Musculoskeletal tissue injuries remain a significant challenge in orthopaedics research. For example, currently, millions of patients are suffering from cartilage injuries, with associated annual financial costs of more than $100 billion dollars. There are several viable clinical options to address these injuries. In this context, human mesenchymal stem cells (hMSCs) are a promising cell source for cartilage tissue engineering as they are capable of differentiating down the chondrogenic pathway, can be obtained from bone marrow in a minimally invasive manner, and are easily grown in culture. Although differentiation of hMSCs is regulated by soluble molecules, insoluble biochemical signals and mechanical cues, the combinatorial effects of mechanical loading and biomaterial signals are largely unknown. Our central hypothesis is that the use of high-throughput systems (HTSs) under mechanical stimulation can be used to elucidate single and synergistic microenvironmental factors for directing the chondrogenic differentiation of hMSCs, and thereby functional engineered cartilage formation in vitro and in a clinically relevant in vivo model. The information obtained from the HTS will help in the development of macroscale constructs with enhanced chondrogenesis, and the performance of these constructs will be validated in vivo by treating critical-sized articular cartilage defects. These goals will be accomplished by achieving the following specific Aims: (1) to develop three dimensional (3D) combinatorial HTS hydrogel-based microarrays, consisting of different extracellular matrix molecules and growth factors, which can be mechanically deformed to mimic the chondrogenic microenvironment of hMSCs, (2) to evaluate quantitatively the chondrogenic differentiation response of hMSCs in 3D combinatorial HTS microarrays and macroscale constructs selected from these microarrays, and (3) to determine the potential of hMSC-laden constructs with HTS-identified compositions and mechanical stimulation regimes to induce cartilage regeneration in vivo. The orthopedic community would benefit from a better understanding of these chondroinductive microenvironments that will ultimately induce neo-tissue formation and will represent a viable alternative to current clinical therapies. While the ultimate objective of this research is to engineer clinically relevant articular cartilage therapies, this HTS can also be applicable to test regeneration strategies for other tissues.

Public Health Relevance

Articular cartilage injuries remain a significant challenge in orthopaedics, as there are few viable treatment options and they affect more than 21 million patients each year in the U.S. In this research proposal, a high- throughput system under mechanical stimulation will be engineered and applied to identify biomaterial microenvironments and soluble growth factor signaling combinations that enhance chondrogenic differentiation of human mesenchymal stem cells and can be used to repair in vivo chondral defects with functional cartilage tissue. The results from this proposal will have great potential to positively impact the large number of patients who undergo cartilage replacement and repair procedures, and this platform high-throughput system will permit rapid future examination of the signals that induce the formation of different types of tissue for other clinically relevant problems.

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)
Program Officer
Wang, Fei
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Case Western Reserve University
Biomedical Engineering
Biomed Engr/Col Engr/Engr Sta
United States
Zip Code
Nguyen, Minh K; Jeon, Oju; Dang, Phuong N et al. (2018) RNA interfering molecule delivery from in situ forming biodegradable hydrogels for enhancement of bone formation in rat calvarial bone defects. Acta Biomater 75:105-114
Miri, Amir K; Nieto, Daniel; Iglesias, Luis et al. (2018) Microfluidics-Enabled Multimaterial Maskless Stereolithographic Bioprinting. Adv Mater 30:e1800242
Miri, Amir K; Khalilpour, Akbar; Cecen, Berivan et al. (2018) Multiscale bioprinting of vascularized models. Biomaterials :
Liu, Wanjun; Zhong, Zhe; Hu, Ning et al. (2018) Coaxial extrusion bioprinting of 3D microfibrous constructs with cell-favorable gelatin methacryloyl microenvironments. Biofabrication 10:024102
Seo, Jungmok; Shin, Jung-Youn; Leijten, Jeroen et al. (2018) High-throughput approaches for screening and analysis of cell behaviors. Biomaterials 153:85-101
Alarçin, Emine; Lee, Tae Yong; Karuthedom, Sobha et al. (2018) Injectable shear-thinning hydrogels for delivering osteogenic and angiogenic cells and growth factors. Biomater Sci 6:1604-1615
McMillan, Alexandra; Nguyen, Minh Khanh; Gonzalez-Fernandez, Tomas et al. (2018) Dual non-viral gene delivery from microparticles within 3D high-density stem cell constructs for enhanced bone tissue engineering. Biomaterials 161:240-255
Lobo, Anderson Oliveira; Afewerki, Samson; de Paula, Mirian Michele Machado et al. (2018) Electrospun nanofiber blend with improved mechanical and biological performance. Int J Nanomedicine 13:7891-7903
Jeon, Oju; Lee, Keewon; Alsberg, Eben (2018) Spatial Micropatterning of Growth Factors in 3D Hydrogels for Location-Specific Regulation of Cellular Behaviors. Small 14:e1800579
Sheikhi, Amir; Afewerki, Samson; Oklu, Rahmi et al. (2018) Effect of ionic strength on shear-thinning nanoclay-polymer composite hydrogels. Biomater Sci 6:2073-2083

Showing the most recent 10 out of 30 publications