Publications by authors named "Clark T Hung"

133 Publications

Attachment of cartilage wear particles to the synovium negatively impacts friction properties.

J Biomech 2021 10 4;127:110668. Epub 2021 Aug 4.

Columbia University, Department of Biomedical Engineering, New York, NY, United States; Columbia University, Department of Orthopedic Surgery, New York, NY, United States. Electronic address:

Cartilage wear particles are released into the synovial fluid by mechanical and chemical degradation of the articular surfaces during osteoarthritis and attach to the synovial membrane. Accumulation of wear particles could alter key tissue-level mechanical properties of the synovium, hindering its characteristically low-friction interactions with underlying articular surfaces in the synovial joint. The present study employs a custom loading device to further the characterization of native synovium friction properties, while investigating the hypothesis that attachment of cartilage wear particles increases friction coefficient. Juvenile bovine synovium demonstrated characteristically low friction coefficients in sliding contact with glass, in agreement with historical measurements. Friction coefficient increased with higher normal load in saline, while lubrication with native synovial fluid maintained low friction coefficients at higher loads. Cartilage wear particles generated from juvenile bovine cartilage attached directly to synovium explants in static culture, with incorporation onto the tissue denoted by cell migration onto the particle surface. In dilute synovial fluid mimicking the decreased lubricating properties during osteoarthritis, wear particle attachment significantly increased friction coefficient against glass, and native cartilage and synovium. In addition to providing a novel characterization of synovial joint tribology this work highlights a potential mechanism for cartilage wear particles to perpetuate the degradative environment of osteoarthritis by modulating tissue-level properties of the synovium that could impact macroscopic wear as well as mechanical stimuli transmitted to resident cells.
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http://dx.doi.org/10.1016/j.jbiomech.2021.110668DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8492547PMC
October 2021

Sustained Delivery of SB-431542, a Type I Transforming Growth Factor Beta-1 Receptor Inhibitor, to Prevent Arthrofibrosis.

Tissue Eng Part A 2021 11 12;27(21-22):1411-1421. Epub 2021 May 12.

Department of Biomedical Engineering, Columbia University, New York, New York, USA.

Fibrosis of the knee is a common disorder resulting from an aberrant wound healing response and is characterized by extracellular matrix deposition, joint contraction, and scar tissue formation. The principal regulator of the fibrotic cascade is transforming growth factor beta-1 (TGF-β1), a factor that induces rapid proliferation and differentiation of resident fibroblasts. In this study, we demonstrate successful inhibition of TGF-β1-driven myofibroblastic differentiation in human fibroblast-like synoviocytes using a small molecule TGF-β1 receptor inhibitor, SB-431542. We also demonstrate successful encapsulation of SB-431542 in poly(D,L-lactide-co-glycolide) (PLGA) as a potential prophylactic treatment for arthrofibrosis and characterize drug release and bioactivity in a three-dimensional collagen gel contraction assay. We assessed the effects of TGF-β1 and SB-431542 on cell proliferation and viability in monolayer cultures. Opposing dose-dependent trends were observed in cell proliferation, which increased in TGF-β1-treated cultures and decreased in SB-431542-treated cultures relative to control ( < 0.05). SB-431542 was not cytotoxic at the concentrations studied (0-50 μM) and inhibited TGF-β1-induced collagen gel contraction in a dose-dependent manner. Specifically, TGF-β1-treated gels contracted to 18% ± 1% of their initial surface area, while gels treated with TGF-β1 and ≥10 μM SB-431542 showed no evidence of contraction ( < 0.0001). Upon removal of the compound, all gels contracted to control levels after 44 h in culture, necessitating sustained delivery for prolonged inhibition. To this end, SB-431542 was encapsulated in PLGA microspheres (SBMS) that had an average diameter of 87.5 ± 24 μm and a loading capacity of 4.3 μg SB-431542 per milligram of SBMS. Functional assessment of SBMS revealed sustained inhibition of TGF-β1-induced gel contraction as well as hallmark features of myofibroblastic differentiation, including α-smooth muscle actin expression and connective tissue growth factor production. These results suggest that SB-431542 may be used to counter TGF-β1-driven events in the fibrotic cascade in the knee cartilage. Impact statement Arthrofibrosis is the most prevalent comorbidity resulting from orthopedic procedures such as total knee arthroplasty that is characterized by excess deposition and accumulation of extracellular matrix. Despite its prevalence, treatments are generally palliative, and there is no effective prophylactic therapy. We report that the small molecule transforming growth factor beta-1 (TGF-β1) receptor inhibitor, SB-431542, can inhibit the TGF-β1-driven myofibroblastic differentiation of fibroblast-like synoviocytes. To provide sustained inhibition, we explored the use of SB-laden microspheres as a prophylactic therapy in a three-dimensional contraction model of fibrosis and propose that such therapies will have the potential to improve the standard of care for arthrofibrosis.
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http://dx.doi.org/10.1089/ten.TEA.2021.0029DOI Listing
November 2021

Direct Osmotic Pressure Measurements in Articular Cartilage Demonstrate Nonideal and Concentration-Dependent Phenomena.

J Biomech Eng 2021 04;143(4)

Department of Mechanical Engineering, Columbia University, New York, NY 10027; Department of Biomedical Engineering, Columbia University, New York, NY 10027.

The osmotic pressure in articular cartilage serves an important mechanical function in healthy tissue. Its magnitude is thought to play a role in advancing osteoarthritis. The aims of this study were to: (1) isolate and quantify the magnitude of cartilage swelling pressure in situ; and (2) identify the effect of salt concentration on material parameters. Confined compression stress-relaxation testing was performed on 18 immature bovine and six mature human cartilage samples in solutions of varying osmolarities. Direct measurements of osmotic pressure revealed nonideal and concentration-dependent osmotic behavior, with magnitudes approximately 1/3 those predicted by ideal Donnan law. A modified Donnan constitutive behavior was able to capture the aggregate behavior of all samples with a single adjustable parameter. Results of curve-fitting transient stress-relaxation data with triphasic theory in febio demonstrated concentration-dependent material properties. The aggregate modulus HA increased threefold as the external concentration decreased from hypertonic 2 M to hypotonic 0.001 M NaCl (bovine: HA=0.420±0.109 MPa to 1.266±0.438 MPa; human: HA=0.499±0.208 MPa to 1.597±0.455 MPa), within a triphasic theory inclusive of osmotic effects. This study provides a novel and simple analytical model for cartilage osmotic pressure which may be used in computational simulations, validated with direct in situ measurements. A key finding is the simultaneous existence of Donnan osmotic and Poisson-Boltzmann electrostatic interactions within cartilage.
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http://dx.doi.org/10.1115/1.4049158DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7872001PMC
April 2021

Immature bovine cartilage wear by fatigue failure and delamination.

