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Label-free mapping of microstructural organisation in self-aligning cellular collagen hydrogels using image
Kathleen Sanen1, Rik Paesen1, Sander Luyck1
1Biophysics Group, Biomedical Research Institute, Hasselt University, Agoralaan Building C, 3590 Diepenbeek, Belgium.
Acta Biomaterialia
|November 6, 2015
Summary
This study introduces an advanced image correlation spectroscopy (ICS) model to analyze collagen fibril orientation in hydrogels for regenerative medicine. The method quanties cell-induced changes in hydrogel structure, aiding tissue engineering research.
Area of Science:
- Biomaterials Science
- Regenerative Medicine
- Microscopy and Imaging
Background:
- Hydrogels are crucial biomaterials in regenerative medicine, but understanding cell-mediated remodeling remains challenging.
- Second harmonic generation (SHG) microscopy visualizes collagen, but quantifying fibril orientation requires advanced methods.
- Existing models struggle to capture dynamic changes in fibril alignment crucial for anisotropic tissue constructs.
Purpose of the Study:
- To extend an image correlation spectroscopy (ICS) model for quantifying collagen fibril orientation in cellular hydrogels.
- To demonstrate the model's utility in analyzing cell-induced hydrogel modifications and microstructural changes.
- To assess the model's sensitivity and potential for high-throughput screening in tissue engineering.
Main Methods:
- Developed an extended image correlation spectroscopy (ICS) model incorporating fibril orientation distribution.
- Utilized second harmonic generation (SHG) microscopy for label-free, non-invasive imaging of collagen type I hydrogels.
- Applied the model to a collagen hydrogel contraction assay to monitor cell-induced changes over time.
Main Results:
- The extended ICS model successfully quantified collagen fibril orientation and density changes in cellular hydrogels.
- Observed a threefold increase in collagen density and corresponding decrease in hydrogel area within 24 hours.
- Demonstrated that cellular processes accelerated local fibril alignment, with regions axial to processes aligning 1.5 times faster.
Conclusions:
- The developed automated ICS model accurately characterizes microstructural organization in cellular hydrogels.
- The technique is sensitive to minor temporal and spatial changes in collagen density and fibril orientation.
- This method offers potential for fundamental cell-matrix research and high-throughput screening of hydrogel scaffolds in tissue engineering.
