Delivery of Human Adipose Stem Cells Spheroids into Lockyballs.
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.
Laboratory of Tissue Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), Duque de Caxias, Rio de Janeiro, Brazil.|Nucleus of Multidisciplinary Research in Biology (Numpex-Bio), Federal University of Rio de Janeiro-Xerém, Duque de Caxias, Rio de Janeiro, Brazil.
Karina R
KR
Silva
Division of 3D Technologies, Renato Archer Center for Information Technology (CTI), Campinas, São Paulo, Brazil.
Rodrigo A
RA
Rezende
Division of 3D Technologies, Renato Archer Center for Information Technology (CTI), Campinas, São Paulo, Brazil.
Frederico D A S
FD
Pereira
Institute of Materials Science and Technology, TU Wien (Technische Universität Wien), Vienna, Austria.
Peter
P
Gruber
Laboratory of Tissue Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), Duque de Caxias, Rio de Janeiro, Brazil.
Mellannie P
MP
Stuart
Institute of Materials Science and Technology, TU Wien (Technische Universität Wien), Vienna, Austria.
Aleksandr
A
Ovsianikov
Mathematics Department, Susquehanna University, Selinsgrove, Pennsylvania, United States of America.
Ken
K
Brakke
Riga Stradins University and Riga Technical University, Riga, Latvia.
Vladimir
V
Kasyanov
Division of 3D Technologies, Renato Archer Center for Information Technology (CTI), Campinas, São Paulo, Brazil.
Jorge V L
JV
da Silva
Laboratory of Tissue Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), Duque de Caxias, Rio de Janeiro, Brazil.|Bioengineering Laboratory, Fluminense Federal University, Niterói, Rio de Janeiro, Brazil.
José M
JM
Granjeiro
Laboratory of Tissue Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), Duque de Caxias, Rio de Janeiro, Brazil.|Nucleus of Multidisciplinary Research in Biology (Numpex-Bio), Federal University of Rio de Janeiro-Xerém, Duque de Caxias, Rio de Janeiro, Brazil.
Leandra S
http://orcid.org/0000-0001-9998-8044
LS
Baptista
Laboratory of Tissue Bioengineering, National Institute of Metrology, Quality and Technology (Inmetro), Duque de Caxias, Rio de Janeiro, Brazil.|Division of 3D Technologies, Renato Archer Center for Information Technology (CTI), Campinas, São Paulo, Brazil.
Vladimir
V
Mironov
D000273:Adipose Tissue / Q000166:cytology*; D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D055119:Elastic Modulus; D005260:Female; D015870:Gene Expression; D006801:Humans; D020100:Hydrogels / Q000737:chemistry; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D044968:Regenerative Medicine / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000235:genetics; D012685:Sepharose / Q000737:chemistry; D018874:Spheroids, Cellular / Q000737:chemistry / Q000166:cytology / Q000637:transplantation*; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*; D055815:Young Adult
D050659:Core Binding Factor Alpha 1 Subunit; D020100:Hydrogels; C493709:RUNX2 protein, human; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D012685:Sepharose