Effect of construct properties on encapsulated chondrocyte expression of insulin-like growth factor-1

Yoon, Diana M.; Hawkins, Emily C.; Francke-Carroll, Sabine; Fisher, John P.

doi:10.1016/j.biomaterials.2006.08.039

Cited by 31 publications

(27 citation statements)

References 21 publications

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“…[26][27][28] Briefly, alginate solution was prepared by mixing and heating alginic acid sodium salt from brown algae (Sigma-Aldrich, St. Louis, MO, molecular weight 80,000-120,000 Da, M:G ratio $1.56, viscosity !2,000 cP), 0.15 M sodium chloride (Sigma-Aldrich), and 0.025 M HEPES, sodium salt (J.T. Baker, Phillipsburg, NJ) into deionized water (pH 7.4), and then autoclaved for sterilization.…”

Section: Bone Marrow Stromal Cell Isolationmentioning

confidence: 99%

Matrix molecule influence on chondrocyte phenotype and proteoglycan 4 expression by alginate‐embedded zonal chondrocytes and mesenchymal stem cells

Coates

Riggin

Fisher

2012

Journal Orthopaedic Research

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Articular cartilage resists load and provides frictionless movement at joint surfaces. The tissue is organized into the superficial, middle, deep, and calcified zones throughout its depth, each which serve distinct functions. Proteoglycan 4 (PRG4), found in the superficial zone, is a critical component of the joint's lubricating mechanisms. Maintenance of both the chondrocyte and zonal chondrocyte phenotype remain challenges for in vitro culture and tissue engineering. Here we investigate the expression of PRG4 mRNA and protein by primary bovine superficial zone chondrocytes, middle/deep zone chondrocytes, and mesenchymal stem cells encapsulated in alginate hydrogels with hyaluronic acid (HA) and chondroitin sulfate (CS) additives. Chondrogenic phenotype and differentiation markers are evaluated by mRNA expression, histochemical, and immunohistochemical staining. Results show middle/ deep cells express no measurable PRG4 mRNA by day 7. In contrast, superficial zone cells express elevated PRG4 mRNA throughout culture time. This expression can be significantly enhanced up to 15-fold by addition of both HA and CS to scaffolds. Conversely, PRG4 mRNA expression is downregulated (up to 5-fold) by CS and HA in differentiating MSCs, possibly due to build up of entrapped protein.HA and CS demonstrate favorable effects on chondrogenesis by upregulating transcription factor Sox9 mRNA (up to 4.6-fold) and downregulating type I collagen mRNA (up to 18-fold). Results highlight the important relationship between matrix components and expression of critical lubricating proteins in an engineered cartilage scaffold.

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Section: Bone Marrow Stromal Cell Isolationmentioning

confidence: 99%

Matrix molecule influence on chondrocyte phenotype and proteoglycan 4 expression by alginate‐embedded zonal chondrocytes and mesenchymal stem cells

Coates

Riggin

Fisher

2012

Journal Orthopaedic Research

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“…Alginate concentration and cell density were chosen based on previous results from the lab indicating high cell viability. 27 Control beads were cultured in serum-free chondrogenic media made of highglucose MEMa (Gibco/Invitrogen) + 110 mg/mL sodium pyruvate, 40 mg/mL proline, 50 mg/mL ascorbate 2-phosphate, 0.1 mM dexamethasone, 1% ITS + premix (BD Biosciences), and 10 ng/mL TGF-b3 treatment (R&D Systems). Coculture groups were set up with either superficial or middle/deep explants chips in the bottom of a six-well plate, a transwell membrane placed in the well, and alginate beads containing MSCs on top of the membrane.…”

