Human Septal Chondrocyte Redifferentiation in Alginate, Polyglycolic Acid Scaffold, and Monolayer Culture

Homicz, Mark R.; Chia, Stanley H.; Schumacher, Barbara L.; Masuda, Koichi; Thonar, Eugene J-M.A.; Sah, Robert L.; Watson, Deborah

doi:10.1097/00005537-200301000-00005

Cited by 92 publications

(63 citation statements)

References 31 publications

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“…This suggests that the initial extracellular matrix scaffold does not require all components of the final basement membrane and that the UB itself might synthesize any necessary supplementary proteins in an isolated system. In addition, two inert ECM molecules, alginate, which has been used extensively in cartilage tissue engineering (42)(43)(44), and Puramatrix, which was successfully used to support neuronal migration (45) and to promote osteoblast differentiation (46), did not support UB branching. This may be due to the inability of the UB to remodel the artificial matrix to allow room for new branches.…”

Section: Discussionmentioning

confidence: 99%

Staged in vitro reconstitution and implantation of engineered rat kidney tissue

Rosines

Sampogna²,

Johkura³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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A major hurdle for current xenogenic-based and other approaches aimed at engineering kidney tissues is reproducing the complex three-dimensional structure of the kidney. Here, a stepwise, in vitro method of engineering rat kidney-like tissue capable of being implanted is described. Based on the fact that the stages of kidney development are separable into in vitro modules, an approach was devised that sequentially induces an epithelial tubule (the Wolffian duct) to undergo in vitro budding, followed by branching of a single isolated bud and its recombination with metanephric mesenchyme. Implantation of the recombined tissue results in apparent early vascularization. Thus, in principle, an unbranched epithelial tubular structure (potentially constructed from cultured cells) can be induced to form kidney tissue such that this in vitro engineered tissue is capable of being implanted in host rats and developing glomeruli with evidence of early vascularization. Optimization studies (of growth factor and matrix) indicate multiple suitable combinations and suggest both a most robust and a minimal system. A whole-genome microarray analysis suggested that recombined tissue recapitulated gene expression changes that occur in vivo during later stages of kidney development, and a functional assay demonstrated that the recombined tissue was capable of transport characteristic of the differentiating nephron. The approach includes several points where tissue can be propagated. The data also show how functional, 3D kidney tissue can assemble by means of interactions of independent modules separable in vitro, potentially facilitating systems-level analyses of kidney development. kidney development ͉ systems biology ͉ tissue engineering

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Section: Discussionmentioning

confidence: 99%

Staged in vitro reconstitution and implantation of engineered rat kidney tissue

Rosines

Sampogna²,

Johkura³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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“…Droplets of the alginate-chondrocyte suspension were polymerized in 102 mM CaCl 2 for 5 min to form beads. 3 The volume of each bead was approximately 10 mm 3 (40,000 cells). After washing with 0.9% saline, the beads were transferred to culture medium (DMEM/F-12, 25 mg/mL ascorbate, 0.4 mM L-proline, 2 mM L-glutamine, 0.1 mM nonessential amino acids, 10 mM HEPES, 100 U/mL penicillin G, 100 mg/mL streptomycin sulfate, and 0.25 mg/ mL amphotericin B).…”

Section: Culture In Alginatementioning

confidence: 99%

“…1,2 Expanded cells are then cultured in a three-dimensional (3D) scaffold such as alginate, agarose, or polyglycolic acid, inducing redifferentiation and production of ECM. [3][4][5] The capacity of cells to redifferentiate and form cartilaginous tissue becomes impaired with increasing levels of expansion. [2][3][4][5] Variables such as medium composition, growth factors, cell seeding density, 3D scaffold properties, and physical stimulation influence the ability of expanded cells to redifferentiate and produce functional cartilaginous ECM.…”

mentioning

confidence: 99%

Insulin-like Growth Factor-I and Growth Differentiation Factor-5 Promote the Formation of Tissue-Engineered Human Nasal Septal Cartilage

