USE OF VARIOUS RESEARCH METHODS IN FORENSIC PRACTICE TO DETERMINE LIFETIME AND AGE OF INJURY: LITERATURE REVIEW



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Abstract: The problem of the prescription of damage formation has been of interest to forensic physicians since the XIX century to the present. After analyzing the literature sources on the problem of the prescription of traumatic hemorrhages, it becomes clear that there is no single position, approach to the study of this issue, an unambiguous time frame that accurately determines the period from injury to the moment of death of the victim. Both in the nineteenth century and in the twenty-first, forensic doctors initially focused on the macroscopic picture of the injury followed by microscopic examination. When studying micropreparations, they are guided by: the cellular composition of hemorrhage, based on already known data, on their own experience and knowledge in this matter, on the brief circumstances of the incident (if any are indicated in the direction of forensic histological examination), on the age of the victim. Over the years, with the acquired experience and accumulated knowledge, researchers have determined approximate, developed approximate time frames for the formation of damage, now taking into account not only the above parameters, but also the condition of the victim during and after the injury. But scientists have not come to a consensus. Medicine, achievements, progress and perseverance of scientists strive forward, involving related sciences in the study of antiquity. As a result, such methods as: molecular genetic, biophysical, biochemical, immunohistochemical, etc., have found their application, allowing to study not only the cellular composition, but also enzymatic, hormonal changes in damaged tissues. Instrumental methods are being developed: ultrasound. When using both laboratory and instrumental methods, as well as their combination, this problem remains completely unresolved.

