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| Repairing Larynx Cartilage Defects with Prefabricated PLAG Scaffold Compounded with BMSCs Cell Sheets Transfected by GDF5 |
| YAO Mei1, CUI Ying2, HU Jing1, LI Jun-xia1, WANG Yu1 |
1. The Graduate School of Liaoning Medical College, Jinzhou 121000, China;
2. The First Affiliated Hospital of Liaoning Medical College, Jinzhou 121000, China |
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Abstract Cell sheet technology is a new skill for cell harvesting and transferring in medicine tissue engineering, which makes the cultured cells separate from the dish surface to form a layer of complete lamellar structure containing extracellular matrix. It has circumvent these limitations, such as insufficient biological activity, low cell-attachment efficiency, which being in traditional tissue engineering. So a compound which was constituted with mesenchymal stem cell sheet transfected with GDF5 by adenovirus vector and PLGA scaffold prefabricated with GDF5 gene-transferred BMSCs was structured, explore the clinical application and make a preliminary assessment on the repairing effect.Isolated and cultured rabbit bone marrow mesenchymal stem cells in vitro, tansfected the passage 4 cells with exogenous recombinant growth differentiation factor 5 by adenovirus vector. Harvested the transgenic bone marrow mesenchymal stem cell sheet through temperature-responsive culture dish after culturing 7~14 days. Using real time PCR to detect the express of GDF5 in transgenic cell sheet. Preparing a prefabricated PLAG scaffold through injecting GDF5 gene-transferred BMSCs to PLGA scaffold, then co-culture the compound constituted with the cell sheet and prefabricated PLGA scaffold for 2 days. Transplant the compound to rabbit larynx cartilage defects and discuss the repair capacity after 4 or 8 weeks at the gross level as well as at the histological level. The experiment divided into three groups:(A)the compound constituted with the transgenic cell sheet and prefabricated PLGA scaffold;(B)prefabricated PLGA scaffold;(C)PLGA scaffold loaded BMSCs. The Results showed the whole transgenic cell sheet have been harvested successfully, real time PCR can detect the express of GDF5 in transgenosis cell sheets. The results of immunohistochemistry and Alcian Blue staining showed the expression of collagen Ⅱ and GAG in group A and group B, furthermore, there was more collagen Ⅱ and GAG in group A than that in group B with statistical difference (P<0.05).Thus, the compound constituted with the transgenosis BMSCs cell sheet and PLGA scaffold prefabricated with transgenosis BMSCs has the more superior chondrogenic capacity than traditional PLGA scaffold loaded with cells, it can promote the repair of cartilage effectively.
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Received: 15 November 2013
Published: 25 March 2014
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