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In-vivo evaluation of subcutaneously implanted cell-loaded apatite microcarriers for osteogenic potency

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Abstract

Cell-loaded apatite microcarriers present as potential scaffolds for direct in-vivo delivery of cells post-expansion to promote bone regeneration. The objective of this study was to evaluate the osteogenic potency of human foetal mesenchymal stem cells (hfMSC)-loaded apatite microcarriers when implanted subcutaneously in a mouse model. This was done by examining for ectopic bone formation at 2 weeks, 1 month and 2 months, which were intended to coincide with the inflammation, healing and remodelling phases, respectively. Three histological examinations including haematoxylin and eosin staining to examine general tissue morphology, Masson’s trichrome staining to identify tissue type, and Von Kossa staining to examine extent of tissue mineralisation were performed. In addition, immunohistochemistry assay of osteopontin was conducted to confirm active bone formation by the seeded hfMSCs. Results showed a high level of tissue organisation and new bone formation, with active bone remodelling being observed at the end of 2 months, and an increase in tissue density, organisation, and mineralisation could also be observed for hfMSC-loaded apatite microcarriers. Various cell morphology resembling that of osteoblasts and osteoclasts could be seen on the surfaces of the hfMSC-loaded apatite microcarriers, with presence of woven bone tissue formation being observed at the intergranular space. These observations were consistent with evidence of ectopic bone formation, which were absent in group containing apatite microcarriers only. Overall, results suggested that hfMSC-loaded apatite microcarriers retained their osteogenic potency after implantation, and provided an effective platform for bone tissue regeneration.

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Acknowledgements

This work was supported by the Singapore Ministry of Health’s National Medical Research Council under its NMRC New Investigator Grant NIG10nov032.

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    Authors and Affiliations

    1. Department of Mechanical Engineering, National University of Singapore, Singapore, 117 576, Singapore

      Poon Nian Lim, Jason Feng, Zuyong Wang, Toshiisa Konishi & Eng San Thian

    2. College of Materials Science and Engineering, Hunan University, Changsha, 410 082, China

      Zuyong Wang

    3. Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore, 637 459, Singapore

      Mark Chong

    4. Graduate School of Natural Science and Technology, Okayama University, Okayama, 700 8530, Japan

      Toshiisa Konishi

    5. Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 119 228, Singapore

      Lay Geok Tan & Jerry Chan

    6. Department of Reproductive Medicine, Division of Obstetrics & Gynaecology, KK Women’s and Children’s Hospital, Singapore, 229 899, Singapore

      Jerry Chan

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    1. Poon Nian Lim
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    Correspondence to Eng San Thian.

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    Poon Nian Lim and Jason Feng contributed equally to this work.

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    Lim, P., Feng, J., Wang, Z. et al. In-vivo evaluation of subcutaneously implanted cell-loaded apatite microcarriers for osteogenic potency. J Mater Sci: Mater Med 28, 86 (2017). https://doi.org/10.1007/s10856-017-5897-4

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