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Light-mediated Formation and Patterning of Hydrogels for Cell Culture Applications

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Network Formation Dynamics in Thiol-ene Crosslinked Hyaluronic Acid Hydrogels: Design Principles for In Vitro Tissue

Kyley Burkey^1,2, Yiwen Zheng³, Kinsey Drake⁴

¹Bioengineering Graduate Program, University of Kansas, Lawrence, KS, United States.

Biorxiv : the Preprint Server for Biology

|June 4, 2026

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This summary is machine-generated.

This study characterizes thiol-ene hydrogel formation for tissue engineering, optimizing network properties like mesh size and mechanics. Molecular Dynamics simulations predict reaction efficiency, validating the system with encapsulated cells.

Area of Science:

Biomaterials Science
Tissue Engineering
Polymer Chemistry

Background:

Hydrogels are crucial 3D cell culture systems for studying mechanotransduction in tissue engineering.
Photoinitiated thiol-ene click chemistry offers control over hydrogel properties but can have unpredictable reaction kinetics.
Thorough characterization of thiol-ene network formation is essential for understanding cell behavior within engineered microenvironments.

Purpose of the Study:

To investigate thiol-ene crosslinking dynamics for hyaluronic acid hydrogels.
To determine hydrogel network formation, mesh size, and mechanical properties.
To establish a predictive method for thiol-ene reaction efficiency using Molecular Dynamics simulations.

Main Methods:

Modulated polymer concentration (4-8 wt%) and thiol:ene molar ratios (0.15:1 to 1:1).

Keywords:

hydrogel mechanotransduction meniscus mesh size thiol-ene

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Assessed hydrogel network properties, including shear storage modulus and relative mesh size.

Employed Molecular Dynamics (MD) simulations to model crosslinking and predict reaction efficiency.

Main Results:

Hydrogel parameters significantly influenced network properties, including mechanical strength and mesh size.
MD simulations provided a method to predict thiol-ene reaction efficiency.
The developed hydrogel system supported the metabolic activity of encapsulated human meniscal cells, confirming its *in vitro* modeling feasibility.

Conclusions:

Optimized thiol-ene chemistry in hyaluronic acid hydrogels allows tunable mechanical properties and mesh sizes for tissue engineering.
Molecular Dynamics simulations are a valuable tool for predicting and understanding thiol-ene hydrogel formation.
This hydrogel system is suitable for *in vitro* modeling of cellular responses in a controlled microenvironment.