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In Vitro Assessment of Fluorine Nanoemulsion-Labeled Hyaluronan-Based Hydrogels for Precise Intrathecal Transplantation of Glial-Restricted Precursors

  • Marcin Piejko
  • Piotr Walczak
  • Xiaowei Li
  • Jeff W. M. Bulte
  • Miroslaw JanowskiEmail author
Research Article

Abstract

Purpose

We studied the feasibility of labeling hydrogel scaffolds with a fluorine nanoemulsion for 19F- magnetic resonance imaging (MRI) to enable non-invasive visualization of their precise placement and potential degradation.

Procedure

Hyaluronan-based hydrogels (activated hyaluronan, HA) with increasing concentrations of fluorine nanoemulsion (V-sense) were prepared to measure the gelation time and oscillatory stress at 1 h and 7 days after the beginning of gelation. All biomechanical measurements were conducted with an ARES 2 rheometer. Diffusion of fluorine from the hydrogel: Three hydrogels in various Vs to HA volumetric ratios (1:50, 1:10, and 1:5) were prepared in duplicate. Hydrogels were incubated at 37 °C. To induce diffusion, three hydrogels were agitated at 1000 rpm. 1H and 19F MRI scans were acquired at 1, 3, 7 days and 2 months after gel preparation on a Bruker Ascend 750 scanner. To quantify fluorine content, scans were analyzed using Voxel Tracker 2.0. Assessment of cell viability in vitro and in vivo: Luciferase-positive mouse glial-restricted progenitors (GRPs) were embedded in 0:1, 1:50, 1:10, and 1:5 Vs:HA mixtures (final cell concentration  =1 × 107/ml). For the in vitro assay, mixtures were placed in 96-wells plate in triplicate and bioluminescence was measured after 1, 3, 7, 14, 21, and 28 days. For in vivo experiments, Vs/HA mixtures containing GRPs were injected subcutaneously in SCID mice and BLI was acquired at 1, 3, 7, and 14 days post-injection.

Results

Mixing of V-sense at increasing ratios of 1:50, 1:10, and 1:5 v/v of fluorine/activated hyaluronan (HA) hydrogel gradually elongated the gelation time from 194 s for non-fluorinated controls to 304 s for 1:5 V-sense:HA hydrogels, while their elastic properties slightly decreased. There was no release of V-sense from hydrogels maintained in stationary conditions over 2 months. The addition of V-sense positively affected in vitro survival of scaffolded GRPs in a dose-dependent manner.

Conclusions

These results show that hydrogel fluorination does not impair its beneficial properties for scaffolded cells, which may be used to visualize scaffolded GRP transplants with 19F MRI.

Key Words

Hydrogel Glial-restricted precursors Fluorine MRI Scaffold 

Notes

Funding Information

This work was supported by R01 EB023647, R56 NS098520, MSCRFD-3899, MSCRFII=2829, R01NS091110, R01NS091100, and R21NS106436. MP was supported by the KNOW program of Jagiellonian University, Cracow, Poland.

Compliance with Ethical Standards

The study was approved by our Institutional Animal Care and Use Committee at the Johns Hopkins University.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Copyright information

© World Molecular Imaging Society 2019

Authors and Affiliations

  1. 1.Russell H. Morgan Department of Radiology and Radiological Science, Division of MR ResearchThe Johns Hopkins University School of MedicineBaltimoreUSA
  2. 2.Cellular Imaging Section and Vascular Biology Program, Institute for Cell EngineeringThe Johns Hopkins University School of MedicineBaltimoreUSA
  3. 3.3rd Department of General SurgeryJagiellonian University Medical CollegeKrakowPoland
  4. 4.Department of Neurology and NeurosurgeryUniversity of Warmia and MazuryOlsztynPoland
  5. 5.Translational Tissue Engineering CenterThe Johns Hopkins University School of MedicineBaltimoreUSA
  6. 6.Mary and Dick Holland Regenerative Medicine Program, Department of Neurological SciencesThe University of Nebraska Medical CenterOmahaUSA
  7. 7.Department of Biomedical EngineeringThe Johns Hopkins University School of MedicineBaltimoreUSA
  8. 8.Department of Chemical and Biomolecular Engineering, Whiting School of EngineeringThe Johns Hopkins UniversityBaltimoreUSA
  9. 9.Department of OncologyThe Johns Hopkins University School of MedicineBaltimoreUSA

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