Enhance Skeletal Muscle Adhesion and Longevity in Microcantilever Platforms

Abstract

The University of Central Florida invention is a system and methods that better integrate myotubes with silicon microcantilevers, enabling longer-term investigations of skeletal muscles for drug efficacy and toxicity. Microcantilever platforms are functional models for studying skeletal muscle force dynamics in vitro. However, the contractile force generated by myotubes can cause them to detach from the cantilevers, especially during long-term experiments. Myotubes are immature muscle cells that form when myoblasts (stem cells) fuse. The UCF invention improves the integration of myotubes with microcantilevers for long-term cultures of mechanically active cells.

In developing the invention, the researchers drew inspiration from the elastomeric proteins elastin and resilin in animals and insects. The spring action of the proteins plays a critical role in force dampening in vivo. In animals, elastin is in the collagenous matrix of the tendon, where muscle attaches to bone. The tendon microenvironment consists of elastin, collagen, and an aqueous jelly-like mass of proteoglycans.

As an example of the invention’s effectiveness, the researchers mimicked the tendon environment by depositing elastin, collagen, heparan sulfate proteoglycan, and hyaluronic acid on a positively charged silane substrate. This resulted in the long-term survival of mechanically active myotubes on glass and silicon microcantilevers for more than 28 days.

Technical Details: Using a biomimetic approach, researchers successfully developed an in vitro extracellular matrix (ECM) composition that enables the long-term survival of mechanically active skeletal muscle myotubes on glass and silicon microcantilevers. The invention enables integration into multiorgan microphysiological platforms.

The UCF system can include a cell culture surface with a silicon oxide first layer and a second layer comprising numerous amphiphilic molecules. Each amphiphilic molecule has a positively charged portion extending from a silane portion that is bound to the silicon oxide first layer. The cell culture surface can also include extracellular matrix components and water molecules dispersed between and bonded to positively charged portions of the amphiphilic molecules.

Example methods of using the system include plating mechanically active cells onto the cell culture surface to form a mechanically active cell culture. With such methods, the mechanically active cells adhere to the cell culture surface for at least 14 days. The mechanically active cell culture can be exposed to an agent, and the cell culture’s responses can be detected.

About the image: As shown in the example image above, the system includes a cell culture surface (102) formed on the exterior surface of a silicon substrate (103), like those used to fabricate micromechanical electrical systems. It has a silicon oxide first layer (104) and a second layer (106) with organosilanes. Each organosilane has a positively charged side (108) extending from a silane (110) covalently bound to the silicon oxide first layer (104). The cell culture surface also includes extracellular matrix components (112: elastin 120, collagen 122, heparan sulfate proteoglycan 124, and hyaluronic acid 126) and water molecules (114) dispersed between and bonded to positively charged portions of the organosilanes.

Partnering Opportunity: The research team is seeking partners for licensing, research collaboration, or both.

Stage of Development: Prototype available.

Benefit

Market Application

Publications

Mimicking the Tendon Microenvironment to Enhance Skeletal Muscle Adhesion and Longevity in a Functional Microcantilever Platform, Himanshi Jangir and James J. Hickman, ACS Biomaterials Science & Engineering 2023 9 (8), 4698-4708, https://doi.org/10.1021/acsbiomaterials.3c00235, Published July 18, 2023. Copyright © 2023 The Authors. Published by the American Chemical Society. The publication is licensed under CC-BY-NC-ND 4.0.