Investigating materials and orientation parameters for the creation of a 3D musculoskeletal interface co-culture model.

Hamad Alsaykhan,Jennifer Z Paxton

doi:10.1093/rb/rbaa018

Hamad Alsaykhan, Jennifer Z Paxton

Open Access

https://doi.org/10.1093/rb/rbaa018

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Abstract

Musculoskeletal tissue interfaces are a common site of injury in the young, active populations. In particular, the interface between the musculoskeletal tissues of tendon and bone is often injured and to date, no single treatment has been able to restore the form and function of damaged tissue at the bone–tendon interface. Tissue engineering and regeneration hold great promise for the manufacture of bespoke in vitro models or implants to be used to advance repair and so this study investigated the material, orientation and culture choices for manufacturing a reproducible 3D model of a musculoskeletal interface between tendon and bone cell populations. Such models are essential for future studies focussing on the regeneration of musculoskeletal interfaces in vitro. Cell-encapsulated fibrin hydrogels, arranged in a horizontal orientation though a simple moulding procedure, were shown to best support cellular growth and migration of cells to form an in vitro tendon–bone interface. This study highlights the importance of acknowledging the material and technical challenges in establishing co-cultures and suggests a reproducible methodology to form 3D co-cultures between tendon and bone, or other musculoskeletal cell types, in vitro.

Highlights

Tissue engineering is defined as the development of tissues or organs by manipulating biological, biophysical and biochemical factors in a laboratory setting(Castells-Sala et al 2013)
4.1 Comparing vertical and horizontal 3D co-culture models to represent a musculoskeletal tissue interface 3D co-culture models were assessed through gross morphology, co-culture feasibility and ease of preparation to compare vertical (Figure 1) to horizontal (Figure 2) orientations
Gross assessment of 3D co-cultures in both the vertical and horizontal systems appeared to show a clear demarcation between two hydrogels, the position of cells was assessed to ensure a clear boundary existed between cell types on formation of the co-culture (Figures 3,4,5)

Summary

Introduction

Tissue engineering is defined as the development of tissues or organs by manipulating biological, biophysical and biochemical factors in a laboratory setting(Castells-Sala et al 2013). The enthesis is the biological and mechanical junction between tendon and bone(Benjamin et al 2006; Apostolakos et al 2014). It is commonly injured in young, active populations e.g. such as anterior cruciate ligament injuries(Gianotti et al 2009) tennis elbow, jumper’s knee, rotator cuff tendon tears and calcaneal tendon avulsion(Benjamin et al 2006; Apostolakos et al 2014; Kim et al 2014). Research into methods to help restore the natural gradation of the enthesis and its mechanical function following injury is much needed

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