Current Optics and Photonics

  • 제1권4호
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  • Pages.412-420
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  • 2017
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  • 2508-7266(pISSN)
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  • 2508-7274(eISSN)
한국광학회 (Optical Society of Korea)
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Label-free Noninvasive Characterization of Osteoclast Differentiation Using Raman Spectroscopy Coupled with Multivariate Analysis

  • Jung, Gyeong Bok (Department of Physics Education, Chosun University) ;
  • Kang, In Soon (Department of Pharmacology, Inha University School of Medicine) ;
  • Lee, Young Ju (Department of Biomedical Engineering & Healthcare Industry Research Institute, College of Medicine, Kyung Hee University) ;
  • Kim, Dohyun (Department of Industrial and Management Engineering, Myongji University) ;
  • Park, Hun-Kuk (Department of Biomedical Engineering & Healthcare Industry Research Institute, College of Medicine, Kyung Hee University) ;
  • Lee, Gi-Ja (Department of Biomedical Engineering & Healthcare Industry Research Institute, College of Medicine, Kyung Hee University) ;
  • Kim, Chaekyun (Department of Pharmacology, Inha University School of Medicine)
  • 투고 : 2016.12.22
  • 심사 : 2017.06.27
  • 발행 : 2017.08.25
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초록

Multinucleated bone resorptive osteoclasts differentiate from bone marrow-derived monocyte/macrophage precursor cells. During osteoclast differentiation, mononuclear pre-osteoclasts change their morphology and biochemical characteristics. In this study, Raman spectroscopy with multivariate techniques such as Principal Component Analysis (PCA) and Linear Discriminant Analysis (LDA) were used to extract biochemical information related to various cellular events during osteoclastogenesis. This technique allowed for label-free and noninvasive monitoring of differentiating cells, and clearly discriminated four different time points during osteoclast differentiation. The Raman band intensity showed significant time-dependent changes that increased up to day 4. The results of Raman spectroscopy agreed with results from atomic force microscopy (AFM) and tartrate-resistant acid phosphatase (TRAP) staining, a conventional biological assay. Under AFM, normal spindle-like mononuclear pre-osteoclasts became round and smaller at day 2 after treatment with a receptor activator of nuclear $factor-{\kappa}B$ ligand and they formed multinucleated giant cells at day 4. Thus, Raman spectroscopy, in combination with PCA-LDA, may be useful for noninvasive label-free quality assessment of cell status during osteoclast differentiation, enabling more efficient optimization of the bioprocesses.

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