The effects of oyster shell/alpha-calcium sulfate hemihydrate/platelet-rich plasma/bone mesenchymal stem cells bioengine
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TISSUE ENGINEERING CONSTRUCTS AND CELL SUBSTRATES Original Research
The effects of oyster shell/alpha-calcium sulfate hemihydrate/ platelet-rich plasma/bone mesenchymal stem cells bioengineering scaffold on rat critical-sized calvarial defects Jinwu Wang1,2 Linzhen Xie1,2 Xingyu Wang1,2 Wenhao Zheng1,2 Hua Chen1,2 Leyi Cai1,2 Long Chen1,2 ●
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Received: 15 February 2020 / Accepted: 24 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Engineering scaffolds combining natural biomineral and artificially synthesized material hold promising potential for bone tissue regeneration. We fabricated a bioengineering scaffold, oyster shell (OS) and alpha-calcium sulfate hemihydrate (αCSH) as scaffold, platelet-rich plasma (PRP) as provider of growth factors and bone mesenchymal stem cells (BMSCs) as seed cells, and determined it could be applied as a new type of bone graft substitutes by rat calvarial defects repairing experiment in vitro and in vivo. SEM showed that the mean diameter of the pores was about 150 μm with a range of 50–200 μm, and scaffold’s porosity was ~27.4% by Archimedes’ Principle. In vitro, Scaffold + BMSCs + PRP group presented a higher ALP activity compared with other groups by ELISA (P < 0.05). But the expression of OC was not detectable on day 4 or 8. The MTT assay showed that the relative cell number of BMSCs+PRP group increased significantly (P < 0.05). In vivo, the smallest defect area of skull and highest volume of regenerated new bone were observed in Scaffold + PRP + BMSCs group by X-ray and Micro-CT analysis (P < 0.05). And the similar results also were observed in HE and Masson staining. The immunohistochemistry staining for osteogenic marker proteins ALP and OC showed that the most obvious positive staining was observed in Scaffold + PRP + BMSCs group (P < 0.05). The expression of inflammatory markers IL-6 and TNF-α was the lowest in control group (P < 0.05). In conclusion, a bioengineering scaffold based on OS, created by simply combining α-CSH and PRP and implanting with BMSCs, could be clinically useful and has marked advantages as a targeted, off-the-shelf, cell-loaded treatment option for the bone healing of critical-size calvarial defects. Abbreviation oyster shell alpha-calcium sulfate hemihydrate platelet-rich plasma bone mesenchymal stem cell fetal bovine serum phosphate buffer saline alkaline phosphatase
(OS) (α-CSH) (PRP) (BMSC) (FBS) (PBS) (ALP)
These authors contributed equally: Jinwu Wang, Linzhen Xie Jinwu Wang and Linzhen Xie should be considered co-first authors. * Long Chen [email protected] 1
Department of Orthopaedics Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, NO.109, XueYuan West Road, Luheng District, Wenzhou, Zhejiang Province 325000, P.R. China
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Wenzhou Medical University, Wenzhou, Zhejiang, China
osteocalcin enzyme linked immunosorbent assay hethylenediaminetetraacetic acid calcium sulphate dihydrate
(OC) (ELISA)
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