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

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.