J Biomech 2020 06 19;107:109852. Epub 2020 May 19.

Department of Biomedical Engineering, Columbia University, New York, NY, USA; Department of Mechanical Engineering, Columbia University, New York, NY, USA. Electronic address:

This study investigated wear damage of immature bovine articular cartilage using reciprocal sliding of tibial cartilage strips against glass or cartilage. Experiments were conducted in physiological buffered saline (PBS) or mature bovine synovial fluid (SF). A total of 63 samples were tested, of which 47 exhibited wear damage due to delamination of the cartilage surface initiated in the middle zone, with no evidence of abrasive wear. There was no difference between the friction coefficient of damaged and undamaged samples, showing that delamination wear occurs even when friction remains low under a migrating contact area configuration. No difference was observed in the onset of damage or in the friction coefficient between samples tested in PBS or SF. The onset of damage occurred earlier when testing cartilage against glass versus cartilage against cartilage, supporting the hypothesis that delamination occurs due to fatigue failure of the collagen in the middle zone, since stiffer glass produces higher strains and tensile stresses under comparable loads. The findings of this study are novel because they establish that delamination of the articular surface, starting in the middle zone, may represent a primary mechanism of failure. Based on preliminary data, it is reasonable to hypothesize that delamination wear via subsurface fatigue failure is similarly the primary mechanism of human cartilage wear under normal loading conditions, albeit requiring far more cycles of loading than in immature bovine cartilage.
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http://dx.doi.org/10.1016/j.jbiomech.2020.109852DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7809915PMC
June 2020

Pulsed electromagnetic fields promote repair of focal articular cartilage defects with engineered osteochondral constructs.

Biotechnol Bioeng 2020 05 5;117(5):1584-1596. Epub 2020 Feb 5.

Department of Biomedical Engineering, Columbia University, New York, NY.

Articular cartilage injuries are a common source of joint pain and dysfunction. We hypothesized that pulsed electromagnetic fields (PEMFs) would improve growth and healing of tissue-engineered cartilage grafts in a direction-dependent manner. PEMF stimulation of engineered cartilage constructs was first evaluated in vitro using passaged adult canine chondrocytes embedded in an agarose hydrogel scaffold. PEMF coils oriented parallel to the articular surface induced superior repair stiffness compared to both perpendicular PEMF (p = .026) and control (p = .012). This was correlated with increased glycosaminoglycan deposition in both parallel and perpendicular PEMF orientations compared to control (p = .010 and .028, respectively). Following in vitro optimization, the potential clinical translation of PEMF was evaluated in a preliminary in vivo preclinical adult canine model. Engineered osteochondral constructs (∅ 6 mm × 6 mm thick, devitalized bone base) were cultured to maturity and implanted into focal defects created in the stifle (knee) joint. To assess expedited early repair, animals were assessed after a 3-month recovery period, with microfracture repairs serving as an additional clinical control. In vivo, PEMF led to a greater likelihood of normal chondrocyte (odds ratio [OR]: 2.5, p = .051) and proteoglycan (OR: 5.0, p = .013) histological scores in engineered constructs. Interestingly, engineered constructs outperformed microfracture in clinical scoring, regardless of PEMF treatment (p < .05). Overall, the studies provided evidence that PEMF stimulation enhanced engineered cartilage growth and repair, demonstrating a potential low-cost, low-risk, noninvasive treatment modality for expediting early cartilage repair.
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http://dx.doi.org/10.1002/bit.27287DOI Listing
May 2020

Sustained low-dose dexamethasone delivery via a PLGA microsphere-embedded agarose implant for enhanced osteochondral repair.

Acta Biomater 2020 01 2;102:326-340. Epub 2019 Dec 2.

Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 1210 Amsterdam Avenue, New York 10027, NY United States. Electronic address:

Articular cartilage defects are a common source of joint pain and dysfunction. We hypothesized that sustained low-dose dexamethasone (DEX) delivery via an acellular osteochondral implant would have a dual pro-anabolic and anti-catabolic effect, both supporting the functional integrity of adjacent graft and host tissue while also attenuating inflammation caused by iatrogenic injury. An acellular agarose hydrogel carrier with embedded DEX-loaded poly(lactic-co-glycolic) acid (PLGA) microspheres (DLMS) was developed to provide sustained release for at least 99 days. The DLMS implant was first evaluated in an in vitro pro-inflammatory model of cartilage degradation. The implant was chondroprotective, as indicated by maintenance of Young's modulus (E) (p = 0.92) and GAG content (p = 1.0) in the presence of interleukin-1β insult. In a subsequent preliminary in vivo experiment, an osteochondral autograft transfer was performed using a pre-clinical canine model. DLMS implants were press-fit into the autograft donor site and compared to intra-articular DEX injection (INJ) or no DEX (CTL). Functional scores for DLMS animals returned to baseline (p = 0.39), whereas CTL and INJ remained significantly worse at 6 months (p < 0.05). DLMS knees were significantly more likely to have improved OARSI scores for proteoglycan, chondrocyte, and collagen pathology (p < 0.05). However, no significant improvements in synovial fluid cytokine content were observed. In conclusion, utilizing a targeted DLMS implant, we observed in vitro chondroprotection in the presence of IL-1-induced degradation and improved in vivo functional outcomes. These improved outcomes were correlated with superior histological scores but not necessarily a dampened inflammatory response, suggesting a primarily pro-anabolic effect. STATEMENT OF SIGNIFICANCE: Articular cartilage defects are a common source of joint pain and dysfunction. Effective treatment of these injuries may prevent the progression of osteoarthritis and reduce the need for total joint replacement. Dexamethasone, a potent glucocorticoid with concomitant anti-catabolic and pro-anabolic effects on cartilage, may serve as an adjuvant for a variety of repair strategies. Utilizing a dexamethasone-loaded osteochondral implant with controlled release characteristics, we demonstrated in vitro chondroprotection in the presence of IL-1-induced degradation and improved in vivo functional outcomes following osteochondral repair. These improved outcomes were correlated with superior histological cartilage scores and minimal-to-no comorbidity, which is a risk with high dose dexamethasone injections. Using this model of cartilage restoration, we have for the first time shown the application of targeted, low-dose dexamethasone for improved healing in a preclinical model of focal defect repair.
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http://dx.doi.org/10.1016/j.actbio.2019.11.052DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6956850PMC
January 2020

Cartilage Wear Particles Induce an Inflammatory Response Similar to Cytokines in Human Fibroblast-Like Synoviocytes.