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For centuries, forensic doctors have been looking for ways to solve the problem in determining the lifetime of the formation of traumatic hemorrhages, the most urgent and difficult question remains the timing of damage formation from the moment of injury to the death of the victim. Over time, the relevance of the issue, in determining the lifetime and prescription, increased, was repeatedly emphasized at All-Russian Scientific and Practical conferences (1973-2022), congresses, plenums of the Society of forensic physicians, and already at the initial stages of the development of forensic medical practice, researchers began to study this problem.
Shishkin I.P. (1895), analyzing the achievements of previous authors and the actual research carried out, comes to the conclusion that: bruises are accompanied by certain tissue reactions that increase over time; tissues have "survivability". An experimental study (Berg O.Yu. et al., 2008) testifies to the same, about the "survivability" of tissues and insufficiently pronounced phenomena of the initial period of reparative processes in the injury zone, showing the similarity of changes in tissues, according to their morphological indicators, in lifetime and posthumously inflicted injuries. By determining the expression of proinflammatory cytokines (interleukins 6 and 20; IL-6 and IL-20), which is elevated to 72 hours of the postmortem period in damaged tissues (Ye MY, Xu D, Liu JC, Lyu HP, Xue Y, He JT, Huang HY, Zhang KK, Xie XL, Wang Q., 2018) and a simultaneous increase in IL-6, tumor necrosis factor-alpha (TNF-α) can indicate that the damage is less than 30 minutes (Birincioğlu I, Akbaba M, Alver A, Kul S, Özer E, Turan N, Şentürk A, Ince I., 2016). Also, the main inflammatory mediators (cytokines, growth factor) (Kondo T, Ishida Y., 2010), collagen in the study of injuries can be reliable indicators and markers for determining the viability or age of the wound (Kondo T., 2007).
Some authors have drawn attention to the peculiarities of the course, the traumatic process, depending on the action of the external environment. Thus, the authors, analyzing the data of previous researchers, come to the conclusion that a sequence of morphological processes is observed in damaged tissues, changing under numerous conditions that can affect their severity and cause significant difficulties in diagnosis (Osminkin V.A., Kuznetsova A.V., Vaseva M.A., 2004).
When studying injuries on corpses exposed to water, there are no clear ideas about the methods of studying injuries exposed to such environmental factors (Konoreva A.K., Khasanyanova S.V. 2010).
However, the use of a monoclonal anti-human antibody to glycophorin A (anti–GPA) in order to assess the presence of erythrocytes and their decay products has shown that extravasated erythrocyte residues can be detected in hemorrhages for at least up to 15 days outdoors and one week in water (Taborelli A, Andreola S, Di Giancamillo A, Gentile G, Domeneghini C, Grandi M, Cattaneo C., 2011). The importance and reliability of studying anti-GPA, immunohistochemical staining, in order to assess the viability of damage from corpses in various states of decomposition (Baldari B, Vittorio S, Sessa F, Cipolloni L, Bertozzi G, Neri M, Cantatore S, Fineschi V, Aromatario M., 2021) has been confirmed. The study of aquaporins 1 and 3 (APQ1 and APQ3) in various types of mechanical and thermal injuries did not give unambiguous both positive and negative results, which indicates the need for further research and establishment of sufficient sensitivity and specificity of this method (Prangenberg J, Doberentz E, Witte A-, Madea B., 2021). The Ponceau/Victoria blue B (P/VB) staining technique for collagen fibers and muscles was applied on a severely decomposed corpse (1.5 years after death), the method declared itself simple and alternative for the diagnosis of lifetime trauma (Sun T, Pan M, Zhu W, Liu H, Yang C, Dong H., 2022). Biophysical parameters (electrical resistance and electrical capacitance) make it possible to differentiate bruising in putrefactively altered tissues with a high degree of reliability (Amineva G.M., Khalikov A.A., Vavilov A.Yu., Naidenova T.V., 2017). Markers: tryptase, glycophorin, IL15, CD 15, CD 45 and MMP9 showed a high degree of expression in putrefactively altered skin for 15 days (Bertozzi G, Ferrara M, La Russa R, Pollice G, Gurgoglione G, Frisoni P, Alfieri L, De Simone S, Neri M, Cipolloni L., 2021). In the study of frozen corpses, algorithms should be followed (Zagryadskaya A.P., Chikun V.I., Bald V.I., 2001), based on the already studied signs (Tenkov A.A., Luneva Z.M., Syedin M.S., 2019), it should also be remembered that with rapid freezing of the corpse (during the first hours after death), the hemorrhages formed in soft tissues microscopically practically do not differ from those in vivo (Shigeev V.B., Shigeev S.V., 2016) and to date, no solution to this problem has been found (Kaplunovsky P. A., Olkhovsky V. O., Gubin M. V., Pershina L. V., 2019).
