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The utility of human dedifferentiated fat cells in bone tissue engineering in vitro

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Abstract

We compared the osteoblastic differentiation abilities of dedifferentiated fat cells (DFATs) and human bone marrow mesenchymal stem cells (hMSCs) as a cell source for bone regeneration therapies. In addition, the utility of DFATs in bone tissue engineering in vitro was assessed by an alpha-tricalcium phosphate (α-TCP)/collagen sponge (CS). Human DFATs were isolated from the submandibular of a patient by ceiling culture. DFATs and hMSCs at passage 3 were cultured in control medium or osteogenic medium (OM) for 14 days. Runx2 gene expression, alkaline phosphatase (ALP) activity, as well as osteocalcin (OCN) and calcium contents were analyzed to evaluate the osteoblastic differentiation ability of both cell types. DFATs seeded in a α-TCP/CS and cultured in OM for 14 days were analyzed by scanning electron microscopy (SEM) and histologically. Compared with hMSCs, DFATs cultured in OM generally underwent superior osteoblastogenesis by higher Runx2 gene expression at all days tested, as well as higher ALP activity at day 3 and 7, OCN expression at day 14, and calcium content at day 7. In SEM analyses, DFATs seeded in a α-TCP/CS were well spread and covered the α-TCP/CS by day 7. In addition, numerous spherical deposits were found to almost completely cover the α-TCP/CS on day 14. Von Kossa staining showed that DFATs differentiated into osteoblasts in the α-TCP/CS and formed cultured bone by deposition of a mineralized extracellular matrix. The combined use of DFATs and an α-TCP/CS may be an attractive option for bone tissue engineering.

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Acknowledgments

We thank Dr. Noboru Sasaki (Department of Oral and Maxillofacial Surgery, Amagasaki Chuo Hospital) for his cooperation with collecting the adipose tissue. This study was supported by a Grant-in-Aid for Scientific Research (B) No. 24792263 from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

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Authors and Affiliations

  1. Department of Orthodontics, Osaka Dental University, 8-1 Hanazonocho, Kuzuha, Hirakata, 573-1121, Japan

    Fumito Sakamoto & Naoyuki Matsumoto

  2. Department of Biomaterials, Osaka Dental University, 8-1 Hanazonocho, Kuzuha, Hirakata, 573-1121, Japan

    Yoshiya Hashimoto

  3. Department of Anesthesiology, Osaka Dental University, 8-1 Hanazonocho, Kuzuha, Hirakata, 573-1121, Japan

    Naotaka Kishimoto

  4. Institute of Dental Research, Osaka Dental University, 8-1 Hanazonocho, Kuzuha, Hirakata, 573-1121, Japan

    Yoshitomo Honda

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  1. Fumito Sakamoto
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  2. Yoshiya Hashimoto
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  3. Naotaka Kishimoto
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  4. Yoshitomo Honda
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  5. Naoyuki Matsumoto
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Correspondence to Fumito Sakamoto.

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Sakamoto, F., Hashimoto, Y., Kishimoto, N. et al. The utility of human dedifferentiated fat cells in bone tissue engineering in vitro. Cytotechnology 67, 75–84 (2015). https://doi.org/10.1007/s10616-013-9659-y

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  • DOI: https://doi.org/10.1007/s10616-013-9659-y

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