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冻干富血小板纤维蛋白(Ly-PRF)在组织再生中的临床和免疫组织化学表现。

Clinical and immunohistochemical performance of lyophilized platelet-rich fibrin (Ly-PRF) on tissue regeneration.

机构信息

Department of Stomatology, General Hospital, Tianjin Medical University, Tianjin, 300052, People's Republic of China.

School of Dentistry, Hospital of Stomatology, Tianjin Medical University, Tianjin, 300070, People's Republic of China.

出版信息

Clin Implant Dent Relat Res. 2017 Jun;19(3):466-477. doi: 10.1111/cid.12473. Epub 2017 Feb 13.

Abstract

BACKGROUND

Platelet-rich fibrin (PRF) has been widely used in oral implantology and other fields, but benefits of the fresh PRF (FPRF (fresh platelet-rich fibrin)) were consequently limited because of its short-term application. Thus, a protocol for the combination of PRF and lyophilization comes up in the present study to address the issue of PRF storage and delayed clinical application, which has little been reported in this field at home and abroad by now.

PURPOSE

The aim of the present study was to evaluate the applicability of lyophilized platelet-rich fibrin (Ly-PRF) used as the scaffold material for craniofacial tissue regeneration and to compare its biochemical properties with commonly used fresh PRF.

MATERIALS AND METHODS

Two volunteers with both genders were selected as the source of PRF and Ly-PRF samples. Macro- and micro-scopic appearance evaluation as well as immunohistochemical comparison were performed on PRF samples before and after freeze-drying at -196°C. The second experimental phase was to observe clinical performance when fresh and lyophilized PRF were applied in guided bone regeneration (GBR) operations in 39 patients losing teeth in the anterior maxillary region who required an oral implantation followed by labial bone grafting.

RESULTS

The conventional histological and transmission electron microscopy images showed the microstructure of Ly-PRF, which resembled a mesh containing apparently irregularly shaped platelets with less alpha-granule than fresh PRF in micro and a translucent membrane with less elasticity than fresh PRF in macro. Simultaneous immunohistological staining results showed positive expression of PDGF-BB, IL-1, IL-4, TNF, TGF-β in both fresh and lyophilized PRF, while the expression of PDGF-BB, IL-1, TNF, TGF-β has no statistical difference between them (P > .05) but that of IL-4 in Ly-PRF is statistically higher than in fresh PRF (P < .05). When applied in GBR operations, there were no significant differences between Ly-PRF and FPRF in factors of histological and clinical evaluations (i.e., color, swelling, bleeding of the mucosa, pain leveland, and remodeling of hard tissue) performed 3 days, 7 days, and 4 months after the surgery (P > .05).

CONCLUSIONS

This study strongly supports that lyophilization at -196°C does not largely influence the expression of bioactive factors, the microstructure of fibrinogen or the clinical effects of PRF.

摘要

背景

富血小板纤维蛋白(PRF)已广泛应用于口腔种植学及其他领域,但由于其短期应用,新鲜 PRF(FPRF(新鲜血小板富纤维蛋白))的益处因此受到限制。因此,本研究提出了 PRF 与冻干相结合的方案,以解决 PRF 储存和延迟临床应用的问题,目前国内外对此领域的报道甚少。

目的

本研究旨在评估冻干富血小板纤维蛋白(Ly-PRF)用作颅面组织再生支架材料的适用性,并将其生物化学特性与常用的新鲜 PRF 进行比较。

材料和方法

选择两名性别均为男性和女性的志愿者作为 PRF 和 Ly-PRF 样本的来源。对 -196°C 冷冻干燥前后的 PRF 样本进行宏观和微观外观评估以及免疫组织化学比较。第二阶段实验是观察 39 例上颌前牙缺失患者在引导骨再生(GBR)手术中应用新鲜和冻干 PRF 的临床效果,这些患者需要进行口腔种植体植入和唇侧骨移植。

结果

常规组织学和透射电子显微镜图像显示 Ly-PRF 的微观结构,类似于含有明显不规则形状血小板的网格,与新鲜 PRF 相比,α-颗粒较少,宏观上呈半透明膜,弹性较小。同时免疫组织化学染色结果显示,新鲜和冻干 PRF 均呈 PDGF-BB、IL-1、IL-4、TNF、TGF-β 阳性表达,两者之间 PDGF-BB、IL-1、TNF、TGF-β 表达无统计学差异(P>.05),但 Ly-PRF 中 IL-4 的表达明显高于新鲜 PRF(P<.05)。在 GBR 手术中,Ly-PRF 和 FPRF 在术后 3 天、7 天和 4 个月的组织学和临床评价(即黏膜颜色、肿胀、出血、疼痛程度和硬组织重塑)因素方面无显著差异(P>.05)。

结论

本研究强烈支持 -196°C 冻干对生物活性因子的表达、纤维蛋白原的微观结构或 PRF 的临床效果没有太大影响。

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