Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Renovating Neural Networks With Viral‐Mediated Gene Transfer From A Tissue Contacting Matrix Mimic

doi: 10.1002/smll.202510539

Figure Lengend Snippet: In vitro transduction of Primary Cortical Neurons (CTX), and striatal neurons (lateral Ganglionic Eminence (LGE)) using rAAVDJ‐CMV‐BDNF‐P2A‐GFP (AAVDJ‐BDNF) viral vector. (A) Schematic of the AAVDJ‐BDNF construct. A representative image of (B,C) transduced CTX and (D) LGE cells stained with transduction efficiency marker GFP (green), Map2/ β3‐Tubulin neuronal markers (red), GABA (magenta) and Hoechst (Blue). CTX were transduced with 10 4 viral genomes (vg)/cell, and LGE cells underwent transduction with 10 3 vg/cell of AAV‐BDNF. (E) Flow Cytometry data indicating the proportion of cells positive for both GFP and neural markers (β3‐tubulin) as well as GABA in the AAVDJ‐BDNF viral vector system. (F) Quantification of the percentage of transduced cells corresponding to the representative images, showing co‐expression with neural and striatal markers. The data is presented as the mean ± SD. Analysis was conducted using ANOVA followed by Tukey's post‐hoc test for statistical differentiation between groups, with **** indicating p <0.0001. Scale bar = 100 µm.

Article Snippet: Following the manufacturer's instructions, BDNF protein release was quantified using a Human/Mouse BDNF DuoSet ELISA kit (R&D Systems).

Techniques: In Vitro, Transduction, Plasmid Preparation, Construct, Staining, Marker, Flow Cytometry, Expressing

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Renovating Neural Networks With Viral‐Mediated Gene Transfer From A Tissue Contacting Matrix Mimic

doi: 10.1002/smll.202510539

Figure Lengend Snippet: BDNF mRNA and protein expression in transduced cells. (A) RT‐PCR analysis of the hBDNF mRNA expression analysed by agarose gel electrophoresis. Control, non‐transduced mouse primary cortical neurons as well as striatal cells; AAVDJ‐BDNF viral vector. According to the results, no hBDNF was detected in non‐transduced primary cortical neurons and striatal neuron cells (control). However, hBDNF was detected in transduced primary cortical neuron cells and striatal neurons by the AAVDJ‐BDNF viral vector tool. (B) Assessment of BDNF protein release via ELISA. ELISA results indicate a notable release of BDNF protein in supernatant samples from primary cortical and striatal neurons infected with AAVDJ‐BDNF viral vector, in comparison to non‐transduced cells (control). The data is shown as mean ± SD. Statistical analysis was performed using ANOVA followed by Tukey's post‐hoc test to identify significant differences between groups, with **** indicating p <0.0001 and ** signifying p <0.01.

Article Snippet: Following the manufacturer's instructions, BDNF protein release was quantified using a Human/Mouse BDNF DuoSet ELISA kit (R&D Systems).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Control, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Infection, Comparison

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Renovating Neural Networks With Viral‐Mediated Gene Transfer From A Tissue Contacting Matrix Mimic

doi: 10.1002/smll.202510539

Figure Lengend Snippet: Fmoc‐DDIKVAV hydrogel characterization. (A) HPLC analysis of Fmoc‐DDIKVAV. The chromatogram shows a distinct peak, indicating the sample's purity. (B) MS profile of Fmoc‐DDIKVAV hydrogel system. The spectrum highlights a predominant single component along with minor degradation products. (C) Optical image of generated Fmoc‐DDIKVAV. (D) The FTIR spectra of Fmoc‐DDIKVAV hydrogel system show a major peak around 1630 cm − 1 and a minor peak at 1690 cm − 1 , indicating anti‐parallel β‐sheet arrangements. (E) CD spectra of Fmoc‐DDIKVAV reveal β‐sheet structures, with significant transitions below 220 nm. (F) The TEM image reveals that the nanofibers are intertwined, creating fine fibrils; the scale bar represents 200 nm. (G) cryoSEM image of the nanofibers demonstrating the nano and microstructural network formed by the fibres. (H) The AFM image demonstrates the nanofibrous architecture of Fmoc‐DDIKVAV hydrogel. (I) The mesh size (ξ) (nm) of Fmoc‐DDIKVAV was determined using the equation from the theory of rubber elasticity. (J) Assessment of surface ζ‐potential in Fmoc‐DDIKVAV system using zetasizer. (K) Rheological analysis verifies that the fabricated hydrogel exhibits viscoelastic properties, as indicated by a storage modulus (G′) that exceeds the loss modulus (G″). (L) Conducting an oscillatory rheological test to observe the variations in the hydrogel's modulus over time, further illustrating its shear‐thinning characteristics. (M) Analysis of the SAP hydrogel's recovery process. This involved initially applying a low shear rate of 0.01 s −1 for 30 s, followed by a high shear rate of 100 s −1 for another 30 s, and concluding with a return to the initial low shear rate for 15 min. This procedure helps to understand how the hydrogel regains its original viscosity after the cessation of stress, (N) AAVDJ‐ release characteristic from the Fmoc‐SAP hydrogel biomaterial. The release pattern of the AAVDJ‐BDNF was analyzed through qPCR, quantifying vector‐encoded DNA in the Fmoc‐DDIKVAV hydrogel at specified intervals (1, 4, 8, 24, 48, 72, and 120 h). The presented data reflect the mean + SD, with three replicates at each timepoint. Statistical analysis was performed using ANOVA followed by Tukey's post‐hoc test to identify significant differences between groups, with **** indicating p < 0.0001 and ** signifying p < 0.01.

