Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.4.5-13
A. P. Sysoeva, N. Makarova, E. E. Kraevaya
For a long time, the role of seminal plasma during human fertilization remained underestimated. Numerous studies related to the development of different methods for human embryo in vitro cultivation were gener-ally concerned with the quality of male and female gametes. However, in recent years, the development of Omix technologies provided a new insight into great seminal plasma influence on the morphofunctional characteristics of spermatozoa. This is especially true for the regulatory function of extracellular vesicles secreted by male reproductive tract cells. In this work, we attempted to analyze current data on the influence of extracellular seminal plasma vesicles on the morphofunctional characteristics of spermatozoa to solve male infertility topical issues. The review includes studies by foreign and Russian research groups that werу conducted within the past 5 years and found in PubMed, Google Scholar, and Cochrane Library databases. Very few studies demonstrate that seminal plasma vesicles act as functional regulators of male fertility and their dysfunction may lead to infertility. The use of seminal plasma extracellular vesicles in clinical practice may significantly increase the success of IVF programs, especially in impaired spermatogenesis. Keywords: extracellular vesicles, exosomes, biomarkers, seminal plasma, spermatozoa, assisted reproductive technology, cell biology, morphology
{"title":"The role of seminal plasma extracellular vesicles in changes in the morphofunctional characteristics of human spermatozoa","authors":"A. P. Sysoeva, N. Makarova, E. E. Kraevaya","doi":"10.31088/cem2021.10.4.5-13","DOIUrl":"https://doi.org/10.31088/cem2021.10.4.5-13","url":null,"abstract":"For a long time, the role of seminal plasma during human fertilization remained underestimated. Numerous studies related to the development of different methods for human embryo in vitro cultivation were gener-ally concerned with the quality of male and female gametes. However, in recent years, the development of Omix technologies provided a new insight into great seminal plasma influence on the morphofunctional characteristics of spermatozoa. This is especially true for the regulatory function of extracellular vesicles secreted by male reproductive tract cells. In this work, we attempted to analyze current data on the influence of extracellular seminal plasma vesicles on the morphofunctional characteristics of spermatozoa to solve male infertility topical issues. The review includes studies by foreign and Russian research groups that werу conducted within the past 5 years and found in PubMed, Google Scholar, and Cochrane Library databases. Very few studies demonstrate that seminal plasma vesicles act as functional regulators of male fertility and their dysfunction may lead to infertility. The use of seminal plasma extracellular vesicles in clinical practice may significantly increase the success of IVF programs, especially in impaired spermatogenesis. Keywords: extracellular vesicles, exosomes, biomarkers, seminal plasma, spermatozoa, assisted reproductive technology, cell biology, morphology","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/CEM2021.10.1.33-40
A. Balkanov, I. D. Rozanov, A. Golanov, L. Gaganov, V. Chernikov
Introduction. Despite the treatment methods’ improvement, the brain glioblastoma (GB) patient survival remains at the level of 12–14 months. In this regard, it is very relevant to optimize the choice of the treat-ment of recurrent GB, including using stereotactic radiosurgery (SRS). Materials and methods. Comparative analysis of the endothelial cells in the capillaries of blood vessels of the brain peritumoral zone (PZ) before (1 group – 32 capillaries) and 12 – 14 months after (2 group – 75 capillaries) adjuvant external radiation therapy (AERT) was performed using transmission electron micros-copy (TEM). Results. Apoptosis of endotheliocytes was observed in the capillaries of the brain PZ from 2 group much more often than from 1 group (60% and 12.6% of capillaries, respectively, p<0.001). Capillary death and thrombosis occurred in 4% and 1.4% of 2 group capillaries. An important finding was that only 69.3 of capillaries from 2 group revealed thickening of the basement membrane (BM), and 26.9% of them also revealed its peak deformation. Conclusion. 12–14 months after AERT, the brain PZ remains ischemic due to apoptosis of the capillary endothelium, thickening and peak deformation of the BM. The detected signs of revascularization of the brain PZ after previously performed AERT suggest that it is possible to consider SRS as a method of treat-ment for recurrent brain GB. Keywords: brain glioblastoma, peritumoral zone, endotheliocyte apoptosis, thickening of the basement membrane, stereotactic radiosurgery, glioblastoma relapse
