Raman spectroscopy for grading of live osteosarcoma cells.
BACKGROUND
Osteosarcoma is the most common primary malignant bone tumor, and the grading of osteosarcoma cells relies on traditional histopathology and molecular biology methods, which require RNA extraction, protein isolation and immunohistological staining. All these methods require cell isolation, lysis or fixation, which is time-consuming and requires certain amount of tumor specimen. In this study, we report the use of Raman spectroscopy for grading of malignant osteosarcoma cells.


METHODS
We demonstrate that, based on the detection of differential production of mineral species, Raman spectroscopy can be used as a live cell analyzer to accurately assess the grades of osteosarcoma cells by evaluating their mineralization levels. Mineralization level was assessed by measuring amount of hydroxyapatite (HA), which is highly expressed in mature osteoblasts, but not in poorly differentiated osteosarcoma cell or mesenchymal stem cells, the putative cell-of-origin of osteosarcoma.


RESULTS
We found that under Raman spectroscopy, the level of HA production was high in MG-63 cells, which are low-grade. Moreover, hydroxyapatite production was low in high-grade osteosarcoma cells such as 143B and SaOS2 cells (p < 0.05). Matrix metalloproteinase MMP2, MMP9 were highly expressed in SaOS2, 143B and MSCs and decreased in human fetal osteoblast (FOB) and MG-63 cells as expected (p < 0.05). These results may highlight the inverse correlation between HA level and prognosis of osteosarcoma.


CONCLUSIONS
The use of Raman spectroscopy for the measurement of HA production by the protocol reported in this study may serve as a useful tool to rapidly and accurately assess the degree of malignancy in osteosarcoma cells in a label-free manner. Such application may shorten the period of pathological diagnosis and may benefit patients who are inflicted with osteosarcoma.
Institute of Clinical Medicine, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. chiang340@gmail.com.|Department of Orthopaedics, National Yang-Ming University Hospital, No. 152, Xinmin Road, Yi-Lan, 260, Taiwan. chiang340@gmail.com.
Yi-Hung
YH
Chiang
Institute of Biophotonics, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. stewart37@gmail.com.
Stewart H
SH
Wu
Institute of Clinical Medicine, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. carol9596@hotmail.com.
Yi-Chun
YC
Kuo
Institute of Biophotonics, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. hfchen3@ym.edu.tw.
How-Foo
HF
Chen
Institute of Biophotonics, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. aechiou@ym.edu.tw.
Arthur
A
Chiou
Institute of Clinical Medicine, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. DAV47@tpech.gov.tw.|Department of Orthopaedic Surgery, Taipei City Hospital, No. 145, Zhengzhou Road, Taipei, 10341, Taiwan. DAV47@tpech.gov.tw.|Stem Cell Research Center, National Yang-Ming University, No. 155, Sec2, Linong Street, Taipei, 112, Taiwan. DAV47@tpech.gov.tw.|Department of Medical Research, Taipei Veterans General Hospital, No. 201, Sec 2, Shipai Road, Taipei, 11217, Taiwan. DAV47@tpech.gov.tw.
Oscar K
OK
Lee
D051926:Basigin / Q000378:metabolism; D001859:Bone Neoplasms / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation; D002460:Cell Line; D017886:Durapatite / Q000737:chemistry*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000737:chemistry / Q000378:metabolism; D060787:Neoplasm Grading; D010006:Osteoblasts / Q000737:chemistry / Q000378:metabolism; D012516:Osteosarcoma / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D012313:RNA / Q000032:analysis / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013059:Spectrum Analysis, Raman
D051926:Basigin; D012313:RNA; D017886:Durapatite; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9