Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: Neurovascular pathological markers expressed in TBI mice ameliorated with ASC-CCM: Potential role for COL4A1. A Timeline and analysis of mTBI study. B Volcano plot of sham blast compared to blast mice and blast mice compared to blast mice receiving ASC-CCM displaying each gene’s −log10 (p-value) and log2 fold change with the selected covariate. Highly statistically significant genes fall at the top of the plot above the horizontal lines, and highly differentially expressed genes fall to either side. Horizontal lines indicate various False Discovery Rate (FDR) thresholds or p-value thresholds if there is no adjustment to the p-values. Genes are colored if the resulting p-value is below the given FDR or p-value threshold. The 40 most statistically significant genes are labeled in the plot. Data represent combined averages from n = 4–9 animals/group using NanoString Advanced Analysis. C Realtime qPCR confirmation of decreased COL4A1 gene expression but no other collagens in blast injury retinal tissue alleviated with ASC-CCM at 1 month post-blast injury. Normalized gene expression from n = 3–4 animals/group. One-way ANOVA. D . Cytokine-primed ASC-CCM was subjected to COL4A1 ELISA, demonstrating abundant levels of COL4A1. For comparison, unconcentrated and unstimulated ASC-CM with low levels of COL4A1 is shown. Data represents n = 3 donors. One-way ANOVA. *** p < 0.001

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: Labeling, Gene Expression, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: Intravitreal COL4A1 shRNA results in visual deficits in wildtype mice. A Timeline and analysis of the study. B One month post intravitreal injection fundus imaging showed virus integration evidenced by GFP expression in both the groups. Immunohistology confirmed co-localization of COL4A1 with GFP in Con shRNA mice. C . qPCR confirmation of significant reduction in COL4A1 expression in mice receiving COL4A1 shRNA. D Mice receiving COL4A1 shRNA but not scramble (Con shRNA) demonstrated decreased visual acuity and contrast sensitivity in the left eye (LE). Right eye (RE) with no injection looked near normal. Data represents n = 5–6/group. One-way ANOVA. * p < 0.05; *** p < 0.001; ns = non-significant

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: shRNA, Injection, Imaging, Virus, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: Intravitreal COL4A1 shRNA exacerbates visual deficits in blast injury. A Timeline, experimental groups and experimental assessments. B Visual acuity and contrast sensitivity further decreased with blast injury with COL4A1 knockdown ameliorated with ASC-CCM. Data represents n = 5–6/group. One-way ANOVA. C Dark adapted scotopic ERG shows altered b- and a-waves ameliorated with ASC-CCM. Amplitude data represent n = 5–6/group shown at 1 cd.s.m 2 . Representative electroretinogram tracings from each of the groups are shown below. Only blast mice data shown. One-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001; # non-significant

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: shRNA, Knockdown

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: Intravitreal COL4A1 shRNA exacerbates neuroinflammatory markers in blast injury. A Increased glial and ER stress transcripts with blast injury with COL4A1 knockdown ameliorated with ASC-CCM. All gene transcripts were normalized to 18s rRNA and shown relative to Sham-blast mice. Data represents n = 3–4 mice/group. One-way ANOVA. B Immunostaining for GFAP increased with COL4A1 knockdown in blast mice and was nearly normalized with ASC-CCM. Data represents n = 3–4/group. One-way ANOVA. C Immunostaining for synaptophysin decreased with COL4A1 knockdown in blast mice near normalized with ASC-CCM. Quantitative analysis of synaptophysin positive area at the OPL is represented to the right. Data represents n = 3 animals/group. One-way ANOVA * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: shRNA, Knockdown, Immunostaining

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: Intravitreal COL4A1 shRNA decreases blood vessel integrity in blast injury. A Vasodegeneration, observed with decreased perlecan immunostaining in retinal sections, correlated with degenerated vasculature from retinal flat mounts labeled with isolectin-B4 in blast/COL4A1 shRNA mice, appears near normal with ASC-CCM. Data represents n = 3 animals/group. B Increased endothelial activation gene transcripts with blast injury with COL4A1 knockdown ameliorated with ASC-CCM. All gene transcripts were normalized to 18s rRNA and shown relative to Sham-blast mice. Data represents n = 3–4 mice/group. One-way ANOVA. * p < 0.05; ** p < 0.01

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: shRNA, Immunostaining, Labeling, Activation Assay, Knockdown

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: ASC-CCM ameliorates COL4A1 knockdown induced endothelial integrity in-vitro. A COL4A1 siRNA in HRECs shows > 95% knockdown with no changes in cell morphology. B Transmigration of THP-1 monocytes across endothelial monolayer increased with COL4A1 knockdown decreased with ASC-CCM. C Reticulate VE-Cadherin with loss of protein (yellow arrows) in HREC knockdown with COL4A1 is protected by ASC-CCM. D Endothelial dysfunction resulting in poor rates of migration in a scratch wound healing assay after COL4A1 knockdown improved with ASC-CCM. Data represents two independent experiments with technical replicates. One-way ANOVA. *** p < 0.001; ** p < 0.01

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: Knockdown, In Vitro, Transmigration Assay, Migration, Wound Healing Assay

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: COL4A1 secreted by ASCs necessary for microglial and endothelial integrity in-vitro. A Immunodepletion of ASC-CCM with COL4A1 IgG or the isotype IgG demonstrating the specificity of immune depletion by ELISA and immunoblotting. B Nitrite release (upper panel) from activated microglial cells as assessed by Griess assay improved only with ASC-CCM +COL4A1 . Increased CD86 and IL1b gene expression (lower panel) in activated microglia improved only with ASC-CCM +COL4A1 . C TNF-α induced loss in endothelial resistance as assessed by ECIS improved only with ASC-CCM +COL4A1 . Data represents two independent experiments with technical replicates. One-way ANOVA. *** p < 0.001; ** p < 0.01; * p < 0.05; ns = non-significant

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: In Vitro, Immunodepletion, Enzyme-linked Immunosorbent Assay, Western Blot, Griess Assay, Gene Expression

Journal: Stem Cell Research & Therapy

Article Title: Neurotrauma induced retinal basement membrane COL4A1 defects are restored by adipose tissue derived mesenchymal stem cell concentrated conditioned medium

doi: 10.1186/s13287-025-04804-6

Figure Lengend Snippet: Potential pathophysiological effects of COL4A1 in mTBI. Under physiological state, COL4A1 associated with extracellular matrix (ECM) via integrins maintains neurovascular integrity, while on the other hand, TBI results in defective COL4A1, leading to endothelial cell (EC) activation, microglial polarization, synaptic defects, and neurodegeneration. Created in Microsoft PowerPoint with artwork from Servier under Creative Commons Attribution 3.0 Unported license (CC BY 3.0; http://www.servier.com/Powerpoint-image-bank )

Article Snippet: Collagen, type IV, alpha 1 (Col4a1) , Mm01210125_m1 , NM_009931.2.

Techniques: Activation Assay