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Cell Biology JoVE (Open Access) Citable · DOI

Designing Porous Silicon Films as Carriers of Nerve Growth Factor

DOI: 10.3791/58982-v
What you'll learn
  • ✓ Fabricate oxidized porous silicon films as NGF delivery carriers
  • ✓ Load and quantify nerve growth factor release from PSi matrices
  • ✓ Assess neuronal cell viability and differentiation with NGF-loaded carriers
Protocol

Here, we present a protocol to design and fabricate nanostructured porous silicon (PSi) films as degradable carriers for the nerve growth factor (NGF). Neuronal differentiation and outgrowth of PC12 cells and mice dorsal root ganglion (DRG) neurons are characterized upon treatment with the NGF-loaded PSi carriers.

Difficulty
advanced
Total time
~3–5 days (including PSi fabrication, NGF loading, release kinetics, and cell culture assays)
Model organism
PC12 cells; mouse dorsal root ganglion (DRG) neurons
Biosafety
BSL-1

Steps

1
Fabricate oxidized porous silicon carrier films

Design and produce nanostructured PSiO2 films using established electrochemical etching or oxidation protocols. Characterize pore morphology and surface properties for NGF loading.

▶ 00:57
2
Load NGF onto PSiO2 and measure release kinetics

Apply nerve growth factor to PSiO2 carriers, quantify loaded NGF concentration, and monitor in vitro release profiles over time using standard analytical methods.

▶ 02:10
3
Evaluate cell viability with NGF-loaded carriers

Culture PC12 cells and DRG neurons in presence of NGF-loaded PSiO2 carriers and assess cell survival and metabolic activity using viability assays.

▶ 04:42
4
Analyze neuronal differentiation and outgrowth

Characterize differentiation markers and neurite outgrowth morphology in treated cells using immunofluorescence, microscopy, or quantitative morphological analysis.

▶ 06:00
5
Present representative neuronal growth outcomes

Demonstrate and interpret key results showing effects of NGF-loaded PSiO2 carriers on neuronal development and comparative control responses.

▶ 07:27
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