标准化的重力允许制备类似的富含血小板的纤维蛋白质量,而不管转子的角度如何。

Tissue Engineering Part A Pub Date : 2022-04-01 Epub Date: 2022-02-02 DOI:10.1089/ten.TEA.2021.0113
Sarah Al-Maawi, Eva Dohle, Winfried Kretschmer, James Rutkowski, Robert Sader, Shahram Ghanaati
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引用次数: 4

摘要

富血小板纤维蛋白(PRF)是一种支持组织再生的自体浓缩血。离心转子角度对PRF制备的影响尚未完全阐明。本研究的假设是:在施加相同的重力(相对离心力[RCF])和离心时间的情况下,无论是采用摆出转子还是固定角度转子都不改变PRF组分和生物活性。为此,使用三种不同的离心机(一种固定角度作为对照,两种不同的旋转式离心机),使用五位献血者的外周血样本获得固体(710 ×g, 8分钟)和液体(44 ×g, 8分钟)PRF基质。测量固体PRF的物理特性以评估凝块形成和细胞分布。用液体PRF评价细胞数量、生物活性以及对体外培养的原代人成骨细胞(pob)和原代人成纤维细胞(phf)的影响。与两种评估的旋转式离心机相比,固定转子离心机组的固体PRF凝块明显更大。在评估固体PRF内的细胞分布时,没有观察到差异。在不同获得的PRF样品中,液体PRF样品中的细胞密度(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)没有统计学上的显著差异。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β 1)无统计学差异。经PRF条件培养基处理后的pob和phf活力在评估组之间无统计学差异。然而,使用固定角度转子获得的PRF处理的贴壁细胞数量明显高于使用摆动转子获得的PRF处理的细胞数量。结果表明,无论使用何种离心机转子,固体和液体PRF基质的成分和生物活性都受到施加的RCF和离心时间的影响。这些发现对于强调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允许可重复的结果具有重要意义。富血小板纤维蛋白(PRF)是从自体外周血中制备的,在研究和临床治疗中有着广泛的应用。制备PRF时使用的离心参数直接影响其成分和生物活性。通过使用标准化协议,本研究表明,使各种离心机适应标准化的相对离心力和离心协议,无论离心机转子角度如何,都可以获得具有相似生物活性的可重复PRF基质。这些发现强调了仔细调整和正确报告科学研究中使用的离心机和离心方案的必要性,以允许可重复的结果。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A Standardized g-Force Allows the Preparation of Similar Platelet-Rich Fibrin Qualities Regardless of Rotor Angle.

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 g-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 ×g, 8 min) and liquid (44 ×g, 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) in vitro. 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.

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Tissue Engineering Part A
Tissue Engineering Part A CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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