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 enhanced expression of MAP-2 kinase and β-tubulin III. Immunohistochemistry studies have also demonstrated the improved neuronal differentiation of BM-MSCs. BM-MSCs grown on electro-conductive 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) and CD163 (M2) and low CD86 (M1) 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.

使用冷冻凝胶法制备了具有合适的电信号（3.8±0.2 mSiemens / cm）和机械信号（具有100-347 kPa的年轻模量值）的新型氨基官能化石墨烯交联胶原蛋白神经导管，用于干细胞移植和神经组织再生。发达的导管显示出足够高的孔隙度，且孔隙之间具有互连性。拉曼光谱分析显示，由于掺入了氨基官能化的石墨烯，所开发的冷冻凝胶中胶原分子的有序性和交联性增加。生长在石墨烯胶原蛋白冷冻凝胶上的BM-MSC在电刺激（100 mV / mm）后显示出CD90和CD73基因的表达增强，有助于维持其干性。此外，在电刺激下，还观察到生长在石墨烯胶原冷冻凝胶上的BM-MSC的ATP分泌增加，这可能有助于神经元的再生和免疫调节。在电刺激下，石墨烯胶原冷冻凝胶上的BM-MSC的神经元分化显示MAP-2激酶和β-微管蛋白III的表达增强。免疫组织化学研究还证明了BM-MSC的神经元分化得到改善。炎性微环境下在导电胶原蛋白凝胶上生长的BM-MSC 免疫组织化学研究还证明了BM-MSC的神经元分化得到改善。炎性微环境下在导电胶原蛋白凝胶上生长的BM-MSC 免疫组织化学研究还证明了BM-MSC的神经元分化得到改善。炎性微环境下在导电胶原蛋白凝胶上生长的BM-MSC体外显示出高吲哚胺2，3双加氧酶活性。此外，生长在石墨烯胶原蛋白冷冻凝胶上的巨噬细胞已经显示出高CD206（M2）和CD163（M2）和低CD86（M1）基因表达，表明巨噬细胞M2极化，这可能有助于组织修复。在器官型培养中，发达的冷冻凝胶导管支持了成年大鼠脊髓的细胞生长和迁移。因此，这种新颖的导电石墨烯胶原蛋白冷冻凝胶具有抑制神经炎症和促进神经元细胞迁移和增殖的潜力，这是脊髓再生期间的主要障碍。