Construction of a decellularized spinal cord matrix/GelMA composite scaffold and its effects on neuronal differentiation of neural stem cells

Abstract

Spinal cord injury (SCI) leads to severe loss of motor and sensory functions, and the rehabilitation of SCI is a worldwide problem. Tissue-engineered scaffolds offer new hope for SCI patients, while the newly developed materials encountered a challenge in modeling the microenvironment around the lesion site. We constructed a new composite scaffold by mixing decellularized spinal cord extracellular matrix (dECM) with gelatin methacryloyl (GelMA). The dECM, as a natural biological material, retained a large number of proteins and growth factors related to neurogenesis. GelMA was a photopolymerizable material, harbored a polymer network structure, soft texture, certain shape and plenty of water. The viability, proliferation, and differentiation of neural stem cells (NSCs) on the composite scaffold were evaluated by cell count kit-8 (CCK8), Live/Dead assay, phalloidin staining, 5-Ethynyl-2’-deoxyurdine (EdU), immunofluorescence staining and western blot. The Live/Dead assay, phalloidin staining, EdU, and CCK8 assay showed that the composite scaffold had good biocompatibility and provided better support for proliferation of NSCs. Results of immunocytochemistry and western blot showed that the composite scaffolds promoted the specific differentiation of NSCs into neuron cells. Together, this dECM/GelMA composite scaffold can be used as a cell culture coating, the isolated NSCs seeded on the surface of composite scaffold expressed neuronal markers and assumed neuronal morphology. Our work provided a new method that would be widely used in tissue engineering of SCI.

Keywords:

• Neural stem cells
• GelMA hydrogel
• decellularized spinal cord
• extracellular matrix
• spinal cord injury

Authors’ contributions

Wenhua He, Hui Wang, Xuanxuan Zhang: Conception, Design, Collection, Data analysis, Interpretation, Manuscript writing, Yan Lu, Yu Gu, Dingyue Ju: Data analysis, Interpretation, Tiantian Mao, Longju Qi: Formal analysis, Chuanming Dong, Qinghua Wang: Conceptualization, Methodology, Resources, Writing-review & editing, Supervision, Project administration, Funding acquisition.

Data availability statement

The raw/processed data required to reproduce these findings are available from the corresponding author by request.

Disclosure statement

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Additional information

Funding

This work was supported by the National Natural Science Foundation of China (82171380), Postgraduate Research & Practice Innovation Program of Jiangsu Province (KYCX22_3360), Jiangsu Students’ Platform for innovation and entrepreneurship training program (202110304098Y).