Alexander

Sage²,

Chen

et al. 2010

Tissue Engineering Part C: Methods

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Introduction: Tissue engineering of human nasal septal chondrocytes offers the potential to create large quantities of autologous material for use in reconstructive surgery of the head and neck. Culture with recombinant human growth factors may improve the biochemical and biomechanical properties of engineered tissue. The objectives of this study were to (1) perform a high-throughput screen to assess multiple combinations of growth factors and (2) perform more detailed testing of candidates identified in part I. Methods: In part I, human nasal septal chondrocytes from three donors were expanded in monolayer with pooled human serum (HS). Cells were then embedded in alginate beads for 2 weeks of culture in medium supplemented with 2% or 10% HS and 1 of 90 different growth factor combinations. Combinations of insulin-like growth factor-I (IGF-1), bone morphogenetic protein (BMP)-2, BMP-7, BMP-13, growth differentiation factor-5 (GDF-5), transforming growth factor b (TGFb)-2, insulin, and dexamethasone were evaluated. Glycosaminoglycan (GAG) accumulation was measured. A combination of IGF-1 and GDF-5 was selected for further testing based on the results of part I. Chondrocytes from four donors underwent expansion followed by threedimensional alginate culture for 2 weeks in medium supplemented with 2% or 10% HS with or without IGF-1 and GDF-5. Chondrocytes and their associated matrix were then recovered and cultured for 4 weeks in 12 mm transwells in medium supplemented with 2% or 10% HS with or without IGF-1 and GDF-5 (the same medium used for alginate culture). Biochemical and biomechanical properties of the neocartilage were measured. Results: In part I, GAG accumulation was highest for growth factor combinations including both IGF-1 and GDF-5. In part II, the addition of IGF-1 and GDF-5 to 2% HS resulted in a 12-fold increase in construct thickness compared with 2% HS alone ( p < 0.0001). GAG and type II collagen accumulation was significantly higher with IGF-1 and GDF-5. Confined compression modulus was greatest with 2% HS, IGF-1, and GDF-5. Conclusion: Supplementation of medium with IGF-1 and GDF-5 during creation of neocartilage constructs results in increased accumulation of GAG and type II collagen and improved biomechanical properties compared with constructs created without the growth factors.

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“…On encapsulation in alginate, the cells adopt a three dimensional rounded phenotype normally associated with the chondrocyte. With time it has been shown that a return to an expression pattern associated with cartilage is also achieved [2][3][4]34]. Following encapsulation, treatment with IGF1 (Gr.…”

Section: A B C Dmentioning

confidence: 99%

Generation of a scaffold free cartilage-like implant from a small amount of starting material

Stoddart

Ettinger

Häuselmann

2006

Journal of Cellular and Molecular Medicine

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Introduction: An autologous cellular based treatment of a traumatic cartilage injury requires a procedure whereby a biopsy of healthy cartilage is removed from the patient and the cells isolated and expanded by monolayer passage. This increases the cell number to required levels but also leads to a de-differentiation of the cells. We aim to produce a scaffold-free, de-novo implant from a biopsy of cartilage. Methods: Bovine chondrocytes were isolated from a small biopsy and expanded. The chondrocytic phenotype of the monolayer expanded cells was recovered during a period of culture in alginate and the effect of factors such as IGF1, TFGβ1 and dexamethasone was investigated. Results: During the alginate culture period a pre-treatment with IGF1 and dexamethasone was shown to have little effect. IGF1 however increased the glycosaminoglycan/DNA (GAG/DNA) content on day 14 to 84.95±5ng/ng compared with 37.3±1.8ng/ng in the controls (P <0.001). 35S labeling demonstrated an increased GAG synthesis in the presence of IGF1 (P < 0.001). IGF1 also induced a increase of DNA content 1383±314ng/bead compared to 512±19ng/bead in the controls (P < 0.001).The cells were released from the alginate and cultured in a silicon mould for a further 14 days to obtain a three dimensional implant. Releasing the cells from the alginate and casting in a mould produced an implant of defined shape which contained no foreign material. After 31 days of culture the implants contained 152.4±13.14ng/ng GAG/DNA and 42.93±10.23ng/ng collagen II. Discussion: We believe alginate released chondrocytes provide a real alternative to artificial scaffolds.

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Human Septal Chondrocyte Redifferentiation in Alginate, Polyglycolic Acid Scaffold, and Monolayer Culture

Cited by 92 publications

References 31 publications

Staged in vitro reconstitution and implantation of engineered rat kidney tissue

Staged in vitro reconstitution and implantation of engineered rat kidney tissue

Insulin-like Growth Factor-I and Growth Differentiation Factor-5 Promote the Formation of Tissue-Engineered Human Nasal Septal Cartilage

Generation of a scaffold free cartilage-like implant from a small amount of starting material

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