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The Evaluation of Meniscus Regenerative Effects of LIPUS-Induced CCN Proteins: Induction by LIPUS of CCN2 and Meniscus-Related Genes in Cultured Meniscus Cells and Meniscus Tissues

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CCN Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2582))

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Abstract

Menisci are a pair of crescent-shaped fibrocartilages and composed primarily of type I collagen. Inner region of the meniscus has similar characteristics to articular cartilage. Low-intensity pulsed ultrasound (LIPUS) has been reported to have chondroprotective effects on chondrocytes by inducing the expression of chondrocyte differentiation markers and CCN2/CTGF production. Here, we describe an experimental approach that investigates the distinct cellular behavior of human inner and outer meniscus cells in response to LIPUS stimulation. Our experimental model can analyze the relationships between LIPUS-induced CCN2 and its repairing role in the meniscus.

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References

  1. Tanaka T, Fujii K, Kumagae Y (1999) Comparison of biochemical characteristics of cultured fibrochondrocytes isolated from the inner and outer regions of human meniscus. Knee Surg Sports Traumatol Arthrosc 7(2):75–80. https://doi.org/10.1007/s001670050125

    Article  CAS  PubMed  Google Scholar 

  2. Messner K, Gao J (1998) The menisci of the knee joint. Anatomical and functional characteristics, and a rationale for clinical treatment. J Anat 193(Pt 2):161–178. https://doi.org/10.1046/j.1469-7580.1998.19320161.x

    Article  Google Scholar 

  3. Arnoczky SP, Warren RF (1982) Microvasculature of the human meniscus. Am J Sports Med 10(2):90–95. https://doi.org/10.1177/036354658201000205

    Article  CAS  PubMed  Google Scholar 

  4. Makris EA, Hadidi P, Athanasiou KA (2011) The knee meniscus: structure-function, pathophysiology, current repair techniques, and prospects for regeneration. Biomaterials 32(30):7411–7431. https://doi.org/10.1016/j.biomaterials.2011.06.037

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Furumatsu T, Kanazawa T, Yokoyama Y, Abe N, Ozaki T (2011) Inner meniscus cells maintain higher chondrogenic phenotype compared with outer meniscus cells. Connect Tissue Res 52(6):459–465. https://doi.org/10.3109/03008207.2011.562061

    Article  CAS  PubMed  Google Scholar 

  6. Leighton R, Watson JT, Giannoudis P, Papakostidis C, Harrison A, Steen RG (2017) Healing of fracture nonunions treated with low-intensity pulsed ultrasound (LIPUS): a systematic review and meta-analysis. Injury 48(7):1339–1347. https://doi.org/10.1016/j.injury.2017.05.016

    Article  PubMed  Google Scholar 

  7. Harrison A, Lin S, Pounder N, Mikuni-Takagaki Y (2016) Mode & mechanism of low intensity pulsed ultrasound (LIPUS) in fracture repair. Ultrasonics 70:45–52. https://doi.org/10.1016/j.ultras.2016.03.016

    Article  CAS  PubMed  Google Scholar 

  8. Uddin SM, Richbourgh B, Ding Y, Hettinghouse A, Komatsu DE, Qin YX et al (2016) Chondro-protective effects of low intensity pulsed ultrasound. Osteoarthr Cartil 24(11):1989–1998. https://doi.org/10.1016/j.joca.2016.06.014

    Article  CAS  Google Scholar 

  9. Cheng K, Xia P, Lin Q, Shen S, Gao M, Ren S et al (2014) Effects of low-intensity pulsed ultrasound on integrin-FAK-PI3K/Akt mechanochemical transduction in rabbit osteoarthritis chondrocytes. Ultrasound Med Biol 40(7):1609–1618. https://doi.org/10.1016/j.ultrasmedbio.2014.03.002

    Article  PubMed  Google Scholar 

  10. Nishida T, Kubota S, Aoyama E, Yamanaka N, Lyons KM, Takigawa M (2017) Low-intensity pulsed ultrasound (LIPUS) treatment of cultured chondrocytes stimulates production of CCN family protein 2 (CCN2), a protein involved in the regeneration of articular cartilage: mechanism underlying this stimulation. Osteoarthr Cartil 25(5):759–769. https://doi.org/10.1016/j.joca.2016.10.003

