Speaker
Description
Tendon tissue engineering remains a critical challenge due to the need for biomaterials that simultaneously support mechanical load bearing and guide lineage specific cellular differentiation. To address this, we designed a hybrid scaffold system that spatially integrates mechanical reinforcement and tenogenic bioactivity, aiming to mimic native tendon properties more closely than conventional uniform scaffolds.
We developed a dual bioink system engineered to deliver region specific functionality. The first bioink was formulated to enhance mechanical integrity through molecular-level interactions with extracellular proteins, while the second bioink incorporated human adipose derived stem cells (ASCs) and tendon derived extracellular matrix (tECM) components to promote tenogenic differentiation. The two inks were co-delivered using a custom collector system to enable parallel deposition, ensuring microstructural anisotropy. After scaffold fabrication, constructs were cultured in vitro under static conditions.
Cell viability was assessed using live/dead staining and MTT assays at multiple timepoints. Tenogenic differentiation was evaluated by RT-PCR for tendon specific markers including SCX, TNMD, and COL1A1. Mechanical testing was conducted to quantify elastic modulus and ultimate tensile strength, benchmarking against the native tendon range.
The dual bioink scaffold exhibited clear spatial heterogeneity in both mechanical and biological responses. Live/dead staining confirmed high cell viability throughout the biologically active regions, while MTT assays showed sustained metabolic activity. Gene expression analysis revealed a significant upregulation of tenogenic markers in the ASC-tECM regions compared to control hydrogels lacking ECM supplementation. Mechanical analysis demonstrated a significant increase in tensile strength in the reinforced compartment compared to standard gelatin-based constructs, approaching values characteristic of early-stage regenerating tendon.
Our strategy highlights the importance of spatially controlled scaffold design in tendon regeneration. By combining mechanically supportive and biologically inductive regions within a single construct, we achieved both structural resilience and lineage specific differentiation cues. The observed anisotropy is expected to play a crucial role in guiding aligned ECM depositions and long-term tendon remodeling. Importantly, the method avoids reliance on growth factors instead, ECM cues for differentiation, enhancing translational relevance.
This study presents a promising step toward fabricating functional tendon scaffolds with tunable regions tailored to biological and mechanical demands. Further in vivo studies are underway to validate long-term integration and remodeling.
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