Nell Ginley | St. Mary Seminary and Graduate School of Theology (original) (raw)
Papers by Nell Ginley
CARTILAGE, 2012
Objective: To develop a tissue culture expansion method for rabbit chondrocytes that promotes rob... more Objective: To develop a tissue culture expansion method for rabbit chondrocytes that promotes robust expansion while preserving chondrogenic potential. Design: Rabbit chondrocytes isolated from articular or auricular chondrocytes were assessed for chondrogenic differentiation potential versus population doubling using different expansion and differentiation conditions. Expansion conditions included serum alone, serum plus basic fibroblast growth factor 2 (FGF-2), and serum plus insulin-like growth factor 1 (IGF-1) and FGF-2. Differentiation conditions consisted of defined medium with and without bone morphogenetic protein 2 (BMP-2). Results: Nonsupplemented chondrocytes showed limited expandability, whereas supplementation with FGF-2 allowed articular chondrocytes to be expanded past 10 population doublings (PDs) and allowed auricular chondrocytes to expand past 15 population doublings. Differentiation, as measured by glycosaminoglycan production in aggregate cultures, was minimal i...
Osteoarthritis and Cartilage, 2007
The purpose of this study is to develop and test a method to fabricate sheets of cartilage that c... more The purpose of this study is to develop and test a method to fabricate sheets of cartilage that can be used to produce replacement cartilage in a rabbit trachea model. Methods and Materials: Auricular cartilage was harvested from white New Zealand Rabbits sequentially digested with trypsin, testicular hyaluronidase and collagenase type II, passed through a 70 µm nitex trypsinized stored in liquid nitrogen until needed. To form scaffoldfree cartilage sheets, 50-100 million chondrocytes were injected into a 6 x 7 cm bioreactor (composed of two polystyrene gas-permeable membranes) and cultured in serum-free medium containing ITS,1 mM sodium pyruvate, 50 µM ascorbate-2-phosphate, 10-7 M dexamethasone and non-essential amino acids, and cultured for 3 weeks. Results: The results showed that the implanted cartilage survived intact, integrated well with the surrounding fascia, which was well vascularized, and had a well-formed lumen of 4-5 mm in diameter, and there was no sign of an immune reaction even in tracheas formed from allo-cartilage sheets. The cartilage stained positively for collagens II and X, much like the native auricular cartilage, but showed no signs of mineralization. Conclusions: These results show that chondrocytes sheets can be used to fabricate vascularized neo-trachea that retains a lumen and may be used as a model to repair segmental defects in rabbits.
Rheumatology (Sunnyvale, Calif.), Jan 26, 2012
Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory r... more Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory response pathways in arthritic joints where inflammation may be accompanied by the loss of chondrocyte vitality via apoptosis. To address this possibility in vitro, chondrocyte cultures were employed to determine the extent to which monosodium urate and recombinant TNF-α altered the frequency of apoptotic chondrocytes. Apoptosis as a function of the activation of p38 kinase, C-Jun-terminal kinase, signal transducer and activator of transcription-3 and/or the activity of xanthine oxidase was also studied. Using normal human chondrocytes, monosodium urate or recombinant tumor necrosis factor-α increased the frequency of apoptosis and activity of xanthine oxidase. However, the xanthine oxidase-specific inhibitor, febuxostat, failed to blunt this response. Monosodium urate, tumor necrosis factor-α or the Janus kinase inhibitor, AG-490, increased the frequency of apoptotic nuclei in macroaggre...
Rheumatology (Sunnyvale, Calif.), Jan 26, 2012
Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory r... more Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory response pathways in arthritic joints where inflammation may be accompanied by the loss of chondrocyte vitality via apoptosis. To address this possibility in vitro, chondrocyte cultures were employed to determine the extent to which monosodium urate and recombinant TNF-α altered the frequency of apoptotic chondrocytes. Apoptosis as a function of the activation of p38 kinase, C-Jun-terminal kinase, signal transducer and activator of transcription-3 and/or the activity of xanthine oxidase was also studied. Using normal human chondrocytes, monosodium urate or recombinant tumor necrosis factor-α increased the frequency of apoptosis and activity of xanthine oxidase. However, the xanthine oxidase-specific inhibitor, febuxostat, failed to blunt this response. Monosodium urate, tumor necrosis factor-α or the Janus kinase inhibitor, AG-490, increased the frequency of apoptotic nuclei in macroaggre...
