Analysis and modification of biosilica-forming sponge (Suberites domuncula) primmorph cells and hydroxyapatite-forming human SaOS-2 cells
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Abstract
The marine sponge Suberites domuncula (Demospongiae) produces biosilica spicules by enzymatically controlled biomineralization and is a potential natural source for bioactive sponge compounds. S. domuncula cell culture is possible by generating three‐dimensional cell aggregates termed primmorphs.
In this thesis, as an initial step, a comparative transcriptome analysis was performed. A new bioreactor for S. domuncula cell culture was developed. It yields a significant improvement and upscaling of primmorph production. With different analytical approaches it was shown that most if not all primmorph cells engulfed large numbers of nanoparticles and microparticles, respectively. By quantitative PCR, it was determined that uptake of silica core-shell microparticles enhanced the expression of proteins involved in spiculogenesis. It was also found that the biosilica spicules were specifically coated by nano‐screenMAG‐CMX nanoparticles in vivo, whereas such coating of extracted spicules in vitro required prior chemical silanization. Magnetic and fluorescent properties to primmorph individuals, their cells, and their spicules were introduced by using nanoparticles.
Human bone-forming SaOS‐2 cells with the gene encoding S. domuncula silicatein‐α were stably transfected, which significantly enhanced their bone‐forming capacity.
The present experiments with sponge cells and human SaOS‐2 cells open new possibilities in biology, material science and biomedicine in the context of research, production, modification and application of biosilica.