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Title: Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates

Abstract

Cell-derived extracellular matrix (ECM) provides a niche to promote osteogenic differentiation, cell adhesion, survival, and trophic factor secretion. To determine whether osteogenic preconditioning would improve the bone-forming potential of unfractionated bone marrow aspirate (BMA), we perfused cells on ECM-coated scaffolds to generate naïve and preconditioned constructs, respectively. The composition of cells selected from BMA was distinct on each scaffold. Naïve constructs exhibited robust proangiogenic potential in vitro, while preconditioned scaffolds contained more mesenchymal stem/stromal cells (MSCs) and endothelial cells (ECs) and exhibited an osteogenic phenotype. Upon implantation into an orthotopic calvarial defect, BMA-derived ECs were present in vessels in preconditioned implants, resulting in robust perfusion and greater vessel density over the first 14 days compared to naïve implants. After 10 weeks, human ECs and differentiated MSCs were detected in de novo tissues derived from naïve and preconditioned scaffolds. These results demonstrate that bioreactor-based preconditioning augments the bone-forming potential of BMA.

Authors:
ORCiD logo [1];  [1];  [2];  [3]; ORCiD logo [4];  [3]; ORCiD logo [5]
  1. Univ. of California, Davis, CA (United States). Dept. of Biomedical Engineering
  2. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Physical and Life Sciences
  3. Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Physical and Life Sciences
  4. Univ. of California, Davis, CA (United States). Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine
  5. Univ. of California, Davis, CA (United States). Dept. of Biomedical Engineering; UC Davis Health, Sacramento, California (United States). Department of Orthopaedic Surgery, School of Medicine
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1626036
Grant/Contract Number:  
AC52-07NA27344
Resource Type:
Accepted Manuscript
Journal Name:
Science Advances
Additional Journal Information:
Journal Volume: 6; Journal Issue: 7; Journal ID: ISSN 2375-2548
Publisher:
AAAS
Country of Publication:
United States
Language:
English
Subject:
Science & Technology - Other Topics

Citation Formats

Harvestine, J. N., Gonzalez-Fernandez, T., Sebastian, A., Hum, N. R., Genetos, D. C., Loots, G. G., and Leach, J. K. Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates. United States: N. p., 2020. Web. doi:10.1126/sciadv.aay2387.
Harvestine, J. N., Gonzalez-Fernandez, T., Sebastian, A., Hum, N. R., Genetos, D. C., Loots, G. G., & Leach, J. K. Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates. United States. doi:10.1126/sciadv.aay2387.
Harvestine, J. N., Gonzalez-Fernandez, T., Sebastian, A., Hum, N. R., Genetos, D. C., Loots, G. G., and Leach, J. K. Sat . "Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates". United States. doi:10.1126/sciadv.aay2387. https://www.osti.gov/servlets/purl/1626036.
@article{osti_1626036,
title = {Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates},
author = {Harvestine, J. N. and Gonzalez-Fernandez, T. and Sebastian, A. and Hum, N. R. and Genetos, D. C. and Loots, G. G. and Leach, J. K.},
abstractNote = {Cell-derived extracellular matrix (ECM) provides a niche to promote osteogenic differentiation, cell adhesion, survival, and trophic factor secretion. To determine whether osteogenic preconditioning would improve the bone-forming potential of unfractionated bone marrow aspirate (BMA), we perfused cells on ECM-coated scaffolds to generate naïve and preconditioned constructs, respectively. The composition of cells selected from BMA was distinct on each scaffold. Naïve constructs exhibited robust proangiogenic potential in vitro, while preconditioned scaffolds contained more mesenchymal stem/stromal cells (MSCs) and endothelial cells (ECs) and exhibited an osteogenic phenotype. Upon implantation into an orthotopic calvarial defect, BMA-derived ECs were present in vessels in preconditioned implants, resulting in robust perfusion and greater vessel density over the first 14 days compared to naïve implants. After 10 weeks, human ECs and differentiated MSCs were detected in de novo tissues derived from naïve and preconditioned scaffolds. These results demonstrate that bioreactor-based preconditioning augments the bone-forming potential of BMA.},
doi = {10.1126/sciadv.aay2387},
journal = {Science Advances},
number = 7,
volume = 6,
place = {United States},
year = {2020},
month = {2}
}

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Works referenced in this record:

Quantifying Mesenchymal Stem Cells in the Mononuclear Cell Fraction of Bone Marrow Samples Obtained for Cell Therapy
journal, January 2013


Preconditioning influences mesenchymal stem cell properties in vitro and in vivo
journal, February 2018

  • Hu, Chenxia; Li, Lanjuan
  • Journal of Cellular and Molecular Medicine, Vol. 22, Issue 3
  • DOI: 10.1111/jcmm.13492