One of the first Russian histologists who began to observe cellular, vascular reactions in response to mechanical damage, including acute blood loss, focused on the speed of death after injury, on the general condition of the body at the time of injury and after it, with conventional light microscopy, were Gromov L.I., Mityaeva N.A.., (1965-66). Subsequently, Kireeva E.A. (2008), in her dissertation research, relying on the blood filling of vessels, capillaries, cellular composition, reactive and proliferative changes in hemorrhages in soft tissues from the area of rib fracture, derived qualitative morphological signs of the prescription of the formation of rib fractures.
The probability of postmortem chemotaxis of leukocytes was studied based on known data that neutrophils, detected after 6-24 hours, tend to be the first to the focus of inflammation; a little later – monocytes and lymphocytes, after 24-48 hours, and it was not possible to identify such a process in a scientific study (Bihert E.A. Demchuk O.N. Vlasyuk I.V., 2018).
In the study of hemorrhages in soft tissues, to assess morphological changes in them, the histologist is based not only on the already available research results obtained by different authors, but also on his own experience.
"Reactive changes of cells and tissues in the wound process are based on the patterns of embryonic and postembryonic histogenesis; activation and proliferation of low-grade cells, their differentiation and interaction with subsequent adaptive restructuring of the regenerate" (Chepurnenko M.N., Chepurnenko D.A., 2020).
As can be seen from the data analysis (Tables 1, appendix) there is no single approach and precise, uniform time boundaries that speak about the specific time that has passed from the moment of injury to the moment of death of the victim. The data obtained by the authors for such a long period of study in this area do not always coincide, and sometimes radically differ in the morphological assessment of changes and cellular composition in injuries of both different and the same localization. These data are known, a number of methodological recommendations have been compiled based on them, written by the teams of authors Bogomolov D.V., Bogomolova I.N. and co-authors. (based on the RCME, 2010); Yankovsky V.E., Sarkisyan B.A., Malinina E.I. (based on the KGUZ "Altai Regional Bureau of Forensic Medical Examination", 2008); based on the OBZ "Bureau of SME" of the Kursk region (Lyskova S.V., 2014), which are based on the data of current regulatory documents and literature; informational letters (Belikov V.K. Mazurenko M.D., 1990; Frolova I.A., Asmolova N.D., Nazarova R.A., 2007). Original scientific articles, works of Russian scientists have been written, analyzing and summarizing literary sources on the problem of prescription and lifetime of hemorrhages; showing the state, prospects and problems in this matter (Kontsevich I.A. Kidraliev S.K. Gaibov A.G., 1977; Khalikov A.A., Enikeev D.A., Sharafutdinov A.N., 2011); the need to comply with the principles of a systematic approach in the study of damage (Khalikov A.A., Vavilov A.Yu., 2004; Khalikov A.A., Viter V.I., 2009).
In addition to the standard histological staining, histochemical techniques are used, for example, showing the state of fibrin as an important factor in the comprehensive assessment of determining the prescription of hemorrhages in soft tissues with rib fractures (Saenko A.V., 1998); with mechanical injuries of soft tissues of various localization (Karpenko T.A., 2018); also for histological assessment of fibrin maturity in blood clots, thrombi and thrombotic emboli based on age-dependent differential fibrin staining (Khismatullin, Rafael & Shakirova, Asia & Weisel, John & Litvinov, Rustem., 2020) indicate that fibrin filaments are dyed in different colors over time. At the moment, in the routine practice of histological laboratories, the method of tricolor staining on fibrin is used to determine its "age", the MSB (OKG) method. Which was modified by Zerbino D.D. and Lukasevich L.L. (1984), and subsequently modified by Gadetskaya M.D. and T.G. Abramova (1998). This method of staining is recommended to be compared with cellular reactions occurring in the tissues of the material under study, due to the fact that fibrin of various ages is determined in the material submitted for study, according to the authors (Zerbino D.D. and Lukasevich L.L.; Godetskaya M.D., Abramova T.G.) this is due to the sequence of blood coagulation processes.
