TY - JOUR
T1 - Delivery of human adipose stem cells spheroids into lockyballs
AU - Silva, Karina R.
AU - Rezende, Rodrigo A.
AU - Pereira, Frederico D.A.S.
AU - Gruber, Peter
AU - Stuart, Mellannie P.
AU - Ovsianikov, Aleksandr
AU - Brakke, Ken
AU - Kasyanov, Vladimir
AU - Da Silva, Jorge V.L.
AU - Granjeiro, José M.
AU - Baptista, Leandra S.
AU - Mironov, Vladimir
N1 - Funding Information:
This work was funded by CNPq - 457541/2013-0 - Chamada No 47/2013 MCTI/CNPq/CT-Sa?de/CT-Biotecnologia/MS/SCTIE/DECIT - Novas Terapias Portadoras de Futuro to JMG; European Research Council (Starting Grant-307701) to AO; CNPq Regenerative Medicine project 467643/2014-8 to JVLS; and Brazilian Institute of Biofabrication (INCT-BIOFABRIS) 08/57860-3 to JVLS. National Institute of Metrology, Quality and Technology (INMETRO), Duque de Caxias, is acknowledged for supplying facilities for Microscopic analyzes. Authors would like to thank Prof. C.P. Heisenberg from Austrian Institute of Science and Technology for kind permission to use Microsquisher? for measurement of material properties of lockyballs in his laboratory and Dr. Gabriel Krenz from his lab for training and assisting in performing measurements; Prof. Jurgen Stampfl from Technische Universit?t Wien (TU Wien), Austria, for valuable advices and support. We also would like to thank Brunno Ver?oza from Numpex-bio and Ricardo Vilela from Inmetro for technical assistance on the microscopic facility and Prof. Cesar Claudio-da-Silva, MD, PhD from Federal University of Rio de Janeiro and Marcelo Aniceto, MD, MSc for providing the human lipoaspirate samples. Authors also would like to thank Prof. Estevam Augusto Bonfante from Department of Prosthodontics (FOB-USP) for English Language revision.
Publisher Copyright:
© 2016 Silva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2016/11/1
Y1 - 2016/11/1
N2 - Adipose stem cells (ASCs) spheroids show enhanced regenerative effects compared to single cells. Also, spheroids have been recently introduced as building blocks in directed selfassembly 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.
AB - Adipose stem cells (ASCs) spheroids show enhanced regenerative effects compared to single cells. Also, spheroids have been recently introduced as building blocks in directed selfassembly 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.
UR - http://www.scopus.com/inward/record.url?scp=84994430938&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0166073
DO - 10.1371/journal.pone.0166073
M3 - Article
C2 - 27829016
AN - SCOPUS:84994430938
SN - 1932-6203
VL - 11
JO - PLoS ONE
JF - PLoS ONE
IS - 11
M1 - e0166073
ER -
