Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (14): 2223-2227.doi: 10.3969/j.issn.2095-4344.1674

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Fabrication and biocompatibility assessment of the scaffold with biomimetic interconnected macropore structure

Li Zhi1, Tan Chunhua1, Cai Xianhua1, Wang Huasong1, Ding Xiaoming2, Zhao Yanhong3   

  1.  赵艳红,副主任医师,天津医科大学口腔医院正畸科,天津市 300070
  • Received:2018-12-26
  • Contact: 1Department of Orthopedics, General Hospital of Chinese PLA Central Theater Command & Hubei Provincial Center for Bone Trauma Treatment Clinical Medicine, Wuhan 430070, Hubei Province, China; 2Fifth Department of Orthopedics, Rizhao Hospital of Traditional Chinese Medicine, Rizhao 276800, Shandong Province, China; 3Department of Orthodontics, Stomatological Hospital of Tianjin Medical University, Tianjin 300070, China
  • About author:Li Zhi, MD, Associate chief physician, Department of Orthopedics, General Hospital of Chinese PLA Central Theater Command & Hubei Provincial Center for Bone Trauma Treatment Clinical Medicine, Wuhan 430070, Hubei Province, China
  • Supported by:

    the Natural Science Foundation of Hubei Province, No. 2017CFB569 (to LZ); the National Natural Science Foundation of China, No. 31470937 (to DXM) and 31300798 (to ZYH); the Foundation of Health and Family Planning Commission of Tianjin, No. 16KG114 and 15KG125 (both to ZYH)

Abstract:

BACKGROUND: Development of bone tissue engineering provides a new method to solve bone defect repair. Scaffold material and structure construction are issues of concern.

OBJECTIVE: To fabricate a silk fibroin and hydroxyapatite scaffold with biomimetic interconnected macropores structure using the paraffin-sphere leaching technology and to evaluate its possibility of cytocompatibility.
METHODS: The scaffold with biomimetic interconnected macropores structure was made by the paraffin-sphere leaching technology. The structure of scaffold was observed by the stereomicroscope and scanning electron microscope. The pore size and elasticity modulus were calculated. Passage 3 rabbit adipose-derived mesenchymal stem cells were seeded into the scaffold. The cell viability was detected by live/dead staining at 48 hours after culture. The cell adhesion was observed by hematoxylin-eosin staining at 1 week of culture. The scaffold-cell composite was observed under scanning electron microscope at 3 days of culture. The cell proliferation was detected by the cell counting-kit 8 assay at 1, 3, 5 and 7 days of culture. Those cells cultured alone were considered as control group.
RESULTS AND CONCLUSION: (1) Stereomicroscope showed the ivory silk fibroin/hydroxyapatite scaffold. Scanning electron microscope revealed pore structures in cross-section and longitudinal-section with good connectivity. The scaffold pore size was (362.23±26.52) μm and the elasticity modulus was (54.93±5.44) kPa. (2) Scanning electron microscope showed that adipose-derived mesenchymal stem cells adhered and stretched well in the pore wall and connected pore, secreted abundant extracellular matrix, and filled in the pores of silk fibroin/hydroxyapatite scaffold. (3) Hematoxylin-eosin staining results found that adipose-derived mesenchymal stem cells evenly adhered onto the inner wall of silk fibroin/hydroxyapatite scaffold, and proliferated well. (4) Live/dead staining revealed a good viability of adipose-derived mesenchymal stem cells. (5) Cell counting-kit 8 assay results showed the good proliferation of adipose-derived mesenchymal stem cells on the scaffold. (6) To conclude, the silk fibroin/hydroxyapatite scaffold possesses good pore size and cytocompatibility.

Key words: Silk, Hydroxyapatites, Cell Adhesion, Biocompatible Materials, Tissue Engineering

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