Multilayer Electrospun-Aligned Fibroin/Gelatin Implant for Annulus Fibrosus Repair: An In Vitro and In Vivo Evaluation

Ming-Hsiao Hu; Kai-Chiang Yang; Chih-Wei Chen; Po-Han Chu; Yun-Liang Chang; Yuan-Hui Sun; Feng-Huei Lin; Shu-Hua Yang

doi:10.3390/biomedicines10092107

Multilayer Electrospun-Aligned Fibroin/Gelatin Implant for Annulus Fibrosus Repair: An In Vitro and In Vivo Evaluation

Biomedicines. 2022 Aug 29;10(9):2107. doi: 10.3390/biomedicines10092107.

Authors

Ming-Hsiao Hu¹, Kai-Chiang Yang², Chih-Wei Chen^{1

3}, Po-Han Chu³, Yun-Liang Chang¹, Yuan-Hui Sun¹, Feng-Huei Lin³, Shu-Hua Yang¹

Affiliations

¹ Department of Orthopedics, National Taiwan University Hospital, National Taiwan University College of Medicine, Taipei 100225, Taiwan.
² Department of Dental Technology, College of Oral Medicine, Taipei Medical University, Taipei 11031, Taiwan.
³ Department of Biomedical Engineering, National Taiwan University, Taipei 106216, Taiwan.

Abstract

Annulus fibrosus (AF) damage is proven to prompt intervertebral disc (IVD) degeneration, and unrepaired AF lesions after surgical discectomy may boost herniation of the nucleus pulposus (NP) which may lead to further compression of neural structures. Moreover, vascular and neural ingrowth may occur within the defect which is known as a possible reason for discogenic pain. Due to a limited healing capacity, an effective strategy to repair and close the AF defect is necessary. In this study, using electrospinning technology, two nature polymers, silk fibroin and gelatin, were linked to imitate the unique lamellae structure of native AF. Our findings revealed that a multilayer electrospun-aligned fibroin/gelatin scaffold with mechanical and morphological properties mimicking those of native AF lamellae have been developed. The average diameter of the nanofiber is 162.9 ± 38.8 nm. The young’s modulus is around 6.70 MPa with an ultimate tensile strength of around 1.81 MP along preferred orientation. The in vitro test confirmed its biocompatibility and ability to maintain cell viability and colonization. Using a porcine model, we demonstrated that the multilayer-aligned scaffold offered a crucial microenvironment to induce collagen fibrous tissue production within native AF defect. In the implant-repaired AF, H&E staining showed homogeneous fibroblast-like cell infiltration at the repaired defect with very little vascular ingrowth, which was confirmed by magnetic resonance imaging findings. Picrosirius red staining and immunohistochemical staining against type I collagen revealed positively stained fibrous tissue in an aligned pattern within the implant-integrated site. Relative to the intact control group, the disc height index of the serial X-ray decreased significantly in both the injury control and implant group at 4 weeks and 8 weeks (p < 0.05) which indicated this scaffold may not reverse the degenerative process. However, the results of the discography showed that the effectiveness of annulus repair of the implant group is much superior to that of the untreated group. The scaffold, composed with nature fibroin/gelatin polymers, could potentially enhance AF healing that could prevent IVD recurrent herniation, as well as neural and neovascular ingrowth after discectomy surgeries.

Keywords: annulus fibrosis; electrospinning; fibroin; intervertebral disc; multilayer scaffold.

Grants and funding

This work was supported by the Ministry of Science and Technology, Taiwan, grant No. [107-2314-B-002-105 & 108-2314-B-002-123-MY2] and National Taiwan University, grant No. [NTU- 111L7484].