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  • A novel method to apply osteogenic potential of adipose derived stem cells in orthopaedic surgery.

A novel method to apply osteogenic potential of adipose derived stem cells in orthopaedic surgery.

PloS one (2014-03-04)
Xiang Fang, Hideki Murakami, Satoru Demura, Katsuhiro Hayashi, Hidenori Matsubara, Satoshi Kato, Katsuhito Yoshioka, Kei Inoue, Takashi Ota, Kazuya Shinmura, Hiroyuki Tsuchiya
ABSTRACT

A number of publications have reported that adipose derived stem cells (ADSCs) have the capacity to be induced to differentiate into osteoblasts both in vitro and in vivo. However, it has been difficult to use separate ADSCs for cortical bone regeneration and bone reconstruction so far. Inspired by the research around stromal stem cells and cell sheets, we developed a new method to fabricate ADSCs sheets to accelerate and enhance the bone regeneration and bone reconstruction. To fabricate ADSCs sheets and evaluate their capacity to be induced to differentiate to osteoblasts in vitro. Human adipose derived stem cells (hADSCs) were employed in this research. The fabricating medium containing 50 µM ascorbate-2-phosphate was used to enhance the secretion of collagen protein by the ADSCs and thus to make the cell sheets of ADSCs. As the separate ADSCs were divided into osteo-induction group and control group, the ADSCs sheets were also divided into two groups depending on induction by osteogenesis medium or no induction. The osteogenic capacity of each group was evaluated by ALP staining, Alizarin Red staining and ALP activity. The ADSCs sheets were fabricated after one-week culture in the fabricating medium. The ALP staining of ADSCs sheets showed positive results after 5 days osteo-induction and the Alizarin Red staining of ADSCs sheets showed positive results after 1 week osteo-induction. The ALP activity showed significant differences between these four groups. The ALP activity of ADSCs sheets groups showed higher value than that of separate ADSCs. The experiments demonstrated that ADSCs sheets have better capacity than separate ADSCs to be induced to differentiate into osteoblasts. This indicates that it is possible to use the ADSCs sheets as a source of mesenchymal stem cells for bone regeneration and bone reconstruction.

MATERIALS
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Sigma-Aldrich
Osteogenesis Quantitation Kit, The Osteogenesis Quantitation Kit provides all the necessary reagents & a protocol to enable quantification of osteogenesis using a standard plate reader.