{"title":"Endothelium changes of peritumoral zone capillaries after brain glioblastoma\u0000 adjuvant radiation therapy","authors":"A. Balkanov, I. D. Rozanov, A. Golanov, L. Gaganov, V. Chernikov","doi":"10.31088/CEM2021.10.1.33-40","DOIUrl":"https://doi.org/10.31088/CEM2021.10.1.33-40","url":null,"abstract":"Introduction. Despite the treatment methods’ improvement, the brain glioblastoma (GB) patient survival remains at the level of 12–14 months. In this regard, it is very relevant to optimize the choice of the treat-ment of recurrent GB, including using stereotactic radiosurgery (SRS). Materials and methods. Comparative analysis of the endothelial cells in the capillaries of blood vessels of the brain peritumoral zone (PZ) before (1 group – 32 capillaries) and 12 – 14 months after (2 group – 75 capillaries) adjuvant external radiation therapy (AERT) was performed using transmission electron micros-copy (TEM). Results. Apoptosis of endotheliocytes was observed in the capillaries of the brain PZ from 2 group much more often than from 1 group (60% and 12.6% of capillaries, respectively, p<0.001). Capillary death and thrombosis occurred in 4% and 1.4% of 2 group capillaries. An important finding was that only 69.3 of capillaries from 2 group revealed thickening of the basement membrane (BM), and 26.9% of them also revealed its peak deformation. Conclusion. 12–14 months after AERT, the brain PZ remains ischemic due to apoptosis of the capillary endothelium, thickening and peak deformation of the BM. The detected signs of revascularization of the brain PZ after previously performed AERT suggest that it is possible to consider SRS as a method of treat-ment for recurrent brain GB. Keywords: brain glioblastoma, peritumoral zone, endotheliocyte apoptosis, thickening of the basement membrane, stereotactic radiosurgery, glioblastoma relapse","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279615","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.2.62-69
E. A. Ponomarenko, M. A. Diatroptova, K. A. Artemyeva, A. Shelkov
Introduction. Obtaining primary cultures of fibroblasts is necessary for conducting experimental studies to investigate the basic cell response mechanisms to various stimuli. Despite the widespread use of fibroblast cultures, methods for obtaining them from the skin are not standardized. The aim of the studywas to find the optimal conditions to obtain a fibroblast culture from rat skin using enzymatic tissue disaggregation. Materials and methods. The fibroblast culture was obtained from the dermis of 18 male Wistar rats (N=12, 5–6-week-old, and 60–65 g body weight; N=6, 8–10-week-old rats, and 160–180 g body weight). A series of experiments was carried out to obtain a fibroblast culture with selecting the collagenase type and defining its concentration and exposure time. Results.An optimized protocol to obtain rat dermis fibroblasts is presented. Moreover, the problems of obtaining a culture and practical aspects of its use are discussed. Conclusion. When obtaining a primary culture of rat skin fibroblasts we should consider the following fac-tors: the type of the enzyme, its concentration, and exposure time; the age of the animals; the area of skin graft collection. In young rats, the optimal result was achieved when the cells were isolated from the axillary zone using collagenase type II at a 1 mg/ml concentration during a 90-minute exposure. In adult animals, the enzymatic effect of collagenase type II on the skin graft obtained from the back was optimal at a 5 mg/ml concentration during a 120-minute exposure. Keywords: fibroblast culture, dermis, preparation protocols, rats
{"title":"Protocol optimization for obtaining the culture of rat dermal fibroblasts","authors":"E. A. Ponomarenko, M. A. Diatroptova, K. A. Artemyeva, A. Shelkov","doi":"10.31088/cem2021.10.2.62-69","DOIUrl":"https://doi.org/10.31088/cem2021.10.2.62-69","url":null,"abstract":"Introduction. Obtaining primary cultures of fibroblasts is necessary for conducting experimental studies to investigate the basic cell response mechanisms to various stimuli. Despite the widespread use of fibroblast cultures, methods for obtaining them from the skin are not standardized. The aim of the studywas to find the optimal conditions to obtain a fibroblast culture from rat skin using enzymatic tissue disaggregation. Materials and methods. The fibroblast culture was obtained from the dermis of 18 male Wistar rats (N=12, 5–6-week-old, and 60–65 g body weight; N=6, 8–10-week-old rats, and 160–180 g body weight). A series of experiments was carried out to obtain a fibroblast culture with selecting the collagenase type and defining its concentration and exposure time. Results.An optimized protocol to obtain rat dermis fibroblasts is presented. Moreover, the problems of obtaining a culture and practical aspects of its use are discussed. Conclusion. When obtaining a primary culture of rat skin fibroblasts we should consider the following fac-tors: the type of the enzyme, its concentration, and exposure time; the age of the animals; the area of skin graft collection. In young rats, the optimal result was achieved when the cells were isolated from the axillary zone using collagenase type II at a 1 mg/ml concentration during a 90-minute exposure. In adult animals, the enzymatic effect of collagenase type II on the skin graft obtained from the back was optimal at a 5 mg/ml concentration during a 120-minute exposure. Keywords: fibroblast culture, dermis, preparation protocols, rats","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.3.29-38