J Orthop Res 2019 09 17;37(9):1979-1987. Epub 2019 May 17.

Department of Biomedical Engineering, Columbia University, New York, New York.

The synovium plays a key role in the development of osteoarthritis, as evidenced by pathological changes to the tissue observed in both early and late stages of the disease. One such change is the attachment of cartilage wear particles to the synovial intima. While this phenomenon has been well observed clinically, little is known of the biological effects that such particles have on resident cells in the synovium. The present work investigates the hypothesis that cartilage wear particles elicit a pro-inflammatory response in diseased and healthy human fibroblast-like synoviocytes, like that induced by key cytokines in osteoarthritis. Fibroblast-like synoviocytes from 15 osteoarthritic human donors and a subset of three non-osteoarthritic donors were exposed to cartilage wear particles, interleukin-1α or tumor necrosis factor-α for 6 days and analyzed for proliferation, matrix production, and release of pro-inflammatory mediators and degradative enzymes. Wear particles significantly increased proliferation and release of nitric oxide, interleukin-6 and -8, and matrix metalloproteinase-9, -10, and -13 in osteoarthritic synoviocytes, mirroring the effects of both cytokines, with similar trends in non-osteoarthritic cells. These results suggest that cartilage wear particles are a relevant physical factor in the osteoarthritic environment, perpetuating the pro-inflammatory and pro-degradative cascade by modulating synoviocyte behavior at early and late stages of the disease. Future work points to therapeutic strategies for slowing disease progression that target cell-particle interactions. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1979-1987, 2019.
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http://dx.doi.org/10.1002/jor.24340DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834361PMC
September 2019

A Functional Tissue-Engineered Synovium Model to Study Osteoarthritis Progression and Treatment.

Tissue Eng Part A 2019 04 31;25(7-8):538-553. Epub 2018 Oct 31.

1 Department of Biomedical Engineering, Columbia University , New York, New York.

Impact Statement: The synovium envelops the diarthrodial joint and plays a key regulatory role in defining the composition of the synovial fluid through filtration and biosynthesis of critical boundary lubricants. Synovium changes often precede cartilage damage in osteoarthritis. We describe a novel tissue engineered model, validated against native synovium explants, to investigate the structure-function of synovium through quantitative solute transport measures. Synovium was evaluated in the presence of a proinflammatory cytokine, interleukin-1, or the clinically relevant corticosteroid, dexamethasone. We anticipate that a better understanding of synovium transport would support efforts to develop more effective strategies aimed at restoring joint health.
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http://dx.doi.org/10.1089/ten.TEA.2018.0142DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6482911PMC
April 2019

Musculoskeletal mechanobiology: A new era for MechanoMedicine.

J Orthop Res 2018 02 6;36(2):531-532. Epub 2018 Feb 6.

Department of Orthopaedic Surgery, Washington University School of Medicine, 660 S. Euclid Avenue, St. Louis, MO 63110.

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http://dx.doi.org/10.1002/jor.23789DOI Listing
February 2018

Fibroblast-like synoviocyte mechanosensitivity to fluid shear is modulated by interleukin-1α.

J Biomech 2017 07 28;60:91-99. Epub 2017 Jun 28.

Columbia University, Department of Biomedical Engineering, New York, NY, United States. Electronic address:

Fibroblast-like synoviocytes (FLS) reside in the synovial membrane of diarthrodial joints and are exposed to a dynamic fluid environment that presents both physical and chemical stimuli. The ability of FLS to sense and respond to these stimuli plays a key role in their normal function, and is implicated in the alterations to function that occur in osteoarthritis (OA). The present work characterizes the response of FLS to fluid flow-induced shear stress via real-time calcium imaging, and tests the hypothesis that this response is modulated by interleukin-1α (IL-1α), a cytokine elevated in OA. FLS demonstrated a robust calcium signaling response to fluid shear that was dose dependent upon stress level and required both external and internal calcium sources. Preconditioning with 10ng/mL IL-1α for 24h heightened this shear stress response by significantly increasing the percent of responding cells and peak magnitude, while significantly decreasing the time for a peak to occur. Intercellular communication via gap junctions was found to account for a portion of the FLS population response in normal conditions, and was significantly increased by IL-1α preconditioning. IL-1α was also found to significantly increase average length and incidence of the primary cilium, an organelle commonly implicated in shear mechanosensing. These findings suggest that the elevated levels of IL-1α found in the OA environment heighten FLS sensitivity to fluid shear by altering both intercellular communication and individual cell sensitivity, which could affect downstream functions and contribute to progression of the disease state.
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http://dx.doi.org/10.1016/j.jbiomech.2017.06.011DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5788292PMC
July 2017

Constrained Cage Culture Improves Engineered Cartilage Functional Properties by Enhancing Collagen Network Stability.

Tissue Eng Part A 2017 08 27;23(15-16):847-858. Epub 2017 Mar 27.

1 Department of Biomedical Engineering, Columbia University , New York, New York.

When cultured with sufficient nutrient supply, engineered cartilage synthesizes proteoglycans rapidly, producing an osmotic swelling pressure that destabilizes immature collagen and prevents the development of a robust collagen framework, a hallmark of native cartilage. We hypothesized that mechanically constraining the proteoglycan-induced tissue swelling would enhance construct functional properties through the development of a more stable collagen framework. To test this hypothesis, we developed a novel "cage" growth system to mechanically prevent tissue constructs from swelling while ensuring adequate nutrient supply to the growing construct. The effectiveness of constrained culture was examined by testing constructs embedded within two different scaffolds: agarose and cartilage-derived matrix hydrogel (CDMH). Constructs were seeded with immature bovine chondrocytes and cultured under free swelling (FS) conditions for 14 days with transforming growth factor-β before being placed into a constraining cage for the remainder of culture. Controls were cultured under FS conditions throughout. Agarose constructs cultured in cages did not expand after the day 14 caging while FS constructs expanded to 8 × their day 0 weight after 112 days of culture. In addition to the physical differences in growth, by day 56, caged constructs had higher equilibrium (agarose: 639 ± 179 kPa and CDMH: 608 ± 257 kPa) and dynamic compressive moduli (agarose: 3.4 ± 1.0 MPa and CDMH 2.8 ± 1.0 MPa) than FS constructs (agarose: 193 ± 74 kPa and 1.1 ± 0.5 MPa and CDMH: 317 ± 93 kPa and 1.8 ± 1.0 MPa for equilibrium and dynamic properties, respectively). Interestingly, when normalized to final day wet weight, cage and FS constructs did not exhibit differences in proteoglycan or collagen content. However, caged culture enhanced collagen maturation through the increased formation of pyridinoline crosslinks and improved collagen matrix stability as measured by α-chymotrypsin solubility. These findings demonstrate that physically constrained culture of engineered cartilage constructs improves functional properties through improved collagen network maturity and stability. We anticipate that constrained culture may benefit other reported engineered cartilage systems that exhibit a mismatch in proteoglycan and collagen synthesis.
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http://dx.doi.org/10.1089/ten.TEA.2016.0467DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5567877PMC
August 2017

Concise Review: Mesenchymal Stem Cells for Functional Cartilage Tissue Engineering: Taking Cues from Chondrocyte-Based Constructs.