Histochemical methods are also possible to trace changes in specific enzymes involved in metabolism and reflecting the level of intracellular energy processes (Kidraliev S.K., 1977), so pronounced changes in dehydrogenases and NAD-diaphorase in damaged muscles were revealed already 10 minutes after mechanical damage. Determination of the lifetime and prescription of the formation of post-traumatic hemorrhages in soft tissues by the biochemical method (Khisamutdinova A.D., 2018), in particular, the study of gemin; the author draws conclusions about the high efficiency of this method in the differential diagnosis of prescription injuries.
The histomolecular method of mRNA research makes a significant contribution to forensic examination (De Simone S, Giacani E, Bosco MA, Vittorio S, Ferrara M, Bertozzi G, Cipolloni L, La Russa R., 2021). The histomolecular approach based on mRNA expression shows that differential mRNA expression was demonstrated in pre- and post-mortem wounds (Manetti AC, Maiese A, Baronti A, Mezzetti E, Frati P, Fineschi V, Turillazzi E., 2021), mRNA-205 and 21 expressions detected by PCR were significantly increased and increased within 24 hours after the wound was inflicted, then decreased sharply (Ibraheem, Samah & Ali, Mona & Basyouni, Hoda & Rashed, Laila & Amer, Eman & El-Kareem, Dalia., 2019). mRNA before and after 48 hours in damaged skin shows an increase in the level of CXCL1 mRNA – up to 48 and an increase in the level of CXCR2 mRNA – after 48 hours (He JT, Huang HY, Qu D, Xue Y, Zhang KK, Xie XL, Wang Q., 2018). miRNAs are involved in differentiation, migration and apoptosis of keratinocytes (Bertozzi G, Ferrara M, La Russa R, Pollice G, Gurgoglione G, Frisoni P, Alfieri L, De Simone S, Neri M, Cipolloni L., 2021). A study of the expression of chitinase 3-like protein 1 (CHI3L1) showed that wounds from 4 to 6 days after injury can be clearly distinguished from other wounds based on a cut-off value of 2.75, sensitivity of 92.31% and specificity of 85.14% (Murase T, Shinba Y, Mitsuma M, Abe Y, Yamashita H, Ikematsu K., 2022).
The analytical method (Western blotting) – showed a marked decrease in LC3-II and a reciprocal increase in p62 compared to intact skin tissues, suggests that the level of autophagy is reduced in damaged tissues (Kimura A, Ishida Y, Nosaka M, Shiraki M, Hama M, Kawaguchi T, Kuninaka Y, Shimada E, Yamamoto H, Takayasu T, Kondo T., 2015).
Also, one of the promising methods of laboratory research is immunohistochemical / immunohistocytochemical.
The range of studied immunohistochemical markers is large, the use of one of them increases the probability of obtaining contradictory results significantly (Ishida Y, Nosaka M, Kondo T., 2022; Mansueto G, Feola A, Zangani P, Porzio A, Carfora A, Campobasso CP., 2022), and their combination increases the accuracy and reliability of the results (Li N, Du Q, Bai R, Sun J., 2018). In the course of experimental studies using the immunohistochemical method, scientists "test" markers in various combinations: MMP-9, MMP-2 and TIMP-1 (Niedecker A, Huhn R, Ritz-Timme S, Mayer F., 2021); CD15, TNFa, IL-6, IL-1β, TGFa or TGFß1 (Casse JM, Martrille L, Vignaud JM, Gauchotte G., 2016), TNFa, IL-6, IL-1β, TGFa or TGFß1 (Gauchotte G, Martrille L, Plénat F, Vignaud JM., 2013), CD15 and myeloperoxidase (MPO) (Gauchotte G, Bochnakian A, Campoli P, Lardenois E, Brix M, Simon E, Colomb S, Martrille L, Peyron PA., 2022); CD68, α-SMA, VEGF and TGFb1 (Khalaf AA, Hassanen EI, Zaki AR, Tohamy AF, Ibrahim MA., 2019).
These research methods, according to Khromova A.M. and Kalinina Yu.P. (2003), are highly informative; Novoselova V.P., Savchenko S.V., Tseluyeva E.A. (2008) make it possible to improve the quality of forensic medical examinations and objectify the expert's conclusions. Bogomolov D.V., Bogomolova I.N., Zavalishina L.E., Kovalev A.V., Kulbitsky B.N., Fedulova M.V. (2014), relying on the analysis of literary sources, argue that the introduction of IHC methods in scientific and practical forensic examination will answer many questions, some of which on today remains unanswered. For a number of objective reasons (Bogomolov D.V., Kochoyan A.L., Martemyanova A.A., 2014), the immunohistochemical method has not yet gained wide application in the practice of forensic medicine. "At the same time, it should be remembered that IHC methods are additional to traditional histological examination and do not replace it. Nevertheless, these methods help to identify pathological processes occurring at a deeper than tissue level and thus increase the accuracy and reliability of forensic histological diagnostics" (Fedulova M.V., Kovalev A.V., Zavalishina L.E., Bogomolov D.V., Kupriyanov D.D., 2022).