    Article  CAS  Google Scholar 

  11. Kamatsuki Y, Aoyama E, Furumatsu T, Miyazawa S, Maehara A, Yamanaka N et al (2019) Possible reparative effect of low-intensity pulsed ultrasound (LIPUS) on injured meniscus. J Cell Commun Signal 13(2):193–207. https://doi.org/10.1007/s12079-018-0496-9

    Article  PubMed  Google Scholar 

  12. Takigawa M (2003) CTGF/Hcs24 as a multifunctional growth factor for fibroblasts, chondrocytes and vascular endothelial cells. Drug News Perspect 16(1):11–21. https://doi.org/10.1358/dnp.2003.16.1.829302

    Article  CAS  PubMed  Google Scholar 

  13. Takigawa M (2018) An early history of CCN2/CTGF research: the road to CCN2 via hcs24, ctgf, ecogenin, and regenerin. J Cell commun Signal 12(1):253–264. https://doi.org/10.1007/s12079-017-0414-6

    Article  PubMed  Google Scholar 

  14. Nishida T, Kubota S, Kojima S, Kuboki T, Nakao K, Kushibiki T et al (2004) Regeneration of defects in articular cartilage in rat knee joints by CCN2 (connective tissue growth factor). J Bone Miner Res 19(8):1308–1319. https://doi.org/10.1359/jbmr.040322

    Article  CAS  PubMed  Google Scholar 

  15. Nishida T, Maeda A, Kubota S, Takigawa M (2008) Role of mechanical-stress inducible protein Hcs24/CTGF/CCN2 in cartilage growth and regeneration: mechanical stress induces expression of Hcs24/CTGF/CCN2 in a human chondrocytic cell line HCS-2/8, rabbit costal chondrocytes and meniscus tissue cells. Biorheology 45(3–4):289–299

    Article  PubMed  Google Scholar 

  16. Kikuchi T, Kubota S, Asaumi K, Kawaki H, Nishida T, Kawata K et al (2008) Promotion of bone regeneration by CCN2 incorporated into gelatin hydrogel. Tissue Eng Part A 14(6):1089–1098. https://doi.org/10.1089/tea.2007.0167

    Article  CAS  PubMed  Google Scholar 

  17. Hara C, Kubota S, Nishida T, Hiasa M, Hattori T, Aoyama E et al (2016) Involvement of multiple CCN family members in platelets that support regeneration of joint tissues. Mod Rheumatol 26(6):940–949. https://doi.org/10.3109/14397595.2016.1155255

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Dr. Satoshi Kubota and Dr. Takashi Nishida for their kind assistance. This work was supported by the programs JSPS KAKENHI Grants-in-Aid for (B) #JP19H03817 (to MT) and Challenging Research (Pioneering) #JP20K20611 (to MT) from the Japan Society for the Promotion of Science, Japan.

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Authors and Affiliations

  1. Department of Orthopedic Surgery, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Kitaku, Okayama, Japan

    Yusuke Kamatsuki, Takayuki Furumatsu & Toshifumi Ozaki

  2. Advanced Research Center for Oral and Craniofacial Sciences (ARCOCS), Okayama University Dental School/Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Kitaku, Okayama, Japan

    Yusuke Kamatsuki, Eriko Aoyama & Masaharu Takigawa

Authors
  1. Yusuke Kamatsuki
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  2. Eriko Aoyama
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  3. Takayuki Furumatsu
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  4. Toshifumi Ozaki
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  5. Masaharu Takigawa
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Corresponding author

Correspondence to Masaharu Takigawa .

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Editors and Affiliations

  1. Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan

    Masaharu Takigawa

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Kamatsuki, Y., Aoyama, E., Furumatsu, T., Ozaki, T., Takigawa, M. (2023). The Evaluation of Meniscus Regenerative Effects of LIPUS-Induced CCN Proteins: Induction by LIPUS of CCN2 and Meniscus-Related Genes in Cultured Meniscus Cells and Meniscus Tissues. In: Takigawa, M. (eds) CCN Proteins. Methods in Molecular Biology, vol 2582. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2744-0_15

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  • DOI: https://doi.org/10.1007/978-1-0716-2744-0_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2743-3

  • Online ISBN: 978-1-0716-2744-0

  • eBook Packages: Springer Protocols

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