Stem Cells Transl Med 2017 04 8;6(4):1295-1303. Epub 2017 Feb 8.

Columbia University, New York, New York, USA.

Osteoarthritis, the most prevalent form of joint disease, afflicts 9% of the U.S. population over the age of 30 and costs the economy nearly $100 billion annually in healthcare and socioeconomic costs. It is characterized by joint pain and dysfunction, though the pathophysiology remains largely unknown. Due to its avascular nature and limited cellularity, articular cartilage exhibits a poor intrinsic healing response following injury. As such, significant research efforts are aimed at producing engineered cartilage as a cell-based approach for articular cartilage repair. However, the knee joint is mechanically demanding, and during injury, also a milieu of harsh inflammatory agents. The unforgiving mechano-chemical environment requires tissue replacements that are capable of bearing such burdens. The use of mesenchymal stem cells (MSCs) for cartilage tissue engineering has emerged as a promising cell source due to their ease of isolation, capacity to readily expand in culture, and ability to undergo lineage-specific differentiation into chondrocytes. However, to date, very few studies utilizing MSCs have successfully recapitulated the structural and functional properties of native cartilage, exposing the difficult process of uniformly differentiating stem cells into desired cell fates and maintaining the phenotype during in vitro culture and after in vivo implantation. To address these shortcomings, here, we present a concise review on modulating stem cell behavior, tissue development and function using well-developed techniques from chondrocyte-based cartilage tissue engineering. Stem Cells Translational Medicine 2017;6:1295-1303.
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http://dx.doi.org/10.1002/sctm.16-0271DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5442836PMC
April 2017

Current strategies for integrative cartilage repair.

Connect Tissue Res 2017 Sep 6;58(5):393-406. Epub 2016 Sep 6.

a Biomaterials and Interface Tissue Engineering Laboratory, Department of Biomedical Engineering , Columbia University , New York , NY , USA.

Osteoarthritis (OA) is a degenerative joint condition characterized by painful cartilage lesions that impair joint mobility. Current treatments such as lavage, microfracture, and osteochondral implantation fail to integrate newly formed tissue with host tissues and establish a stable transition to subchondral bone. Similarly, tissue-engineered grafts that facilitate cartilage and bone regeneration are challenged by how to integrate the graft seamlessly with surrounding host cartilage and/or bone. This review centers on current approaches to promote cartilage graft integration. It begins with an overview of articular cartilage structure and function, as well as degenerative changes to this relationship attributed to aging, disease, and trauma. A discussion of the current progress in integrative cartilage repair follows, focusing on graft or scaffold design strategies targeting cartilage-cartilage and/or cartilage-bone integration. It is emphasized that integrative repair is required to ensure long-term success of the cartilage graft and preserve the integrity of the newly engineered articular cartilage. Studies involving the use of enzymes, choice of cell source, biomaterial selection, growth factor incorporation, and stratified versus gradient scaffolds are therefore highlighted. Moreover, models that accurately evaluate the ability of cartilage grafts to enhance tissue integrity and prevent ectopic calcification are also discussed. A summary and future directions section concludes the review.
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http://dx.doi.org/10.1080/03008207.2016.1231180DOI Listing
September 2017

Nutrient Channels Aid the Growth of Articular Surface-Sized Engineered Cartilage Constructs.

Tissue Eng Part A 2016 09 23;22(17-18):1063-74. Epub 2016 Aug 23.

1 Department of Biomedical Engineering, Columbia University , New York, New York.

Symptomatic osteoarthritic lesions span large regions of joint surfaces and the ability to engineer cartilage constructs at clinically relevant sizes would be highly desirable. We previously demonstrated that nutrient transport limitations can be mitigated by the introduction of channels in 10 mm diameter cartilage constructs. In this study, we scaled up our previous system to cast and cultivate 40 mm diameter constructs (2.3 mm overall thickness); 4 mm diameter and channeled 10 mm diameter constructs were studied for comparison. Furthermore, to assess whether prior results using primary bovine cells are applicable for passaged cells-a more clinically realistic scenario-we cast constructs of each size with primary or twice-passaged cells. Constructs were assessed mechanically for equilibrium compressive Young's modulus (EY), dynamic modulus at 0.01 Hz (G*), and friction coefficient (μ); they were also assessed biochemically, histologically, and immunohistochemically for glycosaminoglycan (GAG) and collagen contents. By maintaining open channels, we successfully cultured robust constructs the size of entire human articular cartilage layers (growing to ∼52 mm in diameter, 4 mm thick, mass of 8 g by day 56), representing a 100-fold increase in scale over our 4 mm diameter constructs, without compromising their functional properties. Large constructs reached EY of up to 623 kPa and GAG contents up to 8.9%/ww (% of wet weight), both within native cartilage ranges, had G* >2 MPa, and up to 3.5%/ww collagen. Constructs also exhibited some of the lowest μ reported for engineered cartilage (0.06-0.11). Passaged cells produced tissue of lower quality, but still exhibited native EY and GAG content, similar to their smaller controls. The constructs produced in this study are, to our knowledge, the largest engineered cartilage constructs to date which possess native EY and GAG, and are a testament to the effectiveness of nutrient channels in overcoming transport limitations in cartilage tissue engineering.
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http://dx.doi.org/10.1089/ten.TEA.2016.0179DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5312614PMC
September 2016

Optimizing nutrient channel spacing and revisiting TGF-beta in large engineered cartilage constructs.

J Biomech 2016 07 21;49(10):2089-2094. Epub 2016 May 21.

