Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (14): 2169-2176.doi: 10.3969/j.issn.2095-4344.1669

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Type I collagen combined titanium dioxide nanotube composite coating modified titanium surface improves osteoblast adhesion and osseointegration

Li Ying1, You Yapeng1, Li Baoe2, Song Yunjia1, Ma Aobo1, Chen Bo1, Han Wen1, Li Changyi1   

  1. 1Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China; 2School of Materials Science and Engineering, Hebei University of Technology, Tianjin 300130, China
  • Received:2018-12-26
  • Contact: Li Changyi, Professor, Doctoral supervisor, Chief physician, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • About author:Li Ying, MD, Associate chief physician, Master’s supervisor, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81870809, 31470920 (to LCY), and 81500886 (to LY); the Natural Science Foundation of Tianjin, No. 16JCYBJC28700 (to LY); the Natural Science Foundation of Hebei Province, No. E2017202032 (to LY)

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Abstract:

BACKGROUND: Simple nanotube surface modification of titanium implant has been shown to promote adhesion, proliferation and differentiation of osteoblasts. Collagen coating can promote osteoblast adhesion and osseointegration in vivo.

OBJECTIVE: To observe the effects of type I collagen combined titanium dioxide nanotube composite coating modified titanium surface on osteoblast adhesion in vitro and osseointegration in vivo.
METHODS: The titanium dioxide nanotube was fabricated on the pure titanium surface, then type I collagen was combined with the nanotube structure to form composite coating. Scanning electron microscope observation was used to characterize the surface topography of the pure titanium, titanium dioxide nanotube and type I collagen combined titanium dioxide nanotube surfaces. Contact angle test was employed to evaluate the hydrophilicity of different samples. MC3T3-E1 murine preosteoblasts were seeded on the three kinds of materials for 4 hours. Cell adhesion morphology was examined by scanning electron microscope. Adherent cell counting was detected under inverted fluorescence microscope. Expression of actin cytoskeleton and vinculin was observed under laser scanning confocal microscope. The gene expression of vinculin and osteoprotegerin mRNA was detected by real-time quantitative PCR. The three kinds of samples were implanted into the tibia of Sprague-Dawley rats (provided by Laboratory Animal Center, Institute of Radiation Medicine, Chinese Academy of Medical Sciences), and tibia samples were removed after 4 weeks of implantation for biological push-out test and histological observation.
RESULTS AND CONCLUSION: (1) Scanning electron microscope: There was mechanical scratch on the pure titanium surface. There was controllable, and uniform vertical arrangement of nanotubular structures with a diameter of approximately 70 nm on the titanium dioxide nanotube surface, and collagen adhered surrounding the nanotubular structures on the type I collagen combined titanium dioxide nanotube substrate, and partial tubule orifices were closed. (2) The hydrophicility of type I collagen combined titanium dioxide nanotube was significantly larger than those of the other two materials (P < 0.05). (3) Compared with the pure titanium and titanium dioxide nanotube surfaces, the type I collagen combined titanium dioxide nanotube substrate displayed increased adherent cell number, much well-organized cytoskeleton, enhanced immunofluorescence intensity of vinculin protein staining, and increased expression levels of vinculin and osteoprotegerin mRNA levels (all P < 0.05). (4) In vivo test revealed that the maximum push-out force in the type I collagen combined titanium dioxide nanotube group was significantly higher than that in the pure titanium and titanium dioxide nanotube groups (P < 0.05). Hematoxylin-eosin staining results showed that there were few bones, but many fibrous connective tissue surrounding the implant in the pure titanium group; there were more newly-born bones, and less fibrous connective tissue surrounding the implant in the titanium dioxide nanotube group; there were dense newly-born bones, and few thin fibrous connective tissue surrounding the implant in the type I collagen combined titanium dioxide nanotube group. (5) These results indicate that type I collagen combined titanium dioxide nanotube surface can facilitate osteoblast cell adhesion and promote osseointegration in vivo.

Key words: Dental Implants, Titanium, Nanotubes, Collagen, Osteoblasts, Cell Adhesion, Synostosis

CLC Number: