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Abstract：
OBJECTIVE: To explore the feasibility of injectable platelet-rich fibrin (i-PRF) in regenerative endodontics by comparing the effect of i-PRF and platelet-rich fibrin (PRF) on the biological behavior and angiogenesis of human stem cells from the apical papilla (SCAPs).
METHODS: i-PRF and PRF were obtained from venous blood by two different centrifugation methods, followed by hematoxylin-eosin (HE) staining and scanning electron microscopy (SEM). Enzyme-linked immunosorbent assay (ELISA) was conducted to quantify the growth factors. SCAPs were cultured with different concentrations of i-PRF extract (i-PRFe) and PRF extract (PRFe), and the optimal concentrations were selected using the Cell Counting Kit-8 (CCK-8) assay. The cell proliferation and migration potentials of SCAPs were then observed using the CCK-8 and Transwell assays. Mineralization ability was detected by alizarin red staining (ARS), and angiogenesis ability was detected by tube formation assay. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to evaluate the expression of genes related to mineralization and angiogenesis. The data were subjected to statistical analysis.
RESULTS: i-PRF and PRF showed a similar three-dimensional fibrin structure, while i-PRF released a higher concentration of growth factors than PRF ( P <.05). 1/4× i-PRFe and 1/4× PRFe were selected as the optimal concentrations. The cell proliferation rate of the i-PRFe group was higher than that of the PRFe group ( P <.05), while no statistical difference was observed between them in terms of cell mitigation ( P >.05). More importantly, our results showed that i-PRFe had a stronger effect on SCAPs than PRFe in facilitating mineralization and angiogenesis, with the consistent result of RT-qPCR ( P <.05).
CONCLUSIONS: This study revealed that i-PRF released a higher concentration of growth factors and was superior to PRF in promoting proliferation, mineralization and angiogenesis of SCAPs, which indicates that i-PRF could be a promising biological scaffold for application in pulp regeneration.
METHODS: i-PRF and PRF were obtained from venous blood by two different centrifugation methods, followed by hematoxylin-eosin (HE) staining and scanning electron microscopy (SEM). Enzyme-linked immunosorbent assay (ELISA) was conducted to quantify the growth factors. SCAPs were cultured with different concentrations of i-PRF extract (i-PRFe) and PRF extract (PRFe), and the optimal concentrations were selected using the Cell Counting Kit-8 (CCK-8) assay. The cell proliferation and migration potentials of SCAPs were then observed using the CCK-8 and Transwell assays. Mineralization ability was detected by alizarin red staining (ARS), and angiogenesis ability was detected by tube formation assay. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to evaluate the expression of genes related to mineralization and angiogenesis. The data were subjected to statistical analysis.
RESULTS: i-PRF and PRF showed a similar three-dimensional fibrin structure, while i-PRF released a higher concentration of growth factors than PRF ( P <.05). 1/4× i-PRFe and 1/4× PRFe were selected as the optimal concentrations. The cell proliferation rate of the i-PRFe group was higher than that of the PRFe group ( P <.05), while no statistical difference was observed between them in terms of cell mitigation ( P >.05). More importantly, our results showed that i-PRFe had a stronger effect on SCAPs than PRFe in facilitating mineralization and angiogenesis, with the consistent result of RT-qPCR ( P <.05).
CONCLUSIONS: This study revealed that i-PRF released a higher concentration of growth factors and was superior to PRF in promoting proliferation, mineralization and angiogenesis of SCAPs, which indicates that i-PRF could be a promising biological scaffold for application in pulp regeneration.
摘要：
目的：通过比较i-PRF和富血小板纤维蛋白(PRF)对人根尖乳头干细胞(SCAPs)生物学行为和血管生成的影响，探讨可注射富血小板纤维蛋白(i-PRF)在再生牙髓治疗中的可行性。
方法：通过两种不同的离心方法从静脉血中获得i-PRF和PRF，随后进行苏木精-伊红(HE)染色和扫描电镜(SEM)。进行酶联免疫吸附测定(ELISA)以定量生长因子。用不同浓度的i-PRF提取物(i-PRFe)和PRF提取物(PRFe)培养SCAP,使用细胞计数试剂盒-8(CCK-8)测定选择最佳浓度。然后使用CCK-8和Transwell测定观察SCAP的细胞增殖和迁移潜力。通过茜素红染色（ARS）检测矿化能力，通过试管形成试验检测血管生成能力。进行实时定量聚合酶链反应（RT-qPCR）以评估与矿化和血管生成相关的基因的表达。对数据进行统计分析。
结果：i-PRF和PRF显示出相似的三维纤维蛋白结构，i-PRF释放的生长因子浓度高于PRF（P<0.05）。选择1/4×i-PRFe和1/4×PRFe作为最佳浓度。i-PRFe组细胞增殖率高于PRFe组(P<0.05),而在细胞缓解方面，它们之间没有观察到统计学差异（P>.05）。更重要的是,我们的结果表明，i-PRFe在促进矿化和血管生成方面对SCAP的作用强于PRFe，与RT-qPCR结果一致(P<0.05)。
结论：这项研究表明，i-PRF释放更高浓度的生长因子，在促进增殖方面优于PRF，SCAP的矿化和血管生成，这表明i-PRF可能是一种有前途的应用于纸浆再生的生物支架。
方法：通过两种不同的离心方法从静脉血中获得i-PRF和PRF，随后进行苏木精-伊红(HE)染色和扫描电镜(SEM)。进行酶联免疫吸附测定(ELISA)以定量生长因子。用不同浓度的i-PRF提取物(i-PRFe)和PRF提取物(PRFe)培养SCAP,使用细胞计数试剂盒-8(CCK-8)测定选择最佳浓度。然后使用CCK-8和Transwell测定观察SCAP的细胞增殖和迁移潜力。通过茜素红染色（ARS）检测矿化能力，通过试管形成试验检测血管生成能力。进行实时定量聚合酶链反应（RT-qPCR）以评估与矿化和血管生成相关的基因的表达。对数据进行统计分析。
结果：i-PRF和PRF显示出相似的三维纤维蛋白结构，i-PRF释放的生长因子浓度高于PRF（P<0.05）。选择1/4×i-PRFe和1/4×PRFe作为最佳浓度。i-PRFe组细胞增殖率高于PRFe组(P<0.05),而在细胞缓解方面，它们之间没有观察到统计学差异（P>.05）。更重要的是,我们的结果表明，i-PRFe在促进矿化和血管生成方面对SCAP的作用强于PRFe，与RT-qPCR结果一致(P<0.05)。
结论：这项研究表明，i-PRF释放更高浓度的生长因子，在促进增殖方面优于PRF，SCAP的矿化和血管生成，这表明i-PRF可能是一种有前途的应用于纸浆再生的生物支架。
