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Related Experiment Video

Updated: Sep 24, 2025

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Engineering a 3D hydrogel system to study optic nerve head astrocyte morphology and behavior.

Ana N Strat1, Alexander Kirschner2, Hannah Yoo2

  • 1Department of Ophthalmology & Visual Sciences, SUNY Upstate Medical University, Syracuse, NY, 13210, USA; Department of Neuroscience and Physiology, SUNY Upstate Medical University, Syracuse, NY, 13210, USA; BioInspired Institute, Syracuse University, Syracuse, NY, 13244, USA.

Experimental Eye Research
|May 7, 2022
PubMed
Summary

Researchers developed a 3D hydrogel to mimic the optic nerve head (ONH) microenvironment, successfully inducing astrocyte reactivity with transforming growth factor beta 2 (TGF β2) for glaucoma research.

Keywords:
Biomechanical StrainCollagen IVExtracellular matrixFibronectinGFAPGlaucomaReactive gliosisTransforming growth factor beta 2

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Area of Science:

  • Biomaterials Science
  • Neuroscience
  • Ophthalmology

Background:

  • Glaucoma involves optic nerve head (ONH) astrocyte reactivity and glial fibrillary acidic protein (GFAP) upregulation.
  • Elevated transforming growth factor beta 2 (TGF β2) is linked to ONH dysfunction in glaucoma.
  • Conventional 2D cell cultures fail to replicate the in vivo ONH microenvironment for astrocyte studies.

Purpose of the Study:

  • To engineer a 3D hydrogel mimicking the mouse ONH astrocyte (MONHA) microenvironment.
  • To assess MONHA morphology, proliferation, and intercellular connections within the 3D hydrogel.
  • To investigate the induction of MONHA reactivity using TGF β2 in the engineered 3D model.

Main Methods:

  • Primary MONHAs were isolated and encapsulated in a photo-crosslinked collagen I and hyaluronic acid hydrogel.
  • Hydrogels were cultured for 3 weeks, followed by 7-day treatment with varying concentrations of TGF β2.
  • Astrocyte reactivity was assessed via F-actin cytoskeleton, GFAP expression, fibronectin, and collagen IV deposition.

Main Results:

  • MONHAs exhibited high viability and proliferation within the 3D hydrogel over 4 weeks.
  • Sholl analysis revealed increased process complexity and length, with intercellular connections and Connexin43 expression.
  • TGF β2 treatment induced astrocyte reactivity, evidenced by cytoskeletal changes and increased extracellular matrix deposition.

Conclusions:

  • A 3D biomimetic hydrogel effectively replicates key aspects of the in vivo ONH astrocyte microenvironment.
  • This 3D model successfully induces astrocyte reactivity in response to TGF β2, mimicking glaucoma-related changes.
  • The engineered hydrogel provides a valuable platform for future studies on astrocyte behavior in ocular injury and disease.