When endogenous repair fails, as is often the case with dense fibrous musculoskeletal tissues (like the knee meniscus), novel strategies and enabling technologies must be developed to enhance tissue regeneration. While numerous tissue engineering therapies have been developed for the engineering of fiber-reinforced tissues, one persistent limitation is the ability to fabricate scaffolds that maintain structural integrity and direct appropriate tissue architecture, while simultaneously promoting cellular infiltration throughout the regeneration period. In this proposal, we address these limitations with the production of a novel multi-polymer composite nanofibrous scaffold that provides a 3-dimensional micropattern for neo-tissue formation. These scaffolds, with fiber diameters on the order of the native extracellular matrix, can be produced with defined fiber anisotropies that mimic the structural arrangement of fiber-reinforced tissues. Further, by introducing flexibility in polymer properties via a library of photocrosslinkable macromers that exhibit a range of mechanical and degradation properties when polymerized, we propose to tailor temporal pore formation within the scaffold through the controlled degradation of individual polymer components. We hypothesize that these nanofibrous multipolymer photocrosslinked meshes will have controlled mechanical properties reflective of the stiffest and slowest degrading component and show a time dependent increase in void space while maintaining their overall mechanical properties. Further, we hypothesize that the controlled increase in void space within these scaffolds, via the erosion of sacrificial elements, will promote cellular infiltration into the multi-polymer mesh and will result in a more uniform functional tissue structure. Additionally, as the mechanical environment of the meniscus is paramount in its maturation and homeostasis, we hypothesize that tailored mechanical preconditioning regimens will likewise promote functional maturation of infiltrated meniscus constructs. To this end, a novel dynamic loading bioreactor that applies compression and tension is developed to promote tissue maturation.
The first Aim of this proposal is to develop technology to electrospin multi-component nanofibrous scaffolds and compare properties to the native tissue using a predictive fiber-reinforced composite model. In the second Aim, the interaction and infiltration of cells and ultimate mechanical properties will be explored in single- and tri-polymer cell-laden nanofibrous scaffolds.
The third Aim i nvolves the development of a bioreactor for pre-conditioning scaffolds and evaluating the impact of mechanical stimulation on tissue formation in the short and long term. If successful, this innovative approach will provide several new enabling technologies for the functional regeneration of damaged fiber-reinforced musculoskeletal tissues. Public Health Relevance Statement (provided by applicant): This project develops a novel multi-polymer nanofiber fabrication system to exert control over polymer chemistry and overall scaffold mechanics and degradation with time to improve cellular infiltration and uniform tissue deposition in fibrous tissue-engineered constructs. If successful, this approach would surmount a major hurdle in the tissue engineering of dense structures of the musculoskeletal system and provide a mechanically functional, structurally anisotropic 3D micro-pattern for directed neo-tissue formation while promoting full cellular colonization and eventual replacement of the polymer structure after complete dissolution. This innovative approach, coupling scaffold fabrication, mechanical loading, and in vivo assessments, will aid in the development of tissue engineered therapies for fiber-reinforced musculoskeletal tissues such as the knee meniscus that otherwise fail to heal and have few clinically viable repair strategies.

Public Health Relevance

This project develops a novel multi-polymer nanofiber fabrication system to exert control over polymer chemistry and overall scaffold mechanics and degradation with time to improve cellular infiltration and uniform tissue deposition in fibrous tissue-engineered constructs. If successful, this approach would surmount a major hurdle in the tissue engineering of dense structures of the musculoskeletal system and provide a mechanically functional, structurally anisotropic 3D micro-pattern for directed neo-tissue formation while promoting full cellular colonization and eventual replacement of the polymer structure after complete dissolution. This innovative approach, coupling scaffold fabrication, mechanical loading, and in vivo assessments, will aid in the development of tissue engineered therapies for fiber-reinforced musculoskeletal tissues such as the knee meniscus that otherwise fail to heal and have few clinically viable repair strategies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR056624-02
Application #
7808890
Study Section
Special Emphasis Panel (ZEB1-OSR-D (J1))
Program Officer
Wang, Fei
Project Start
2009-04-20
Project End
2014-03-31
Budget Start
2010-04-01
Budget End
2011-03-31
Support Year
2
Fiscal Year
2010
Total Cost
$334,184
Indirect Cost
Name
University of Pennsylvania
Department
Orthopedics
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
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Qu, Feini; Holloway, Julianne L; Esterhai, John L et al. (2017) Programmed biomolecule delivery to enable and direct cell migration for connective tissue repair. Nat Commun 8:1780
Heo, Su-Jin; Szczesny, Spencer E; Kim, Dong Hwa et al. (2017) Expansion of mesenchymal stem cells on electrospun scaffolds maintains stemness, mechano-responsivity, and differentiation potential. J Orthop Res :
Cao, Xuan; Ban, Ehsan; Baker, Brendon M et al. (2017) Multiscale model predicts increasing focal adhesion size with decreasing stiffness in fibrous matrices. Proc Natl Acad Sci U S A 114:E4549-E4555
Bansal, Sonia; Keah, Niobra M; Neuwirth, Alexander L et al. (2017) Large Animal Models of Meniscus Repair and Regeneration: A Systematic Review of the State of the Field. Tissue Eng Part C Methods 23:661-672
Szczesny, Spencer E; Mauck, Robert L (2017) The Nuclear Option: Evidence Implicating the Cell Nucleus in Mechanotransduction. J Biomech Eng 139:
Bansal, Sonia; Mandalapu, Sai; Aeppli, CĂ©line et al. (2017) Mechanical function near defects in an aligned nanofiber composite is preserved by inclusion of disorganized layers: Insight into meniscus structure and function. Acta Biomater 56:102-109
Cucchiarini, M; McNulty, A L; Mauck, R L et al. (2016) Advances in combining gene therapy with cell and tissue engineering-based approaches to enhance healing of the meniscus. Osteoarthritis Cartilage 24:1330-9
Heo, Su-Jin; Driscoll, Tristan P; Thorpe, Stephen D et al. (2016) Differentiation alters stem cell nuclear architecture, mechanics, and mechano-sensitivity. Elife 5:
Heo, Su-Jin; Han, Woojin M; Szczesny, Spencer E et al. (2016) Mechanically Induced Chromatin Condensation Requires Cellular Contractility in Mesenchymal Stem Cells. Biophys J 111:864-874

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