Departments of Mechanical Engineering, Biomedical Engineering and Medicine, Columbia University, New York, NY 10027, United States. Electronic address:

Cartilage tissue engineering is a promising approach to treat osteoarthritis. However, current techniques produce tissues too small for clinical relevance. Increasingly close-packed channels have helped overcome nutrient transport limitations in centimeter-sized chondrocyte-agarose constructs, yet optimal channel spacings to recapitulate native cartilage compositional and mechanical properties in constructs this large have not been identified. Transient active TGF-β treatment consistently reproduces native compressive Young׳s modulus (EY) and glycosaminoglycan (GAG) content in constructs, but standard dosages of 10ng/mL exacerbate matrix heterogeneity. To ultimately produce articular layer-sized constructs, we must first optimize channel spacing and investigate the role of TGF-β in the utility of channels. We cultured ∅10mm constructs with 0, 12, 19, or 27 nutrient channels (∅1mm) for 6-8 weeks with 0, 1, or 10ng/mL TGF-β; subsequently we analyzed them mechanically, biochemically, and histologically. Constructs with 12 or 19 channels grew the most favorably, reaching EY=344±113kPa and GAG and collagen contents of 10.8±1.2% and 2.2±0.2% of construct wet weight, respectively. Constructs with 27 channels had significantly less deposited GAG than other groups. Channeled constructs given 1 or 10ng/mL TGF-β developed similar properties. Without TGF-β, constructs with 0 or 12 channels exhibited properties that were indistinguishable, and lower than TGF-β-supplemented constructs. Taken together, these results emphasize that nutrient channels are effective only in the presence of TGF-β, and indicate that spacings equivalent to 12 channels in ∅10mm constructs can be employed in articular-layer-sized constructs with reduced dosages of TGF-β.
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http://dx.doi.org/10.1016/j.jbiomech.2016.05.020DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4922491PMC
July 2016

High seeding density of human chondrocytes in agarose produces tissue-engineered cartilage approaching native mechanical and biochemical properties.

J Biomech 2016 06 10;49(9):1909-1917. Epub 2016 May 10.

Department of Mechanical Engineering, Columbia University, New York, NY 10027, USA; Department of Biomedical Engineering, Columbia University, New York, NY 10027, USA. Electronic address:

Animal cells have served as highly controllable model systems for furthering cartilage tissue engineering practices in pursuit of treating osteoarthritis. Although successful strategies for animal cells must ultimately be adapted to human cells to be clinically relevant, human chondrocytes are rarely employed in such studies. In this study, we evaluated the applicability of culture techniques established for juvenile bovine and adult canine chondrocytes to human chondrocytes obtained from fresh or expired osteochondral allografts. Human chondrocytes were expanded and encapsulated in 2% agarose scaffolds measuring ∅3-4mm×2.3mm, with cell seeding densities ranging from 15 to 90×10(6)cells/mL. Subsets of constructs were subjected to transient or sustained TGF-β treatment, or provided channels to enhance nutrient transport. Human cartilaginous constructs physically resembled native human cartilage, and reached compressive Young's moduli of up to ~250kPa (corresponding to the low end of ranges reported for native knee cartilage), dynamic moduli of ~950kPa (0.01Hz), and contained 5.7% wet weight (%/ww) of glycosaminoglycans (≥ native levels) and 1.5%/ww collagen. We found that the initial seeding density had pronounced effects on tissue outcomes, with high cell seeding densities significantly increasing nearly all measured properties. Transient TGF-β treatment was ineffective for adult human cells, and tissue construct properties plateaued or declined beyond 28 days of culture. Finally, nutrient channels improved construct mechanical properties, presumably due to enhanced rates of mass transport. These results demonstrate that our previously established culture system can be successfully translated to human chondrocytes.
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http://dx.doi.org/10.1016/j.jbiomech.2016.04.039DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4920373PMC
June 2016

Transient expression of the diseased phenotype of osteoarthritic chondrocytes in engineered cartilage.

J Orthop Res 2017 04 29;35(4):829-836. Epub 2016 May 29.

Department of Biomedical Engineering, Columbia University, New York, 1210 Amsterdam Avenue, 351 Engineering Terrace, New York, 10027.

Due to the degradation of osteoarthritic (OA) cartilage in post-traumatic OA (PTOA), these tissues are challenging to study and manipulate in vitro. In this study, chondrocytes isolated from either PTOA (meniscal-release (MR) model) or normal (contralateral limb) cartilage of canine knee joints were used to form micropellets to assess the maintenance of the OA chondrocyte phenotype in vitro. Media samples from the micropellet cultures were used to measure matrix metalloproteinase (MMP), chemokine, and cytokine concentrations. Significant differences in matrix synthesis were observed as a function of disease with OA chondrocytes generally synthesizing more extracellular matrix with increasing time in culture. No donor dependent differences were detected. Luminex multiplex analysis of pellet culture media showed disease and time-dependent differences in interleukin (IL)-8, keratinocyte chemoattractant (KC)-like protein, MMP-1, MMP-2, and MMP-3, which are differentially expressed in OA. This memory of their diseased phenotype persists for the first 2 weeks of culture. These results demonstrate the potential to use chondrocytes from an animal model of OA to study phenotype alterations during the progression and treatment of OA. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:829-836, 2017.
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http://dx.doi.org/10.1002/jor.23301DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5383531PMC
April 2017

Dexamethasone Release from Within Engineered Cartilage as a Chondroprotective Strategy Against Interleukin-1α.

Tissue Eng Part A 2016 Apr 31;22(7-8):621-32. Epub 2016 Mar 31.

1 Department of Biomedical Engineering, Columbia University , New York, New York.

While significant progress has been made toward engineering functional cartilage constructs with mechanical properties suitable for in vivo loading, the impact on these grafts of inflammatory cytokines, chemical factors that are elevated with trauma or osteoarthritis, is poorly understood. Previous work has shown dexamethasone to be a critical compound for cultivating cartilage with functional properties, while also providing chondroprotection from proinflammatory cytokines. This study tested the hypothesis that the incorporation of poly(lactic-co-glycolic acid) (PLGA) (75:25) microspheres that release dexamethasone from within chondrocyte-seeded agarose hydrogel constructs would promote development of constructs with functional properties and protect constructs from the deleterious effects of interleukin-1α (IL-1α). After 28 days of growth culture, experimental groups were treated with IL-1α (10 ng/mL) for 7 days. Reaching native equilibrium moduli and proteoglycan levels, dexamethasone-loaded microsphere constructs exhibited tissue properties similar to microsphere-free control constructs cultured in dexamethasone-supplemented culture media and were insensitive to IL-1α exposure. These findings are in stark contrast to constructs containing dexamethasone-free microspheres or no microspheres, cultured without dexamethasone, where IL-1α exposure led to significant tissue degradation. These results support the use of dexamethasone delivery from within engineered cartilage, through biodegradable microspheres, as a strategy to produce mechanically functional tissues that can also combat the deleterious effects of local proinflammatory cytokine exposure.
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http://dx.doi.org/10.1089/ten.TEA.2016.0018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4841087PMC
April 2016

A puzzle assembly strategy for fabrication of large engineered cartilage tissue constructs.

