Journal
MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS
Volume 118, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.msec.2020.111518
Keywords
Amino-functionalized graphene; Collagen; Nano-crosslinker; BM-MSCS; Neuronal differentiation; Raw 264.7; Macrophage polarization; ATP; CD90 and CD73; Organotypic culture; Spinal explants
Categories
Funding
- National Institute of Pharmaceuticals Education and Research Ahmedabad
- Department of Pharmaceuticals, Ministry of Chemicals and Fertilizers, Government of India
- DST-SERB [ECR/2016/002038]
- DBT supported Ramalingaswami grant [BT/HRD/35/02/2006]
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The novel electro-conductive graphene collagen cryogels fabricated for nerve conduits exhibit appropriate electrical and mechanical cues, promoting the expression of specific genes in BM-MSCs and enhancing neuronal differentiation under electric stimulation. Furthermore, the cryogels can stimulate ATP secretion from BM-MSCs, aiding in neuronal regeneration and immune modulation.
Novel amino-functionalized graphene crosslinked collagen based nerve conduit having appropriate electric (3.8 +/- 0.2 mSiemens/cm) and mechanical cues (having young modulus value of 100-347 kPa) for stem cell transplantation and neural tissue regeneration was fabricated using cryogelation. The developed conduit has shown sufficiently high porosity with interconnectivity between the pores. Raman spectroscopy analysis revealed the increase in orderliness and crosslinking of collagen molecules in the developed cryogel due to the incorporation of amino-functionalized graphene. BM-MSCs grown on graphene collagen cryogels have shown enhanced expression of CD90 and CD73 gene upon electric stimulation (100 mV/mm) contributing towards maintaining their stemness. Furthermore, an increased secretion of ATP from BM-MSCs grown on graphene collagen cryogel was also observed upon electric stimulation that may help in regeneration of neurons and immuno-modulation. Neuronal differentiation of BM-MSCs on graphene collagen cryogel in the presence of electric stimulus showed an enhanced expression of MAP-2 kinase and beta-tubulin III. Immunohistochemistry studies have also demonstrated the improved neuronal differentiation of BM-MSCs. BM-MSCs grown on electroconductive collagen cryogels under inflammatory microenvironment in vitro showed high indoleamine 2,3 dioxygenase activity. Moreover, macrophages cells grown on graphene collagen cryogels have shown high CD206 (M2 polarization marker) and CD163 (M2 polarization marker) and low CD86 (M1 polarization marker) gene expression demonstrating M2 polarization of macrophages, which may aid in tissue repair. In an organotypic culture, the developed cryogel conduit has supported cellular growth and migration from adult rat spinal cord. Thus, this novel electro-conductive graphene collagen cryogels have potential for suppressing the neuro-inflammation and promoting the neuronal cellular migration and proliferation, which is a major barrier during the spinal cord regeneration.
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