TY - JOUR
T1 - A bioartificial and vasculomorphic bone matrix-based organoid mimicking microanatomy of flat and short bones
AU - Toni, Roberto
AU - Barbaro, Fulvio
AU - Di Conza, Giusy
AU - Zini, Nicoletta
AU - Remaggi, Giulia
AU - Elviri, Lisa
AU - Spaletta, Giulia
AU - Quarantini, Enrico
AU - Quarantini, Marco
AU - Mosca, Salvatore
AU - Caravelli, Silvio
AU - Mosca, Massimiliano
AU - Ravanetti, Francesca
AU - Sprio, Simone
AU - Tampieri, Anna
N1 - Funding Information:
As summarized in Figure 8 , replication stability of all mononuclear cell phenotypes was confirmed at 80% subconfluence, showing a statistically significant increase in the total number of cells at each split, also when theoretically constrained by the assumption of cell homogeneity. Consistently, no statistically significant difference was observed in the mean number of culture days to reach subconfluence at each passage, suggesting constancy in cell replication rate during each culture interval. Finally, flow cytometric analysis revealed a percentage of CD45‐, CD73‐, and CD90‐positive cells consistent with an immunophenotype typical for BMSCs of a strain and age of rats as we used, and of P2 subculture passage. Retainment of homogeneity in cell phenotypes was also supported by the constancy in distribution of cell size and granularity at forward and side scatter analysis (Data courtesy of Alessandra Zamparelli, Grant FIRB RBAP10MLK7_004 postdoctoral Fellowship 2010–2015, UNIPR, Parma, Italy and Luca Cattini, Laboratories of Immunorheumatology and Tissue Regeneration, RAMSES, IOR, Bologna, Italy).
Funding Information:
The authors are particularly grateful to Ivan Martin, Head Department of Biomedicine, University of Basel, CH for key suggestions on the general presentation of the experimental material and research data. RT is under the tenure of the DIMEC UNIPR–GMC Collaboration Agreement 2020–2023 to develop the research program: Outpatient Diagnostic–Therapeutic Quality and National and International Guidelines on Endocrine-Metabolic Disorders. Part of these studies have been presented at the National Continuing Medical Education (CME) Program “Osteoporosis: novelties and perspectives” held at CMG on January 18, 2020. The odontostomatological and densitometric data were obtained in the context of the ORTODENT transversal observational clinical study, jointly promoted by GMC and the Municipality of Galliera (BO, Italy) on the resident population (see at: https://www.centromedicogalliera.com/progettoosteonet-comunedigalliera-universit%C3%A0diparma). GS developed the mathematical basis for cell splitting assessment, and supervised statistical analysis procedures; EQ, MQ, SM, SC, and MM followed the dental and orthopedic data on osteoporosis. GR is recipient of a Research Fellowship in the PhD program of Drug Sciences at UNIPR, Italy under the tenure of the EU Research Grant Horizon 2020 SCREENED #825745, and received an S3 PhD Video Contest Award by the ART–EROI Platform, Emilia Romagna, Italy. FR provided technical support for light microscopy; SS and AT gave suggestions and advices for the development of innovative procedures for the use of biomaterials and their application. # The work is dedicated to the memory of Davide Dallatana, for his invaluable scientific and technical contribution.
Funding Information:
The authors are particularly grateful to Ivan Martin, Head Department of Biomedicine, University of Basel, CH for key suggestions on the general presentation of the experimental material and research data. RT is under the tenure of the DIMEC UNIPR–GMC Collaboration Agreement 2020–2023 to develop the research program: . Part of these studies have been presented at the National Continuing Medical Education (CME) Program held at CMG on January 18, 2020. The odontostomatological and densitometric data were obtained in the context of the ORTODENT transversal observational clinical study, jointly promoted by GMC and the Municipality of Galliera (BO, Italy) on the resident population (see at: https://www.centromedicogalliera.com/progettoosteonet-comunedigalliera-universit%C3%A0diparma . GS developed the mathematical basis for cell splitting assessment, and supervised statistical analysis procedures; EQ, MQ, SM, SC, and MM followed the dental and orthopedic data on osteoporosis. GR is recipient of a Research Fellowship in the PhD program of Drug Sciences at UNIPR, Italy under the tenure of the EU Research Grant Horizon 2020 SCREENED #825745, and received an S3 PhD Video Contest Award by the ART–EROI Platform, Emilia Romagna, Italy. FR provided technical support for light microscopy; SS and AT gave suggestions and advices for the development of innovative procedures for the use of biomaterials and their application. Outpatient Diagnostic–Therapeutic Quality and National and International Guidelines on Endocrine‐Metabolic Disorders “Osteoporosis: novelties and perspectives” )
Publisher Copyright:
© 2023 The Authors. Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals LLC.
