The Broad Aim of this proposal is to advance the field of Bone Tissue Engineering by defining general principles and techniques that can be applied to the rapid harvest and intra- operative transplantation of bone marrow derived connective tissue progenitors (CTPs). Work in the initial funding period has defined methods by which CTPs can be rapidly concentrated and positively selected from human bone marrow using an implantable matrix as a surface for selective cell attachment. We have also shown that composite cellular grafts prepared in this way significantly improve the efficacy of bone grafting procedures. This rapid and simple method for concentration and delivery of CTPs offers promise to improve many clinical grafting procedures and other tissue engineering applications through controlled preparation and delivery of cellular composite grafts. Realizing this promise requires a improved understanding and control over the interaction between cells matrix surfaces and the metabolic environment created within a cellular graft. Experimental control over surface properties modulating CTP attachment and selection, as well as survival, proliferation and differentiation is obtained using a molecular surface design approach. A set of small peptide ligands have been strategically selected from the literature based on evidence of interaction with CTPs. These peptides are be presented on a chemically stable but degradable substrate, polycaprolactone(PCL), and screened for specific dose dependent effects on these biologic parameters. Because hypoxia at the site of implantation will have profound effects on the biologic performance of transplanted cells, and the effects of ligands are expected to modulate cellular response to hypoxia, the effect of hypoxia on CTP survival and biologic performance is assessed. The selective attachment of CTPs and other subsets of marrow cells are evaluated in 3D porous matrices providing precise control over ligand presentation, structure and porosity. Selected cellular 3D composite grafts will then be evaluated in vivo to refine effective strategies for optimizing graft efficacy, through control of CTP concentration, cell density, cell composition, and matrix porosity. Quantitative in vivo assessment is made of oxygen tension, metabolic demand, revascularization in the graft site. Quantitative microCT and histology are used to assess bone and tissue formation. Using this unique combination of technologies, we will define fundamental biologic principles. related to cell-matrix interaction, matrix surface chemistry, matrix architecture and effects of local hypoxia that can be applies to the design of cellular implants containing marrow derived cells in a broad range of materials and tissue engineering applications.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Research Project (R01)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-SSS-M (01))
Program Officer
Panagis, James S
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Cleveland Clinic Lerner
United States
Zip Code
Kim, Eun Jung; Fleischman, Aaron J; Muschler, George F et al. (2013) Response of bone marrow derived connective tissue progenitor cell morphology and proliferation on geometrically modulated microtextured substrates. Biomed Microdevices 15:385-96
Kim, Eun Jung; Boehm, Cynthia A; Mata, Alvaro et al. (2010) Post microtextures accelerate cell proliferation and osteogenesis. Acta Biomater 6:160-9
Marcantonio, Nicholas A; Boehm, Cynthia A; Rozic, Richard J et al. (2009) The influence of tethered epidermal growth factor on connective tissue progenitor colony formation. Biomaterials 30:4629-38
Kim, Eun Jung; Boehm, Cynthia A; Fleischman, Aaron J et al. (2009) Modulating human connective tissue progenitor cell behavior on cellulose acetate scaffolds by surface microtextures. J Biomed Mater Res A 90:1198-205
Mata, Alvaro; Kim, Eun Jung; Boehm, Cynthia A et al. (2009) A three-dimensional scaffold with precise micro-architecture and surface micro-textures. Biomaterials 30:4610-7
Villarruel, Sandra M; Boehm, Cynthia A; Pennington, Mark et al. (2008) The effect of oxygen tension on the in vitro assay of human osteoblastic connective tissue progenitor cells. J Orthop Res 26:1390-7
Au, Ada; Boehm, Cynthia A; Mayes, Anne M et al. (2007) Formation of osteogenic colonies on well-defined adhesion peptides by freshly isolated human marrow cells. Biomaterials 28:1847-61
Fan, Vivian H; Tamama, Kenichi; Au, Ada et al. (2007) Tethered epidermal growth factor provides a survival advantage to mesenchymal stem cells. Stem Cells 25:1241-51
Powell, Kimerly A; Nakamoto, Chizu; Villarruel, Sandra et al. (2007) Quantitative image analysis of connective tissue progenitors. Anal Quant Cytol Histol 29:112-21
Takigami, Hidetake; Kumagai, Ken; Latson, Larry et al. (2007) Bone formation following OP-1 implantation is improved by addition of autogenous bone marrow cells in a canine femur defect model. J Orthop Res 25:1333-42

Showing the most recent 10 out of 20 publications