Tetrandrine identified in a small molecule screen to activate mesenchymal stem cells for enhanced immunomodulation
journal, July 2016

  • Yang, Zijiang; Concannon, John; Ng, Kelvin S.
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep30263

Hypoxic Preconditioning Results in Increased Motility and Improved Therapeutic Potential of Human Mesenchymal Stem Cells
journal, August 2008


Expansion of Bone Marrow Mesenchymal Stromal Cells in Perfused 3D Ceramic Scaffolds Enhances In Vivo Bone Formation
journal, September 2017

  • Hoch, Allison I.; Duhr, Ralph; Di Maggio, Nunzia
  • Biotechnology Journal, Vol. 12, Issue 12
  • DOI: 10.1002/biot.201700071

The Extracellular Matrix: Not Just Pretty Fibrils
journal, November 2009


Mixed extracellular matrix ligands synergistically modulate integrin adhesion and signaling
journal, November 2008

  • Reyes, Catherine D.; Petrie, Timothy A.; García, Andrés J.
  • Journal of Cellular Physiology, Vol. 217, Issue 2
  • DOI: 10.1002/jcp.21512

Enhancing the biological performance of synthetic polymeric materials by decoration with engineered, decellularized extracellular matrix
journal, July 2012


Protein and Mineral Composition of Osteogenic Extracellular Matrix Constructs Generated with a Flow Perfusion Bioreactor
journal, December 2011

  • Thibault, Richard A.; Mikos, Antonios G.; Kasper, F. Kurtis
  • Biomacromolecules, Vol. 12, Issue 12
  • DOI: 10.1021/bm200975a

In vitro biomimetic engineering of a human hematopoietic niche with functional properties
journal, June 2018

  • Bourgine, Paul E.; Klein, Thibaut; Paczulla, Anna M.
  • Proceedings of the National Academy of Sciences, Vol. 115, Issue 25
  • DOI: 10.1073/pnas.1805440115

Cell-secreted extracellular matrix influences cellular composition sequestered from unprocessed bone marrow aspirate for osteogenic grafts
journal, January 2019

  • Harvestine, Jenna N.; Saiz, Jr., Augustine M.; Leach, J. Kent
  • Biomaterials Science, Vol. 7, Issue 5
  • DOI: 10.1039/C8BM01478G

Dlx5, a Positive Regulator of Osteoblastogenesis, is Essential for Osteoblast-Osteoclast Coupling
journal, September 2008

  • Samee, Nadeem; Geoffroy, Valerie; Marty, Caroline
  • The American Journal of Pathology, Vol. 173, Issue 3
  • DOI: 10.2353/ajpath.2008.080243

Stem Cells Combined With Platelet-rich Plasma Effectively Treat Corticosteroid-induced Osteonecrosis of the Hip: A Prospective Study
journal, January 2018


Benefits of small volume and small syringe for bone marrow aspirations of mesenchymal stem cells
journal, July 2013

  • Hernigou, Philippe; Homma, Yasuhiro; Flouzat Lachaniette, Charles Henri
  • International Orthopaedics, Vol. 37, Issue 11
  • DOI: 10.1007/s00264-013-2017-z

Strategies for MSC expansion and MSC-based microtissue for bone regeneration
journal, March 2019


Characterization and cost-benefit analysis of automated bioreactor-expanded mesenchymal stem cells for clinical applications: VALIDATION OF AUTOMATED BIOREACTOR
journal, September 2018

  • Russell, Athena L.; Lefavor, Rebecca C.; Zubair, Abba C.
  • Transfusion, Vol. 58, Issue 10
  • DOI: 10.1111/trf.14805

Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner
journal, September 2002

  • Bancroft, G. N.; Sikavitsas, V. I.; van den Dolder, J.
  • Proceedings of the National Academy of Sciences, Vol. 99, Issue 20, p. 12600-12605
  • DOI: 10.1073/pnas.202296599

Mineralized matrix deposition by marrow stromal osteoblasts in 3D perfusion culture increases with increasing fluid shear forces
journal, December 2003

  • Sikavitsas, V. I.; Bancroft, G. N.; Holtorf, H. L.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 25
  • DOI: 10.1073/pnas.2434367100

Flow rates in perfusion bioreactors to maximise mineralisation in bone tissue engineering in vitro
journal, October 2018


Differentiation-Dependent Secretion of Proangiogenic Factors by Mesenchymal Stem Cells
journal, April 2012


Cell-secreted matrices perpetuate the bone-forming phenotype of differentiated mesenchymal stem cells
journal, January 2016


Flow Perfusion Co-culture of Human Mesenchymal Stem Cells and Endothelial Cells on Biodegradable Polymer Scaffolds
journal, July 2013