Article Snippet: Following the manufacturer's instructions, BDNF protein release was quantified using a Human/Mouse BDNF DuoSet ELISA kit (R&D Systems).

Techniques: Generated, Circular Dichroism, Shear, Viscosity, Plasmid Preparation

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Renovating Neural Networks With Viral‐Mediated Gene Transfer From A Tissue Contacting Matrix Mimic

doi: 10.1002/smll.202510539

Figure Lengend Snippet: Hydrogel encapsulated AAVDJ‐BDNF prolonged payload release while maintaining its biofunctionality. Analysis of AAVDJ‐BDNF viral vector's effectiveness post‐release from Fmoc‐DDIKVAV hydrogel on (A) CTX and (B) LGE over 5 days. The neurons were immunostained with GFP (green) to highlight GFP‐tagged virally transduced cells, and counterstained with Hoechst (blue) for total cell visualization. Additionally, the cortical and striatal neurons were marked with the neural marker MAP2 (red) as well as the GABA striatal marker (magenta), respectively. The intense GFP fluorescence indicates a high transduction efficiency of both cells by AAV, confirming the virus's viability and efficacy after being released from the Fmoc‐SAP system, Scale bar = 50 µm. (C) Representative Flow Cytometry data showing the proportion of GFP and MAP2 positive cells and GFP and GABA co‐positive cells. (D) The analysis of transduction levels, expressed as the percentage of neurons positive for GFP and the designated neuronal and LGE markers. The results are presented as the mean ± SD, with three samples for each timepoint.

Article Snippet: Following the manufacturer's instructions, BDNF protein release was quantified using a Human/Mouse BDNF DuoSet ELISA kit (R&D Systems).

Techniques: Plasmid Preparation, Marker, Fluorescence, Transduction, Virus, Flow Cytometry

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Renovating Neural Networks With Viral‐Mediated Gene Transfer From A Tissue Contacting Matrix Mimic

doi: 10.1002/smll.202510539

Figure Lengend Snippet: The synergistic effect of AAVDJ‐BDNF and SAP hydrogel reduces tissue atrophy. (A) Schematic overview of the designed experiment. (B) Quantification of striatal tissue atrophy corresponding to the representative images (C), shown as the volume (mm 3 ) of NeuN‐negative tissue surrounding the injection site in mice implanted with the indicated materials. The analysis was conducted using ANOVA complemented by Tukey's post‐hoc test to discern statistical variances between groups, with significance levels indicated by **** p <0.0001, *** p <0.001, ** p <0.01. Notably, the least amount of tissue degeneration was observed in animals that received QA + AAVDJ‐BDNF+SAP post QA lesioning. (C) In vivo implants of different viral vectors with or without Fmoc‐SAP system in the mouse striatum 8 weeks post‐implantation illustrate the effects of different materials on Huntington's Disease (HD) mouse models. Immunostaining highlights the impact of QA‐induced lesions in the striatum (right hemisphere), evident by the absence of NeuN‐positive cells (green) in the core lesion area. Contrastingly, in mice receiving QA lesions followed by AAVDJ‐BDNF and Fmoc‐DDIKVAV implantation, there is a notable reduction in lesion size and increased neuroprotection, as seen by the denser population of NeuN‐positive cells near the injection site. Scale bars in the images are set at 500 µm.

