BM-MSC-Loaded Graphene-Collagen Cryogels Ameliorate Neuroinflammation in a Rat Spinal Cord Injury Model

A major obstacle to axonal regeneration following spinal cord injury (SCI) is neuroinflammation mediated by astrocytes and microglial cells. We previously demonstrated that graphene-based collagen hydrogels alone can decrease neuroinflammation in SCI. Their regenerative potential, however, is poorly understood and incomplete. Furthermore, stem cells have demonstrated both neuroprotective and regenerative properties in spinal cord regeneration, although there are constraints connected with the application of stem cell-based therapy. In this study, we have analyzed the regeneration capability of human bone marrow mesenchymal stem cell (BM-MSC)-loaded graphene-cross-linked collagen cryogels (Gr-Col) in a thoracic (T10-T11) hemisection model of SCI. Our study found that BM-MSC-loaded Gr-Col improves axonal regeneration, reduces neuroinflammation by decreasing astrocyte reactivity, and promotes M2 macrophage polarization. BM-MSC-loaded-Gr-Col demonstrated enhanced regenerative potential compared to Gr-Col and the injury group control. Next-generation sequencing (NGS) analysis revealed that BM-MSC-loaded-Gr-Col modulates the JAK2-STAT3 pathway, thus decreasing the reactive and scar-forming astrocyte phenotype. The decrease in neuroinflammation in the BM-MSC-loaded-Gr-Col group is attributed to the modulation of Notch/Rock and STAT5a/b and STAT6 signaling. Overall, Gene Set Enrichment Analysis suggests the promising role of BM-MSC-loaded-Gr-Col in promoting axonal regeneration after SCI by modulating molecular pathways such as the PI3/Akt pathway, focal adhesion kinase, and various inflammatory pathways

Boletín de Segovia: Número 3 - 1913 enero 6

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200

Additional file 8: Figure S6. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

(A) Histogram depicting the genomic proximity (localization) of P300 binding sites identified by ChIP-seq in relation to those identified by using ChIA-PET self-ligation PETs. Identical comparison is performed for both DMSO-treated and TA + TNFα-treated data sets. (B) P300 ChIP-seq signal at P300 binding sites commonly identified by ChIP-seq and ChIA-PET and those binding sites that were uniquely detected in the ChIP-seq data set. (C) P300 ChIP-seq signal at P300 binding sites that were either involved (anchor) or not involved (non-anchor) in long-range interaction as identified by ChIA-PET analysis. (D) An example screenshot depicting the P300 interaction subdomains, P300 ChIP-seq binding sites in relation to topological domains as defined by replication timing data ( www.replicationdomain.org ). (E) Localization of all the interaction subdomains identified by ChIA-PET analysis (P300 and POLII) in relation to topological domains. (PDF 1041 kb

Additional file 2: Figure S2. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

Activated p65 induces de novo P300 depositions to latent genomic loci. (A) Pile-up heat map depicting the H3K4me1 and H3K4me3 signal around (±12 kb) all p65-bound promoters and DBSs. (B) Pile-up heat map depicting the p65 and P300 signal at all p65-bound enhancers upon vehicle, DMSO (−), and TNFα (+) treatment. (C) Example screenshot depicting TNFα-induced P300 recruitment at genomic regions (red box) and recruitment of p65 at genomic loci that are pre-marked by P300. (D) Motif occurrence at all p65-bound DBS presented as a function of TNFα-dependent P300 recruitment (x-axis) (top-panel). Level of shared binding of p65 and other TFs at all p65-bound DBS, presented as a function of TNFα-dependent P300 recruitment (bottom panel). (E) Level of H3K27ac, DNase I hypersensitivity, and H3K4me1 at all p65-bound DBSs (induced and constitutive P300 sites). (PDF 909 kb

Additional file 3: Figure S3. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

Large numbers of P300-bound loci are not co-occupied by GR or p65. (A) Average ChIP-seq signal of GR and P300 around (Âą10 kb) all the P300 binding sites that do not show a significant GR occupancy. (B) Average ChIP-seq signal of p65 and P300 around (Âą10 kb) all the P300 binding sites that do not show a significant p65 occupancy. (PDF 168 kb

Additional file 11: Figure S8. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

Direct comparison of long-range interactions identified by P300 ChIA-PET and 4C-seq analyses at FKBP5, DUSP1, and BIRC3 loci. (PDF 1037 kb

Additional file 9: Table S2. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

List of all significant long-range interactions identified by P300 and POLII ChIA-PET analysis. (XLS 2165 kb

Additional file 12: Figure S9. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

Direct comparison of long-range interactions identified by P300 ChIA-PET and 4C-seq analyses at PTPN1, SULF1 and ZFHX3 loci. (PDF 1147 kb

Additional file 15: Table S5. of Glucocorticoid receptor and nuclear factor kappa-b affect three-dimensional chromatin organization

Overview of sequenced and mapped reads of 4C-seq libraries. (XLS 31 kb