Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (16): 2473-2479.doi: 10.12307/2023.464

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Preparation and osteoinductive properties of tricalcium phosphate ceramics with submicron topology

Lu Di1, Wan Xinyu1, Yang Jinxin1, Ding Kexin1, Zhang Cheng1, Duan Rongquan1, Liu Zongxiang1, 2   

  1. 1School of Stomatology, Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China; 2Affiliated Stomatological Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu Province, China
  • Received:2022-05-26 Accepted:2022-07-21 Online:2023-06-08 Published:2022-11-10
  • Contact: Liu Zongxiang, Chief physician, Professor, School of Stomatology, Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China; Affiliated Stomatological Hospital of Xuzhou Medical University, Xuzhou 221002, Jiangsu Province, China Duan Rongquan, MD, Associate professor, School of Stomatology, Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China
  • About author:Lu Di, Master candidate, Physician, School of Stomatology, Xuzhou Medical University, Xuzhou 221004, Jiangsu Province, China
  • Supported by:
    the Transformation Project of Sichuan Provincial Department of Science and Technology, No. 2019ZYZF0081 (to DRQ); Excellent Talents Research Start-up Fund of Xuzhou Medical University, No. D2020005 (to DRQ)

Abstract: BACKGROUND: Synthetic calcium phosphate ceramic materials are similar to the inorganic components of natural bone tissue. Functional design through surface morphology and chemical composition can endow them with excellent osteoconductive and osteoinductive properties. The development of calcium phosphate ceramic materials with osteoinductive properties is the current research hotspot.
OBJECTIVE: To endow the submicron topology tricalcium phosphate ceramics with osteoinductive properties through material morphology regulation and functional design, and test their physicochemical properties and osteoinductive properties. 
METHODS: Novel tricalcium phosphate ceramics were prepared by high temperature sintering method. Clinical available Bio-oss bone meal was set as the control group. The sample surface morphology, the protein adsorption capacity, and the in vitro mineralizability of the two materials were characterized. Human periodontal membrane stem cells at passage 3 were co-cultured with the extracts of the two materials. Cell proliferation was detected by CCK-8 assay. Alizarin red staining was used to detect cell mineralization performance. The human periodontal ligament stem cells at passage 3 were seeded on the surfaces of the two materials. Early osteogenesis was detected by alkaline phosphatase staining. The expression of osteogenesis-related factors was detected by qRT-PCR.
RESULTS AND CONCLUSION: (1) Scanning electron microscope exhibited that both materials had microporous surfaces with granular texture. Bio-Oss particles were significantly smaller than that of tricalcium phosphate ceramics. The total porosity, macroporosity, and microporosity were similar for both materials. Tricalcium phosphate ceramics were mainly submicron pores, with a grain size of 100 nm-1.0 μm. Bio-Oss bone meal was mainly nano-scale pores. In vitro mineralization experiments displayed that the ability of the surface of tricalcium phosphate ceramics to induce bone apatite deposition was stronger than that of Bio-Oss bone meal. Compared with Bio-Oss bone meal, tricalcium phosphate ceramics could adsorb more proteins from fetal bovine serum and bovine serum albumin solution (P < 0.05). (2) CCK-8 assay demonstrated that both materials promoted cell proliferation, and there was no significant difference between the two groups (P > 0.05). (3) The alkaline phosphatase activity in the tricalcium phosphate ceramic group was higher than that in the Bio-Oss group at 4 and 7 days of culture (P < 0.05), and the number of mineralized nodules at 21 days of culture was more than that in the Bio-Oss group; the mRNA expression levels of alkaline phosphatase, osteocalcin, osteopontin and Runx-2 were higher than those in the Bio-Oss group on 7 and 14 days of culture (P < 0.05). (4) The results showed that the new calcium phosphate ceramics have superior in vitro osteoinductive properties. 

Key words: bone defect, calcium phosphate, bone repair, osteoinductive, osteoconductivity, surface topography, human periodontal ligament stem cell, factor

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