Funktionalisierung von PLLA-Nanofasern mittels Kollagen Typ 1 und BMP-2 – Einfluss auf die osteogene Differenzierung von mesenchymalen Stammzellen

Neuartige Knochenersatzmaterialien gewinnen in der Orthopädie und der Mund-Kiefer-Gesichtschirurgie einen immer höheren Stellenwert bei der Be-handlung von unüberbrückbaren Knochendefekten. Standard bei der Knochentransplantation ist der Einsatz von autogener Spongiosa aus dem Beckenkamm. Die darübe...

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Bibliographic Details
Main Author: Veltum, Anne
Contributors: Schofer, Markus (Prof. Dr.) (Thesis advisor)
Format: Doctoral Thesis
Published: Philipps-Universität Marburg 2012
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New bone substitute materials take a great role in the treatment of bone defects in orthopedic, oral and maxillofacial surgery. Standard procedure for bone grafting is the use of autogenous cancellous bone from the iliac crest. Synthetic bone replacement materials consist of metals, polymers, ceramics and composites, and should require a three-dimensional skeleton and are, if at all, mixed with growth factors and stem cells. These are the properties of tissue engineerung, which polymeric material is often used for. For example, spun nanofibers of polymers show several advantages. These imitate the extracellular matrix, which stimulates the cultured cells to differentiate, migrate and proliferate. The biological effect of this stimulation depends on the physical and chemical properties of the fibers. In this context, fibers that were spun from PLLA, after cultivation with mesenchymal stem cells, possess a high cell density and a low cell differentiation. A better cell differentiation is obtained by functionalizing of the cells with collagen type 1 or the growth factor BMP-2. Objective of this dissertation is to show a possible cooperative effect between type 1 collagen and BMP-2. For this purpose, scaffolds made of PLLA, PLLA-coll1-blend and PLLA-coll1-blend-BMP-2 were cultured with human mesenchymal stem cells under growth and osteoinductive conditions for 22 days. In the course of this process the gene expression of alkaline phosphatase, osteocalcin, and collagen type 1 was determined. In addition, the gene expression of the signal transduction molecules FAK and Smad 5 was analyzed. The cell density and the deposition of calcium and collagen type 1 in the extracellular space were shown by fluorescence microscopy. The insertion of BMP-2 in the PLLA-coll1-blend fiber caused a decrease in the diameter and the pore size. The BMP-2 containing scaffolds showed a better adhesion but a lower proliferation of mesenchymal stem cells. In addition, the cells on these scaffolds showed an increase in gene expression of alkaline phosphatase, osteocalcin and collagen type 1. The fluorescene microscopy showed a higher deposition of collagen type 1 and calcium. The presence of BMP-2 in the fibers resulted in an upregulation of gene expression of FAK. The influence of collagen type 1 revealed the gene regulation primarily in up-regulation of Smad 5. This means that by cultivating mesenchymal stem cells on PLLA fibers a combined effect of BMP-2 and collagen type 1 can be realized. This activates different signaling pathways leading to an increasing differentiation of stem cells. Thus the scaffold can be used for tissue engineering.