and provided surfaces that significantly enhanced the MSC phenotype during cell population expansion phases all associated with the defined and controlled contextual presentation of the underlying chemistry.
Keywords. Stem cells, cell adhesion, nano-arrays and silane modification
(48.O6) CHONDROGENIC AND OSTEOGENIC
DIFFERENTIATION OF HMSCS CULTURED ON 3D PEOT/PBT SCAFFOLDS
Leferink AM (1), Karperien M (1), Blitterswijk CA van (1), Moroni L (1)
1. Department of Tissue Regeneration, University of Twente
Recent approaches to regenerate tissues involve the use of human mesenchymcal stem cells (hMSC) either alone or in combination with soluble factors and/or three-dimensional (3D) scaffolds. It is known that scaffold material properties can influence the behavior of hMSCs and their fate in vitro. Less is known about the influence of scaffolds on retaining the multipotency of hMSCs. This study focuses on the differentiation potential of hMSC cultured on rapid prototyped 3D scaffolds towards osteogenic and chondrogenic lineages. When cell fate can be controlled with soluble factors only, the culture system could be transferred to a perfusion bioreactor in which the metabolic activity of the different cell state and the resulting extra cellular matrix (ECM) formation can be monitored real-time. Ultimately, a better understanding of cell metabolic state and ECM production could lead to a model predicting tissue development and to the design of instructive properties in 3D scaffolds.
First hMSCs were seeded on PEOT/PBT scaffolds and cultured for 7 days in expansion medium. Subsequently the expansion medium was replaced with basic or osteogenic and chondrogenic differentiation medium. Biochemical, histological and molecular assays were performed after 7 days and 21 days of differentiation. hMSCs cultured in chondrogenic medium showed a significant increase in glycosaminoglycans (GAGs) production and a significant up-regulation of Sox9, collagen type II and aggrecan gene expression after 21 days. Cells cultured in osteogenic differentiation medium showed a significant increase in alkaline phosphatase (ALP) production and up-regulation of ALP and collagen type I gene expression after respectively 7 and 21 days. No considerable morphological differences were found between the cells of both groups. The produced ECM appeared of fibrillar nature as observed by scanning electron microscopy. Future studies will be performed to further understand ECM formation and development during hMSCs-based cartilage and bone regeneration in 3D scaffolds.
Keywords. Differentiation hMSCs chondrogenic osteogenic
(48.O7) IMPROVEMENT OF VOCAL FOLD WOUND HEALING BY BONE MARROW-DERIVED STEM CELLS USING A XENOGRAFT ANIMAL MODEL
Lim JY (1), Kwon SK (2), Kim HS (3), Yi T (4), Song SU (4), Kim YM (1)
1. Inha University, Department of Otorhinolaryngology; 2. Dongguk University Ilsan Hospital; 3. Ewha University; 4. Inha University
Introduction. Vocal fold scarring can be caused by trauma, inflammation, or surgery and commonly result in severe dysphonia. Cell based therapies to minimize scarring and enhance healing have been growing interest. This study was aimed to investigate the effect of bone-marrow derived stem cells (BMSCs) for prevention of vocal fold scarring by the use of xenograft animal model. Methods. Vocal fold scarring was induced in New Zealand white rabbits by a direct injury. BMSCs were isolated from bone marrow aspirates of GFP transgenic mice and were injected into the vocal folds of rabbits immediately after scarring. PBS was injected into the vocal folds in the same manner for the sham group. Endoscopic, histologic, and biomechanical evaluations of vocal folds were performed after 2 months of the injection.
Results. The BMSCs-treated vocal folds showed decreased collagen bundles compared to the sham group and the relative hyaluronic acid content in BMSCs-treated vocal folds was higher than the sham group. The BMSCs reduced expression of procollgen I. The mean dynamic viscosity in the BMSCs was improved compared to the sham group. A large number of the injected BMSCs were detected in the vocal folds of rabbits at 2 months after injection.
Conclusion. This study suggests that the BMSCs could prevent the scarring-induced dysphonia and be developed as a cell-based therapy for vocal folds regeneration. This xenograft animal model also might be a useful tool for research of vocal fold regeneration. Keywords. vocal fold, scarring, mesenchymal stem cells (48.O8) OPTIMISING EXTRACELLULAR MATRIX COATINGS FOR MSC ATTACHMENT AND PROLIFERATION IN SERUM-FREE CULTURE CONDITIONS
Mooney E (1), Ansboro S (2), Shaw G (1), Coleman C (1), Barron V (1), Barry F (1), Murphy M (1),
1. PurStem, REMEDI, NCBES, National University of Ireland, Galway, Ireland; 2. REMEDI, NCBES, NAtional University of Ireland, Galway, Ireland
Therapeutic protocols using human mesenchymal stem cells (MSC) are being tested clinically for a number of disorders including osteoarthritis, bone fractures,
