Abstract
The menisci are intraarticular fibrocartilaginous structures essential to the normal function of the knee that lack the ability to self-repair. Human meniscal fibrochondrocytes may respond to beneficial genes like human insulin growth factor-1 (hIGF-1) and the meniscal cell may be a feasible donor for gene therapy. To explore this possibility, we amplified the hIGF-1 gene sequence in full length and cloned it into a bicistronic plasmid. This gene was then transfected into cultured human meniscal fibrochondrocytes by the liposome FuGene 6. Green fluorescence was expressed in part of the cells 6 hours after transfection and increased gradually, with a peak concentration of the hIGF-1 in the supernatants to 22.68 ng/mL 56 hours after transfection. Phenotypes of some cells changed and the proliferation accelerated after transfection. Flow cytometry analysis demonstrated upregulation of cell numbers in the G2 and S stages after hIGF-1 gene introduction. We conclude the hIGF-1 gene can be transfected into the human meniscal cell efficiently by liposome and it causes accelerated proliferation and differentiation. Within 10 days after transfection, the cytokine appears to be secreted into supernatants with the bioactivity and promotes the proliferation of the NIH 3T3 cell line.
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Acknowledgments
We thank Dr. Xiaokui Hou for assistance with the manuscript. We also thank Dr. Guanzhen Yang and Dr. Xiangfu Wu for technical assistance.
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Three of the authors (Hai-ning Zhang, Ping Leng and Ying-zhen Wang) received funding from the National Natural Science Foundation of China (No. 30801167).
Each author certifies that he or she has no commercial associations (eg, consultancies, stock ownership, equity interest, patent/licensing arrangements, etc) that might pose a conflict of interest in connection with the submitted article.
Each author certifies that his or her institution has approved the human protocol for this investigation, that all investigations were conducted in conformity with ethical principles of research, and that informed consent for participation in the study was obtained.
This work was performed at The Affiliated Hospital of Tsingtao University, China.
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Zhang, Hn., Leng, P., Wang, Yz. et al. Treating Human Meniscal Fibrochondrocytes with hIGF-1 Gene by Liposome. Clin Orthop Relat Res 467, 3175–3182 (2009). https://doi.org/10.1007/s11999-009-0870-2
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DOI: https://doi.org/10.1007/s11999-009-0870-2