skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization

Abstract

Bone tissue, by definition, is an organic–inorganic nanocomposite, where metabolically active cells are embedded within a matrix that is heavily calcified on the nanoscale. Currently, there are no strategies that replicate these definitive characteristics of bone tissue. Here we describe a biomimetic approach where a supersaturated calcium and phosphate medium is used in combination with a non-collagenous protein analog to direct the deposition of nanoscale apatite, both in the intra- and extrafibrillar spaces of collagen embedded with osteoprogenitor, vascular, and neural cells. This process enables engineering of bone models replicating the key hallmarks of the bone cellular and extracellular microenvironment, including its protein-guided biomineralization, nanostructure, vasculature, innervation, inherent osteoinductive properties (without exogenous supplements), and cell-homing effects on bone-targeting diseases, such as prostate cancer. Ultimately, this approach enables fabrication of bone-like tissue models with high levels of biomimicry that may have broad implications for disease modeling, drug discovery, and regenerative engineering.

Authors:
ORCiD logo [1];  [1];  [1];  [1]; ORCiD logo [1];  [2]; ORCiD logo [1];  [1];  [3]; ORCiD logo [4]; ORCiD logo [5];  [5];  [1];  [1]
  1. Oregon Health and Science Univ., Portland, OR (United States)
  2. Shriners Hospital for Children, Portland, OR (United States)
  3. Portland State Univ., Portland, OR (United States)
  4. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  5. Univ. of Florida, Gainesville, FL (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1571781
Report Number(s):
PNNL-SA-147563
Journal ID: ISSN 2041-1723
Grant/Contract Number:  
AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Thrivikraman, Greeshma, Athirasala, Avathamsa, Gordon, Ryan, Zhang, Limin, Bergan, Raymond, Keene, Douglas R., Jones, James M., Xie, Hua, Chen, Zhiqiang, Tao, Jinhui, Wingender, Brian, Gower, Laurie, Ferracane, Jack L., and Bertassoni, Luiz E. Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization. United States: N. p., 2019. Web. doi:10.1038/s41467-019-11455-8.
Thrivikraman, Greeshma, Athirasala, Avathamsa, Gordon, Ryan, Zhang, Limin, Bergan, Raymond, Keene, Douglas R., Jones, James M., Xie, Hua, Chen, Zhiqiang, Tao, Jinhui, Wingender, Brian, Gower, Laurie, Ferracane, Jack L., & Bertassoni, Luiz E. Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization. United States. doi:10.1038/s41467-019-11455-8.
Thrivikraman, Greeshma, Athirasala, Avathamsa, Gordon, Ryan, Zhang, Limin, Bergan, Raymond, Keene, Douglas R., Jones, James M., Xie, Hua, Chen, Zhiqiang, Tao, Jinhui, Wingender, Brian, Gower, Laurie, Ferracane, Jack L., and Bertassoni, Luiz E. Tue . "Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization". United States. doi:10.1038/s41467-019-11455-8. https://www.osti.gov/servlets/purl/1571781.
@article{osti_1571781,
title = {Rapid fabrication of vascularized and innervated cell-laden bone models with biomimetic intrafibrillar collagen mineralization},
author = {Thrivikraman, Greeshma and Athirasala, Avathamsa and Gordon, Ryan and Zhang, Limin and Bergan, Raymond and Keene, Douglas R. and Jones, James M. and Xie, Hua and Chen, Zhiqiang and Tao, Jinhui and Wingender, Brian and Gower, Laurie and Ferracane, Jack L. and Bertassoni, Luiz E.},
abstractNote = {Bone tissue, by definition, is an organic–inorganic nanocomposite, where metabolically active cells are embedded within a matrix that is heavily calcified on the nanoscale. Currently, there are no strategies that replicate these definitive characteristics of bone tissue. Here we describe a biomimetic approach where a supersaturated calcium and phosphate medium is used in combination with a non-collagenous protein analog to direct the deposition of nanoscale apatite, both in the intra- and extrafibrillar spaces of collagen embedded with osteoprogenitor, vascular, and neural cells. This process enables engineering of bone models replicating the key hallmarks of the bone cellular and extracellular microenvironment, including its protein-guided biomineralization, nanostructure, vasculature, innervation, inherent osteoinductive properties (without exogenous supplements), and cell-homing effects on bone-targeting diseases, such as prostate cancer. Ultimately, this approach enables fabrication of bone-like tissue models with high levels of biomimicry that may have broad implications for disease modeling, drug discovery, and regenerative engineering.},
doi = {10.1038/s41467-019-11455-8},
journal = {Nature Communications},
number = 1,
volume = 10,
place = {United States},
year = {2019},
month = {8}
}