D. Pavlush, I. Dyuizen
Introduction. To date, chronic rhinosinusitis with nasal polyps (CRSwNP) has not yet been extensively studied: the molecular factors and mechanisms involved in the initiation of polypous transformations in nasal mucosa (NM) and sustaining their recurrence probability are still to be determined. Simultaneously, it is necessary to understand the molecular rearrangement in NM tissues to make clinical prognosis and choose an adequate therapeutic or surgical strategy for CRSwNP treatment. The aim of the study was to identify the features of how inflammatory markers localize and are distributed in the NM and polyps in various morphological CRSwNP types. Materials and methods. We studied morphological and chemical structure of nasal polyps and mucosa of the inferior turbinates. The material was obtained during surgical management of patients diagnosed with CRSwNP. The comparison group involved the patients with a deviated septum who underwent septorhi-noplasty and had neither polyposis nor concomitant inflammatory/allergic pathology. The NM removed in surgeries was used to compare morphological and chemical changes. Immunohistochemistry was applied to determine the localization and distribution of SP, NK1, nNOS, iNOS, and IL1b in the tissues. Results. The formation of nasal polyps was found to be accompanied by morphological and chemical altera-tions in the mucous membrane of the inferior turbinates. In polyps of different morphological types, the changes in the activity of inflammatory markers were specific. Conclusion. The data obtained indicate that changes in the NM of the inferior turbinates, which accompany polyposis development, give certain pathological causes that induce and maintain the pathological process. We have revealed the features of the specific signaling microenvironment in the nasal cavity, which provide special conditions for the formation of polyps of various types. The specificity of the activity and distribu-tion of inflammatory markers in the polyps of different morphological types may serve as a prerequisite for the development of personalized therapy for the disease. Keywords: chronic rhinosinusitis with nasal polyps, inflammation, neurokinin receptors, substance P, nitric oxide
{"title":"Inflammatory markers in the nasal mucosa and polyp-modified tissues in chronic rhinosinusitis with nasal polyps","authors":"D. Pavlush, I. Dyuizen","doi":"10.31088/cem2021.10.3.29-38","DOIUrl":"https://doi.org/10.31088/cem2021.10.3.29-38","url":null,"abstract":"Introduction. To date, chronic rhinosinusitis with nasal polyps (CRSwNP) has not yet been extensively studied: the molecular factors and mechanisms involved in the initiation of polypous transformations in nasal mucosa (NM) and sustaining their recurrence probability are still to be determined. Simultaneously, it is necessary to understand the molecular rearrangement in NM tissues to make clinical prognosis and choose an adequate therapeutic or surgical strategy for CRSwNP treatment. The aim of the study was to identify the features of how inflammatory markers localize and are distributed in the NM and polyps in various morphological CRSwNP types. Materials and methods. We studied morphological and chemical structure of nasal polyps and mucosa of the inferior turbinates. The material was obtained during surgical management of patients diagnosed with CRSwNP. The comparison group involved the patients with a deviated septum who underwent septorhi-noplasty and had neither polyposis nor concomitant inflammatory/allergic pathology. The NM removed in surgeries was used to compare morphological and chemical changes. Immunohistochemistry was applied to determine the localization and distribution of SP, NK1, nNOS, iNOS, and IL1b in the tissues. Results. The formation of nasal polyps was found to be accompanied by morphological and chemical altera-tions in the mucous membrane of the inferior turbinates. In polyps of different morphological types, the changes in the activity of inflammatory markers were specific. Conclusion. The data obtained indicate that changes in the NM of the inferior turbinates, which accompany polyposis development, give certain pathological causes that induce and maintain the pathological process. We have revealed the features of the specific signaling microenvironment in the nasal cavity, which provide special conditions for the formation of polyps of various types. The specificity of the activity and distribu-tion of inflammatory markers in the polyps of different morphological types may serve as a prerequisite for the development of personalized therapy for the disease. Keywords: chronic rhinosinusitis with nasal polyps, inflammation, neurokinin receptors, substance P, nitric oxide","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.4.14-24
V. V. Litvinov, L. Lemkina, G. Freynd, V. Korobov
