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Biological reaction control using topography regulation of nanostructured titanium.

ABSTRACT: The micro- and nanosize surface topography of dental implants has been shown to affect the growth of surrounding cells. In this study, standardized and controlled periodic nanopatterns were fabricated with nanosized surface roughness on titanium substrates, and their influence on bone marrow stromal cells investigated. Cell proliferation assays revealed that the bare substrate with a 1.7?nm surface roughness has lower hydrophilicity but higher proliferation ability than that with a 0.6?nm surface roughness. Further, with the latter substrate, directional cell growth was observed for line and groove patterns with a width of 100?nm and a height of 50 or 100?nm, but not for those with a height of 10 or 25?nm. With the smooth substrate, time-lapse microscopic analyses showed that more than 80% of the bone marrow cells on the line and groove pattern with a height of 100?nm grew and divided along the lines. As the nanosized grain structure controls the cell proliferation rate and the nanosized line and groove structure (50-100?nm) controls cell migration, division, and growth orientation, these standardized nanosized titanium structures can be used to elucidate the mechanisms by which surface topography regulates tissue responses to biomaterials.

SUBMITTER: Shiozawa M

PROVIDER: S-EPMC7016147 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Biological reaction control using topography regulation of nanostructured titanium.

Shiozawa Mayuko M Takeuchi Haruka H Akiba Yosuke Y Eguchi Kaori K Akiba Nami N Aoyagi Yujin Y Nagasawa Masako M Kuwae Hiroyuki H Izumi Kenji K Uoshima Katsumi K Mizuno Jun J

Scientific reports 20200212 1

The micro- and nanosize surface topography of dental implants has been shown to affect the growth of surrounding cells. In this study, standardized and controlled periodic nanopatterns were fabricated with nanosized surface roughness on titanium substrates, and their influence on bone marrow stromal cells investigated. Cell proliferation assays revealed that the bare substrate with a 1.7 nm surface roughness has lower hydrophilicity but higher proliferation ability than that with a 0.6 nm surfac ...[more]

PMID: 32051472

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Project description:Rational: Titanium is the most widely used alloy family in dental and orthopedic implants due to its natural ability to integrate into bone, but cytotoxic alloying elements are required in titanium for its mechanical properties to match the functionality of natural bone in high-load bearing applications. Recent advances in nanostructuring, such as Continuous Equal Channel Angular Pressing (C-ECAP), reduce the grain size and increase the strength of pure grades of titanium, thereby eliminating the need for cytotoxic elements and increasing cytocompatibilty as measured by traditional cell biology techniques. Transcriptomic profiling of cells grown on conventional coarse grain (CG) versus nanostructured ultrafine grain (UG) surfaces can simultaneously enhance our understanding of genomics and biomaterials, facilitating the development of a new generation of implantable materials. Objective: We initiated this pilot project to develop a workflow for transcriptional profiling of both protein coding and non-coding RNAs (ncRNAs) in cells grown on Ti discs that are characterized for grain size, 12.1 μm for CG Ti and 0.28 μm for UG Ti. Methods: Two Ti substrates were designed for cell growth in 24-well culture dishes, CG and UG. Mouse pre-osteoblasts were cultured on Ti disc for 72 hours. Cells were harvested and total-RNA extracted. Ribosomal RNA subtraction was performed prior to Illumina sequencing library construction. Single-end 50 base pairs reads were subjected to Illumina quality control and the resulting reads were mapped to the mouse genome and annotations with Genomic Short-read Nucleotide Alignment Program (GSNAP) and HTseq-count. Read count files were normalized and statistically analyzed with empirical analysis of digital gene expression data in R (EdgeR) and filtered for low expression levels using Zipf’s Power Law. Transcripts with adequate coverage and significant expression differences were subjected to GO Biological Process term enrichment and network analyses with the Cytoscape App ClueGO. Results: Gene expression differences with an FDR≤0.05 are limited to six genes, five of which are likely ncRNAs. Of the 432 transcripts with significant differences at p≤0.05 117 form an interconnected network of genes and 12 GO biological processes (P(adj) ≤ 0.05). Biological processes that connect to predominately upregulated genes include cell division, mRNA metabolic processes, chromatin modification and protein localization to the cytoskeleton, while downregulated processes include regulation of apoptosis, of p38MAK cascade, and response to molecules of bacterial origin. Conclusions: After 72 hours, the transcriptional profile of cells on UG Ti confirms more rapid cell division than on CG. These results demonstrate that informative transcriptional profiling of cells growing on substrates that differ only in their crystallographic structure can be accomplished by a workflow that includes standard cell biology techniques, transcriptomic profiling, data mining, and network analyses.

Dataset Information

Biological reaction control using topography regulation of nanostructured titanium.

Publications

Biological reaction control using topography regulation of nanostructured titanium.

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