Evaluation of the Usefulness of Human Adipose-Derived Stem Cell Spheroids Formed Using SphereRing® and the Lethal Damage Sensitivity to Synovial Fluid In Vitro.

Atsushi Fuku ,Terutsugu Koya,Yoshiyuki Tachi,Hironori Kitajima,Satoshi Osawa,Hiroshi Sunami,Ikuhiro Tanida,Yasuhito Ishigaki,Ayumi Kaneuji,Takashi Takaki,Naoki Yamamoto,Yasuhiko Taki,Yuka Nakamura,Norio Kawahara,Yusuke Shimizu,Kaori Nozaki,Toru Ichiseki,Shigetaka Shimodaira,Hiroaki Harai ,Togen Masauji

doi:10.3390/cells11030337

Abstract

Osteoarthritis (OA) is an irreversible degenerative condition causing bone deformation in the joints and articular cartilage degeneration with chronic pain and impaired movement. Adipose-derived stem cell (ADSC) or crushed adipose tissue injection into the joint cavity reportedly improve knee function and symptoms, including pain. Stem cell spheroids may be promising treatment options due to their anti-inflammatory and enhanced tissue regeneration/repair effects. Herein, to form human ADSC spheroids, we used first SphereRing® (Fukoku Co., Ltd., Ageo, Japan), a newly developed rotating donut-shaped tube and determined their characteristics by DNA microarray of mRNA analysis. The variable gene expression cluster was then identified and validated by RT-PCR. Gene expression fluctuations were observed, such as COL15A1 and ANGPTL2, related to vascular endothelial cells and angiogenesis, and TNC, involved in tissue formation. In addition, multiplex cytokine analysis in the medium revealed significant cytokines and growth factors production increase of IL-6, IL-10, etc. However, ADSC administration into the joint cavity involves their contact with the synovial fluid (SF). Therefore, we examined how SF collected from OA patient joint cavities affect 2D-culture ADSCs and ADSC spheroids and observed SF induced cell death. ADSC spheroids could become promising OA treatment options, although studying the administration methods and consider their interaction with SF is essential.

Highlights

Osteoarthritis (OA) is an irreversible degenerative condition that causes deformation of the bones that make up the joints, as well as degeneration of the articular cartilage, causing chronic pain and impaired movement of the extremities
The state of the cultured spheroids was observed under a phase-contrast microscope following swirling speeds of 25, 35, 45, 55, and 65 rpm, which led to successful spheroid formation
Uniform and large spheroids were observed under the condition of 35 rpm

Summary

Introduction

Osteoarthritis (OA) is an irreversible degenerative condition that causes deformation of the bones that make up the joints, as well as degeneration of the articular cartilage, causing chronic pain and impaired movement of the extremities. Inflammation of the synovial membrane surrounding the joints occurs, resulting in its degeneration. This may be accompanied by proliferative changes, such as osteochondrogenesis, around the joints, and these changes are thought to cause fibrosis of the joint capsule accompanied by angiogenesis and nerve fiber proliferation, leading to pain [1,2,3]. OA is considered a serious disease that is associated with great socioeconomic impact and a heavy burden on individuals [7,8]. Aging and obesity are risk factors for its onset and progression; the impact of OA on society is expected to increase in the future [9,10,11]

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Conclusion