The Effect of Electromagnetic Field Stimulation on Chondrogenic Differentiation of Human Adipose-Derived Stem Cells

• Main Authors: Yi-Shan Lin, 林怡珊
• Other Authors: Mei-Ling Ho
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/67357410116116932725

碩士 === 高雄醫學大學 === 生理及分子醫學研究所 === 97 === Articular cartilage defect is usually a consequence of mechanical or biological damage that destabilizes tissue homeostasis in cartilage. Cell-based tissue engineering has been indicated to be one of the ideal treatments to regenerate articular cartilage. Adipose derived stem cells (ADSCs) were reported to be obtained with relatively little discomforts and lower donor site morbidity, and can be expanded in larger number in vitro. The well-known factors for inducing chondrogenesis of mesenchymal stem cells are transforming growth factor-βs (TGF-βs), etc. The pulse electromagnetic field (PEMF) has been approved by FDA, USA for treating non-union bone fracture. Several studies have been reported that PEMF also increased proliferation and proteoglycan synthesis in human chondrocytes. Furthermore, PEMF stimulation was indicated to up-regulated TGF-β and also had a chondroprotective effect during osteoarthritis progression in the knee joint of guinea pigs. However, it remains unclear whether PEMF affects chondrogenic differentiation of human adipose-derived stem cells (hADSCs) for regenerating articular cartilage. In this study, PEMF and an innovative single pulsed electromagnetic field (SPEMF) stimulation are hypothesized to enhance chondrogenic differentiation of hADSCs. We investigated whether PEMF or SPEMF can further enhance the chondrogenesis in hADSCs culture where there the environments are favor for chondrogenesis including pellet cultured system and hyaluronan -coated dishes, without the induction of chondrogenesis. The stimulation module of PEMF was pulsed period 5 ms repeated in 15 Hz, magnetic magnitude in 20 Gauss, stimulated for 8 hrs per day.Module of SPEMF was magnitude in 0.8 Tesla (8*103 Gauss) per pulse, pulsed period 5 ms for 30 times per day. The daily stimulation period was less than 3 min. The chondrogenesis in hADSCs were evaluated by Q-PCR to quantify mRNA expressions of chondrogeneic related genes by and by DMMB assay and Alcian blue stain to quantify glycosaminoglycans (GAGs) synthesis. The results showed that in both pellet cultured system and hyaluronan-coated dishes, the expressions of chondrogenic marker genes including, SOX-9, type II collagen and aggrecan, of hADSCs were significantly increased on day 1, 3, 5, and 7 upon PEMF or SPEMF treatment in comparison with non-treated control cultured. This effect was not significantly difference among PEMF, SPEMF, and chondroinduction groups. For hADSCs cultured in pellet cultured system and hyaluronan-coated dishes, the GAG formed significantly more than that of control cultures after 10 days of either PEMF or SPEMF. This chondrogeneic effect was also similar to that in the chondroinduction medium Our results also showed that the chondrogeneic related gene expressions on hADSCs were similar in the pellet cultured system by PEMF or SPEMF stimulation in the hyaluronan-coated dishes. More importantly, there was no obviously enhanced expressions of osteogenic marker genes including, osteocalcin and collagen type I by SPEMF treatments on hADSCs in pellet cultured system or in the hyaluronan-coated dishes, while PEMF stimulation significantly increased osteoclcin expression (p<0.01). From these results, we suggest that SPEMF might be an alternative method to enhance chondrogeneic differentiation of hADSCs for articular cartilage regeneration.