Abstract
Adipose stem cells (ASCs) spheroids show enhanced regenerative effects compared to single cells. Also, spheroids have been recently introduced as building blocks in directed self-assembly strategy. Recent efforts aim to improve long-term cell retention and integration by the use of microencapsulation delivery systems that can rapidly integrate in the implantation site. Interlockable solid synthetic microscaffolds, so called lockyballs, were recently designed with hooks and loops to enhance cell retention and integration at the implantation site as well as to support spheroids aggregation after transplantation. Here we present an efficient methodology for human ASCs spheroids biofabrication and lockyballs cellularization using micro-molded non-adhesive agarose hydrogel. Lockyballs were produced using two-photon polymerization with an estimated mechanical strength. The Young’s modulus was calculated at level 0.1362 +/-0.009 MPa. Interlocking in vitro test demonstrates high level of loading induced interlockability of fabricated lockyballs. Diameter measurements and elongation coefficient calculation revealed that human ASCs spheroids biofabricated in resections of micro-molded non-adhesive hydrogel had a more regular size distribution and shape than spheroids biofabricated in hanging drops. Cellularization of lockyballs using human ASCs spheroids did not alter the level of cells viability (p › 0,999) and gene fold expression for SOX-9 and RUNX2 (p › 0,195). The biofabrication of ASCs spheroids into lockyballs represents an innovative strategy in regenerative medicine, which combines solid scaffold-based and directed self-assembly approaches, fostering opportunities for rapid in situ biofabrication of 3D building-blocks.
Highlights
The correct delivery of mesenchymal stem cells (MSCs) to injured sites is mandatory to promote tissue repair due to their secretory capacity [1]
Recent studies aimed to improve long-term cell retention and integration by using microencapsulation delivery systems with tailored biomechanical properties and that could rapidly integrate in the implantation site [14,15,16]
Lockyball structures were produced by two-photon polymerization (2PP) of Zr-based hybrid photopolymer
Summary
The correct delivery of mesenchymal stem cells (MSCs) to injured sites is mandatory to promote tissue repair due to their secretory capacity [1]. MSCs and ASCs spheroids or even cell aggregates show enhanced secretion of trophic, anti-apoptotic and anti-inflammatory factors, enhancing their regenerative effects [3,4,5,6]. Spheroids are formed based on self-assembly capacity of cells through molecules recognition process. Cell spheroid assembly can be successively achieved based on spheroid fusion capacity to construct structures at tissue level. Recent studies aimed to improve long-term cell retention and integration by using microencapsulation delivery systems with tailored biomechanical properties and that could rapidly integrate in the implantation site [14,15,16]
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