Modifying oxygen tension affects bone marrow stromal cell osteogenesis for regenerative medicine

AIM To establish a hypoxic environment for promoting osteogenesis in rat marrow stromal cells（MSCs） using osteogenic matrix cell sheets（OMCSs）.METHODS Rat MSCs were cultured in osteogenic media under one of four varying oxygen conditions: Normoxia（21% O2） for 14 d（NN）, normoxia for 7 d followed by h...

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Published inWorld journal of stem cells Vol. 9; no. 7; pp. 98 - 106
Main Authors Inagaki, Yusuke, Akahane, Manabu, Shimizu, Takamasa, Inoue, Kazuya, Egawa, Takuya, Kira, Tsutomu, Ogawa, Munehiro, Kawate, Kenji, Tanaka, Yasuhito
Format Journal Article
LanguageEnglish
Published United States Baishideng Publishing Group Inc 26.07.2017
Subjects
Basic Study
Hypoxia
Marrow stromal cells
Regenerative medicine
Tissue engineering
Osteogenesis
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Summary:AIM To establish a hypoxic environment for promoting osteogenesis in rat marrow stromal cells（MSCs） using osteogenic matrix cell sheets（OMCSs）.METHODS Rat MSCs were cultured in osteogenic media under one of four varying oxygen conditions: Normoxia（21% O2） for 14 d（NN）, normoxia for 7 d followed by hypoxia（5% O2） for 7 d（NH）, hypoxia for 7 d followed by normoxia for 7 d（HN）, or hypoxia for 14 d（HH）. Osteogenesis was evaluated by observing changes in cell morphology and calcium deposition, and by measuring osteocalcin secretion（ELISA） and calcium content. In vivo syngeneic transplantation using OMCSs and β-tricalcium phosphate discs, preconditioned under NN or HN conditions, was also evaluated by histology, calcium content measurements,and real-time quantitative PCR.RESULTS In the NN and HN groups, differentiated, cuboidal-shaped cells were readily observed, along with calcium deposits. In the HN group, the levels of secreted osteocalcin increased rapidly from day 10 as compared with the other groups, and plateaued at day 12（P < 0.05）. At day 14, the HN group showed the highest amount of calcium deposition. In vivo, the HN group showed histologically prominent new bone formation, increased calcium deposition, and higher collagen type Ⅰ?messenger RNA expression as compared with the NN group.CONCLUSION The results of this study indicate that modifying oxygen tension is an effective method to enhance the osteogenic ability of MSCs used for OMCSs.
Bibliography:Yusuke Inagaki;Manabu Akahane;Takamasa Shimizu;Kazuya Inoue;Takuya Egawa;Tsutomu Kira;Munehiro Ogawa;Kenji Kawate;Yasuhito Tanaka;Department of Orthopedic Surgery,Nara Medical University;Department of Public Health,Health Management and Policy,Nara Medical University,Faculty of Medicine;Departments of Artificial Joint and Regenerative Medicine for Bone and Cartilage,Nara Medical University
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Author contributions: Inagaki Y and Akahane M designed the study; Inagaki Y, Akahane M, Shimizu T, Inoue K, Egawa T and Kira T performed the study; Inagaki Y and Akahane M drafted the manuscript; Ogawa M, Kawate K and Tanaka Y critically reviewed the draft; all authors read and approved the final manuscript.
Correspondence to: Yusuke Inagaki, MD, PhD, Assisstant Professor, Department of Orthopedic Surgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara 634-8522, Japan. yinagaki@naramed-u.ac.jp
ISSN:1948-0210
1948-0210
DOI:10.4252/wjsc.v9.i7.98