Construction of Dual-Biofunctionalized Chitosan/Collagen Scaffolds for Simultaneous Neovascularization and Nerve Regeneration

Guicai Li; Qi Han; Panjian Lu; Liling Zhang; Yuezhou Zhang; Shiyu Chen; Ping Zhang; Luzhong Zhang; Wenguo Cui; Hongkui Wang; Hongbo Zhang

doi:10.34133/2020/2603048

Construction of Dual-Biofunctionalized Chitosan/Collagen Scaffolds for Simultaneous Neovascularization and Nerve Regeneration

Research (Wash D C). 2020 Aug 10:2020:2603048. doi: 10.34133/2020/2603048. eCollection 2020.

Authors

Guicai Li^{1

2}, Qi Han^{1

2}, Panjian Lu^{1

2}, Liling Zhang^{1

2}, Yuezhou Zhang³, Shiyu Chen^{1

2}, Ping Zhang^{1

2}, Luzhong Zhang^{1

2}, Wenguo Cui⁴, Hongkui Wang^{1

2}, Hongbo Zhang^{4

5}

Affiliations

¹ Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Nantong University, 226001 Nantong, China.
² Co-Innovation Center of Neuroregeneration, Nantong University, 226001 Nantong, China.
³ Frontiers Science Center for Flexible Electronics, Xi'an Institute of Flexible Electronics (IFE) & Xi'an Institute of Biomedical Materials and Engineering (IBME), Northwestern Polytechnical University, 127 West Youyi Road, Xi'an 710072, China.
⁴ Shanghai Institute of Traumatology and Orthopaedics, Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 197 Ruijin 2nd Road, Shanghai 200025, China.
⁵ Pharmaceutical Sciences Laboratory and Turku Bioscience Centre, Åbo Akademi University, 20520 Turku, Finland.

Abstract

Biofunctionalization of artificial nerve implants by incorporation of specific bioactive factors has greatly enhanced the success of grafting procedures for peripheral nerve regeneration. However, most studies on novel biofunctionalized implants have emphasized the promotion of neuronal and axonal repair over vascularization, a process critical for long-term functional restoration. We constructed a dual-biofunctionalized chitosan/collagen composite scaffold with Ile-Lys-Val-Ala-Val (IKVAV) and vascular endothelial growth factor (VEGF) by combining solution blending, in situ lyophilization, and surface biomodification. Immobilization of VEGF and IKVAV on the scaffolds was confirmed both qualitatively by staining and quantitatively by ELISA. Various single- and dual-biofunctionalized scaffolds were compared for the promotion of endothelial cell (EC) and Schwann cell (SC) proliferation as well as the induction of angiogenic and neuroregeneration-associated genes by these cells in culture. The efficacy of these scaffolds for vascularization was evaluated by implantation in chicken embryos, while functional repair capacity in vivo was assessed in rats subjected to a 10 mm sciatic nerve injury. Dual-biofunctionalized scaffolds supported robust EC and SC proliferation and upregulated the expression levels of multiple genes and proteins related to neuroregeneration and vascularization. Dual-biofunctionalized scaffolds demonstrated superior vascularization induction in embryos and greater promotion of vascularization, myelination, and functional recovery in rats. These findings support the clinical potential of VEGF/IKVAV dual-biofunctionalized chitosan/collagen composite scaffolds for facilitating peripheral nerve regeneration, making it an attractive candidate for repairing critical nerve defect. The study may provide a critical experimental and theoretical basis for the development and design of new artificial nerve implants with excellent biological performance.