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Journal of Peking University (Health Sciences) ›› 2022, Vol. 54 ›› Issue (1): 31-39. doi: 10.19723/j.issn.1671-167X.2022.01.006

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Effect of porous zirconia ceramics on proliferation and differentiation of osteoblasts

WANG Zheng1,DING Qian1,2,△(),GAO Yuan1,MA Quan-quan1,ZHANG Lei1,△(),GE Xi-yuan3,SUN Yu-chun1,4,XIE Qiu-fei1   

  1. 1. Department of Prosthodontics, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology & NHC Research Center of Engineering and Technology for Computerized Dentistry & NMPA Key Laboratory for Dental Materials, Beijing 100081, China
    2. Foshan (Southern China) Institute for New Materials, Foshan 528000, Guangdong, China
    3. Central Laboratory, Peking University School and Hospital of Stomatology, Beijing 100081, China
    4. Center for Digital Dentistry, Peking University School and Hospital of Stomatology, Beijing 100081, China
  • Received:2021-10-07 Online:2022-02-18 Published:2022-02-21
  • Contact: Qian DING,Lei ZHANG E-mail:dingqian.623@163.com;drzhanglei@yeah.net
  • Supported by:
    Capital Health Development Research Special Fund(2020-2-4104);Beijing Natural Science Foundation(7192233);Guangdong Basic and Applied Basic Research Foundation(2019A1515110889)

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Abstract:

Objective: To investigate the effect of porous surface morphology of zirconia on the proliferation and differentiation of osteoblasts. Methods: According to different manufacturing and pore-forming methods, the zirconia specimens were divided into 4 groups, including milled sintering group (M-Ctrl), milled porous group (M-Porous), 3D printed sintering group (3D-Ctrl) and 3D printed porous group (3D-Porous). The surface micromorphology, surface roughness, contact angle and surface elements of specimens in each group were detected by scanning electron microscope (SEM), 3D laser microscope, contact angle measuring device and energy-dispersion X-ray analysis, respectively. MC3T3-E1 cells were cultured on 4 groups of zirconia discs. The cell morphology of MC3T3-E1 cells on zirconia discs was eva-luated on 1 and 7 days by SEM. The cell proliferation was detected on 1, 3 and 5 days by cell counting kit-8 (CCK-8). After osteogenic induction for 14 days, the relative mRNA expression of alkaline phosphatase (ALP), type Ⅰ collagen (Colla1), Runt-related transcription factor-2 (Runx2) and osteocalcin (OCN) in MC3T3-E1 cells were detected by real-time quantitative polymerase chain reaction. Results: The pore size [(419.72±6.99) μm] and pore depth [(560.38±8.55) μm] of 3D-Porous group were significantly larger than the pore size [(300.55±155.65) μm] and pore depth [(69.97±31.38) μm] of M-Porous group (P<0.05). The surface of 3D-Porous group appeared with more regular round pores than that of M-Porous group. The contact angles of all the groups were less than 90°. The contact angles of 3D-Ctrl (73.83°±5.34°) and M-Porous group (72.7°±2.72°) were the largest, with no significant difference between them (P>0.05). Cells adhered inside the pores in M-Porous and 3D-Porous groups, and the proliferation activities of them were significantly higher than those of M-Ctrl and 3D-Ctrl groups after 3 and 5 days’ culture (P<0.05). After 14 days’ incubation, ALP, Colla1, Runx2 and OCN mRNA expression in 3D-Porous groups were significantly lower than those of M-Ctrl and 3D-Ctrl groups (P<0.05). Colla1, Runx2 and OCN mRNA expressions in M-Porous group were higher than those of 3D-Porous group (P<0.05). Conclusion: The porous surface morphology of zirconia can promote the proliferation and adhesion but inhibit the differentiation of MC3T3-E1 cells.

Key words: Zirconia, Porous surface, Osteoblasts, Osseointegration

CLC Number: 

  • R781.3

Figure 1

Zirconia disks M-Ctrl, milled sintering group; M-Porous, milled porous group; 3D-Ctrl, 3D printed sintering group; 3D-Porous, 3D printed porous group."

Table 1

Real-time PCR primer sequence"

Gene Forward primer (5'-3') Reverse primer (5'-3')
GAPDH TGGTGAAGGTCGGTGTGAAC GCTCCTGGAAGATGGTGATGG
ALP TGCCTACTTGTGTGGCGTGAA TCACCCGAGTGGTAGTCACAATG
Colla1 TGAAGAACTGGACTGTCCC TTTGGTGATACGTATTCTTCCG
Runx2 GACTGTGGTTACCGTCATGGC ACTTGGTTTTTCATAACAGCGGA
OCN CCAAGCAGGAGGGCAATAAGGT CTCGTCACAAGCAGGGTTAAGC

Table 2

Surface pore size of porous zirconia disks"

Items M-Porous 3D-Porous
Diameter of pores/μm, $\bar{x}\pm s$ 300.55±155.65 419.72±6.99*
Depth of pores/μm, $\bar{x}\pm s$ 69.97±31.38 560.38±8.55*
Pitch of pores/μm, $\bar{x}\pm s$ 349.87±12.43

Figure 2

Scanning electron microscope images of zirconia disks M-Ctrl, milled sintering group; M-Porous, milled porous group; 3D-Ctrl, 3D printed sintering group; 3D-Porous, 3D printed porous group."

Figure 3

Scanning electron microscope images of MC3T3-E1 cells on zirconia discs after incubation for 1 and 7 days M-Ctrl, milled sintering group; M-Porous, milled porous group; 3D-Ctrl, 3D printed sintering group; 3D-Porous, 3D printed porous group."

Figure 4

Cell proliferation of MC3T3-E1 cells after 1, 3 and 5 days M-Ctrl, milled sintering group; M-Porous, milled porous group; 3D-Ctrl, 3D printed sintering group; 3D-Porous, 3D printed porous group. Four groups comparison: 1 d, H=31.834, P < 0.001; 3 d, F=54.633, P < 0.001; 5 d, H=44.859, P < 0.001. *P<0.05, △P < 0.001."

Figure 5

Relative mRNA expression of ALP, Colla1, Runx2 and OCN in MC3T3-E1 cells Relative gene expression of osteogenic differentiation proteins ALP, Colla1, Runx2 and OCN in MC3T3-E1 cells. M-Ctrl, milled sintering group; M-Porous, milled porous group; 3D-Ctrl, 3D printed sintering group; 3D-Porous, 3D printed porous group. Four groups comparison: ALP, H=61.635, P<0.001; Colla1, H=49.518, P<0.001; Runx2, H=55.728, P<0.001; OCN, H=38.236, P<0.001. #P<0.01, △P < 0.001."

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