The importance of
multi-omic-based approaches to better understand
diverse pathological mechanisms including neurodegenerative diseases
has emerged. Spatial information can be of great help in understanding
how biomolecules interact pathologically and in elucidating target
biomarkers for developing therapeutics. While various analytical methods
have been attempted for imaging-based biomolecule analysis, a multi-omic
approach to imaging remains challenging due to the different characteristics
of biomolecules. Time-of-flight secondary ion mass spectrometry (ToF-SIMS)
is a powerful tool due to its sensitivity, chemical specificity, and
high spatial resolution in visualizing chemical information in cells
and tissues. In this paper, we suggest a new strategy to simultaneously
obtain the spatial information of various kinds of biomolecules that
includes both labeled and label-free approaches using ToF-SIMS. The
enzyme-assisted labeling strategy for the targets of interest enables
the sensitive and specific imaging of large molecules such as peptides,
proteins, and mRNA, a task that has been, to date, difficult for any
MS analysis. Together with the strength of the analytical performance
of ToF-SIMS in the label-free tissue imaging of small biomolecules,
the proposed strategy allows one to simultaneously obtain integrated
information of spatial distribution of metabolites, lipids, peptides,
proteins, and mRNA at a high resolution in a single measurement. As
part of the suggested strategy, we present a sample preparation method
suitable for MS imaging. Because a comprehensive method to examine
the spatial distribution of multiple biomolecules in tissues has remained
elusive, our strategy can be a useful tool to support the understanding
of the interactions of biomolecules in tissues as well as pathological
mechanisms.
In this study, we designed bisphosphonate-conjugated polyanionic hyaluronic acid (HA) microbeads (MBs) for the controlled delivery of bone morphogenetic protein 2 (BMP2). MBs were prepared via the photo-crosslinking of bisphosphonate (alendronate)-conjugated methacrylated HA (Alen-MHA). The polyanionic Alen-MHA MBs actively absorbed cationic BMP2 up to 91.0% of the loading efficacy and displayed a sustained release of BMP2 for 10 days. BMP2/Alen-MHA MBs induced osteogenicrelated genes in cellular experiments and showed the highly increased bone formation efficacy in thigh muscle injection and rat spinal fusion animal models. Thus, BMP2/Alen-MHA MBs provide a promising opportunity to improve the delivery efficiency of BMP2.
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