Tissue Engineering Part A, 2010
To determine whether low oxygen (O 2 ) tension during expansion affects the matrix density, as we... more To determine whether low oxygen (O 2 ) tension during expansion affects the matrix density, as well as quantity, of cartilage formed, and to determine whether application of low O 2 tension during incubation periods alone is sufficient to modulate chondrogenic expression, rabbit chondrocytes expanded at either 21% O 2 or 5% O 2 were analyzed for glycosaminoglycan (GAG) and DNA content, total collagen, and gene expression during expansion and postexpansion aggregate cultures. When cultured as aggregates at 21% O 2 , chondrocytes expanded at 5% O 2 produced cartilage aggregates that contained more total GAG, GAG per wet weight, GAG per DNA, and total collagen than chondrocytes expanded at 21% O 2 . Less of an effect on GAG and collagen content was observed when aggregate culture was performed at 5% O 2 . Real-time polymerase chain reaction analysis of COL2A1 expression showed upregulated levels of type IIA (an early marker) and IIB (a late marker) during expansion and elevated levels of type IIB during aggregate culture in chondrocytes expanded in low O 2 . The application of low O 2 tension during incubation periods of chondrocyte expansion enhances the ultimate cartilage matrix density and quantity, and this enhancement can be achieved through the use of an O 2 control incubator.
Osteoarthritis and Cartilage, 2007
Purpose: The purpose of this study is to develop and test a method to fabricate sheets of cartila... more Purpose: The purpose of this study is to develop and test a method to fabricate sheets of cartilage that can be used to produce replacement cartilage in a rabbit trachea model. Methods and Materials: Auricular cartilage was harvested from white New Zealand Rabbits sequentially digested with trypsin, testicular hyaluronidase and collagenase type II, passed through a 70 µm nitex trypsinized stored in liquid nitrogen until needed. To form scaffoldfree cartilage sheets, 50-100 million chondrocytes were injected into a 6 x 7 cm bioreactor (composed of two polystyrene gas-permeable membranes) and cultured in serum-free medium containing ITS,1 mM sodium pyruvate, 50 µM ascorbate-2-phosphate, 10 -7 M dexamethasone and non-essential amino acids, and cultured for 3 weeks.
CARTILAGE, 2012
Objective: To develop a tissue culture expansion method for rabbit chondrocytes that promotes rob... more Objective: To develop a tissue culture expansion method for rabbit chondrocytes that promotes robust expansion while preserving chondrogenic potential. Design: Rabbit chondrocytes isolated from articular or auricular chondrocytes were assessed for chondrogenic differentiation potential versus population doubling using different expansion and differentiation conditions. Expansion conditions included serum alone, serum plus basic fibroblast growth factor 2 (FGF-2), and serum plus insulin-like growth factor 1 (IGF-1) and FGF-2. Differentiation conditions consisted of defined medium with and without bone morphogenetic protein 2 (BMP-2). Results: Nonsupplemented chondrocytes showed limited expandability, whereas supplementation with FGF-2 allowed articular chondrocytes to be expanded past 10 population doublings (PDs) and allowed auricular chondrocytes to expand past 15 population doublings. Differentiation, as measured by glycosaminoglycan production in aggregate cultures, was minimal i...
Osteoarthritis and Cartilage, 2007
The purpose of this study is to develop and test a method to fabricate sheets of cartilage that c... more The purpose of this study is to develop and test a method to fabricate sheets of cartilage that can be used to produce replacement cartilage in a rabbit trachea model. Methods and Materials: Auricular cartilage was harvested from white New Zealand Rabbits sequentially digested with trypsin, testicular hyaluronidase and collagenase type II, passed through a 70 µm nitex trypsinized stored in liquid nitrogen until needed. To form scaffoldfree cartilage sheets, 50-100 million chondrocytes were injected into a 6 x 7 cm bioreactor (composed of two polystyrene gas-permeable membranes) and cultured in serum-free medium containing ITS,1 mM sodium pyruvate, 50 µM ascorbate-2-phosphate, 10-7 M dexamethasone and non-essential amino acids, and cultured for 3 weeks. Results: The results showed that the implanted cartilage survived intact, integrated well with the surrounding fascia, which was well vascularized, and had a well-formed lumen of 4-5 mm in diameter, and there was no sign of an immune reaction even in tracheas formed from allo-cartilage sheets. The cartilage stained positively for collagens II and X, much like the native auricular cartilage, but showed no signs of mineralization. Conclusions: These results show that chondrocytes sheets can be used to fabricate vascularized neo-trachea that retains a lumen and may be used as a model to repair segmental defects in rabbits.
