The self-assembling process in articular cartilage is emerging as a potentially robust approach for engineering large and small cartilage constructs. The objective of this proposal is to evaluate self-assembled articular cartilage in resurfacing the patella by quantifying the biologic response (i.e., host response to the construct), dose response (i.e., identifying the minimum number of necessary cells), and durability (i.e., stability, integriy, and maturation over a year), as defined by the FDA. Statistical optimization to improve the functionality of self- assembled cartilage constructs has yielded a powerful combination of externally applied stimuli that result in constructs with biomechanical and biochemical properties on par with those of native cartilage. Among a multitude of helpful stimuli, three have emerged as quite potent: hydrostatic pressure (10MPa at 0Hz during days 10-14), TGF-1 (30 ng/ml for 2 wks), and chondroitinase ABC (applied at 2 wks), applied in combination. Mechanisms linking these stimuli to the engineered tissues' biomechanical properties have also been elucidated to explain their synergisms and to consolidate them into simple culture protocols. A functionality index (FI) allowing the establishment of quantitative success criteria and validated for the comparison of constructs to native tissue showed that construct properties have attained FI values approaching 1, the value of native tissue. Based on these promising results and additional in vivo murine, leporine, and ovine data, this proposal will investigate the global hypothesis that constructs will show durability without an adverse host response via three aims: The objective of Aim 1 is to use a short-term (12wks), leporine patella resurfacing model to examine the hypotheses that: 1) not only will constructs retain stability and integrity in vivo, thir FI values will be improved by implantation, and 2) neither the allogeneic cells nor the culture products will elicit adverse host responses (local/systemic).
Aim 2 employs the murine model to validate that implant scale-up would not alter neocartilage biomechanical properties.
Aim 2 will also address certain challenges that are common across diverse cartilage regeneration strategies, namely initial fixation, subsequent integration, and durability against wear; these issues will be tackled using a chondroconductive glue, exogenous lysyl oxidase, and the chondrotuning method that yields robust and lubricious implants. Finally, Aim 3 will test the hypothesis that durable healing can be achieved for up to 12 months in an ovine model.
This aim will also identify a minimum cell number that can be employed to achieve effective healing at 1 year. By following FDA's guidance document (Preparation of IDEs and INDs for products intended to repair or replace knee cartilage), and if the proposed study's hypotheses are proven, the results will provide exciting validation of the clinical translatability of self-assembed articular cartilage constructs.

Public Health Relevance

Current treatment options for cartilage injury such as microfracture, osteochondral grafts, and autologous chondrocyte implantation do not effectively regenerate articular cartilage. This proposal seeks to conduct a series of in vivo studies, informed by FDA guidelines, to advance cartilage tissue engineering toward regeneration of large defects on articular surfaces. Core innovations include 1) developing biomimetic cartilage, engineered via a self-assembling process, for articular cartilage resurfacing of the patella to address the patellofemoral indication, 2) incorporating small and large animal models, 3) employing potent stimuli to not only render large neocartilage constructs mechanically robust but also exhibiting ability to integrate within defects, and 4) basing studies on FDA guidance recommendations. Conducted under Good Laboratory Practice (GLP), the successful completion of this proposal will lay a solid foundation for subsequent clinical studies.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Research Project (R01)
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Musculoskeletal Tissue Engineering Study Section (MTE)
Program Officer
Wang, Fei
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University of California Irvine
Biomedical Engineering
Biomed Engr/Col Engr/Engr Sta
United States
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Huwe, Le W; Brown, Wendy E; Hu, Jerry C et al. (2018) Characterization of costal cartilage and its suitability as a cell source for articular cartilage tissue engineering. J Tissue Eng Regen Med 12:1163-1176
Huwe, Le W; Sullan, Gurdeep K; Hu, Jerry C et al. (2018) Using Costal Chondrocytes to Engineer Articular Cartilage with Applications of Passive Axial Compression and Bioactive Stimuli. Tissue Eng Part A 24:516-526
Lee, Jennifer K; Link, Jarrett M; Hu, Jerry C Y et al. (2017) The Self-Assembling Process and Applications in Tissue Engineering. Cold Spring Harb Perspect Med 7:
Cissell, Derek D; Link, Jarrett M; Hu, Jerry C et al. (2017) A Modified Hydroxyproline Assay Based on Hydrochloric Acid in Ehrlich's Solution Accurately Measures Tissue Collagen Content. Tissue Eng Part C Methods 23:243-250
Hadidi, Pasha; Cissell, Derek D; Hu, Jerry C et al. (2017) Temporal development of near-native functional properties and correlations with qMRI in self-assembling fibrocartilage treated with exogenous lysyl oxidase homolog 2. Acta Biomater 64:29-40
Lee, Jennifer K; Huwe, Le W; Paschos, Nikolaos et al. (2017) Tension stimulation drives tissue formation in scaffold-free systems. Nat Mater 16:864-873
Huang, Brian J; Hu, Jerry C; Athanasiou, Kyriacos A (2016) Effects of passage number and post-expansion aggregate culture on tissue engineered, self-assembled neocartilage. Acta Biomater 43:150-159
Huang, Brian J; Hu, Jerry C; Athanasiou, Kyriacos A (2016) Cell-based tissue engineering strategies used in the clinical repair of articular cartilage. Biomaterials 98:1-22
Kwon, Heenam; Paschos, Nikolaos K; Hu, Jerry C et al. (2016) Articular cartilage tissue engineering: the role of signaling molecules. Cell Mol Life Sci 73:1173-94
Brown, Wendy E; Hu, Jerry C; Athanasiou, Kyriacos A (2016) Ammonium-Chloride-Potassium Lysing Buffer Treatment of Fully Differentiated Cells Increases Cell Purity and Resulting Neotissue Functional Properties. Tissue Eng Part C Methods 22:895-903

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