Articular cartilage tensile integrity: Modulation by matrix depletion is maturation-dependent

Asanbaeva, Anna; Tam, Johnny; Schumacher, Barbara L.; Klisch, Stephen M.; Masuda, Koichi; Sah, Robert L.

doi:10.1016/j.abb.2008.03.012

Cited by 38 publications

(33 citation statements)

References 62 publications

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“…The physical, chemical and biological regulation of the cellular microenvironment is essential to maintain the spatial and temporal behavior of chondrocytes. Cartilage is avascular and aneural; thus, chondrocytes require nourishment by the slow diffusion of synovial fluid (12). Although the traditional two-dimensional (2D) cell culturing technique was typically adopted to explore the optimal expansion environment for chondrocytes in vitro, it remains controversial whether chondrocytes grown in vitro are morphologically identical to those present in vivo (13).…”

Section: Introductionmentioning

confidence: 99%

Effects of insulin-like growth factor 1 and basic fibroblast growth factor on the morphology and proliferation of chondrocytes embedded in Matrigel in a microfluidic platform

Fan

Jiang

et al. 2017

Experimental and Therapeutic Medicine

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Abstract. An integrated microfluidic device was utilized in the present study to investigate the morphology and proliferation of rabbit articular chondrocytes embedded in Matrigel in the presence of insulin-like growth factor 1 (IGF-1) and/or basic fibroblast growth factor (bFGF). The microfluidic device was composed of two parallel channels and a central perfusion-based three-dimensional cell culture module. The rabbit chondrocytes were cultured for 2 weeks at series of concentration gradients of IGF-1 and/or bFGF, which were generated through a diffusion process. At the end of the experiment, the morphology and quantity of cells were measured. Since high expression of collagen II is essential to the function of hyaline cartilage, immunofluorescent images of collagen II expression prior to and after the experiments were gathered for each group. The mean fluorescence intensity ratio (MIR) of collagen II in each group was calculated. The MIRs of collagen II in chondrocytes treated with IGF-1 ranged from 0.6-0.81, those in the cells treated with bFGF ranged from 0.47-0.52, and those in cells treated with a combination of IGF-1 and bFGF ranged from 0.63-0.83. Chondrocyte aggregations were observed in the group treated with 75-100 ng/ml IGF-1 (3.46-fold proliferation ratio). Similarly, a 3.83-fold proliferation ratio was identified in chondrocytes treated with 2.5-5.0 ng/ml bFGF. The group treated with 50-75 ng/ml IGF-1 and 2.5-5.0 ng/ml bFGF exhibited the optimum increase in proliferation (4.83-fold proliferation ratio). The microfluidic device used in the present study can be easily adapted to investigate other growth factors at any concentration gradient. In addition, parallel experiments can be performed simultaneously with a small quantity of cells, making it an attractive platform for the high-throughput screening of cell culture parameters. This platform will aid in the optimization of culture conditions for the in vitro expansion of chondrocytes while maintaining their in vivo morphology, which will improve autologous chondrocyte implantation capabilities for the treatment of cartilage injury.

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Section: Introductionmentioning

confidence: 99%

Effects of insulin-like growth factor 1 and basic fibroblast growth factor on the morphology and proliferation of chondrocytes embedded in Matrigel in a microfluidic platform

Fan

Jiang

et al. 2017

Experimental and Therapeutic Medicine

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“…Previous studies with cultured chondrocytes encapsulated in agarose have shown higher compressive stress with increasing sGAG content with time in culture. 30,50 Depletion of PG in immature cartilage has been shown to increase tensile strength 21 and may alter biomechanical properties in tissue-engineered constructs as well. 22 Studies of in vitro collagen fibril assembly have shown that the presence of PG, such as CS or AGC, alters size and rate of fibril formation and the mechanical properties of the resulting collagen-PG material as compared to collagen alone.…”

Section: Figmentioning

confidence: 99%

“…The samples were incubated with 1 mM 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride for 2 h at 378C to inhibit proteinase K and assayed using HA test kit (Corgenix). 21 HA was analyzed in PBS pooled from day 1 to 3, because the amount released was small in comparison to sGAG release as determined from a pilot study (Supplemental Material, available online at www.liebertonline.com/ten).…”