J Biomech 2016 Mar 9;49(5):668-677. Epub 2016 Feb 9.

Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 1210 Amsterdam Avenue, Mail Code: 8904, New York, NY 10027, USA. Electronic address:

Engineering of large articular cartilage tissue constructs remains a challenge as tissue growth is limited by nutrient diffusion. Here, a novel strategy is investigated, generating large constructs through the assembly of individually cultured, interlocking, smaller puzzle-shaped subunits. These constructs can be engineered consistently with more desirable mechanical and biochemical properties than larger constructs (~4-fold greater Young׳s modulus). A failure testing technique was developed to evaluate the physiologic functionality of constructs, which were cultured as individual subunits for 28 days, then assembled and cultured for an additional 21-35 days. Assembled puzzle constructs withstood large deformations (40-50% compressive strain) prior to failure. Their ability to withstand physiologic loads may be enhanced by increases in subunit strength and assembled culture time. A nude mouse model was utilized to show biocompatibility and fusion of assembled puzzle pieces in vivo. Overall, the technique offers a novel, effective approach to scaling up engineered tissues and may be combined with other techniques and/or applied to the engineering of other tissues. Future studies will aim to optimize this system in an effort to engineer and integrate robust subunits to fill large defects.
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http://dx.doi.org/10.1016/j.jbiomech.2016.01.023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4907770PMC
March 2016

Continuum theory of fibrous tissue damage mechanics using bond kinetics: application to cartilage tissue engineering.

Interface Focus 2016 Feb;6(1):20150063

Department of Biomedical Engineering, Columbia University, 500 West 120th Street, MC4703, New York, NY 10027, USA; Department of Mechanical Engineering, Columbia University, 500 West 120th Street, MC4703, New York, NY 10027, USA.

This study presents a damage mechanics framework that employs observable state variables to describe damage in isotropic or anisotropic fibrous tissues. In this mixture theory framework, damage is tracked by the mass fraction of bonds that have broken. Anisotropic damage is subsumed in the assumption that multiple bond species may coexist in a material, each having its own damage behaviour. This approach recovers the classical damage mechanics formulation for isotropic materials, but does not appeal to a tensorial damage measure for anisotropic materials. In contrast with the classical approach, the use of observable state variables for damage allows direct comparison of model predictions to experimental damage measures, such as biochemical assays or Raman spectroscopy. Investigations of damage in discrete fibre distributions demonstrate that the resilience to damage increases with the number of fibre bundles; idealizing fibrous tissues using continuous fibre distribution models precludes the modelling of damage. This damage framework was used to test and validate the hypothesis that growth of cartilage constructs can lead to damage of the synthesized collagen matrix due to excessive swelling caused by synthesized glycosaminoglycans. Therefore, alternative strategies must be implemented in tissue engineering studies to prevent collagen damage during the growth process.
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http://dx.doi.org/10.1098/rsfs.2015.0063DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4686240PMC
February 2016

High intensity focused ultrasound as a tool for tissue engineering: Application to cartilage.

Med Eng Phys 2016 Feb 24;38(2):192-8. Epub 2015 Dec 24.

Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 1210 Amsterdam Avenue, Mail Code: 8904, New York, NY 10027, USA. Electronic address:

This article promotes the use of High Intensity Focused Ultrasound (HIFU) as a tool for affecting the local properties of tissue engineered constructs in vitro. HIFU is a low cost, non-invasive technique used for eliciting focal thermal elevations at variable depths within tissues. HIFU can be used to denature proteins within constructs, leading to decreased permeability and potentially increased local stiffness. Adverse cell viability effects remain restricted to the affected area. The methods described in this article are explored through the scope of articular cartilage tissue engineering and the fabrication of osteochondral constructs, but may be applied to the engineering of a variety of different tissues.
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http://dx.doi.org/10.1016/j.medengphy.2015.11.016DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4876027PMC
February 2016

Hedgehog signalling does not stimulate cartilage catabolism and is inhibited by Interleukin-1β.

Arthritis Res Ther 2015 Dec 24;17:373. Epub 2015 Dec 24.

Institute of Bioengineering and School of Engineering and Material Sciences, Queen Mary University of London, Mile End Road, London, E1 4NS, UK.

Background: In osteoarthritis, chondrocytes adopt an abnormal hypertrophic morphology and upregulate the expression of the extracellular matrix-degrading enzymes, MMP-13 and ADAMTS-5. The activation of the hedgehog signalling pathway has been established in osteoarthritis and is thought to influence both of these processes. However, the role of this pathway in the initiation and progression of osteoarthritis is unclear as previous studies have been unable to isolate the effects of hedgehog pathway activation from other pathological processes. In this study we test the hypothesis that hedgehog pathway activation causes cartilage degradation in healthy cartilage and in an in vitro model of inflammatory arthritis.

Methods: Isolated articular chondrocytes from the bovine metacarpal-phalangeal joint were stimulated for up to 24 hours with the agonist, recombinant Indian hedgehog (r-Ihh). ADAMTS-5 and MMP-13 gene expression was quantified by real-time PCR. In addition, healthy bovine cartilage explants were treated with r-Ihh or the hedgehog antagonist, cyclopamine, and sGAG release into the media was measured over 72 hours. Studies were repeated using chondrocytes and cartilage explants from human knee joint. Finally, studies were conducted to determine the effect of hedgehog pathway activation on matrix catabolism in the presence of the pro-inflammatory cytokine, IL-1β.

Results: Addition of r-Ihh activated hedgehog signalling, confirmed by upregulation of Gli1 and Ptch1 expression, but did not increase ADAMTS-5 or MMP-13 expression in bovine or human chondrocytes. Furthermore, r-Ihh did not induce sGAG release in healthy bovine or human cartilage explants. IL-1β treatment induced sGAG release, but this response was not altered by the stimulation or inhibition of hedgehog signalling. Hedgehog pathway activation was downregulated by IL-1β. Conversely, r-Ihh weakly suppressed IL-1β-induced ADAMTS-5 expression.

Conclusion: Our results show for the first time that Indian hedgehog does not cause extracellular matrix degradation in healthy ex vivo cartilage or in the presence of IL-1β and that IL-1β downregulates Indian hedgehog induced signalling. Thus, we suggest reported hedgehog induced matrix catabolism in osteoarthritis must be due to its interaction with pathological factors other than IL-1β. Hence, hedgehog signalling and its downstream effects are highly context-dependent.
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http://dx.doi.org/10.1186/s13075-015-0891-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4718026PMC
December 2015

Grading of osteoarthritic cartilage: Correlations between histology and biomechanics.

J Orthop Res 2016 Jan 22;34(1):8-9. Epub 2015 Dec 22.

