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Orthopaedic Proceedings
Vol. 105-B, Issue SUPP_16 | Pages 42 - 42
17 Nov 2023
ADVANCING THE FABRICATION OF 3D MINI-TISSUE ENTHESIS CONSTRUCTS: CHARACTERIZATION OF BONE, TENDON, AND BMSC SPHEROIDS
Prabhakaran V Sobrattee A Melchels FP Paxton JZ
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Abstract

Objectives

The enthesis is a specialised structure at the interface between bone and tendon with gradual integration to maintain functionality and integrity. In the process of fabricating an in-vitro model of this complex structure, this study aims to investigate growth and maturation of bone, tendon and BMSC spheroids followed by 3D mini-tissue production.

Methods

Cell spheroids Spheroids of differentiated rat osteoblasts (dRObs), rat tendon fibroblasts (RTFs) and bone marrow stem cells (BMSC) were generated by culturing in 96 well U bottom cell repellent plates. With dROb spheroids previously analysed [1], RTF spheroids were examined over a duration of up to 28 days at different seeding densities 1×104, 5×104, 1×105, 2×105 in different media conditions with and without FBS (N=3). Spheroid diameter was analysed by imageJ/Fiji; Cell proliferation and viability was assessed by trypan blue staining after dissociating with accutase + type II collagenase mix; necrotic core by H&E staining; and extracellular matrix by picro-sirius red (RTFs) staining to visualise collagen fibres under bright-field and polarised light microscope.

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