Native extracellular matrix probes to target patient- and tissue-specific cell-microenvironment interactions by force...

Area of Science:

• Biophysics
• Cell Biology
• Materials Science

Background:

• Atomic Force Microscopy (AFM) is crucial for studying cellular mechanosensing.
• Current AFM probes functionalized with single extracellular matrix (ECM) components poorly replicate the native microenvironment's complexity.
• This limitation hinders accurate investigation of cell-ECM interactions.

Purpose of the Study:

• To develop novel AFM probes that faithfully retain the structural and biochemical complexity of native ECM.
• To utilize these probes for quantitative analysis of cell-ECM interactions using force spectroscopy.
• To explore applications in personalized medicine by analyzing patient-specific tissue interactions.

Main Methods:

• Fabrication of AFM probes by attaching laser microdissected slices of native decellularized ECM to AFM cantilevers.
• Characterization of ECM probes for morphological, mechanical, and chemical heterogeneity.
• Application of force spectroscopy to measure cell-ECM adhesion forces at the pN level, using rat bladder ECM probes and AY-27 cancer cells.

Main Results:

• The novel native ECM probes successfully preserved the ECM's inherent heterogeneity.
• Force spectroscopy experiments yielded reproducible results, distinguishing adhesion patterns across different bladder ECM regions (submucosa, detrusor, adventitia).
• The study demonstrated the feasibility of dissecting mechanotransductive cell-ECM interactions with high sensitivity (10 pN range).

Conclusions:

• Native ECM probes offer a more biologically relevant tool for studying cell-microenvironment interactions compared to traditional functionalized probes.
• These probes enable detailed investigation of mechanotransduction at the cellular level.
• The technology holds significant promise for advancing personalized medicine through patient-specific tissue analysis.