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标准化力可制备出质量相似的富血小板纤维蛋白,而不受转子角度的影响。

A Standardized -Force Allows the Preparation of Similar Platelet-Rich Fibrin Qualities Regardless of Rotor Angle.

作者信息

Al-Maawi Sarah, Dohle Eva, Kretschmer Winfried, Rutkowski James, Sader Robert, Ghanaati Shahram

机构信息

Frankfurt Oral Regenerative Medicine (FORM), Clinic for Maxillofacial and Plastic Surgery, Goethe University, Frankfurt Am Main, Germany.

Medius Clinic Ostfildern-Ruit, Ostfildern, Germany.

出版信息

Tissue Eng Part A. 2022 Apr;28(7-8):353-365. doi: 10.1089/ten.TEA.2021.0113. Epub 2022 Feb 2.

Abstract

Platelet-rich fibrin (PRF) is an autologous blood concentrate that supports tissue regeneration. The effect of the centrifuge rotor angle in the fabrication of PRF is still not fully elucidated. The hypothesis of this study is: When applying the same -force (relative centrifugal force [RCF]) and centrifugation time, PRF components and bioactivity are not modified using either a swing-out rotor or a fixed angle rotor. For this purpose, peripheral blood samples (from five donors) were used to gain solid (710 ×, 8 min) and liquid (44 ×, 8 min) PRF matrices using three different centrifuges (one fixed angle as a control and two different swing-out rotor centrifuges). The physical characteristics of the solid PRF were measured to evaluate the clot formation and cellular distribution. The liquid PRF was used to evaluate the cell number, bioactivity, and influence on primary human osteoblasts (pOBs) and primary human fibroblasts (pHFs) . Solid PRF clots were significantly larger in the group of fixed rotor centrifuges compared with either of the two evaluated swing-out rotor centrifuges. No differences were observed when evaluating the cellular distribution within the solid PRF. No statistically significant differences were documented in the cell's density in liquid PRF samples (platelets, lymphocytes, neutrophils, eosinophils, and basophils) among the differently gained PRF samples. No statistically significant differences were documented for the released growth factors (vascular endothelial growth factor, epidermal growth factor, and transforming growth factor beta 1) over 7 days. pOBs and pHFs viability after treatment with PRF conditioned media showed no statistically significant differences between the evaluated groups. However, the number of adherent cells treated with PRF obtained with the use of the fixed angle rotor was significantly higher when compared with those treated with PRF obtained by using the swing-out rotors. The presented results confirm that regardless of the centrifuge rotor used, the components and bioactivity of solid and liquid PRF matrices are modified by the applied RCF and centrifugation time. These findings are of great importance for highlighting the essential role of adapting the centrifugation protocols when using different centrifuges and to correctly report the used centrifugation protocols in scientific research to allow for reproducible results. Impact statement Platelet-rich fibrin (PRF) is prepared from autologous peripheral blood and is widely applied in research and clinical treatments. The centrifugation parameters used during the preparation of PRF directly influence its components and bioactivity. By using a standardized protocol, the present study demonstrated that adapting various centrifuges to a standardized relative centrifugal force and centrifugation protocol resulted in reproducible PRF matrices with similar bioactivity, regardless of the centrifuge rotor angle. These findings underline the necessity to carefully adapt and correctly report the used centrifuge and centrifugation protocols in scientific research to allow reproducible results.

摘要

富血小板纤维蛋白(PRF)是一种支持组织再生的自体血液浓缩物。离心机转子角度对PRF制备的影响仍未完全阐明。本研究的假设是:当施加相同的力(相对离心力[RCF])和离心时间时,使用甩平式转子或固定角度转子不会改变PRF的成分和生物活性。为此,使用外周血样本(来自五名供体),通过三台不同的离心机(一台固定角度离心机作为对照,两台不同的甩平式转子离心机)制备固态(710×,8分钟)和液态(44×,8分钟)PRF基质。测量固态PRF的物理特性以评估凝块形成和细胞分布。液态PRF用于评估细胞数量、生物活性以及对原代人成骨细胞(pOBs)和原代人成纤维细胞(pHFs)的影响。与两台评估的甩平式转子离心机中的任何一台相比,固定转子离心机组中的固态PRF凝块明显更大。在评估固态PRF内的细胞分布时未观察到差异。在不同获得的PRF样本中,液态PRF样本(血小板、淋巴细胞、中性粒细胞、嗜酸性粒细胞和嗜碱性粒细胞)中的细胞密度没有统计学上的显著差异。在7天内释放的生长因子(血管内皮生长因子、表皮生长因子和转化生长因子β1)没有统计学上的显著差异。用PRF条件培养基处理后,pOBs和pHFs的活力在评估组之间没有统计学上的显著差异。然而,与用甩平式转子获得的PRF处理的细胞相比,用固定角度转子获得的PRF处理的贴壁细胞数量明显更高。呈现的结果证实,无论使用何种离心机转子,固态和液态PRF基质的成分和生物活性都受所施加的RCF和离心时间的影响。这些发现对于突出在使用不同离心机时调整离心方案的重要作用以及在科学研究中正确报告所使用的离心方案以获得可重复的结果具有重要意义。影响声明富血小板纤维蛋白(PRF)由自体外周血制备,广泛应用于研究和临床治疗。PRF制备过程中使用的离心参数直接影响其成分和生物活性。通过使用标准化方案,本研究表明,无论离心机转子角度如何,将各种离心机调整到标准化的相对离心力和离心方案可产生具有相似生物活性的可重复PRF基质。这些发现强调了在科学研究中仔细调整并正确报告所使用的离心机和离心方案以获得可重复结果的必要性。

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