Page 380 - IJB-9-2

International Journal of Bioprinting 3D gel-printed β-TCP/TiO2 porous scaffolds for cancellous bone tissue engineering

Figure 1. (A) Ceramic samples of β-TCP (a), β-TCP/1-TiO (b), β-TCP/3-TiO (c), and β-TCP/5-TiO (d). (B) XRD diagram of TiO , β-TCP, and β-TCP/5-
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TiO ceramic and details of the XRD diagram of the figure at 24.5°–26.5°. (C) Infrared spectrum of TiO , β-TCP, and β-TCP/5-TiO ceramic. (D) Micro-
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morphology of β-TCP/TiO ceramics detected by SEM: β-TCP/TiO (a), β-TCP/1-TiO (b), β-TCP/3-TiO (c), and β-TCP/5-TiO (d). (E) The shrinkage
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of β-TCP/TiO ceramic samples with 0% TiO , 1% TiO , 3% TiO and 5% TiO . (F) Average diameter of micropores of β-TCP and β-TCP/TiO ceramics.
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(G) The compressive strength of β-TCP and β-TCP/TiO ceramics with different TiO content.
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by field emission scanning electron microscopy (FE- 2.8. Statistical analysis
SEM) equipped with energy dispersive spectroscopy All statistical data were calculated by Origin 2018 and
(EDS). GraphPad Prism 8. One-way analysis of variance (ANOVA)
was conducted (**** P < 0.0001; *** P < 0.001; ** P < 0.01;
2.7.2. Cell biocompatibility and proliferation of β-TCP/ * P < 0.05; NS, no significant difference).
TiO ceramic scaffolds
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Samples of ceramic scaffolds were sterilized by gamma-
ray diffraction and were put into a 96-well cell plate 3. Results and discussion
cultured with MEMα medium containing 1% penicillin 3.1. Characterization, morphology, and mechanical
and streptomycin. 1 mL of mouse preosteoblast cells property of β-TCP/TiO ceramic
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(MC3T3-E1) at a density of 6000 cells/well were seeded Figure 1B revealed the XRD diagram of TiO , β-TCP and
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in each well and then cultured in a CO incubator at 37°C β-TCP/5-TiO ceramic in detail. The XRD diffraction
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for 1 and 3 days. After each incubation period, specimens pattern of TiO showed that the typical phase structure
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were rinsed twice with phosphate-buffered saline, and of the rutile phase, confirmed by the JCPDS card (21-
then 10 μL MTT (5 mg/mL) was added to each well and 1276) at 25.25°, 37.96°, 48.20°, 54.143°, and 62.87° . The
[35]
incubated for another 3 h. The cell viability was detected prominent peak of TiO was found in the XRD diagram
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by the optical density value, and the cell survival rate was of β-TCP/5-TiO , indicating that rutile TiO existed in
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counted. the structure of β-TCP/5-TiO ceramic. Besides, the
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The cells on the scaffold surface were stained and addition of TiO slightly affected the diffraction pattern
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observed to investigate the proliferation and adhesion of β-TCP in the range in diffraction angle from 25° to
of osteoblasts on scaffolds. Briefly, after the end of each 25.5°, indicating that TiO was indeed incorporated into
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incubation period of 1, 3, and 7 days, osteoblasts were the β-TCP matrix.
fixed in 5% paraformaldehyde and subjected to gradient The infrared spectrum of TiO , β-TCP and β-TCP/TiO
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dehydration using 30%, 50%, 80%, 90%, and 100% alcohol, ceramic is shown in Figure 1C. The vibration absorption
followed by SEM observation. Meanwhile, after 1, 3, and peak of PO was close to 1120 cm , 1042cm , 607cm ,
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3-
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7 days of incubation in a cell incubator, the osteoblasts on and 552cm -1[36] . Briefly, 1120 cm and 1042 cm belonged
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scaffolds were also dyed with calcein AM/propidium iodide to the asymmetric stretching vibration of the P=O group;
(PI) solution, and the adhesion of cells on the surface was 607 cm was the symmetric stretching vibration peak of
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observed by an inverted fluorescence microscope (Nikon PO ; 552 cm corresponded to the bending vibration
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Eclipse Ti-E, Japan). absorption. For TiO particle, characteristic absorption
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Volume 9 Issue 2 (2023) 372 https://doi.org/10.18063/ijb.v9i2.673