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

Save / Share:

Works referenced in this record:

Collagen intrafibrillar mineralization as a result of the balance between osmotic equilibrium and electroneutrality
journal, November 2016

  • Niu, Li-na; Jee, Sang Eun; Jiao, Kai
  • Nature Materials, Vol. 16, Issue 3
  • DOI: 10.1038/nmat4789

Fractal-like hierarchical organization of bone begins at the nanoscale
journal, May 2018


Osteocyte differentiation is regulated by extracellular matrix stiffness and intercellular separation
journal, December 2013

  • Mullen, C. A.; Haugh, M. G.; Schaffler, M. B.
  • Journal of the Mechanical Behavior of Biomedical Materials, Vol. 28
  • DOI: 10.1016/j.jmbbm.2013.06.013

A calcium-induced signaling cascade leading to osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells
journal, April 2012


Mechanical properties of mineralized collagen fibrils as influenced by demineralization
journal, June 2008


The role of intracellular calcium phosphate in osteoblast-mediated bone apatite formation
journal, August 2012

  • Boonrungsiman, S.; Gentleman, E.; Carzaniga, R.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 35
  • DOI: 10.1073/pnas.1208916109

In vitro mineralization of dense collagen substrates: A biomimetic approach toward the development of bone-graft materials
journal, August 2011


Review: Mineralization of Synthetic Polymer Scaffolds for Bone Tissue Engineering
journal, May 2007


Micropatterning of bioactive self-assembling gels
journal, January 2009

  • Mata, Alvaro; Hsu, Lorraine; Capito, Ramille
  • Soft Matter, Vol. 5, Issue 6
  • DOI: 10.1039/b819002j

The material and biological characteristics of osteoinductive calcium phosphate ceramics
journal, September 2017

  • Tang, Zhurong; Li, Xiangfeng; Tan, Yanfei
  • Regenerative Biomaterials, Vol. 5, Issue 1
  • DOI: 10.1093/rb/rbx024

Degrading Devices: Invadosomes in Proteolytic Cell Invasion
journal, November 2011


Extracellular Matrix Mineralization Promotes E11/gp38 Glycoprotein Expression and Drives Osteocytic Differentiation
journal, May 2012


Self-Assembly and Mineralization of Peptide-Amphiphile Nanofibers
journal, November 2001

  • Hartgerink, Jeffrey D.; Beniash, Elia; Stupp, Samuel I.
  • Science, Vol. 294, Issue 5547, p. 1684-1688
  • DOI: 10.1126/science.1063187

The relationship between serum calcium and phosphate levels and cardiac valvular procedures in the hemodialysis population
journal, February 2003

  • Rubel, Jeffrey Rand; Milford, Edgar Louis
  • American Journal of Kidney Diseases, Vol. 41, Issue 2
  • DOI: 10.1053/ajkd.2003.50050

Intrafibrillar Collagen Mineralization Produced by Biomimetic Hierarchical Nanoapatite Assembly
journal, December 2010


The role of collagen in bone apatite formation in the presence of hydroxyapatite nucleation inhibitors
journal, October 2010

  • Nudelman, Fabio; Pieterse, Koen; George, Anne
  • Nature Materials, Vol. 9, Issue 12
  • DOI: 10.1038/nmat2875

Osteocytes as Dynamic Multifunctional Cells
journal, November 2007


A materials science vision of extracellular matrix mineralization
journal, June 2016