Scientists have developed and put into practice research methods showing their expediency of use. Khalikov A.A. et al. (2004-2011) a number of scientific articles were written reflecting the course of his dissertation research, as a result of which a multifactorial regression formula was developed that allows determining the prescription of injury by a set of biophysical characteristics of tissue damage in the period of 10-60 hours, taking into account the influence of the age factor and the concentration of alcohol in the blood of the victim at the time of death. When studying the prescription of bruises using the impedance measurement method, an original technique was developed using the values of the electrical resistance of the skin measured at various alternating current frequencies using an original measuring device. A more intensive increase in the heat-conducting properties of the skin in the area of bruising in the elderly has been established, this is determined by biophysical methods, indicating the expediency of taking into account the age characteristics of the body when determining the prescription of bruising (Babushkina K.A., 2008). When objectifying the assessment of bruises by determining the coefficient of their thermal conductivity, the thermophysical properties of human skin were evaluated (Akbashev V.A., Viter V.I., 2002; Akbashev V.A., Vavilov A.Yu., Ledyankina I.A., 2001), the data obtained required further research and refinement. The study of the chemiluminescence method for determining the lifetime and duration of mechanical injury to skeletal muscles (Pashinyan G.A. Prutov V.V., 1978) showed that the parameters of induced chemiluminescence of homogenates of injured muscles naturally change as the post-traumatic period increases. The study of the prescription of bodily injuries by non-contact thermometric method in living persons testifies to the high accuracy and prospects of its use (Kononova S.A., 2010). Sonography reliably determines a hematoma in soft tissues regardless of its prescription; the "age" of hemorrhage is determined up to one month and depends on the localization; the use of ultrasound histography allows us to speak with high probability about the timing of bruising in vivo (Klyushkin I.V., Harin G.M., Gazizyanova R.M., 2004). Examination of victims with post–traumatic hemorrhages up to 1 and more than 1 month old, by ultrasonography, shows a high degree of reliability of this method, using parameters such as sensitivity, specificity, accuracy (Gazizyanova R.M., 2013). The study of the features of thermodynamics in the area of bruising and in intact areas of symmetrical localizations, in living persons using the original objective methodology and the original device, suggests that this research method is accessible and objective, but requires taking into account the individual characteristics of the examined person (Evstafiev A.A., 2001).
To properly assess the changes occurring in the body after injury, it is necessary to know and understand the mechanism of the response and its dynamics. The speed, dynamics, sequence and nature of the reactive changes determined not only by the time the post-traumatic period, but the severity of injury, location of injury (Frolova and Soave., 2007; Novoselov V. P., Savchenko S. V., Makovchuk O. A., V. A. Gritzinger, 2013; Karpenko, T. A., 2018; Spiridonov, V. A., and Soave., 2019), as well as concomitant diseases, the presence of chronic diseases (Khalikov A., Vavilov, A., 2004, 2009; Vinay J, Harish S, Mangala GSR, Hugar BS., 2017), gender and age (Schneider CP, Schwacha MG, Chaudry IH., 2006; Bösch F, Angele MK, Chaudry IH., 2018; Serre-Miranda C, Roque S, Barreira-Silva P, Nobrega C, Vieira N, Costa P, Almeida Palha J, Correia-Neves M., 2022) altering the response of the immune system, intoxications of various genesis (Akbashev V.A., Vavilov A.Yu., 2001; Greiffenstein P, Mathis KW, Stouwe CV, Molina PE., 2007; Wagner N, Franz N, Dieteren S, Perl M, Mörs K, Marzi I, Relja B., 2017; Franz N, Dieteren S, Köhler K, Mörs K, Sturm R, Marzi I, Perl M, Relja B, Wagner N., 2019; Haag F, Janicova A, Xu B, Powerski M, Fachet M, Bundkirchen K, Neunaber C, Marzi I, Relja B, Sturm R., 2022), it is important to take into account the volume of surgical interventions and drug therapy, etc.
With all of the above, at the present stage there are no so-called “standards of microscopic examination” (Savchenko S.V., 2015); the lack of a standardized protocol complicates the interpretation of the data obtained (Manetti AC, Maiese A, Baronti A, Mezzetti E, Frati P, Fineschi V, Turillazzi E., 2021).
From the above it can be seen that some progress has been made in the study of lifetime and prescription in damaged tissues, new research methods, both laboratory and instrumental, have been developed and put into practice, which significantly influenced the understanding of the reactive changes in tissues. The use of the latest techniques will make a significant contribution to the development of forensic medicine. However, some of these methods are complex, time-consuming, which require appropriate material and technical equipment, which is not possible in all forensic histology departments and laboratories.
Summing up, it is important to note that the problem of lifetime and prescription of damage formation cannot be considered completely solved and involves further study.