PY - 2024/1
Y1 - 2024/1
N2 - We engineered an in vitro model of bioartificial 3D bone organoid consistent with an anatomical and vascular microenvironment common to mammalian flat and short bones. To achieve this, we chose the decellularized–decalcified matrix of the adult male rat scapula, implemented with the reconstruction of its intrinsic vessels, obtained through an original intravascular perfusion with polylevolactic (PLLA), followed by coating of the PLLA-fabricated vascularization with rat tail collagen. As a result, the 3D bone and vascular geometry of the native bone cortical and cancellous compartments was reproduced, and the rat tail collagen–PLLA biomaterial could in vitro act as a surrogate of the perivascular extracellular matrix (ECM) around the wall of the biomaterial-reconstituted cancellous vessels. As a proof-of-concept of cell compatibility and site-dependent osteoinductive properties of this bioartificial 3D construct, we show that it in vitro leads to a time-dependent microtopographic positioning of rat mesenchymal stromal cells (MSCs), initiating an osteogenic fate in relation to the bone compartment. In addition, coating of PLLA-reconstructed vessels with rat tail collagen favored perivascular attachment and survival of MSC-like cells (mouse embryonic fibroblasts), confirming its potentiality as a perivascular stroma for triggering competence of seeded MSCs. Finally, in vivo radiographic topography of bone lesions in the human jaw and foot tarsus of subjects with primary osteoporosis revealed selective bone cortical versus cancellous involvement, suggesting usefulness of a human 3D bone organoid engineered with the same principles of our rat organoid, to in vitro investigate compartment-dependent activities of human MSC in flat and short bones under experimental osteoporotic challenge. We conclude that our 3D bioartificial construct offers a reliable replica of flat and short bones microanatomy, and promises to help in building a compartment-dependent mechanistic perspective of bone remodeling, including the microtopographic dysregulation of osteoporosis.
AB - We engineered an in vitro model of bioartificial 3D bone organoid consistent with an anatomical and vascular microenvironment common to mammalian flat and short bones. To achieve this, we chose the decellularized–decalcified matrix of the adult male rat scapula, implemented with the reconstruction of its intrinsic vessels, obtained through an original intravascular perfusion with polylevolactic (PLLA), followed by coating of the PLLA-fabricated vascularization with rat tail collagen. As a result, the 3D bone and vascular geometry of the native bone cortical and cancellous compartments was reproduced, and the rat tail collagen–PLLA biomaterial could in vitro act as a surrogate of the perivascular extracellular matrix (ECM) around the wall of the biomaterial-reconstituted cancellous vessels. As a proof-of-concept of cell compatibility and site-dependent osteoinductive properties of this bioartificial 3D construct, we show that it in vitro leads to a time-dependent microtopographic positioning of rat mesenchymal stromal cells (MSCs), initiating an osteogenic fate in relation to the bone compartment. In addition, coating of PLLA-reconstructed vessels with rat tail collagen favored perivascular attachment and survival of MSC-like cells (mouse embryonic fibroblasts), confirming its potentiality as a perivascular stroma for triggering competence of seeded MSCs. Finally, in vivo radiographic topography of bone lesions in the human jaw and foot tarsus of subjects with primary osteoporosis revealed selective bone cortical versus cancellous involvement, suggesting usefulness of a human 3D bone organoid engineered with the same principles of our rat organoid, to in vitro investigate compartment-dependent activities of human MSC in flat and short bones under experimental osteoporotic challenge. We conclude that our 3D bioartificial construct offers a reliable replica of flat and short bones microanatomy, and promises to help in building a compartment-dependent mechanistic perspective of bone remodeling, including the microtopographic dysregulation of osteoporosis.
KW - bone remodeling
KW - mesenchymal stromal cell
KW - microtopography
KW - organoid
KW - osteoporosis
KW - polylevolactic acid
KW - scaffold
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U2 - 10.1002/jbm.b.35329
DO - 10.1002/jbm.b.35329
M3 - Article
C2 - 37898921
AN - SCOPUS:85171449396
SN - 1552-4973
VL - 112
SP - e35329
JO - Journal of Biomedical Materials Research - Part B Applied Biomaterials
JF - Journal of Biomedical Materials Research - Part B Applied Biomaterials
IS - 1
M1 - e35329
ER -