Introduction. Catheter-associated infection is caused by microorganism colonization of the surface of the implanted catheter with a biofilm formation that significantly increases their resistance to antiseptics and antibiotics, especially in immunosuppression. Low-molecular antibacterial peptides are compounds capable of combating biofilm formation. The aim of the study was to describe morphological characteristics of a catheter-associated infection model on laboratory mice secondary to immunosuppression and to assess the efficacy of the low-molecular cationic antibacterial peptide (warnerin). Materials and methods. An experiment included white outbred mice (25–30 g body weight) under ether anesthesia that received 1.0-cm fragments of intravascular catheters under the skin of the backs. The animals underwent preliminary immunosuppression with cyclophosphamide. We used Staphylococcus epidermidis 33 (in the form of suspensions or biofilms previously grown on catheter segments) and low-molecular cationic peptide warnerin. All animals were sacrificed by ether overdose on days 1, 2, and 3 after the manipulation. We took the tissues surrounding the catheter for histological and immunohistochemical studies with antibodies to CD34, vimentin, CD68, CD3, and CD20. Results. The warnerin administration at the site of the catheter implantation led to disappearance of or a significant decrease in the number of bacterial. In the infiltrate, the number of neutrophils significantly increased, whereas that of fibroblasts decreased. Immunohistochemistry confirmed the features of the cellular reactions around the catheters with bacterial contamination with warnerin administration. Conclusion. In a model of catheter-associated infection in immunosuppressed mice, the warnerin antibacterial manifests in characteristic histological alterations in the inflammatory infiltrate composition. Keywords: catheter-associated infection model, morphology of inflammation, warnerin antibacterial cationic peptide
{"title":"Morphological characteristics of an experimental catheter-associated infection following immunosuppression and the use of a low-molecular cationic peptide of the lantibiotic family – warnerin","authors":"V. V. Litvinov, L. Lemkina, G. Freynd, V. Korobov","doi":"10.31088/cem2021.10.4.14-24","DOIUrl":"https://doi.org/10.31088/cem2021.10.4.14-24","url":null,"abstract":"Introduction. Catheter-associated infection is caused by microorganism colonization of the surface of the implanted catheter with a biofilm formation that significantly increases their resistance to antiseptics and antibiotics, especially in immunosuppression. Low-molecular antibacterial peptides are compounds capable of combating biofilm formation. The aim of the study was to describe morphological characteristics of a catheter-associated infection model on laboratory mice secondary to immunosuppression and to assess the efficacy of the low-molecular cationic antibacterial peptide (warnerin). Materials and methods. An experiment included white outbred mice (25–30 g body weight) under ether anesthesia that received 1.0-cm fragments of intravascular catheters under the skin of the backs. The animals underwent preliminary immunosuppression with cyclophosphamide. We used Staphylococcus epidermidis 33 (in the form of suspensions or biofilms previously grown on catheter segments) and low-molecular cationic peptide warnerin. All animals were sacrificed by ether overdose on days 1, 2, and 3 after the manipulation. We took the tissues surrounding the catheter for histological and immunohistochemical studies with antibodies to CD34, vimentin, CD68, CD3, and CD20. Results. The warnerin administration at the site of the catheter implantation led to disappearance of or a significant decrease in the number of bacterial. In the infiltrate, the number of neutrophils significantly increased, whereas that of fibroblasts decreased. Immunohistochemistry confirmed the features of the cellular reactions around the catheters with bacterial contamination with warnerin administration. Conclusion. In a model of catheter-associated infection in immunosuppressed mice, the warnerin antibacterial manifests in characteristic histological alterations in the inflammatory infiltrate composition. Keywords: catheter-associated infection model, morphology of inflammation, warnerin antibacterial cationic peptide","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.s4.68-76
Y. Kirillov, M. A. Kozlova, L. Makartseva, I. Chernov, D. Areshidze
