Basic Fibroblast Growth Factor and Insulinlike Growth Factor I Support the Growth of Human Septal Chondrocytes in a Serum-Free Environment

Bp, Dunham; Rj, Koch

doi:10.1001/archotol.124.12.1325

Cited by 39 publications

(28 citation statements)

References 20 publications

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“…[25][26][27][28] Richmon et al found increased GAG accumulation during monolayer culture of human nasal septal chondrocytes with IGF-1. 29 van Osch et al reported increased GAG accumulation and type II collagen during alginate bead culture with a combination of IGF-1 and TGFb-2.…”

Section: Figmentioning

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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Section: Figmentioning

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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“…A number of supplementary factors working in cohort seem to be required to rescue cells from SWinduced apoptosis, because rescue was not observed with ECGS alone and because only partial rescue was observed with IGF-1 and bFGF, even at concentrations higher than would normally be used for supplementing growth media (Dunham and Koch, 1998;Saltau and McLaughlin, 1993). Addition of all CGM components, however, successfully rescued cells from undergoing SW-induced apoptosis.…”

Section: Sw-induced Ec Apoptosismentioning

confidence: 99%

A Role for Caspase-1 in Serum Withdrawal-Induced Apoptosis of Endothelial Cells

King

Francis

Bridgeman

et al. 2003

Laboratory Investigation

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SUMMARY:Mouse lung endothelial cells (MLEC) and HUVEC were used under serum withdrawal (SW) conditions as a model of endothelial cell (EC) apoptosis. Apoptosis was quantified by time-lapse video microscopy. Mouse lung ECs from caspase-1 Ϫ/Ϫ mice had significantly reduced rates of SW-induced apoptosis compared with wild-type mice, specifically implicating caspase-1 in proapoptotic signaling in ECs. SW conditions induced HUVEC apoptosis with concomitant activation of caspase-1. Further studies demonstrated that the caspase-1 inhibitors z-VAD and z-YVAD significantly reduced the rate of SW-induced HUVEC apoptosis. HUVEC, when transfected with caspase-1, showed a highly significant increase in apoptosis. SW was associated with increases in reactive oxygen species production that were significantly inhibited by the antioxidant N-acetyl-L-cysteine, although rates of apoptosis and caspase-1 activation were unaffected. These results demonstrate the involvement of caspase-1 in SW-induced EC apoptosis, independently of reactive oxygen species production. (Lab Invest 2003, 83:1497-1508.A poptosis, the process of innate cellular death, is an accepted part of cell biology. The cellular mechanisms of apoptosis are complex and diverse, with considerable cell-type specificity.The activation of intracellular cysteine proteases (caspases) that have a unique cleavage site involving an aspartate residue is crucial to the process of apoptosis (Whyte, 1996;Wolf and Green, 1999). Once activated, caspases cleave cellular substrates, leading to characteristic morphologic hallmarks of apoptosis (Nicholson and Thornberry, 1997). There is a "hierarchy" of caspases in the control of apoptosis, with regulatory caspases (eg, caspase-2, -8, -9, and -10), which have long prodomains, initiating cleavage of downstream executioner caspases (eg, caspase-3, -6, and -7). The prototypic caspase, caspase-1, however, was originally identified as the enzyme responsible for processing IL-1␤ to its 17-kDa mature form (Cerretti et al, 1992). Overexpression of caspase-1, as with most other caspases, produces apoptosis (Miura et al, 1993). However, caspase-1 is not generally regarded as a component of the cell death machinery; together with caspases-4 and -5, it is regarded as an "inflammatory caspase" that functions only in cytokine processing (Martinon et al, 2002;Wolf and Green, 1999).The vascular endothelium has a central role in inflammation, the response to infection, and the pathogenesis of vascular disease. Endothelial cells (EC) can be induced to undergo apoptosis by a number of treatments, including TNF-␣ in the presence of protein synthesis inhibitors (Polunovsky et al, 1994), radiation (Haimovitz-Freidman et al, 1994), certain snake venoms (Araki et al, 1993), lipopolysaccharide (LPS) (Choi et al, 1998), and oxidized low-density lipoprotein (Harada-Shiba et al, 1998;Sata and Walsh, 1998). EC apoptosis occurs as a widespread event in thrombotic thrombocytopenic purpura (Laurence et al, 1996) and may occur in systemic sclerosis induced by anti-EC a...

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“…Our laboratory has already successfully expanded human septal chondrocytes in a serum-free in vitro model. 17 As a prelude to in vitro fabrication of hyaline cartilage in a completely serum-free process, we studied the feasibility of ex vivo generation of cartilage tissue equivalents from fresh and cryogenically preserved septal chondrocytes. Because fabrication of implants for human use requires highfidelity tissue regeneration, our studies combined the use of cell-PGA scaffolding constructs for efficient cell delivery with bioreactors.…”

Section: Construct Analysismentioning

confidence: 99%

Growth of Tissue-Engineered Human Nasoseptal Cartilage in Simulated Microgravity

Falsafi

Koch²

2000

Arch Otolaryngol Head Neck Surg

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Basic Fibroblast Growth Factor and Insulinlike Growth Factor I Support the Growth of Human Septal Chondrocytes in a Serum-Free Environment

Abstract: The combination of IGF-I and bFGF in a serum-free and a serum-supplemented environment supports the growth and viability of human septal chondrocytes in short-term culture. In an SFM, the results obtained approximate those produced in a medium enhanced with 10% fetal calf serum.

Cited by 39 publications

References 20 publications

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

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

A Role for Caspase-1 in Serum Withdrawal-Induced Apoptosis of Endothelial Cells

Growth of Tissue-Engineered Human Nasoseptal Cartilage in Simulated Microgravity

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