APPENDIX Table 1. Study of cell composition over time, data from various researchers

 

Domestic authors and co-authors.

Timing of the appearance of morphological changes/cellular and tissue reactions

the First few minutes to 1 hour

To 1-1,5 hours

To 3 hours

4-6 hours

6-8 hours more than 12 hours

after 12-24 hours

On the second or third day

after 3-6 days

7-10 days

From 2 weeks or more

Belikov V. K. Yamanaka, M. D., 1990

in the bleeding red blood cells are well stained with hedemi contours with the presence of small focal ritrovamento in the perifocal zone

 

 

hin 4 hours along the vessels are identified clusters of segmented leukocytes;

6 hours in the area of hemorrhage gradually leaching, hemolysis of red blood cells, white blood cells are found at the edge of hemorrhage can be detected in cells of macrophage type, isolated fat cells

 

after 12 hours-papacitos products of hemolysis leukocytes, white blood cells are found around the edge, deep in a heap formation of foci accumulation and discharge;

after 12-15 hours - appears mitosis in cells

24 hours - on the edge, thicker ends hemorrhage, hemolysis and vidlakovation of red blood cells, found isolated fat cells, the cells histiocytoma series, methemoglobin;

48 hours - expressed macrophagal-cell response with a predominance of cells histiocytoma number of edges bleeding

3-5 days to the existing cellular infiltration detected fibroblasts revealed biliverdin

7-10 days in the abundance of macrophage cell eof elements is dominated by fibroblast, revealed hemosiderin

10-15 days - the former indicates hemorrhage, hemosiderin in macrophages, extracellular

 

Belyanin V. L., 1996

the first 10 minutes arteriospasm;

in 10 minutes - vasodilation;

in 30 minutes - the phenomenon of the near-wall location (marginalia) white blood cells in the vessels, emigration of white blood cells through the vascular wall, perivascular accumulation of white blood cells;

after 1 hour - foci of a heap accumulation of white blood cells (focal reaction)

 

 

an increase in the number of white blood cells around / near the vessels (up to 60 neutrophils in the focus)

in the period from 3-6 hours-pycnosis and rexis of white blood cell nuclei; after 6 hours-an increase, in two

 

after 12-14 hours-gradual leaching of red blood cells;

after 18-24 hours-macrophage cells , macrophage cells are detected

are detected after 48 hours-identification of macrophage groups and the first lymphocytes

after 72 hours               -lymphocytes, plasma cells, proliferation of a few fibroblasts, vascular kidneys, proliferation of pericytes with the formation of thin-walled capillaries are detected;

from the 4th day-hemosiderophages;

4-6 days - slowing down of reactive processes, changes in the cellular composition of hemorrhage, not noted ("silent days")

 

Day 7-growth of connective tissue, capillaries, growth of young granulation tissue, the presence of numerousfibroblasts, no fibrocytes; the appearance of extracellular hemosiderin

Day 14 - formation of collagen and fibrous tissue in the focus of inflammation; after 1 month-a scar

is formed Permyakov A.V., Viter V. I., Nevolin N. I., 2003

30-40 min - white blood cells are detected in the lumen of the vessels;

after 1 hour - emigration of leukocytes through the vascular wall into the perivascular space, detection of leukocytes along the edge and in the hemorrhage

 

 

itself, the severity of reactive leukocytosis increases sharply, the appearance of lymphoid cells among the leukocytes

, hyperemia decreases, leukostases in the vessels, a pronounced leukocyte reaction around the hemorrhage

, a pronounced demarcation shaft with the presence of macrophages

on the 2nd day-magal-histiocytic reaction increases, the number of fibroblasts increases, mainly around the vessels

 

up to 7-10 days - fibroblast proliferation (before the period of active resorption of hemorrhage);

during 9-10 days - hemosiderin lumps (brown fine-grained pigment) actively phagocytic

on 17-18 days - phagocytes break up, free-lying hemosiderin

is detected Gridasov E. V. Vinogradov O. M. 2005

hemodynamic disorders (arteriolar spasm, hyperemia, blood clots, unchanged red blood cells in hemorrhage); reactive changes (traumaticёedema with swelling of tissue elements; violation of striated striation in myocytes; polymorphism of small vessels metachromasia - violation of the tinctorial properties of the damaged tissue (staining it in a different color)

arteriolospasm; plethora of venules and capillaries, marginallocation of leukocytes in vessels, leukodiapedesis, tissueёк

edema pronounced venular-capillary plethora, intravascular leukocytosis; plasmorrhagia, plasmorrhea, perivascular clusters of polymorphonuclear leukocytes, the presence of their peripheral hemorrhage; widespreadённый отedema, foci ofnecrosis in the muscle tissue. tissues, partial hemolysis of white blood cells, the beginning of leukocyte decay

the presence of a leukocyte shaft around the zones of formed necrosis; maximum diffuseleukocyteinfiltration, leukocyte decay, macrophage detection, fibroblast appearance; signs

 

 

 

of epithelial regeneration groups of macrophages are detected, their predominance over leukocytes is noted; lymphoid infiltrates, histiocytes, fibroblasts, lipophages, erythrophages are detected; multilayer epithelium regenerates;

 from 4-6 days availability extracellular hemosiderin, epithelialization, development of young granulation tissue

 

the term injury period from 2 weeks to 2 months " Collagenization and fibrosis of the inflammatory focus. Formation of scar tissue of various maturity".

