Page 474 - IJB-10-1

International Journal of Bioprinting TPMS bone scaffold

releasing at an average rate of 7.56% per day. On the 10th migration capacity of hBMSCs without inducing cellular
day, the release rate of I-PRF-coated methylene blue almost toxicity. Moreover, the expression of osteogenic markers
reached 100%. As Miron et al. (2017) have stated, I-PRF and the capacity to form mineralized nodules of hBMSCs
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presents a potential advantage in terms of the releasing of were promoted by 20% I-PRF. However, our results show
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quantity and duration of growth factors, including TGF-β1, that concentrations ranging from 1% to 20% of I-PRF
VEGF, EGF, IGF-1, PDGF-AA, PDGF-AB, and PDGF-BB. promoted MC3T3 cell growth. Particularly, a 5% I-PRF
concentration exhibited the highest cell viability. The
3.2. Exploration of the optimum loading differential findings observed between these studies are
concentration of I-PRF likely due to the difference in the species and proliferation
The process of extracting I-PRF from rabbit ear artery capacity of hBMSCs and MC3T3 cells.
and preparing an I-PRF-loaded bone scaffold is shown in
Figure 3A. To explore the optimum loading concentration Next, we evaluated the effect on cell migration of extracts
of I-PRF on the bone scaffold, MC3T3 cells were treated from the different components after 24 h (Figure 3D and
with different concentrations of I-PRF, then its effect on cell E). The findings showed that all test groups significantly
proliferation and migration was evaluated. Figure 3B shows enhanced the migration rate compared to the untreated
the live-dead staining of MC3T3 cells with and without control (P < 0.0001). However, the TPMS bone scaffold with
I-PRF taken by a fluorescence microscope (Olympus, I-PRF exhibited the highest migration rate compared to the
IX73, Japan). The green fluorescence in the image shows I-PRF, TPMS, and the control (P < 0.0001). Collectively,
live cells, indicating that the cells in the scaffold without these findings demonstrate the tremendous potential of
I-PRF were concentrated in the pores and clustered into utilizing I-PRF in conjunction with TPMS scaffolds as a
clusters. In contrast, the scaffolds with I-PRF were filled semi-synthetic autologous bone grafting material.
with filamentous structures between the pore walls, and
the cells were dispersed evenly in the filamentous structure, 3.3. Biocompatibility analysis of the bone
indicating that they could promote cell proliferation and scaffold in vitro
adhesion. The staining results of different concentrations of According to previous research on the osteogenic
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I-PRF showed that the 5% I-PRF group exhibited the highest effect of I-PRF, it positively influenced cell adhesion,
ratio of live cells (green staining) to dead cells (red staining), differentiation, 59,60 migration, 61 proliferation, and
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indicating its enhanced therapeutic potency (Figure 3C). The mineralization potential. In order to evaluate the
experiment confirmed that the scaffold added with I-PRF biocompatibility of 3D printing TPMS scaffold with
possesses a reticular structure, which increases the surface biological materials, MC3T3 cells were inoculated onto
area for cell adhesion. The cells were observed to adhere to each scaffold composite. From the SEM imaging (Figure 4),
the reticular structure and the side wall of the scaffold. The a large number of cells with the typical morphology can be
cell activity in the IT group was significantly higher than observed adhering to all four groups of scaffolds, reflecting
that of the blank scaffold group, further confirming the role that all materials exhibited good cell adhesion. Moreover,
of I-PRF in promoting cell proliferation. following coating the base scaffold with I-PRF and SDF-1,
no significant negative effect on the cell adhesion was
The optimum concentration for subsequent I-PRF
loading was selected based on the results of the evaluation observed, indicating that I-PRF and SDF-1 bound on the
surface of the scaffold had no cytotoxicity to the cells.
of the biological effect of I-PRF on MC3T3 cells. CCK-8 These findings indicate the potential of this material to
tests were performed on cells incubated with I-PRF at 1%, accelerate the healing and reconstruction of injured tissue
5%, 10%, and 20%, respectively, to evaluate its effect on the and promote the healing and regeneration of bone.
proliferation of MC3T3 cells on the scaffolds (Figure 3F).
As expected, the results showed that I-PRF could promote The effects of the different scaffold compositions on
MC3T3 cell proliferation. On day 1, 10% (P < 0.05) and 20% MC3T3 cell proliferation were evaluated using the CCK-8
(P < 0.01) I-PRF significantly promoted cell proliferation assay. Our findings showed that the SIT group exhibited
when compared to the untreated control. On the 2nd the highest cell viability, followed by the ST, IT, T, and NC
day, 5% I-PRF substantially enhanced the proliferation of groups (Figure 5E). These results indicated that SDF-1 and
MC3T3 cells compared to the untreated control. While I-PRF had synergistic effects in promoting cell growth. In
on day 3, 1%, 5%, and 10% I-PRF significantly increased addition, each group was subjected to live-dead staining to
proliferation when compared to the untreated cells (P < determine the impact on MC3T3 cell viability. As shown
0.05); however, no significance was observed from the in Figure 5A, the cells in SIT group showed the highest
20% group (P > 0.05). In the review by Wang et al. (2023), average fluorescence intensity, while the ST group showed
the CCK-8 and wound-healing assay showed that 10% the lowest, followed by the IT and SIT groups with slight
and 20% I-PRF improved the proliferation rate and the differences (Figure 5C).

Volume 10 Issue 1 (2024) 466 https://doi.org/10.36922/ijb.0153