DOI: 10.5061/dryad.vmcvdnczx
title: Data from: Sustained neurotrophin-3 delivery from hyaluronic acid hydrogels for neural tissue regeneration

description: The goal of this work was to design a polymer-based platform capable of
 localized, long-term delivery of biologically active neurotropic factors
 using an affinity-based approach. Here, we synthesized hyaluronic
 acid-methylfuran (HA-mF) hydrogels that provide sustained, affinity-based
 release of neurotrophin-3 (NT-3), a growth factor that promotes axon
 growth for 28 days. A Diels-Alder crosslinking reaction between HA-mF and
 polyethylene glycol (PEG)-dimaleimide occurs within 15 minutes under
 physiological conditions, resulting in injectable hydrogels that can be
 polymerized in the presence of cells and growth factors. We also tuned the
 hydrogel’s storage modulus to match that of native rat spinal cord tissue,
 providing a platform not only for localized drug delivery but also a
 suitable vehicle for cellular transplantation. The NT-3 released from the
 HAmF hydrogels remains bioactive for at least 14 days, promoting axonal
 growth from primary sensory neurons as well as stem cell-derived V2a
 interneurons and motoneurons in vitro. The hydrogels also supported cell
 growth allowing for 3-dimensional axonal extensions within the scaffold
 matrix. Here we confirm the protective role of HA-mF on matrix-bound NT-3
 activity and show that these hydrogels are an excellent platform for drug
 delivery for neural applications.
description: Data for NMR was exported as a .fid file.
 Storage modulus values were extracted from a rheology time sweep,
 the values reported are the final stiffness of each run.
 Data for protein loading and release experiment was collected
 using either an ELISA kit or BCA kit and the measurements were converted
 into concentrations using a standard curve. Axonal
 growth data is reported as the normalized average radius extension to the
 control for each experiment. PCR data is reported as
 the ΔΔCt values for each condition and replicate. Cell
 viability was measured using a Live/Dead assay kit, values reported are
 the percentage of live cells for each replicate and condition as measured
 by fluorescence microscopy and the area quantified.
description: # Data from: Sustained neurotrophin-3 delivery from hyaluronic acid
 hydrogels for neural tissue regeneration Raw data for all figures
 presented in the paper was collected and compiled into one Excel file
 (with the exception of the NMR data). For more details regarding the
 experimental conditions, please refer to the methods section of the
 manuscript. Information regarding the data collection methods is included
 below. ## Description of the Data and file structure Dataset Provided:
 Paper Data Excel File 1. NT-3 Loading and equilibrium. NT-3 concentration
 was measured using a commercially available ELISA kit from R&D
 Systems. The data was normalized using standard curves. Values reported
 are: a) Fraction of NT-3 retained at 24 hours for 10, 50, 100 or 250 ng/mL
 loading concentrations of NT-3 b) Fraction of NT-3 retained (in) and
 released (out) for each condition: Phosphate buffer saline (PBS), PBS with
 1M sodium chloride (NaCl), or PBS with 2M sodium chloride 2. Storage
 Modulus. Stiffness data was collected using a rheometer. Values reported
 are: a) Final storage modulus values in Pa, after a 3 hour time sweep on
 hydrogels with a 1:1.5, 1:3 or 1:5 maleimide to methylfuran molar ratio.
 Pre and post refer to measurements taken for freshly prepared gels (pre)
 or previously lyophilized and rehydrated gels (post). 1:1.5 + ECM are gels
 supplemented with astroctye-derived extracellular matrix. b) Full loss and
 storage modulus time curve over a 3-hour time sweep shown in Pa vs time in
 minutes. 3. 28-day Release. NT-3 concentration was measured using a
 commercially available ELISA kit from R&D Systems. The data was
 normalized using standard curves. Values reported are percentage of NT-3
 released (%) at every time point (in days) in the presence of buffer with
 0.1% BSA or elution buffer. 4. DRG Bioactivity. Average neurite extension
 from DRG cell bodies was measured on ImageJ. Values reported are average
 neurite radius length and normalized to the 10 ng/mL NT-3 positive
 control. Each measurement represents one DRG. 0, 10 and 20 ng/mL refer to
 the concentration of NT-3 and 24h, 7 day and 14 day are release samples of
 NT-3 from each time point, respectively. 5. Aggregate growth. Average
 neurite extension from Hb9 motoneurons or V2a interneurons in the presence
 of 0, 5, 10 or 20 ng/mL NT-3. Values are normalized to the positive
 control in each experiment. 6. TrkC qPCR. Expression of TrkC receptor for
 each cell type and time point, qPCR data was collected using a TaqMan
 assay and QuantStudio. Values reported are DeltaDeltaCt for each condition
 and experiment, Hb9 motoneurons and V2a interneurons before puromycin
 selection (pre-sel) or one (d1) or two (d2) days after selection. Agg
 refers to Hb9 aggregates instead of single cells. 7. Hydrogel Cell
 viability. Data was obtained using a Live/Dead kit. Data reported
 represents each survival measurement as a percentage (%) of the total cell
 population. HAmF refers to Hyaluronic acid-methylfuran. NMR spectra for
 hyaluronic-acid methylfuran are presented as a .mnova file, which can be
 opened in MestreNova software. The .fid proton spectra file is also
 included. ## Sharing/access Information Links to other publicly accessible
 locations of the data: N/A Was data derived from another source? If yes,
 list source(s): N/A