Gene Delivery into Chondrocytes by Liposome

Song, Eun Kyoo; Hong, Keum Ran; Seol, Jong Yoon

Abstract

By transfectting genes of the IGF which increases the expression of type 2 collagen proper phenotype of chondrocyte and inducing the expression of genes, we intend to prevent the change of the expression which is the problem in culture of chondrocyte. By comparing the rate of chondrocyte proliferation and expression of type 2 collagen and proteoglycan, we are to take a look at the value of a hereafter study for joint cartilage regeneration of gene transfection method into chondrocyte using liposome. From the patient having articular cartilage defect, hyaline cartilage was collected aseptically. After pre-culture with RPMI-1640 culture solution for 2 weeks, pcDNA3-IGF-II gene was transferred into chondrocytes by mixing with lipid complex. To affirm the transfection, beta galactosidase was co-transfected with IGF-II and X-gal staining was used. Reverse transcription-polymerized chain reaction was used to compare the rate of synthesis of the type 2 collagen. Also, proteoglycan synthesis was compared by DMB (1,9-dimethyl- methylene) method. By X-gal staining, The absorbance at 420 nm was 0.45 in the control group, which was 1.095 in transfected group and 1.108 in positive control enzyme. In the synthesis of proteoglycan, The absorbance at 560 nm was 0.236 in control group, 0.605 in transfected group. In transfection group, the expression of type 2 collagen was significantly higher than in the control group. We confirmed that transfecting the IGF gene using liposomes to chondrocytes increased the proliferation of chondrocyte, synthesis of type II collagen and proteoglycan, suggesting that liposome may be useful in transfecting the IGF gene to chondrocytes.