Data from: Sustained neurotrophin-3 delivery from hyaluronic acid hydrogels for neural tissue regeneration
Data files
Sep 13, 2023 version files 909.62 KB
-
fid
205.19 KB
-
NMR_HAmF_dec19.mnova
599.89 KB
-
Paper_Data.xlsx
101.21 KB
-
README.md
3.34 KB
Abstract
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.
README: 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
- 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
- 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.
- 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.
- 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.
- 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.
- 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.
- 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
Methods
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.