Biomimetic Mineralization of Biomaterials Using Simulated Body Fluids for Bone Tissue Engineering and Regenerative Medicine
journal, October 2017


Matrix Mineralization as a Trigger for Osteocyte Maturation
journal, December 2007

  • Irie, Kazuharu; Ejiri, Sadakazu; Sakakura, Yasunori
  • Journal of Histochemistry & Cytochemistry, Vol. 56, Issue 6
  • DOI: 10.1369/jhc.2008.950527

Enzyme Directed Templating of Artificial Bone Mineral
journal, January 2009

  • Spoerke, Erik D.; Anthony, Shawn G.; Stupp, Samuel I.
  • Advanced Materials, Vol. 21, Issue 4
  • DOI: 10.1002/adma.200802242

Nucleation of apatite crystals in vitro by self-assembled dentin matrix protein 1
journal, July 2003

  • He, Gen; Dahl, Tom; Veis, Arthur
  • Nature Materials, Vol. 2, Issue 8
  • DOI: 10.1038/nmat945

Bone structure and formation: A new perspective
journal, November 2007

  • Olszta, Matthew J.; Cheng, Xingguo; Jee, Sang Soo
  • Materials Science and Engineering: R: Reports, Vol. 58, Issue 3-5
  • DOI: 10.1016/j.mser.2007.05.001

Osteocyte RANKL: New insights into the control of bone remodeling
journal, February 2012

  • Xiong, Jinhu; O'Brien, Charles A.
  • Journal of Bone and Mineral Research, Vol. 27, Issue 3
  • DOI: 10.1002/jbmr.1547

The role of the amorphous phase on the biomimetic mineralization of collagen
journal, January 2012

  • Nudelman, Fabio; Bomans, Paul H. H.; George, Anne
  • Faraday Discussions, Vol. 159
  • DOI: 10.1039/c2fd20062g

Molecular mechanisms for intrafibrillar collagen mineralization in skeletal tissues
journal, January 2015


Cement lines of secondary osteons in human bone are not mineral-deficient: New data in a historical perspective
journal, January 2005

  • Skedros, John G.; Holmes, Jennifer L.; Vajda, Eric G.
  • The Anatomical Record Part A: Discoveries in Molecular, Cellular, and Evolutionary Biology, Vol. 286A, Issue 1
  • DOI: 10.1002/ar.a.20214

Cell-scaffold interactions in the bone tissue engineering triad
journal, September 2013

  • Murphy, Cm; O’Brien, Fj; Little, Dg
  • European Cells and Materials, Vol. 26
  • DOI: 10.22203/eCM.v026a09

Mineralization-driven bone tissue evolution follows from fluid-to-solid phase transformations in closed thermodynamic systems
journal, October 2013


Nano-mechanical properties of individual mineralized collagen fibrils from bone tissue
journal, October 2010

  • Hang, Fei; Barber, Asa H.
  • Journal of The Royal Society Interface, Vol. 8, Issue 57
  • DOI: 10.1098/rsif.2010.0413

Comparative materials differences revealed in engineered bone as a function of cell-specific differentiation
journal, July 2009

  • Gentleman, Eileen; Swain, Robin J.; Evans, Nicholas D.
  • Nature Materials, Vol. 8, Issue 9
  • DOI: 10.1038/nmat2505

The Role of Nanoscale Architecture in Supramolecular Templating of Biomimetic Hydroxyapatite Mineralization
journal, May 2012


Matrix-embedded cells control osteoclast formation
journal, September 2011

  • Xiong, Jinhu; Onal, Melda; Jilka, Robert L.
  • Nature Medicine, Vol. 17, Issue 10
  • DOI: 10.1038/nm.2448

Protein disorder–order interplay to guide the growth of hierarchical mineralized structures
journal, June 2018


Molecular mechanics of mineralized collagen fibrils in bone
journal, April 2013

  • Nair, Arun K.; Gautieri, Alfonso; Chang, Shu-Wei
  • Nature Communications, Vol. 4, Issue 1
  • DOI: 10.1038/ncomms2720

