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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); USDOE National Nuclear Security Administration (NNSA)
OSTI Identifier:
1626036
Alternate Identifier(s):
OSTI ID: 1755844
Report Number(s):
LLNL-JRNL-789381
Journal ID: ISSN 2375-2548
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:
59 BASIC BIOLOGICAL SCIENCES; extracellular matrix; bone marrow aspirate; bioreactor; osteogenesis; vascularization

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. https://doi.org/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. https://doi.org/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.. Wed . "Osteogenic preconditioning in perfusion bioreactors improves vascularization and bone formation by human bone marrow aspirates". United States. https://doi.org/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}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Figures / Tables:

Fig. 1 Fig. 1: Schematic of experimental design. (A) Medium containing cell-secreted ECM was mixed with (B) mononucleated cells from BMA and seeded onto (C) HA– poly(lactide- co-glycolide) (PLG) scaffolds using a perfusion bioreactor. After 20 hours of perfusion, (D) naïve scaffolds were retrieved, and (E) preconditioned scaffolds were perfused with culturemore » medium containing osteogenic supplements for 13 additional days. (F) Scaffolds were implanted into 3.5-mm bilateral calvarial defects.« less

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