Andrew Walz Professor of Mechanical Engineering & Professor of Biomedical Engineering, Columbia University.

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http://dx.doi.org/10.1002/jor.23067DOI Listing
January 2016

Cytokine preconditioning of engineered cartilage provides protection against interleukin-1 insult.

Arthritis Res Ther 2015 Dec 14;17:361. Epub 2015 Dec 14.

Department of Biomedical Engineering, Columbia University, 1210 Amsterdam Avenue, New York, NY, USA.

Background: During osteoarthritis and following surgical procedures, the environment of the knee is rich in proinflammatory cytokines such as IL-1. Introduction of tissue-engineered cartilage constructs to a chemically harsh milieu may limit the functionality of the implanted tissue over long periods. A chemical preconditioning scheme (application of low doses of IL-1) was tested as a method to prepare developing engineered tissue to withstand exposure to a higher concentration of the cytokine, known to elicit proteolysis as well as rapid degeneration of cartilage.

Methods: Using an established juvenile bovine model system, engineered cartilage was preconditioned with low doses of IL-1α (0.1 ng/mL, 0.5 ng/mL, and 1.0 ng/mL) for 7 days before exposure to an insult dose (10 ng/mL). The time frame over which this protection is afforded was investigated by altering the amount of time between preconditioning and insult as well as the time following insult. To explore a potential mechanism for this protection, one set of constructs was preconditioned with CoCl2, a chemical inducer of hypoxia, before exposure to the IL-1α insult. Finally, we examined the translation of this preconditioning method to extend to clinically relevant adult, passaged chondrocytes from a preclinical canine model.

Results: Low doses of IL-1α (0.1 ng/mL and 0.5 ng/mL) protected against subsequent catabolic degradation by cytokine insult, preserving mechanical stiffness and biochemical composition. Regardless of amount of time between preconditioning scheme and insult, protection was afforded. In a similar manner, preconditioning with CoCl2 similarly allowed for mediation of catabolic damage by IL-1α. The effects of preconditioning on clinically relevant adult, passaged chondrocytes from a preclinical canine model followed the same trends with low-dose IL-1β offering variable protection against insult.

Conclusions: Chemical preconditioning schemes have the ability to protect engineered cartilage constructs from IL-1-induced catabolic degradation, offering potential modalities for therapeutic treatments.
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http://dx.doi.org/10.1186/s13075-015-0876-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4704536PMC
December 2015

Biphasic Analysis of Cartilage Stresses in the Patellofemoral Joint.

J Knee Surg 2016 Feb 7;29(2):92-8. Epub 2015 Dec 7.

Department of Mechanical Engineering, Columbia University, New York, New York.

The objective of this study was to examine the state of stress within the solid matrix of articular cartilage in the patellofemoral joint, using anatomically faithful biphasic models of the articular layers, with the joint subjected to physiologic muscle force magnitudes. Finite element models of five joints were created from human cadaver knees. Biphasic sliding contact analyses were performed using FEBio software to analyze the response of the joint from 30 to 60 degrees of knee flexion. Results demonstrated that the collagen matrix always sustains tensile stresses, despite the fact that the articular layers are loaded in compression. The principal direction of maximum solid stresses was consistent with the known orientation of collagen fibrils in cartilage. The magnitudes of these tensile stresses under muscle forces representative of activities of daily living were well below tensile failure stresses reported in the prior literature. Results also hinted that solid matrix stresses were higher in the patellar versus femoral superficial zone. These anatomically correct finite element models predicted outcomes consistent with our understanding of structure-function relationships in articular cartilage, while also producing solid matrix stress estimates not observable from experiments alone, yet highly relevant to our understanding of tissue degeneration.
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http://dx.doi.org/10.1055/s-0035-1568989DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4747677PMC
February 2016

Heterogeneous engineered cartilage growth results from gradients of media-supplemented active TGF-β and is ameliorated by the alternative supplementation of latent TGF-β.

Biomaterials 2016 Jan 18;77:173-185. Epub 2015 Nov 18.

Department of Biomedical Engineering, Columbia University, New York, NY 10027.

Transforming growth factor beta (TGF-β) has become one of the most widely utilized mediators of engineered cartilage growth. It is typically exogenously supplemented in the culture medium in its active form, with the expectation that it will readily transport into tissue constructs through passive diffusion and influence cellular biosynthesis uniformly. The results of this investigation advance three novel concepts regarding the role of TGF-β in cartilage tissue engineering that have important implications for tissue development. First, through the experimental and computational analysis of TGF-β concentration distributions, we demonstrate that, contrary to conventional expectations, media-supplemented exogenous active TGF-β exhibits a pronounced concentration gradient in tissue constructs, resulting from a combination of high-affinity binding interactions and a high cellular internalization rate. These gradients are sustained throughout the entire culture duration, leading to highly heterogeneous tissue growth; biochemical and histological measurements support that while biochemical content is enhanced up to 4-fold at the construct periphery, enhancements are entirely absent beyond 1 mm from the construct surface. Second, construct-encapsulated chondrocytes continuously secrete large amounts of endogenous TGF-β in its latent form, a portion of which undergoes cell-mediated activation and enhances biosynthesis uniformly throughout the tissue. Finally, motivated by these prior insights, we demonstrate that the alternative supplementation of additional exogenous latent TGF-β enhances biosynthesis uniformly throughout tissue constructs, leading to enhanced but homogeneous tissue growth. This novel demonstration suggests that latent TGF-β supplementation may be utilized as an important tool for the translational engineering of large cartilage constructs that will be required to repair the large osteoarthritic defects observed clinically.
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http://dx.doi.org/10.1016/j.biomaterials.2015.10.018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4968414PMC
January 2016

The friction coefficient of shoulder joints remains remarkably low over 24 h of loading.

J Biomech 2015 Nov 9;48(14):3945-9. Epub 2015 Oct 9.

