The Effect of the Duration of Mechanical Stimulation and Post-Stimulation Culture on the Structure and Properties of Dynamically Compressed Tissue-Engineered Menisci

Puetzer, Jennifer L.; Ballyns, Jeffrey J.; Bonassar, Lawrence J.

doi:10.1089/ten.tea.2011.0589

Cited by 43 publications

(32 citation statements)

References 56 publications

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“…A possibility for further maturation of these larger constructs is mechanical stimulation [Puetzer et al, 2012] or in vivo implantation [Freymann et al, 2012]. Future research will be focused on the generation of larger constructs via bioplotting (bottom-up approach).…”

Section: Discussionmentioning

confidence: 99%

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Berneel¹,

Philips²,

Declercq³

et al. 2016

Cells Tissues Organs

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In meniscus tissue engineering strategies, enhancing the matrix quality of the neomeniscal tissue is important. When the differentiated phenotype of fibrochondrocytes is lost, the quality of the matrix becomes compromised. The objective of this study was to produce uniform fibrochondrocyte micro-aggregates with desirable phenotype and tissue homogeneity in large quantities using a simple and reproducible method. Furthermore, we investigated if hypoxia could enhance the matrix quality. Porcine fibrochondrocytes were expanded at 21% oxygen until passage 3 (P3) and a gene expression profile was determined. P3 fibrochondrocytes were cultivated in chondrogenic medium at 5 and 21% oxygen in high-throughput agarose chips containing 2,865 microwells 200 µm in diameter. Evaluation included live/dead staining, histological examination, immunohistochemistry, dimethylmethylene blue assay and real-time reverse transcriptase quantitative polymerase chain reaction of the micro-aggregates. Gene expression analysis showed a drastic decline in collagen II and high expression of collagen I during monolayer culture. After 4 days, uniform and stable micro-aggregates could be produced. The redifferentiation and matrix quality of the hypoxic cultured micro-aggregates were enhanced relative to the normoxic cultures. Sulfated glycosaminoglycan synthesis was significantly higher, and collagen II expression and the collagen II/collagen I ratio were significantly upregulated in the hypoxic cultures. High-throughput production of uniform microtissues holds promise for the generation of larger-scale tissue engineering constructs or optimization of redifferentiation mechanisms for clinical applications.

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Section: Discussionmentioning

confidence: 99%

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Berneel¹,

Philips²,

Declercq³

et al. 2016

Cells Tissues Organs

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“…These results were consistent with a number of other studies that have utilized chondrocytes encapsulated within alginate for the formation of three-dimensional molded cartilage constructs. [36][37][38] In the present study, the formation of collagen and elastin were observed to be localized to the periphery of the construct, suggesting that there may be limitations in mass transport within these large-scale constructs. However, GAG formation was observed throughout the construct thickness, which suggests that GAG formation may occur more rapidly than does collagen and elastin formation.…”

Section: Discussionmentioning

confidence: 50%

“…[36][37][38] For example, dynamic mechanical stimulation of alginate constructs in the first 2 weeks of culture followed by static culture was shown to increase accumulation of both GAG and collagen with a concurrent increase in mechanical properties. 36,37 However, due to the complex geometry and large size of the samples in the present study, dynamic stimulation of the image-guided constructs would likely require development of a highly complex bioreactor system. Controlled media mixing is a simple method of mechanical conditioning that has been shown in a number of studies to increase ECM production of chondrocyte-loaded constructs.…”

Section: Discussionmentioning

confidence: 99%

Computed Tomography-Guided Tissue Engineering of Upper Airway Cartilage

Brown

Siebenlist²,

Cheetham³

et al. 2014

Tissue Engineering Part C: Methods

Self Cite

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Normal laryngeal function has a large impact on quality of life, and dysfunction can be life threatening. In general, airway obstructions arise from a reduction in neuromuscular function or a decrease in mechanical stiffness of the structures of the upper airway. These reductions decrease the ability of the airway to resist inspiratory or expiratory pressures, causing laryngeal collapse. We propose to restore airway patency through methods that replace damaged tissue and improve the stiffness of airway structures. A number of recent studies have utilized image-guided approaches to create cell-seeded constructs that reproduce the shape and size of the tissue of interest with high geometric fidelity. The objective of the present study was to establish a tissue engineering approach to the creation of viable constructs that approximate the shape and size of equine airway structures, in particular the epiglottis. Computed tomography images were used to create three-dimensional computer models of the cartilaginous structures of the larynx. Anatomically shaped injection molds were created from the three-dimensional models and were seeded with bovine auricular chondrocytes that were suspended within alginate before static culture. Constructs were then cultured for approximately 4 weeks post-seeding and evaluated for biochemical content, biomechanical properties, and histologic architecture. Results showed that the three-dimensional molded constructs had the approximate size and shape of the equine epiglottis and that it is possible to seed such constructs while maintaining 75%+ cell viability. Extracellular matrix content was observed to increase with time in culture and was accompanied by an increase in the mechanical stiffness of the construct. If successful, such an approach may represent a significant improvement on the currently available treatments for damaged airway cartilage and may provide clinical options for replacement of damaged tissue during treatment of obstructive airway disease.

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“…Last, the third hypothesis, that the inner and outer meniscus zones will show integration, was validated through histology and tensile testing. Previous work in the field has successfully produced tissue-engineered meniscus constructs, 32,33 but to our knowledge, this is the first study to engineer menisci with both inner and outer zones by integrating the two together.…”

Section: Figmentioning

confidence: 99%

Building an Anisotropic Meniscus with Zonal Variations

Higashioka

Chen

et al. 2014

Tissue Engineering Part A

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Toward addressing the difficult problems of knee meniscus regeneration, a self-assembling process has been used to re-create the native morphology and matrix properties. A significant problem in such attempts is the recapitulation of the distinct zones of the meniscus, the inner, more cartilaginous and the outer, more fibrocartilaginous zones. In this study, an anisotropic and zonally variant meniscus was produced by self-assembly of the inner meniscus (100% chondrocytes) followed by cell seeding the outer meniscus (coculture of chondrocytes and meniscus cells). After 4 weeks in culture, the engineered, inner meniscus exhibited a 42% increase in both instantaneous and relaxation moduli and a 62% increase in GAG/DW, as compared to the outer meniscus. In contrast, the circumferential tensile modulus and collagen/DW of the outer zone was 101% and 129% higher, respectively, than the values measured for the inner zone. Furthermore, there was no difference in the radial tensile modulus between the control and zonal engineered menisci, suggesting that the inner and outer zones of the engineered zonal menisci successfully integrated. These data demonstrate that not only can biomechanical and biochemical properties be engineered to differ by the zone, but they can also recapitulate the anisotropic behavior of the knee meniscus.

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The Effect of the Duration of Mechanical Stimulation and Post-Stimulation Culture on the Structure and Properties of Dynamically Compressed Tissue-Engineered Menisci

Cited by 43 publications

References 56 publications

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Redifferentiation of High-Throughput Generated Fibrochondrocyte Micro-Aggregates: Impact of Low Oxygen Tension

Computed Tomography-Guided Tissue Engineering of Upper Airway Cartilage

Building an Anisotropic Meniscus with Zonal Variations

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