Optimalization of engineered bone constructs by cell selection, cell loading and cell culture (NGT.6052)

Tissue engineering is a recently developed approach to reconstruct and/or regenerate lost or damaged body tissues. In order to improve the cell-based approach of tissue engineering, three different strategies will be used i.e. cell selection, coatings and cell loading as well as culturing. For selection and expansion of the population of progenitor cells present in bone marrow, various techniques have been developed. However, the now used process of rat bone marrow cells takes a week and for human cells even longer. Therefore, methods must be found to select cells at an earlier time point. In literature, antibodies have been reported to be useful for the enrichment of osteoprogenitor cells (HOP-26, STRO-1 and ALP). However, the above-mentioned studies were used for identification of the different cell types present in a bone marrow aspirate. In this project, we use a new approach in which we want to apply the selected cells for direct seeding into scaffolds. Further, we want to determine the gene expression profiles of the enriched cell populations that we will obtain after selection. For that, we will use real time quantitative RT-PCR (QPCR) and cDNA microarray to identify the expression profiles of bone-specific genes in the different enriched cell populations. Further, ECM proteins are involved during cell adhesion as well as osteoblast differentiation. In view of this, lumican plays an important role in determining the structural phenotype of the mature collagen fibril in various tissues. Also, the surface chemistry of the scaffold material can have an effect on cell selection during cell seeding. In line of the abovementioned, we will use ECM proteins (mixed layered), sol-gel and calcium phosphate as coatings on titanium fiber mesh material to improve the cell seeding efficiency and osteogenic differentiation. We will focus on lumican, a new ECM protein, as coating to improve cell loading and differentiation. Finally, the cell loading method and/or nutrient conditions can be optimized by combining flow perfusion system with ECM proteins. We observed already that a dynamic seeding and culturing method (flow perfusion system) enhanced the proliferation and differentiation of osteogenic cells in titanium meshes. In this project we want to improve the bone construct by using ECM proteins coated meshes in the flow perfusion system.

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Er zijn vier bedrijven en één andere universiteit bij dit project betrokken.