Columbia University, Departments of Mechanical Engineering, Biomedical Engineering, and Orthopaedic Surgery, 500 West 120th Street, MC 4703, New York, NY 10027, USA. Electronic address:

The frictional response of whole human joints over durations spanning activities of daily living has not been reported previously. This study measured the friction of human glenohumeral joints during 24 h of reciprocal loading in a pendulum testing device, at moderate (0.2 mm/s, 4320 cycles) and low (0.02 mm/s, 432 cycles) sliding speeds, under a 200 N load. The effect of joint congruence was also investigated by testing human humeral heads against significantly larger mature bovine glenoids. Eight human joints and six bovine joints were tested in four combinations: human joints tested at moderate (hHCMS, n=6) and low speed (hHCLS, n=3), human humeral heads tested against bovine glenoids at moderate speed (LCMS, n=3), and bovine joints tested at moderate speed (bHCMS, n=3). In the first half hour the mean±standard deviation of the friction coefficient was hHCMS: 0.0016±0.0011, hHCLS: 0.0012±0.0002, LCMS: 0.0008±0.0002 and bHCMS: 0.0024±0.0008; in the last four hours it was hHCMS: 0.0057±0.0025, hHCLS: 0.0047±0.0017, LCMS: 0.0012±0.0003 and bHCMS: 0.0056±0.0016. The initial value was lower than the final value (p<0.0001). The value in LCMS was significantly lower than in hHCMS and bHCMS (p<0.01). No visual damage was observed in any of the specimens. These are the first results to demonstrate that the friction coefficient of natural human shoulders remains remarkably low (averaging as little as 0.0015 and no greater than 0.006) for up to 24 h of continuous loading. The sustained low friction coefficients observed in incongruent joints (~0.001) likely represent rolling rather than sliding friction.
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http://dx.doi.org/10.1016/j.jbiomech.2015.09.029DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4707938PMC
November 2015

Biocompatibility of polysebacic anhydride microparticles with chondrocytes in engineered cartilage.

Colloids Surf B Biointerfaces 2015 Dec 28;136:207-13. Epub 2015 Aug 28.

Earth and Environmental Engineering, 500 W, 120th street, 918 Mudd Columbia University, New York, NY, 10027, United States.

One of main challenges in developing clinically relevant engineered cartilage is overcoming limited nutrient diffusion due to progressive elaboration of extracellular matrix at the periphery of the construct. Macro-channels have been used to decrease the nutrient path-length; however, the channels become occluded with matrix within weeks in culture, reducing nutrient diffusion. Alternatively, microparticles can be imbedded throughout the scaffold to provide localized nutrient delivery. In this study, we evaluated biocompatibility of polysebacic anhydride (PSA) polymers and the effectiveness of PSA-based microparticles for short-term delivery of nutrients in engineered cartilage. PSA-based microparticles were biocompatible with juvenile bovine chondrocytes for concentrations up to 2mg/mL; however, cytotoxicity was observed at 20mg/mL. Cytotoxicity at high concentrations is likely due to intracellular accumulation of PSA degradation products and resulting lipotoxicity. Cytotoxicity of PSA was partially reversed in the presence of bovine serum albumin. In conclusion, the findings from this study demonstrate concentration-dependent biocompatibility of PSA-based microparticles and potential application as a nutrient delivery vehicle that can be imbedded in scaffolds for tissue engineering.
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http://dx.doi.org/10.1016/j.colsurfb.2015.08.040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4698167PMC
December 2015

Porous titanium bases for osteochondral tissue engineering.

Acta Biomater 2015 Nov 28;27:286-293. Epub 2015 Aug 28.

Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 1210 Amsterdam Avenue, Mail Code: 8904, New York, NY 10027, USA. Electronic address:

Unlabelled: Tissue engineering of osteochondral grafts may offer a cell-based alternative to native allografts, which are in short supply. Previous studies promote the fabrication of grafts consisting of a viable cell-seeded hydrogel integrated atop a porous, bone-like metal. Advantages of the manufacturing process have led to the evaluation of porous titanium as the bone-like base material. Here, porous titanium was shown to support the growth of cartilage to produce native levels of Young's modulus, using a clinically relevant cell source. Mechanical and biochemical properties were similar or higher for the osteochondral constructs compared to chondral-only controls. Further investigation into the mechanical influence of the base on the composite material suggests that underlying pores may decrease interstitial fluid pressurization and applied strains, which may be overcome by alterations to the base structure. Future studies aim to optimize titanium-based tissue engineered osteochondral constructs to best match the structural architecture and strength of native grafts.

Statement Of Significance: The studies described in this manuscript follow up on previous studies from our lab pertaining to the fabrication of osteochondral grafts that consist of a bone-like porous metal and a chondrocyte-seeded hydrogel. Here, tissue engineered osteochondral grafts were cultured to native stiffness using adult chondrocytes, a clinically relevant cell source, and a porous titanium base, a material currently used in clinical implants. This porous titanium is manufactured via selective laser melting, offering the advantages of precise control over shape, pore size, and orientation. Additionally, this manuscript describes the mechanical influence of the porous base, which may have applicability to porous bases derived from other materials.
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http://dx.doi.org/10.1016/j.actbio.2015.08.045DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4698168PMC
November 2015

Development of a whole organ culture model for intervertebral disc disease.

J Orthop Translat 2016 Apr 29;5:1-8. Epub 2015 Aug 29.

Comparative Orthopaedic Laboratory, University of Missouri, Columbia, MO, USA.

Background/objective: Whole organ intervertebral disc models have been associated with poor maintenance of cell viability. No previous studies have used a rotating wall vessel bioreactor for intervertebral disc explants culture. The purpose of this study was to develop and validate an model for the assessment of biological and biomechanical measures of intervertebral disc health and disease.

Methods: To this end, endplate-intervertebral disc-endplate whole organ explants were harvested from the tails of rats. For the injured group, the annulus fibrosus was penetrated with a 20G needle to the nucleus pulposus and aspirated. Explants were cultured in a rotating wall vessel bioreactor for 14 days.

Results: Cell viability and histologic assessments were performed at Day 0, Day 1, Day 7, and Day 14. Compressive mechanical properties of the intervertebral disc were assessed at Day 0 and Day 14. In the annulus fibrosus and nucleus pulposus cells, the uninjured group maintained high viability through 14 days of culture, whereas cell viability in annulus fibrosus and nucleus pulposus of the injured intervertebral discs was markedly lower at Day 7 and Day 14. Histologically, the uninjured intervertebral discs maintained cell viability and tissue morphology and architecture through 14 days, whereas the injured intervertebral discs showed areas of cell death, loss of extracellular matrix integrity, and architecture by Day 14. Stiffness values for uninjured intervertebral discs were similar at Day 0 and Day 14, whereas the stiffness for the injured intervertebral discs was approximately 2.5 times greater at Day 14.

Conclusion: These results suggest that whole organ intervertebral discs explants can be successfully cultured in a rotating wall vessel bioreactor to maintain cell viability and tissue architecture in both annulus fibrosus and nucleus pulposus for at least 14 days. In addition, the injury used produced pathologic changes consistent with those seen in degenerative intervertebral disc disease in humans. This model will permit further study into potential future treatments and other mechanisms of addressing intervertebral disc disease.
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http://dx.doi.org/10.1016/j.jot.2015.08.002DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5987001PMC
April 2016
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