期刊
BIOSENSORS & BIOELECTRONICS
卷 105, 期 -, 页码 151-158出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2018.01.018
关键词
Fluorescent probe; Near-infrared; Alkaline phosphatase; Three-dimensional scaffold; Bioimaging
类别
资金
- BioNano Health-Guard Research Center - Ministry of Science and ICT(MSIT) of Korea [H-GUARD_2014M3A6B2060489]
- National Research Council of Science & Technology - MSIT [CAP-15-09-KIMS]
- NRF (National Research Foundation of Korea) - Korean Government [2017H1A2A1042758]
- NRF (National Research Foundation of Korea) - Korean Government (NRF-Fostering Core Leaders of the Future Basic Science Program/Global Ph.D. Fellowship Program)
- KRIBB Initiative Research Program
- National Research Council of Science & Technology (NST), Republic of Korea [KGM1121824] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Foundation of Korea [2017H1A2A1042758] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Alkaline phosphatase (ALP) is a critical biological marker for osteoblast activity during early osteoblast differentiation, but few biologically compatible methods are available for its detection. Here, we describe the discovery of highly sensitive and rapidly responsive novel near-infrared (NIR) fluorescent probes (NIR-Phos-1, NIR-Phos-2) for the fluorescent detection of ALP. ALP cleaves the phosphate group from the NIR skeleton and substantially alters its photophysical properties, therefore generating a large turn-on fluorescent signal resulted from the catalytic hydrolysis on fluorogenic moiety. Our assay quantified ALP activity from 0 to 1.0 U mL(-1) with a 10(-5)-10(-3) U mL(-1) limit of detection (LOD), showing a response rate completed within 1.5 min. A potentially powerful approach to probe ALP activity in biological systems demonstrated real-time monitoring using both concentration- and time-dependent variations of endogenous ALP in live cells and animals. Based on high binding affinity to bone tissue of phosphate moiety, bone-like scaffold-based ALP detection in vivo was accessed using NIR probe-labeled three-dimensional (3D) calcium deficient hydroxyapatite (CDHA) scaffolds. They were subcutaneously implanted into mice and monitored ALP signal changes using a confocal imaging system. Our results suggest the possibility of early-stage ALP detection during neo-bone formation inside a bone defect, by in vivo fluorescent evaluation using 3D CDHA scaffolds.
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