THE MATERIAL BONE: Structure-Mechanical Function Relations
journal, August 1998


The predominant role of collagen in the nucleation, growth, structure and orientation of bone apatite
journal, July 2012

  • Wang, Yan; Azaïs, Thierry; Robin, Marc
  • Nature Materials, Vol. 11, Issue 8
  • DOI: 10.1038/nmat3362

Surface Energetics of the Hydroxyapatite Nanocrystal–Water Interface: A Molecular Dynamics Study
journal, October 2014

  • Zhao, Weilong; Xu, Zhijun; Yang, Yang
  • Langmuir, Vol. 30, Issue 44
  • DOI: 10.1021/la503158p

Water-mediated structuring of bone apatite
journal, November 2013

  • Wang, Yan; Von Euw, Stanislas; Fernandes, Francisco M.
  • Nature Materials, Vol. 12, Issue 12
  • DOI: 10.1038/nmat3787

The amazing osteocyte
journal, January 2011

  • Bonewald, Lynda F.
  • Journal of Bone and Mineral Research, Vol. 26, Issue 2
  • DOI: 10.1002/jbmr.320

Designing materials to direct stem-cell fate
journal, November 2009

  • Lutolf, Matthias P.; Gilbert, Penney M.; Blau, Helen M.
  • Nature, Vol. 462, Issue 7272
  • DOI: 10.1038/nature08602

Tuning the Elastic Modulus of Hydrated Collagen Fibrils
journal, December 2009

  • Grant, Colin A.; Brockwell, David J.; Radford, Sheena E.
  • Biophysical Journal, Vol. 97, Issue 11
  • DOI: 10.1016/j.bpj.2009.09.010

25th Anniversary Article: Rational Design and Applications of Hydrogels in Regenerative Medicine
journal, November 2013

  • Annabi, Nasim; Tamayol, Ali; Uquillas, Jorge Alfredo
  • Advanced Materials, Vol. 26, Issue 1
  • DOI: 10.1002/adma.201303233

Matrix dimensionality and stiffness cooperatively regulate osteogenesis of mesenchymal stromal cells
journal, March 2016


Calcium phosphate-bearing matrices induce osteogenic differentiation of stem cells through adenosine signaling
journal, January 2014

  • Shih, Y. -R. V.; Hwang, Y.; Phadke, A.
  • Proceedings of the National Academy of Sciences, Vol. 111, Issue 3
  • DOI: 10.1073/pnas.1321717111

Biomaterials & scaffolds for tissue engineering
journal, March 2011


Receptor Activator of Nuclear Factor κB Ligand (RANKL)/Osteoprotegerin (OPG) Ratio Is Increased in Severe Osteolysis
journal, November 2003


Organoapatites: Materials for artificial bone. II. Hardening reactions and properties
journal, March 1993

  • Stupp, Samuel I.; Mejicano, George C.; Hanson, Jacqueline A.
  • Journal of Biomedical Materials Research, Vol. 27, Issue 3
  • DOI: 10.1002/jbm.820270303

Biomineralization of bone: a fresh view of the roles of non-collagenous proteins
journal, January 2011

  • Gorski, Jeffrey Paul
  • Frontiers in Bioscience, Vol. 16, Issue 1
  • DOI: 10.2741/3875

Dentin Matrix Protein 1 Is Predominantly Expressed in Chicken and Rat Osteocytes But Not in Osteoblasts
journal, November 2001


Mapping amorphous calcium phosphate transformation into crystalline mineral from the cell to the bone in zebrafish fin rays
journal, March 2010

  • Mahamid, J.; Aichmayer, B.; Shimoni, E.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 14
  • DOI: 10.1073/pnas.0914218107

Osteogenic Differentiation of MSC through Calcium Signaling Activation: Transcriptomics and Functional Analysis
journal, February 2016


Multifunctional role of osteopontin in directing intrafibrillar mineralization of collagen and activation of osteoclasts
journal, January 2014