoid materials secreted from the IVD cells, promote the MSC’s chondrogenesis and maintain the hydration content providing mechanical strength to decelerate the disc degeneration progression; (2) Methods: This study expressed a cartilage-anabolic factor runx1 by a baculoviral vector (BV) transduced MSCs through a Cre/LoxP gene editing and recombination system for sustained recombinant runx1 transcription factor production. The Cre/LoxP BV modified MSCs were encapsulated by hyaluronan hydrogel, due to its’ vital composition in ECM of a healthy disc and transplanted to a punctured coccygeal disc in rats through micro-injection, followed by X-ray radiography and histological analysis at the 4- and 12-weeks post-transplantation; (3) Results: Data reveals the Cre/LoxP BV system-mediated long-termed runx1 gene expression, possessing good biosafety characteristics in the in vitro cell transduction and in vivo MSCs transplantation, and maintained superior hydration content in the disc than that of mock transduced MSCs; (4) Conclusions: This proof-of-concept study fulfills the need of implanting therapeutic cells accompanied with microinjection in the disc, such as a discography and paves a road to manufacture composite hyaluronan, such as peptide modified hyaluronan as an MSC carrier for IVD regeneration in the future study. |
| first_indexed | 2024-03-09T20:05:45Z |
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| id | doaj.art-553a7f248db84da2a447b1bb928dabe4 |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T20:05:45Z |
| publishDate | 2022-02-01 |
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| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-553a7f248db84da2a447b1bb928dabe42023-11-24T00:31:57ZengMDPI AGBiomedicines2227-90592022-02-0110355510.3390/biomedicines10030555Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc DegenerationLong-Yi Chan0Cheng-Chung Chang1Po-Liang Lai2Tomoji Maeda3Horng-Chaung Hsu4Chin-Yu Lin5Shu-Jui Kuo6Institute of New Drug Development, College of Medicine, China Medical University, Taichung 40402, TaiwanInstitute of New Drug Development, College of Medicine, China Medical University, Taichung 40402, TaiwanDepartment of Orthopedic Surgery, Chang Gung Memorial Hospital, Tau-Yuan 333, TaiwanTsuzuki Institute for Traditional Medicine, College of Pharmacy, China Medical University, Taichung 40402, TaiwanSchool of Medicine, China Medical University, Taichung 40402, TaiwanInstitute of New Drug Development, College of Medicine, China Medical University, Taichung 40402, TaiwanSchool of Medicine, China Medical University, Taichung 40402, Taiwan(1) Background: Inexplicable low back and neck pain frequently results from spinal disc degeneration with an imbalanced intervertebral disc (IVD) cell homeostasis. We hypothesize that introducing MSC expressing a sustained cartilage-anabolic factor in the IVD may stimulate the mucoid materials secreted from the IVD cells, promote the MSC’s chondrogenesis and maintain the hydration content providing mechanical strength to decelerate the disc degeneration progression; (2) Methods: This study expressed a cartilage-anabolic factor runx1 by a baculoviral vector (BV) transduced MSCs through a Cre/LoxP gene editing and recombination system for sustained recombinant runx1 transcription factor production. The Cre/LoxP BV modified MSCs were encapsulated by hyaluronan hydrogel, due to its’ vital composition in ECM of a healthy disc and transplanted to a punctured coccygeal disc in rats through micro-injection, followed by X-ray radiography and histological analysis at the 4- and 12-weeks post-transplantation; (3) Results: Data reveals the Cre/LoxP BV system-mediated long-termed runx1 gene expression, possessing good biosafety characteristics in the in vitro cell transduction and in vivo MSCs transplantation, and maintained superior hydration content in the disc than that of mock transduced MSCs; (4) Conclusions: This proof-of-concept study fulfills the need of implanting therapeutic cells accompanied with microinjection in the disc, such as a discography and paves a road to manufacture composite hyaluronan, such as peptide modified hyaluronan as an MSC carrier for IVD regeneration in the future study.https://www.mdpi.com/2227-9059/10/3/555runx1 transcription factorbaculoviral vectorgene therapyCre/LoxP gene editingdisc degeneration |
| spellingShingle | Long-Yi Chan Cheng-Chung Chang Po-Liang Lai Tomoji Maeda Horng-Chaung Hsu Chin-Yu Lin Shu-Jui Kuo Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration Biomedicines runx1 transcription factor baculoviral vector gene therapy Cre/LoxP gene editing disc degeneration |
| title | Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration |
| title_full | Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration |
| title_fullStr | Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration |
| title_full_unstemmed | Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration |
| title_short | Cre/LoxP Genetic Recombination Sustains Cartilage Anabolic Factor Expression in Hyaluronan Encapsulated MSCs Alleviates Intervertebral Disc Degeneration |
| title_sort | cre loxp genetic recombination sustains cartilage anabolic factor expression in hyaluronan encapsulated mscs alleviates intervertebral disc degeneration |
| topic | runx1 transcription factor baculoviral vector gene therapy Cre/LoxP gene editing disc degeneration |
| url | https://www.mdpi.com/2227-9059/10/3/555 |
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