CLINICAL PROTOCOL FOR THE PREPARATION AND ELECTRON MICROSCOPIC ANALYSIS OF THE OBTAINED PRODUCTS OF AUTOLOGOUS MESOCONCENTRATE – PLASMA RICH IN GROWTH FACTORS (PRGF)

A. P. Oshurko, Ihor Yu. Oliinyk, T. Kerimova, E. Pompii
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Abstract

Introduction. Ensuring control over regenerative processes by using the patient's blood is a unique concept of an autogenous transplant product, a universal and safe method of application that contains growth factors and cytokines stored in the fibrin matrix and provides stimulating properties of tissue regeneration through the processes of angiogenesis, proliferation, cell migration, and extracellular matrix synthesis. The aim. To substantiate the effectiveness of the protocol for collecting, preparing, and forming autocellular mesoconcentrate products in the analysis of the results of their electron microscopic examination. Materials and methods. Based on our own clinical experience of using the Endoret-PRGF technique (Human Technology, BTI, Spain) to collect, prepare, and form autocellular transplant products rationally, we used the method of morphological study of objects using a stream of electrons that passed through thin films under high voltage, allowing us to study the structure of these objects at the macromolecular and subcellular levels – transmission electron microscopy, which contributed to the study of the density (number of occurrences in 10 μm2 ), diameter (Ø) of the formed fibrin fibers in the PRGF – F1 and F2 fractions of mesoconcentrate products, namely, isolating membranes (M) and obturating blocks (B). An equally important step in PRGF technology is to follow the step-by-step instructions for preparing and using autocellular transplants. Results. Protocol for the collection, preparation, and formation of autocellular transplants using the KMU15 kit (Human Technology, BTI) for the application of the Endoret – PRGF technique, platelet-rich plasma, as a unique and first scientifically proven technique recognized worldwide and patented by the Institute of Human Biotechnology, Spain. The results of the clinical application of mesoconcentrate products, namely isolating membranes widely used in targeted tissue regeneration and obturating blocks, confirm their biological effectiveness. The results of the median (Me) and interquartile range (IQR) during the statistical analysis were obtained, where F1-M = 0.196 (0.176; 0.286) compared to F1-B = 0.344 (0.325; 0.394); F2-M = 0.180 (0.168; 0.214) - F2-B = 0.254 (0.202; 0.338), which gives us the right to assert the reliability of the data set and confirms the quantitative characterization of interfraction differences, and, accordingly, the quality in the sequence of protocol stages. Conclusions. The protocol for the efficiency of autocellular graft collection and formation is synchronized with the analysis of electron microscopic images characterizing the density and diameter of fibrin fibers, which differ in the fractional products of the mesoconcentrate and are higher in the obturating blocks than in the isolating membranes, indicating its completeness and compliance.
自体中胚层浓缩物--富含生长因子的血浆(PRGF)制备和电子显微镜分析临床方案
简介利用患者的血液确保对再生过程的控制是自体移植产品的一个独特概念,这是一种普遍而安全的应用方法,它含有储存在纤维蛋白基质中的生长因子和细胞因子,通过血管生成、增殖、细胞迁移和细胞外基质合成过程提供刺激组织再生的特性。 目的通过对电子显微镜检查结果的分析,证实收集、制备和形成自细胞间质浓缩物产品的方案的有效性。 材料和方法。根据我们使用 Endoret-PRGF 技术(人类技术公司,BTI,西班牙)合理收集、制备和形成自细胞移植产物的临床经验,我们使用了在高压下通过薄膜的电子流对物体进行形态学研究的方法、透射电子显微镜有助于研究 PRGF - F1 和 F2 中胚层浓缩产物(即分离膜(M)和钝化块(B))中纤维蛋白纤维的密度(10 μm2 中出现的数量)和直径(Ø)。PRGF 技术中同样重要的一步是按照步骤说明制备和使用自细胞移植体。 结果。使用 KMU15 试剂盒(人类技术公司,BTI)收集、制备和形成自细胞移植的规程,该试剂盒用于 Endoret - PRGF 技术(血小板丰富血浆)的应用,该技术是世界公认的独一无二的、首个经过科学验证的技术,并已获得西班牙人类生物技术研究所的专利。中胚层浓缩物产品,即广泛应用于定向组织再生和闭合块的分离膜,其临床应用结果证实了其生物有效性。统计分析期间的中位数(Me)和四分位数间距(IQR)结果得出,F1-M = 0.196 (0.176; 0.286) 与 F1-B = 0.344 (0.325; 0.394) 相比;F2-M = 0.180 (0.168; 0.214) - F2-B = 0.254 (0.202; 0.338),这使我们有权断言数据集的可靠性,并确认了衍射间差异的定量特征,以及相应的方案阶段顺序的质量。 结论自细胞移植物收集和形成效率方案与表征纤维蛋白纤维密度和直径的电子显微镜图像分析同步进行,纤维蛋白纤维密度和直径在中胚层浓缩物的分馏产物中存在差异,在闭合块中高于分离膜,这表明其完整性和合规性。
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