Section: Bone Marrow Stromal Cell Isolationmentioning

confidence: 99%

Engineering Superficial Zone Chondrocytes from Mesenchymal Stem Cells

Coates

Fisher

2014

Tissue Engineering Part C: Methods

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Recent cartilage engineering efforts have focused on development of zonally organized tissue. However, there remains a need for protocols that differentiate progenitor populations into chondrocytes of zonal phenotype. Here, we evaluate the potential of coculture of bovine mesenchymal stem cells (MSCs) and zonal explants of bovine cartilage tissue to drive MSC differentiation to chondrocytes with the superficial zone phenotype. Two coculture systems were set up: one between alginate encapsulated MSCs and superficial zone cartilage explants, and one between MSCs and middle/deep zone cartilage explants. Chondrogenic and superficial zone markers were monitored over a 21-day differentiation period via gene and protein expression. A control conditioned media study was used to determine the impact of communication via soluble factors between cell populations during differentiation. At day 21, results show superficial zone explant coculture without transforming-growth factor b3 supplementation induces upregulation of chondrogenic gene expression markers SOX9 and type II collagen 3.4-fold and 11.4-fold, respectively, over standard chondrogenic control media. Further, coculture of MSCs and superficial zone explants can be used to upregulate mRNA expression of the superficial zone marker proteoglycan-4 in MSCs (1.75-fold over chondrogenic control at day 21), indicating the superficial zone chondrocyte phenotype. Gene expression data show middle/deep zone explant and MSC coculture did not induce the chondrogenesis observed in superficial zone explant coculture. Likewise, poor chondrogenesis was observed in all conditioned media groups. Results highlight the importance of superficial zone cartilage and cells in guiding stem cell fate and regulating differentiation of MSCs to chondrocytes of the superficial zone type.

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“…10 Chondrocytes derived from natural sources or from MSCs differentiating toward chondrocytes are commonly used as a cell source in cartilage tissue engineering; however, there are unmet challenges in expanding and maintaining chondrocytes (e.g., the composition of the construct and the regulation of culture conditions need careful monitoring). 13,14,50,56,57 In addition, chondrogenic growth factors play a key role in chondrocyte differentiation and cartilage generation. 58 The most challenging aspect of cartilage tissue engineering is to replicate the natural compressive behavior of cartilage under mechanical loading conditions and its porous structure for cell nutrients.…”

Section: Cartilage Tissue Engineeringmentioning

confidence: 99%

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

Kotecha

Klatt

Magin

2013

Tissue Engineering Part B: Reviews

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A key technical challenge in cartilage tissue engineering is the development of a noninvasive method for monitoring the composition, structure, and function of the tissue at different growth stages. Due to its noninvasive, three-dimensional imaging capabilities and the breadth of available contrast mechanisms, magnetic resonance imaging (MRI) techniques can be expected to play a leading role in assessing engineered cartilage. In this review, we describe the new MR-based tools (spectroscopy, imaging, and elastography) that can provide quantitative biomarkers for cartilage tissue development both in vitro and in vivo. Magnetic resonance spectroscopy can identify the changing molecular structure and alternations in the conformation of major macromolecules (collagen and proteoglycans) using parameters such as chemical shift, relaxation rates, and magnetic spin couplings. MRI provides high-resolution images whose contrast reflects developing tissue microstructure and porosity through changes in local relaxation times and the apparent diffusion coefficient. Magnetic resonance elastography uses low-frequency mechanical vibrations in conjunction with MRI to measure soft tissue mechanical properties (shear modulus and viscosity). When combined, these three techniques provide a noninvasive, multiscale window for characterizing cartilage tissue growth at all stages of tissue development, from the initial cell seeding of scaffolds to the development of the extracellular matrix during construct incubation, and finally, to the postimplantation assessment of tissue integration in animals and patients.

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Effect of construct properties on encapsulated chondrocyte expression of insulin-like growth factor-1

Cited by 31 publications

References 21 publications

Matrix molecule influence on chondrocyte phenotype and proteoglycan 4 expression by alginate‐embedded zonal chondrocytes and mesenchymal stem cells

Matrix molecule influence on chondrocyte phenotype and proteoglycan 4 expression by alginate‐embedded zonal chondrocytes and mesenchymal stem cells

Engineering Superficial Zone Chondrocytes from Mesenchymal Stem Cells

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

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