浓缩生长因子联合 iRoot BP Plus 促进发炎牙髓修复：一项体内外研究。
Concentrated growth factor combined with iRoot BP Plus promotes inflamed pulp repair: an in vitro and in vivo study.
机构信息
Hospital of Stomatology, Sun Yat-Sen University; Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, 510055, China.
Guanghua School of Stomatology, Sun Yat-Sen University, Guangzhou, 510055, China.
出版信息
BMC Oral Health. 2023 Apr 19;23(1):225. doi: 10.1186/s12903-023-02903-5.
BACKGROUND
Platelet concentrates combined with calcium silicate cements may promote reparative dentin formation. However, few studies have reported their effect on dental pulp inflammation. This study aimed to evaluate the effects of concentrated growth factor (CGF) combined with iRoot BP Plus on inflammatory human dental pulp stem cells (hDPSCs) in vitro and inflamed pulp in rats in vivo.
METHODS
The proliferation of LPS-stimulated hDPSCs treated with 50% CGF with/without 25% iRoot BP Plus was evaluated using Cell Counting Kit-8 on days 1, 4 and 7. The expression of genes associated with inflammation on day 1 and differentiation on day 14 was analysed by real-time polymerase chain reaction. The exposed pulp of rat maxillary molars was injected with 10 mg/mL LPS and directly capped with CGF membrane with/without iRoot BP Plus extract for 1, 7 and 28 days. The teeth were subjected to histologic analyses and immunohistochemistry.
RESULTS
The proliferation rates of the inflammatory hDPSCs after the combination treatment were significantly higher than those after the other treatments on days 4 and 7 (P < 0.05). IL-1β, IL-6, and TNF-α levels were increased in inflammatory hDPSCs but decreased after treatment with CGF combined with iRoot BP Plus extract, whereas IL-4 and IL-10 showed the opposite expression patterns. Expression of the odontogenesis-related genes OCN, Runx2, and ALP was dramatically enhanced by combined treatment with CGF and iRoot BP Plus extract. In rat pulp, the average inflammation scores of the CGF and CGF-iRoot BP Plus groups significantly decreased in comparison with those of the LPS group (P < 0.05), and the CGF-iRoot BP Plus group had more reparative dentin than the CGF and BP groups. Immunohistochemical staining showed fewer M1 macrophages on day 1 and more M2 macrophages on day 7 in the CGF-iRoot BP Plus group than in the other groups.
CONCLUSIONS
The combination of CGF and iRoot BP Plus showed a synergistic effect on anti-inflammatory potential and promoted greater pulp healing than CGF or iRoot BP Plus alone.
背景
血小板浓缩物与硅酸钙水泥结合可能促进修复性牙本质形成。然而，很少有研究报道它们对牙髓炎症的影响。本研究旨在评估浓缩生长因子（CGF）与 iRoot BP Plus 联合应用于体外培养的炎症性人牙髓干细胞（hDPSCs）和体内大鼠炎症牙髓的效果。
方法
使用 Cell Counting Kit-8 在第 1、4 和 7 天分别评估 LPS 刺激的 hDPSCs 用 50%CGF 与/或 25%iRoot BP Plus 处理后的增殖情况。采用实时聚合酶链反应分析第 1 天与炎症相关的基因表达和第 14 天的分化情况。将上颌磨牙暴露的牙髓注入 10mg/mL LPS，并直接用 CGF 膜与/或 iRoot BP Plus 提取物覆盖 1、7 和 28 天。对牙齿进行组织学分析和免疫组织化学分析。
结果
联合治疗后，炎症 hDPSCs 的增殖率在第 4 天和第 7 天均明显高于其他处理组（P<0.05）。IL-1β、IL-6 和 TNF-α水平在炎症 hDPSCs 中升高，但用 CGF 联合 iRoot BP Plus 提取物处理后降低，而 IL-4 和 IL-10 则呈现相反的表达模式。OCN、Runx2 和 ALP 等牙发生相关基因的表达在 CGF 与 iRoot BP Plus 提取物联合处理后显著增强。在大鼠牙髓中，与 LPS 组相比，CGF 组和 CGF-iRoot BP Plus 组的平均炎症评分显著降低（P<0.05），且 CGF-iRoot BP Plus 组的修复性牙本质多于 CGF 组和 BP 组。免疫组织化学染色显示，与其他组相比，CGF-iRoot BP Plus 组在第 1 天的 M1 巨噬细胞较少，在第 7 天的 M2 巨噬细胞较多。
结论
CGF 与 iRoot BP Plus 的联合应用具有协同的抗炎潜力，并促进了比 CGF 或 iRoot BP Plus 单独应用更大的牙髓愈合。
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