Functional tissue engineering (FTE) seeks to enhance tissue engineered (TE) repairs using in vivo loads and strains from normal tissues to condition the constructs to their mechanical environment prior to surgery. FTE is particularly relevant for highly loaded tissues such as menisci in the human knee. In this novel application, we will develop a real-time assessment technology to shorten the development cycle for creating TE products. We will screen, in real time, how mechanical stimuli affect gene expression. Using specialized murine mesenchymal stem cells in a mechanical stimulation system, we will address the following aims:
Aim 1 : Create a source of murine MSCs having dual, mutually exclusive colorimetric indicators for fibroblastic and chondrocytic programs. Expression genes that combine human type I collagen promoter (yellow fluorescent protein-YFP) and cartilage-specific promoter element of CD-RAP (cyan fluorescent protein-CFP) will be constructed and injected separately into mouse blastocysts. Separate transgenic lines will be mated, and MSCs will be harvested from doubly transgenic animals that induce chondrocytic differentiation or fibroblast phenotype.
Aim 2 """""""" Develop test platforms that control mechanical signals delivered to 3-D cell-seeded constructs and that provide real-time feedback of biological and mechanical events. Finalize designs of advanced, multi-specimen testing systems to apply precisely calibrated compressive and/or tensile strains to cell-seeded specimens and that enable real-time recordings of colorimetric changes within them.
Aim 3 """""""" Evaluate gene expression for murine MSCs in translucent gels under controlled strain states. After inducing MSCs to chondrocytic phenotype, test the hypotheses that application of: 1) compressive strains to cell-gel constructs will maintain CFP indicative of a cartilage-specific promoter element, 2) tensile strains will induce dedifferentiation and transition from a cyan fluorescent chondrocyte to a yellow fluorescent fibroblast, and 3) tensile strains to MSC-gel constructs already exposed to compressive strains and already expressing CFP will yield dedifferentiation to fibroblasts resulting in a mixed composition of tissue components and colors. This technology can dramatically reduce time/expense during the development phase of TE fibrocartilage.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21EB002361-01
Application #
6726243
Study Section
Special Emphasis Panel (ZRG1-SSS-M (58))
Program Officer
Kelley, Christine A
Project Start
2003-09-19
Project End
2005-08-31
Budget Start
2003-09-19
Budget End
2004-08-31
Support Year
1
Fiscal Year
2003
Total Cost
$230,250
Indirect Cost
Name
University of Cincinnati
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
041064767
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Butler, David L; Dyment, Nathaniel A; Shearn, Jason T et al. (2013) Evolving strategies in mechanobiology to more effectively treat damaged musculoskeletal tissues. J Biomech Eng 135:020301
Maye, Peter; Fu, Yu; Butler, David L et al. (2011) Generation and characterization of Col10a1-mcherry reporter mice. Genesis 49:410-8
Butler, David L; Gooch, Cynthia; Kinneberg, Kirsten R C et al. (2010) The use of mesenchymal stem cells in collagen-based scaffolds for tissue-engineered repair of tendons. Nat Protoc 5:849-63
Chokalingam, Kumar; Hunter, Shawn; Gooch, Cynthia et al. (2009) Three-dimensional in vitro effects of compression and time in culture on aggregate modulus and on gene expression and protein content of collagen type II in murine chondrocytes. Tissue Eng Part A 15:2807-16
Chokalingam, Kumar; Juncosa-Melvin, Natalia; Hunter, Shawn A et al. (2009) Tensile stimulation of murine stem cell-collagen sponge constructs increases collagen type I gene expression and linear stiffness. Tissue Eng Part A 15:2561-70
Butler, David L; Hunter, Shawn A; Chokalingam, Kumar et al. (2009) Using functional tissue engineering and bioreactors to mechanically stimulate tissue-engineered constructs. Tissue Eng Part A 15:741-9
Ingber, Donald E; Mow, Van C; Butler, David et al. (2006) Tissue engineering and developmental biology: going biomimetic. Tissue Eng 12:3265-83
Hunter, Shawn A; Noyes, Frank R; Haridas, Balakrishna et al. (2005) Meniscal material properties are minimally affected by matrix stabilization using glutaraldehyde and glycation with ribose. J Orthop Res 23:555-61
Butler, David L; Shearn, Jason T; Juncosa, Natalia et al. (2004) Functional tissue engineering parameters toward designing repair and replacement strategies. Clin Orthop Relat Res :S190-9