Gene therapy for meniscal injury: Enhanced synthesis of proteoglycan and collagen by meniscal cells transduced with a TGFβ1 gene

H. Goto, F. D. Shuler, Christopher Niyibizi, F. H. Fu, P. D. Robbins, C. H. Evans

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Objective: To determine whether meniscal cells can express a TGFβ1 transgene delivered by a retroviral vector, and respond to the gene product by increasing matrix synthesis. Methods: Monolayer cultures of human and canine meniscal cells were infected with retroviruses carrying either a human TGFβ1 cDNA or marker genes. Conditioned media were assayed for the presence of TGFβ1. Biosynthesis assays using radiolabeled precursors were employed to determine the effects of the transgenes on the synthesis of proteoglycan, collagen and noncollagenous proteins. Collagen phenotyping was performed by SDS-PAGE. Results: Media conditioned by canine and human meniscal cells transduced with the TGFβ1 gene, accumulated several nanograms/106 cells of TGFβ1 during a 48 h incubation. Media conditioned by control cells contained very little TGFβ1. Transduction with the TGFβ1 gene, but not marker genes, increased the synthesis of collagen and proteoglycan by 8-15-fold. The synthesis of noncollagenous proteins was enhanced more modestly. Monolayers of meniscal cells synthesized types I, III, V and VI collagen. The TGFβ1 gene increased the synthesis of all types of collagen without altering the ratios between them. Conclusions: Meniscal cells are readily transduced by retroviral vectors and respond to the transfer of a TGFβ1 cDNA by greatly increasing matrix synthesis. These findings encourage the further development of genetic approaches to the healing of meniscal lesions. (C) 2000 OsteoArthritis Research Society International.

Original languageEnglish (US)
Pages (from-to)266-271
Number of pages6
JournalOsteoarthritis and Cartilage
Volume8
Issue number4
DOIs
StatePublished - Jan 1 2000

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Gene therapy
Proteoglycans
Collagen
Genetic Therapy
Genes
Wounds and Injuries
Conditioned Culture Medium
Transgenes
Monolayers
Canidae
Proteins
Complementary DNA
Collagen Type VI
Biosynthesis
Collagen Type III
Retroviridae
Collagen Type I
Assays
Cells
Polyacrylamide Gel Electrophoresis

All Science Journal Classification (ASJC) codes

  • Rheumatology
  • Biomedical Engineering
  • Orthopedics and Sports Medicine

Cite this

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title = "Gene therapy for meniscal injury: Enhanced synthesis of proteoglycan and collagen by meniscal cells transduced with a TGFβ1 gene",
abstract = "Objective: To determine whether meniscal cells can express a TGFβ1 transgene delivered by a retroviral vector, and respond to the gene product by increasing matrix synthesis. Methods: Monolayer cultures of human and canine meniscal cells were infected with retroviruses carrying either a human TGFβ1 cDNA or marker genes. Conditioned media were assayed for the presence of TGFβ1. Biosynthesis assays using radiolabeled precursors were employed to determine the effects of the transgenes on the synthesis of proteoglycan, collagen and noncollagenous proteins. Collagen phenotyping was performed by SDS-PAGE. Results: Media conditioned by canine and human meniscal cells transduced with the TGFβ1 gene, accumulated several nanograms/106 cells of TGFβ1 during a 48 h incubation. Media conditioned by control cells contained very little TGFβ1. Transduction with the TGFβ1 gene, but not marker genes, increased the synthesis of collagen and proteoglycan by 8-15-fold. The synthesis of noncollagenous proteins was enhanced more modestly. Monolayers of meniscal cells synthesized types I, III, V and VI collagen. The TGFβ1 gene increased the synthesis of all types of collagen without altering the ratios between them. Conclusions: Meniscal cells are readily transduced by retroviral vectors and respond to the transfer of a TGFβ1 cDNA by greatly increasing matrix synthesis. These findings encourage the further development of genetic approaches to the healing of meniscal lesions. (C) 2000 OsteoArthritis Research Society International.",
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Gene therapy for meniscal injury : Enhanced synthesis of proteoglycan and collagen by meniscal cells transduced with a TGFβ1 gene. / Goto, H.; Shuler, F. D.; Niyibizi, Christopher; Fu, F. H.; Robbins, P. D.; Evans, C. H.

In: Osteoarthritis and Cartilage, Vol. 8, No. 4, 01.01.2000, p. 266-271.

Research output: Contribution to journalArticle

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T2 - Enhanced synthesis of proteoglycan and collagen by meniscal cells transduced with a TGFβ1 gene

AU - Goto, H.

AU - Shuler, F. D.

AU - Niyibizi, Christopher

AU - Fu, F. H.

AU - Robbins, P. D.

AU - Evans, C. H.

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Y1 - 2000/1/1

N2 - Objective: To determine whether meniscal cells can express a TGFβ1 transgene delivered by a retroviral vector, and respond to the gene product by increasing matrix synthesis. Methods: Monolayer cultures of human and canine meniscal cells were infected with retroviruses carrying either a human TGFβ1 cDNA or marker genes. Conditioned media were assayed for the presence of TGFβ1. Biosynthesis assays using radiolabeled precursors were employed to determine the effects of the transgenes on the synthesis of proteoglycan, collagen and noncollagenous proteins. Collagen phenotyping was performed by SDS-PAGE. Results: Media conditioned by canine and human meniscal cells transduced with the TGFβ1 gene, accumulated several nanograms/106 cells of TGFβ1 during a 48 h incubation. Media conditioned by control cells contained very little TGFβ1. Transduction with the TGFβ1 gene, but not marker genes, increased the synthesis of collagen and proteoglycan by 8-15-fold. The synthesis of noncollagenous proteins was enhanced more modestly. Monolayers of meniscal cells synthesized types I, III, V and VI collagen. The TGFβ1 gene increased the synthesis of all types of collagen without altering the ratios between them. Conclusions: Meniscal cells are readily transduced by retroviral vectors and respond to the transfer of a TGFβ1 cDNA by greatly increasing matrix synthesis. These findings encourage the further development of genetic approaches to the healing of meniscal lesions. (C) 2000 OsteoArthritis Research Society International.

AB - Objective: To determine whether meniscal cells can express a TGFβ1 transgene delivered by a retroviral vector, and respond to the gene product by increasing matrix synthesis. Methods: Monolayer cultures of human and canine meniscal cells were infected with retroviruses carrying either a human TGFβ1 cDNA or marker genes. Conditioned media were assayed for the presence of TGFβ1. Biosynthesis assays using radiolabeled precursors were employed to determine the effects of the transgenes on the synthesis of proteoglycan, collagen and noncollagenous proteins. Collagen phenotyping was performed by SDS-PAGE. Results: Media conditioned by canine and human meniscal cells transduced with the TGFβ1 gene, accumulated several nanograms/106 cells of TGFβ1 during a 48 h incubation. Media conditioned by control cells contained very little TGFβ1. Transduction with the TGFβ1 gene, but not marker genes, increased the synthesis of collagen and proteoglycan by 8-15-fold. The synthesis of noncollagenous proteins was enhanced more modestly. Monolayers of meniscal cells synthesized types I, III, V and VI collagen. The TGFβ1 gene increased the synthesis of all types of collagen without altering the ratios between them. Conclusions: Meniscal cells are readily transduced by retroviral vectors and respond to the transfer of a TGFβ1 cDNA by greatly increasing matrix synthesis. These findings encourage the further development of genetic approaches to the healing of meniscal lesions. (C) 2000 OsteoArthritis Research Society International.

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