Frolova I. A., Asmolova N. D., Nazarova R. A., 2007

from the moment of occurrence of damage and within an hour-frometo various degrees of severity, vasospasm, followed by hyperemia;

after an hour from the moment of damage and later-the appearance of white blood cells in the area of damage (the beginning of a white blood cell reaction)

 

 

 

 

, an increase in the number of white blood cells, a change in nuclei in the form of pycnosis and rexis; detection of morphological signs of soft tissue necrosis

is possible; the number of disintegrating leukocytes increases with a gradual decrease in the intensity of the leukocyte reaction, the first macrophages appear

at the end of the 1st day, the beginning of the 2nd day    -leukocytes disappear from the damage zone, the number of macrophages increases, fibroblast proliferation is determined

perivascularly 3-4 days and later - hemosiderophages (beginning of resorption); 4-7 days and later-fibroblasts line up in chains in the area of damage with the formation of connective tissue fibers (beginning of the organization process)

 

 

Novoselov V. P., Savchenko S. V., Sakovchuk O. A., Gritsinger V. A., 2013

after 1 hour - in the area of hemorrhage-frometo, focal leukocyte reaction (perivascular)

 

 

swelling ofёsoft tissue edema, connective tissue fibers loosened; red blood cells, well-colored withёclear contours; diffuse arrangement of white blood cells was detected in the hemorrhage zone

 

pronounced soft tissue edema; gradual hemolysis, leaching of red blood cells; leukocyte infiltration persists, the first macrophages appear

identification of single fibroblasts, thin filaments of fibrin are distinguishable

24 hours   -pale staining of red blood cells, the presence of fibrin, diffuse mixed-cell infiltration (neutrophilic leukocytes, macrophages, lymphocytes and fibroblasts); the appearance of newly formed, thin-walled capillaries is noted;

after 48 hours               -some normalization of blood flow in the microcirculatory

 

bed after 5 days-the development of reparative regeneration

 

 

 

processes continued Frolova I. A., 2016

about an hour-a leukocyte reaction - in the area where the blood flow of the microcirculatory bed is located. peripheral hemorrhages, perivascularly, in the lumen of blood vessels-accumulations of leukocytes with its gradual

 

 

 

 

 

increase the first macrophages in the leukocyte infiltrate with a subsequent decrease in the number of leukocytes and an increase in the number of macrophages

on the second or third day-fibroblasts are detected

 

on the 3rd-4th day-hemosiderophages

 

 

 

Spiridonov V. A., Khromova A.M., Alexandrova L. G., Bibisheva L. R., Nasybullina E. L., 2019

perivascular leukodiapedesis with a gradual increase in their number (up to tens and hundreds when countingёin the field of view)

 

 

 

after 6-8 hours-leukocyte decay; possible necrotic changes in the structural elements of the tissue;

12-20 hours-leukocytes concentrate on the edge of the hemorrhage, limiting the area of damage to the formation of the leukocyte shaft

by the formation of the leukocyte shaft; a day later and later               - a decrease in the number of leukocytes with their gradual disappearance; a decreaseёin tissue edema; on the first day - hemolysis of red blood cells

on the second or third day - histiocytes scattered in the thickness and along the edge of the hemorrhage; macrophages appear;

 

on the third day -- the fourth day - the first hemosiderophages.

 

 

 

Seitova R. R., 2019

 

 

 

 

 

 

 

on day 3-4-single hemosiderophages with pale blue staining of the cytoplasm with an increase in color intensity over time

by 10 days               -single hemosiderophages containing well-colored hemosiderin grains in the cytoplasm are detected

by day 14-single grains of extracellular hemosiderin; from two weeks to one month      - continuation of the process of resorption and organization;

months and years        - presence of lumps of extracellular hemosiderin, formation of foci of hemosiderosis

 

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About the authors

F Frolova

Author for correspondence.
Email: olga.frolog@yandex.ru
ORCID iD: 0000-0002-0785-6819
Russian Federation

Vladimir Aleksandrovich Klevno

Email: vladimir.klevno@yandex.ru
Russian Federation

Fedor Georgievich Zabozlaev

Email: fzab@mail.ru
Russian Federation

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СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
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СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ЭЛ № ФС 77 – 59181 выдано 03.09.2014 г.


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