Chonnam Med J.  2002 Dec;38(4):396-402.

Transfection of Insulin-like Growth Factor II Gene into Human Anterior Cruciate Ligament-derived Fibroblasts

• Song EK
• Shim SD
• Kim MS
• Choi J

Abstract

Gene therapy using growth factors in the treatment of tissue injuries plays a very important role in improving the wound healing process and regeneration of such as bones, cartilages and ligaments. It is well known that the direct injection of IGF improves healing of injured ligaments. We analyzed the effects of transfection of the IGF-II gene into fibroblasts isolated from human anterior cruciate ligament (ACL). The tissue specimen excised under sterile conditions was treated with enzymes and cultured. The gene was obtained by ligating the insulin-like growth factor-II (IGF-II) gene into pcDNA3 and transfected using liposome. Co-transfection of pbeta- gal control vector was done in order to confirm the efficiency of the gene transfer. The type I collagen synthesis was measured with RT-PCR, proteoglycan synthesis with 1,9- dimethylene blue (DMB) and UV-spectrometer, and the amount of DNA, which is the index of fibroblast proliferation, with a microplate fluorescence reader. The results were compared with the control group. X-gal stain in the transfected group was about 20%. At the end of 2 weeks incubation, the number of cells increased to 12 X 10(5) in transfected group, compared 8 X 10(5) in the control group showing 50% faster increase. The cell shape also showed more linear phenomenon in the transfected group. In the transfected group, the synthesis of proteoglycan was increased by two folds, and the synthesis of type I collagen was also increased more than 1.5 times as revealed by a thicker band at 300 bp. The amount of DNA also increased by 1.6 folds in the transfected group. We confirmed that the transfrection of IGF-II gene promoted the proliferation of fibroblasts obtained from human ACL and synthesis of type I collagen and proteoglycan.

Keyword

Fibroblast; Gene therapy; IGF-II; Liposome

MeSH Terms

Anterior Cruciate Ligament
Cartilage
Cell Shape
Collagen Type I
DNA
Fibroblasts*
Fluorescence
Genetic Therapy
Humans*
Insulin-Like Growth Factor II*
Intercellular Signaling Peptides and Proteins
Ligaments
Liposomes
Proteoglycans
Regeneration
Transfection*
Wound Healing
Collagen Type I
DNA
Insulin-Like Growth Factor II
Intercellular Signaling Peptides and Proteins
Liposomes
Proteoglycans