Rheumatology (Sunnyvale, Calif.), Jan 26, 2012
Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory r... more Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory response pathways in arthritic joints where inflammation may be accompanied by the loss of chondrocyte vitality via apoptosis. To address this possibility in vitro, chondrocyte cultures were employed to determine the extent to which monosodium urate and recombinant TNF-α altered the frequency of apoptotic chondrocytes. Apoptosis as a function of the activation of p38 kinase, C-Jun-terminal kinase, signal transducer and activator of transcription-3 and/or the activity of xanthine oxidase was also studied. Using normal human chondrocytes, monosodium urate or recombinant tumor necrosis factor-α increased the frequency of apoptosis and activity of xanthine oxidase. However, the xanthine oxidase-specific inhibitor, febuxostat, failed to blunt this response. Monosodium urate, tumor necrosis factor-α or the Janus kinase inhibitor, AG-490, increased the frequency of apoptotic nuclei in macroaggre...
Rheumatology (Sunnyvale, Calif.), Jan 26, 2012
Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory r... more Monosodium urate and tumor necrosis factor-α, are two potent mediators of separate inflammatory response pathways in arthritic joints where inflammation may be accompanied by the loss of chondrocyte vitality via apoptosis. To address this possibility in vitro, chondrocyte cultures were employed to determine the extent to which monosodium urate and recombinant TNF-α altered the frequency of apoptotic chondrocytes. Apoptosis as a function of the activation of p38 kinase, C-Jun-terminal kinase, signal transducer and activator of transcription-3 and/or the activity of xanthine oxidase was also studied. Using normal human chondrocytes, monosodium urate or recombinant tumor necrosis factor-α increased the frequency of apoptosis and activity of xanthine oxidase. However, the xanthine oxidase-specific inhibitor, febuxostat, failed to blunt this response. Monosodium urate, tumor necrosis factor-α or the Janus kinase inhibitor, AG-490, increased the frequency of apoptotic nuclei in macroaggre...
Tissue Engineering Part A, 2010
To determine whether low oxygen (O 2 ) tension during expansion affects the matrix density, as we... more To determine whether low oxygen (O 2 ) tension during expansion affects the matrix density, as well as quantity, of cartilage formed, and to determine whether application of low O 2 tension during incubation periods alone is sufficient to modulate chondrogenic expression, rabbit chondrocytes expanded at either 21% O 2 or 5% O 2 were analyzed for glycosaminoglycan (GAG) and DNA content, total collagen, and gene expression during expansion and postexpansion aggregate cultures. When cultured as aggregates at 21% O 2 , chondrocytes expanded at 5% O 2 produced cartilage aggregates that contained more total GAG, GAG per wet weight, GAG per DNA, and total collagen than chondrocytes expanded at 21% O 2 . Less of an effect on GAG and collagen content was observed when aggregate culture was performed at 5% O 2 . Real-time polymerase chain reaction analysis of COL2A1 expression showed upregulated levels of type IIA (an early marker) and IIB (a late marker) during expansion and elevated levels of type IIB during aggregate culture in chondrocytes expanded in low O 2 . The application of low O 2 tension during incubation periods of chondrocyte expansion enhances the ultimate cartilage matrix density and quantity, and this enhancement can be achieved through the use of an O 2 control incubator.
Osteoarthritis and Cartilage, 2007
Purpose: The purpose of this study is to develop and test a method to fabricate sheets of cartila... more Purpose: The purpose of this study is to develop and test a method to fabricate sheets of cartilage that can be used to produce replacement cartilage in a rabbit trachea model. Methods and Materials: Auricular cartilage was harvested from white New Zealand Rabbits sequentially digested with trypsin, testicular hyaluronidase and collagenase type II, passed through a 70 µm nitex trypsinized stored in liquid nitrogen until needed. To form scaffoldfree cartilage sheets, 50-100 million chondrocytes were injected into a 6 x 7 cm bioreactor (composed of two polystyrene gas-permeable membranes) and cultured in serum-free medium containing ITS,1 mM sodium pyruvate, 50 µM ascorbate-2-phosphate, 10 -7 M dexamethasone and non-essential amino acids, and cultured for 3 weeks.