  • Dahlin, Rebecca L.; Gershovich, Julia G.; Kasper, F. Kurtis
  • Annals of Biomedical Engineering, Vol. 42, Issue 7
  • DOI: 10.1007/s10439-013-0862-y

Stem Cell-Derived Endochondral Cartilage Stimulates Bone Healing by Tissue Transformation: CARTILAGE TRANSFORMATION STIMULATES BONE HEALING
journal, April 2014

  • Bahney, Chelsea S.; Hu, Diane P.; Taylor, Aaron J.
  • Journal of Bone and Mineral Research, Vol. 29, Issue 5
  • DOI: 10.1002/jbmr.2148

Osteogenesis and Trophic Factor Secretion are Influenced by the Composition of Hydroxyapatite/Poly(Lactide-Co-Glycolide) Composite Scaffolds
journal, January 2010


VEGF Scaffolds Enhance Angiogenesis and Bone Regeneration in Irradiated Osseous Defects
journal, February 2006

  • Kaigler, Darnell; Wang, Zhuo; Horger, Kim
  • Journal of Bone and Mineral Research, Vol. 21, Issue 5
  • DOI: 10.1359/jbmr.060120

Establishment of an Early Vascular Network Promotes the Formation of Ectopic Bone
journal, February 2016

  • Eman, Rhandy M.; Meijer, Henriette A. W.; Öner, F. Cumhur
  • Tissue Engineering Part A, Vol. 22, Issue 3-4
  • DOI: 10.1089/ten.tea.2015.0227

Extracellular Matrix-Coated Composite Scaffolds Promote Mesenchymal Stem Cell Persistence and Osteogenesis
journal, September 2016


Cell-secreted extracellular matrix, independent of cell source, promotes the osteogenic differentiation of human stromal vascular fraction
journal, January 2018

  • Harvestine, Jenna N.; Orbay, Hakan; Chen, Jonathan Y.
  • Journal of Materials Chemistry B, Vol. 6, Issue 24
  • DOI: 10.1039/C7TB02787G

In vitro generated extracellular matrix and fluid shear stress synergistically enhance 3D osteoblastic differentiation
journal, February 2006

  • Datta, N.; Pham, Q. P.; Sharma, U.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 8
  • DOI: 10.1073/pnas.0505661103

Hydrogel biophysical properties instruct coculture‐mediated osteogenic potential
journal, September 2015

  • Murphy, Kaitlin C.; Stilhano, Roberta S.; Mitra, Debika
  • The FASEB Journal, Vol. 30, Issue 1
  • DOI: 10.1096/fj.15-279984

Human endothelial colony-forming cells serve as trophic mediators for mesenchymal stem cell engraftment via paracrine signaling
journal, June 2014

  • Lin, R. -Z.; Moreno-Luna, R.; Li, D.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 28
  • DOI: 10.1073/pnas.1405388111

VEGF: an Essential Mediator of Both Angiogenesis and Endochondral Ossification
journal, October 2007


Analysis of Osteocalcin Expression in Transgenic Mice Reveals a Species Difference in Vitamin D Regulation of Mouse and Human Osteocalcin Genes
journal, October 1997


Design of Experiments Approach to Engineer Cell-Secreted Matrices for Directing Osteogenic Differentiation
journal, December 2010


Transferable cell-secreted extracellular matrices enhance osteogenic differentiation
journal, February 2012


Bioreactor culture duration of engineered constructs influences bone formation by mesenchymal stem cells
journal, November 2017


Oscillating perfusion of cell suspensions through three-dimensional scaffolds enhances cell seeding efficiency and uniformity
journal, September 2003

  • Wendt, D.; Marsano, A.; Jakob, M.
  • Biotechnology and Bioengineering, Vol. 84, Issue 2
  • DOI: 10.1002/bit.10759

Mapping RNA‐seq Reads with STAR
journal, September 2015


featureCounts: an efficient general purpose program for assigning sequence reads to genomic features
journal, November 2013


voom: precision weights unlock linear model analysis tools for RNA-seq read counts
journal, January 2014


ToppGene Suite for gene list enrichment analysis and candidate gene prioritization
journal, May 2009

  • Chen, J.; Bardes, E. E.; Aronow, B. J.
  • Nucleic Acids Research, Vol. 37, Issue Web Server
  • DOI: 10.1093/nar/gkp427

Osteogenic comparison of expanded and uncultured adipose stromal cells
journal, July 2010


Engineered Fibrin Gels for Parallel Stimulation of Mesenchymal Stem Cell Proangiogenic and Osteogenic Potential
journal, December 2014

  • Murphy, Kaitlin C.; Hughbanks, Marissa L.; Binder, Bernard Y. K.
  • Annals of Biomedical Engineering, Vol. 43, Issue 8
  • DOI: 10.1007/s10439-014-1227-x