Article Snippet: Following the manufacturer's instructions, BDNF protein release was quantified using a Human/Mouse BDNF DuoSet ELISA kit (R&D Systems).

Techniques: Injection, In Vivo, Immunostaining

Journal: Small (Weinheim an Der Bergstrasse, Germany)

Article Title: Renovating Neural Networks With Viral‐Mediated Gene Transfer From A Tissue Contacting Matrix Mimic

doi: 10.1002/smll.202510539

Figure Lengend Snippet: Incorporation of AAVDJ‐BDNF within SAP hydrogel attenuates the host inflammatory response. (A) Representative overview of the core section of the brain in the 5 different groups stained for Iba‐1 (red) and GFP (green), Scale bar = 500 µm. (B) Quantification of microglial activation density within the right hemisphere of the core sections corresponding to the images in panel A. (C,D) Quantified measurements of the density of microglial activation within FOV1 and FOV2 in the striatum of mice corresponding to the images in panel G,H. Data are represented as mean ± SD (n = 4 per group). ** p <0.01, *** p <0.001, **** p <0.0001. (E) Schematic diagram of the overview of the images section (shown as magenta) and selected field of view (FOV) for analysis and higher magnification images. (F) Representative images of the core section of the injection site in different groups stained for Iba‐1 (magenta), scale bar = 500 µm. (G,H) High magnification of the fluorescent images of the different FOV1 and FOV2 of different groups stained for Iba‐1, Scale bar = 100 µm.

Article Snippet: Following the manufacturer's instructions, BDNF protein release was quantified using a Human/Mouse BDNF DuoSet ELISA kit (R&D Systems).

Techniques: Staining, Activation Assay, Injection

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Delayed administration of type II ampakines enhance synaptic plasticity and cognitive recovery in juvenile global cerebral ischemia

doi: 10.1177/0271678X251405662

Figure Lengend Snippet: BDNF expression decreases after GCI in juvenile mice. (a) ELISA in juvenile mice 14, 21 days after sham or GCI showing decreased BDNF expression followed by recovery. (b) There is no change in BDNF in adults 14 days after GCI. p values given in figure, one-way ANOVA in the juvenile BDNF expression, t -test in adult BDNF expression.

Article Snippet: Protein extracts were collected and BDNF levels were measured using mouse BDNF ELISA kit (LS-F2404; LifeSpan Biosciences, Seattle, WA, USA) as per manufacture’s instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Delayed administration of type II ampakines enhance synaptic plasticity and cognitive recovery in juvenile global cerebral ischemia

doi: 10.1177/0271678X251405662

Figure Lengend Snippet: Ampakine effect on BDNF expression. (a) Experimental design for injection of ampakines in sham P21 mice. (b) Quantification of hippocampal BDNF showing dose response of LY404187 and IDRA-21 24 h after injection in sham mice. (c) Experimental design for injection of 1 mg/kg of ampakine 13 days after sham or GCI surgery, followed by collection of (d) quantification of hippocampal BDNF following LY404187 (LY) or IDRA-21 (ID) administration is sham or juvenile GCI mice. * p < 0.05 by ANOVA.

Article Snippet: Protein extracts were collected and BDNF levels were measured using mouse BDNF ELISA kit (LS-F2404; LifeSpan Biosciences, Seattle, WA, USA) as per manufacture’s instructions.

Techniques: Expressing, Injection

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Delayed administration of type II ampakines enhance synaptic plasticity and cognitive recovery in juvenile global cerebral ischemia

doi: 10.1177/0271678X251405662

Figure Lengend Snippet: Recombinant BDNF rescues LTP impairment 14 days after juvenile GCI. (a) Experimental design. (b) Time plot of responses to TBS (40 pulses) in paired experiments where slices were treated with vehicle or BDNF. (c) Quantification of LTP experiments when BDNF (5 nM) was applied in paired experiment 14 days after sham or GCI Each dot represents an individual experiment, and the lines indicate the experiments that were paired. TBS: theta-burst stimulation. * p < 0.05, unpaired t -test for sham vehicle versus GCI vehicle; paired t -test for GCI vehicle versus GCI BDNF.

Article Snippet: Protein extracts were collected and BDNF levels were measured using mouse BDNF ELISA kit (LS-F2404; LifeSpan Biosciences, Seattle, WA, USA) as per manufacture’s instructions.

Techniques: Recombinant