Section: Biochemical Analysis Of Released and Retained Pg Componentsmentioning

confidence: 99%

“…19,20 By modulating the assembly of aggregated PG, it may be possible to achieve the desired levels of PG components, which may modulate the mechanical properties of the engineered construct. 21,22 Additionally, the ability to accelerate the formation of PG aggregates in the engineered tissue has the potential to save a significant amount of time compared to the biological replenishment and redistribution of AGC. Thus, AGC may be a useful component to include during the assembly of cartilaginous constructs-alone, þHA, or þHAþLP.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Tissue Engineering by Molecular Disassembly and Reassembly: Biomimetic Retention of Mechanically Functional Aggrecan in Hydrogel

Han

Wilensky

Schumacher

et al. 2010

Tissue Engineering Part C: Methods

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In vitro assembly of key functional extracellular matrix constituents for tissue-engineered constructs may provide a tool to modulate the retention of proteoglycan (PG) aggregates, which are crucial to compressive biomechanical properties of connective tissues. This study tested the hypotheses that (1) biomimetic molecular reassembly of PG aggregates (native aggrecan [AGC] with hyaluronan [HA] AE link protein [LP]) affects AGC retention kinetics in hydrogel constructs, (2) the compressive properties of such hydrogel constructs are related to the content of retained AGC, and (3) the reassembly method is compatible with chondrocytes. Addition of HA to AGC in hydrogel constructs increased AGC retention in a dose-dependent manner, and the addition of LP to AGC þ HA further enhanced AGC retention. The level of AGC retention, in turn, was associated with increased equilibrium compressive stress of the constructs. Chondrocytes could be included in the process, and maintained expression of the chondrogenic phenotype, secreting type II collagen but little type I collagen. Thus, by altering the assembly of PG aggregates with HA AE LP, which affects AGC retention, it may be possible to achieve the targeted levels of PG components to modulate the mechanical properties of the engineered construct for cartilage as well as other tissues containing PG and PG aggregates.

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“…In another method, selective enzymatic degradation of PG in cartilage and engineered constructs resulted in increased COL concentration, COL:PG ratio, and tensile stiffness of the tissue. [14][15][16] These approaches highlight the importance of the content of and balance between COL and PG in the mechanical function of tissue-engineered constructs. However, direct mechanical compaction to increase ECM concentration and mechanical function has not been studied previously.…”

Section: Introductionmentioning

confidence: 99%

Compaction Enhances Extracellular Matrix Content and Mechanical Properties of Tissue-Engineered Cartilaginous Constructs

Han

Ge²,

Chen³

et al. 2012

Tissue Engineering Part A

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Many cell-based tissue-engineered cartilaginous constructs are mechanically softer than native tissue and have low content and abnormal proportions of extracellular matrix (ECM) constituents. We hypothesized that the load-bearing mechanical properties of cartilaginous constructs improve with the inclusion of collagen (COL) and proteoglycan (PG) during assembly. The objectives of this work were to determine (1) the effect of addition of PG, COL, or COL + PG on compressive properties of 2% agarose constructs and (2) the ability of mechanical compaction to concentrate matrix content and improve the compressive properties of such constructs. The inclusion of COL + PG improved the compressive properties of hydrogel constructs compared with PG or COL alone. Mechanical compaction increased the PG and COL concentrations in and compressive stiffness of the constructs. Chondrocytes included in the constructs maintained high viability after compaction. These results support the concepts that the assembly of cartilaginous constructs with COL + PG and application of mechanical compaction enhance the ECM content and compressive properties of engineered cartilaginous constructs.

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Articular cartilage tensile integrity: Modulation by matrix depletion is maturation-dependent

Cited by 38 publications

References 62 publications

Effects of insulin-like growth factor 1 and basic fibroblast growth factor on the morphology and proliferation of chondrocytes embedded in Matrigel in a microfluidic platform

Effects of insulin-like growth factor 1 and basic fibroblast growth factor on the morphology and proliferation of chondrocytes embedded in Matrigel in a microfluidic platform

Tissue Engineering by Molecular Disassembly and Reassembly: Biomimetic Retention of Mechanically Functional Aggrecan in Hydrogel

Compaction Enhances Extracellular Matrix Content and Mechanical Properties of Tissue-Engineered Cartilaginous Constructs

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