Optical imaging of mouse articular cartilage using the glycosaminoglycans binding property of fluorescent-labeled octaarginine

K. Inagawa, Toshitaka Oohashi, Keiichiro Nishida, J. Minaguchi, T. Tsubakishita, K. O. Yaykasli, Aiji Ohtsuka, Toshihumi Ozaki, T. Moriguchi, Y. Ninomiya

Research output: Contribution to journalArticle

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Abstract

Objective: The aim of the current study was to examine the cartilage-specific binding property of polyarginine peptides (R4, 8, 12, and 16) and specifically to test octaarginine peptides for the optical imaging of articular cartilage in experimentally induced arthritis in mice. Methods: Four rhodamine-labeled polyarginine peptides each with a different-length arginine chain (R4, 8, 12, or 16) were injected into the knee joints of C57BL/6J mice (n = 20). The joints were excised 1 h later and the fluorescent signal intensity in cartilage cryosections was compared for the four peptides. To examine the substrate of R8 in cartilage, femoral condyles obtained from another set of mice were treated with chondroitinase ABC (Ch'ase ABC), keratanase or heparitinase then immersed in R8-rhodamine. Fluorescent signals were examined by fluorescent microscopy. Next, R8-rhodamine was injected into the right knee joints of three control and three collagen antibody-induced arthritis (CAIA) mice, and fluorescent intensity in normal and degenerative cartilage was semi-quantitatively analysed on the histological sections using image software. Finally, femoral condyles from normal mice (n = 2) and CAIA mice (n = 2) were immersed in R8-rhodamine and calcein, then imaged using optical projection tomography (OPT). Results: Fluorescent signals were specifically detected in the cartilage pericellular matrix from the surface to the tide mark but were completely absent in the calcified layer or bone marrow. The number of arginine residues significantly influenced peptide accumulation in articular cartilage, with R8 accumulating the most. The fluorescent signal in the femoral condylar cartilage diminished when it was treated with Ch'ase ABC. R8 accumulation was significantly decreased in the degenerative cartilage of CAIA mice, and this was demonstrated both histologically and in three-dimensional (3D)-reconstruction image by OPT. Conclusion: R8 may be a useful new experimental probe for optical imaging of normal and arthritic articular cartilage.

Original languageEnglish
Pages (from-to)1209-1218
Number of pages10
JournalOsteoarthritis and Cartilage
Volume17
Issue number9
DOIs
Publication statusPublished - Sep 2009

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Optical Imaging
Cartilage
Articular Cartilage
Glycosaminoglycans
Rhodamines
Imaging techniques
Experimental Arthritis
Thigh
Optical Tomography
Chondroitin ABC Lyase
Peptides
keratan-sulfate endo-1,4-beta-galactosidase
heparitinsulfate lyase
Knee Joint
Arthritis
Arginine
Collagen
Antibodies
Bone and Bones
Computer-Assisted Image Processing

Keywords

  • Cartilage
  • Chondroitin sulfate
  • Collagen antibody-induced arthritis (CAIA) mouse model
  • Imaging
  • Octaarginine
  • Optical projection tomography

ASJC Scopus subject areas

  • Biomedical Engineering
  • Orthopedics and Sports Medicine
  • Rheumatology

Cite this

Optical imaging of mouse articular cartilage using the glycosaminoglycans binding property of fluorescent-labeled octaarginine. / Inagawa, K.; Oohashi, Toshitaka; Nishida, Keiichiro; Minaguchi, J.; Tsubakishita, T.; Yaykasli, K. O.; Ohtsuka, Aiji; Ozaki, Toshihumi; Moriguchi, T.; Ninomiya, Y.

In: Osteoarthritis and Cartilage, Vol. 17, No. 9, 09.2009, p. 1209-1218.

Research output: Contribution to journalArticle

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AU - Minaguchi, J.

AU - Tsubakishita, T.

AU - Yaykasli, K. O.

AU - Ohtsuka, Aiji

AU - Ozaki, Toshihumi

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