Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (34): 5445-5452.doi: 10.12307/2021.237

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Cartilage composite scaffold loaded with transforming growth factor beta 3 using three-dimensional bioprinting

Yang Zhen1, 2, Li Hao1, 2, Fu Liwei1, 2, Gao Cangjian1, 2, Jiang Shuangpeng2, Wang Fuxin2, Yuan Zhiguo2, Sun Zhiqiang1, 2, Zha Kangkang1, 2, Tian Guangzhao1, 2, Cao Fuyang2, Sui Xiang2, Liu Shuyun2, Guo Quanyi2   

  1. 1Medical College of Nankai University, Tianjin 300071, China; 2Institute of Orthopedics, the First Medical Center, Chinese PLA General Hospital, Beijing Key Laboratory of Regenerative Medicine in Orthopedics, Key Laboratory of Musculoskeletal Trauma & War Injuries, PLA, Beijing 100853, China
  • Received:2020-06-28 Revised:2020-07-03 Accepted:2020-08-04 Online:2021-12-08 Published:2021-07-26
  • Contact: Guo Quanyi, Professor, Institute of Orthopedics, the First Medical Center, Chinese PLA General Hospital, Beijing Key Laboratory of Regenerative Medicine in Orthopedics, Key Laboratory of Musculoskeletal Trauma & War Injuries, PLA, Beijing 100853, China
  • About author:Yang Zhen, Master candidate, Medical College of Nankai University, Tianjin 300071, China; Institute of Orthopedics, the First Medical Center, Chinese PLA General Hospital, Beijing Key Laboratory of Regenerative Medicine in Orthopedics, Key Laboratory of Musculoskeletal Trauma & War Injuries, PLA, Beijing 100853, China
  • Supported by:
    the National Key Research and Development Plan Project, No. 2019YFA0110600 (to GQY); the National Natural Science Foundation of China, No. 81772319 (to GQY)

Abstract: BACKGROUND: The therapeutic strategy of in situ regeneration of cartilage injury by recruiting endogenous stem cells is a new research direction of cartilage tissue engineering in the future.
OBJECTIVE: To construct a tissue engineering cartilage composite scaffold that can not only recruit stem cells, promote cell adhesion and proliferation, but also be beneficial to the maturation of neo-tissue.
METHODS: Acellular cartilage extracellular matrix (ECM) and methacrylate gelatin (GelMA) were mixed to prepare photosensitive bio-ink, and three-dimensional bioprinting technology was used to prepare polycaprolactone (PCL) scaffolds and PCL/GelMA/ECM scaffolds. Transforming growth factor β3 (TGF-β3) was loaded into bio-ink to prepare PCL/GelMA/ECM/TGF-β3 scaffold, and its sustained release performance was tested. The physical and chemical properties of PCL/GelMA/ECM scaffolds were evaluated from the point of view of morphology, histology, biochemistry and biomechanics. CCK-8 assay was used to detect the cytotoxicity of PCL/GelMA/ECM scaffolds. After adipose-derived mesenchymal stem cells were seeded on PCL/GelMA/ECM scaffold for 1, 4 and 7 days, the cell viability was observed by confocal microscope and the cell adhesion was observed by scanning electron microscope. PCL/GelMA/ECM scaffolds were implanted subcutaneously in SD rats, and the infiltration of inflammatory cells and the degradation of scaffolds were observed histologically. The effects of PCL/GelMA/ECM scaffold and PCL/GelMA/ECM/TGF-β3 scaffold on the migration of adipose-derived mesenchymal stem cells were detected by Transwell chamber test, and the cells cultured alone were used as negative control.
RESULTS AND CONCLUSION: (1) The PCL/GelMA/ECM scaffolds had a three-dimensional porous reticular structure, without cellular components, contained cartilage-specific components such as type II collagen and glycosaminoglycan and its elastic modulus was (14.24±2.44) MPa. (2) PCL/GelMA/ECM scaffolds showed no obvious cytotoxicity. (3) Adipose-derived mesenchymal stem cells were adhered closely to the PCL/GelMA/ECM scaffolds, had good cell activity and could secrete extracellular matrix. (4) One week after PCL/GelMA/ECM scaffolds were implanted subcutaneously in rats, there was a mild acute inflammatory reaction, and the inflammatory reaction was alleviated after 3 weeks, and the scaffolds were gradually degraded. (5) PCL/GelMA/ECM/TGF-β3 scaffold had good sustained release performance, and TGF-β3 could be released continuously for 60 days. (6) Compared with the negative control group, PCL/GelMA/ECM scaffolds and PCL/GelMA/ECM/TGF-β3 scaffolds could promote the migration of adipose-derived mesenchymal stem cells, and PCL/GelMA/ECM/TGF-β3 scaffold could promote the migration of adipose-derived mesenchymal stem cells more significantly. (7) The results showed that three-dimensional printing PCL/GelMA/ECM/TGF-β3 scaffolds could promote the proliferation, adhesion and migration of adipose-derived mesenchymal stem cells.

Key words: one, materials, scaffold, transforming growth factor-β3, 3D bioprinting, cartilage injury, adipose derived mesenchymal stem cells, recruitment, migration

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