Introduction. Alcohol abuse is one of major health and social problems of modern society, the essence of which is an increase in morbidity and mortality not only from alcohol misuse and its complications, but also from associated comorbid pathologies. The liver is the organ that is affected most by ethanol and its metabolites. A significant factor leading to melatonin-associated morphological manifestations in the liver is the so-called light pollution, a violation of the light–dark regime. At the same time, heavy consumption of alcohol over a short period of time (alcoholic excesses) and other forms of alcoholic illness are often associated with various sleep disorders. The aim of the study was to investigate the morphological and functional state of the liver of Wistar rats under experimental alcohol intoxication in conditions of a fixed light regime and constant illumination. Materials and methods. Male rats (n=160) were kept under a fixed light regime, constant illumination, and at a combination of both light regimes with simultaneous experimental alcohol intoxication. We performed a pathological examination of the liver and determined morphometric parameters, hepatocyte ploidy, and the proportion of binuclear hepatocytes. Results. The consumption of 15% ethanol solution as a drink ad libitum for 3 weeks at a fixed light regime causes the development of fatty degeneration in the liver of laboratory animals. Secondary to it, changing lighting from fixed to constant is accompanied by the development of alcoholic hepatitis in about half of the animals. The results of micromorphometry indicate significant changes in the studied parameters of hepatocytes, which are expressed least of all in the case of ethanol consumption under fixed lighting condi-tions and affect most the cells of the liver parenchyma of rats with chronic alcohol intoxication secondary to constant lighting. Conclusion. Alcohol intoxication that occurs secondary to a melatonin deficiency caused by constant lighting is accompanied by a much greater severity, prevalence, and intensity of inflammatory tissue reactions and a variety of manifestations of liver pathology. Keywords: hepatocyte, alcohol, lighting regime, stress, fatty degeneration
{"title":"The effect of chronic alcohol intoxication and constant illumination on the morphological and functional state of the liver","authors":"Y. Kirillov, M. A. Kozlova, L. Makartseva, I. Chernov, D. Areshidze","doi":"10.31088/cem2021.10.s4.68-76","DOIUrl":"https://doi.org/10.31088/cem2021.10.s4.68-76","url":null,"abstract":"Introduction. Alcohol abuse is one of major health and social problems of modern society, the essence of which is an increase in morbidity and mortality not only from alcohol misuse and its complications, but also from associated comorbid pathologies. The liver is the organ that is affected most by ethanol and its metabolites. A significant factor leading to melatonin-associated morphological manifestations in the liver is the so-called light pollution, a violation of the light–dark regime. At the same time, heavy consumption of alcohol over a short period of time (alcoholic excesses) and other forms of alcoholic illness are often associated with various sleep disorders. The aim of the study was to investigate the morphological and functional state of the liver of Wistar rats under experimental alcohol intoxication in conditions of a fixed light regime and constant illumination. Materials and methods. Male rats (n=160) were kept under a fixed light regime, constant illumination, and at a combination of both light regimes with simultaneous experimental alcohol intoxication. We performed a pathological examination of the liver and determined morphometric parameters, hepatocyte ploidy, and the proportion of binuclear hepatocytes. Results. The consumption of 15% ethanol solution as a drink ad libitum for 3 weeks at a fixed light regime causes the development of fatty degeneration in the liver of laboratory animals. Secondary to it, changing lighting from fixed to constant is accompanied by the development of alcoholic hepatitis in about half of the animals. The results of micromorphometry indicate significant changes in the studied parameters of hepatocytes, which are expressed least of all in the case of ethanol consumption under fixed lighting condi-tions and affect most the cells of the liver parenchyma of rats with chronic alcohol intoxication secondary to constant lighting. Conclusion. Alcohol intoxication that occurs secondary to a melatonin deficiency caused by constant lighting is accompanied by a much greater severity, prevalence, and intensity of inflammatory tissue reactions and a variety of manifestations of liver pathology. Keywords: hepatocyte, alcohol, lighting regime, stress, fatty degeneration","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69280064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.4.72-80
N. Tikhonova, A. Milovanov, V. Aleksankina, T. V. Fokina, M. N. Boltovskaya, A. Aleksankin
Introduction. Data about the role of components of adipose tissue in the repair of damaged uterine walls are limited, although a number of authors claim that cell-based drugs from adipose tissue have a positive effect on the repair of damaged uterine walls in laboratory rodents. Meanwhile, stem cells derived from adipose tissue are considered to be the most promising type of cells in regenerative medicine. The aim of the studywas to evaluate the adipocyte components in the uterine wall of rats in healing after a full-thickness surgi-cal incision. Materials and methods. We conducted the study on 40 female Sprague Dawley rats. The animals were sub-jected to a full-thickness longitudinal incision in the wall of the right uterine horn, with the left one serving as an intact control. We carried out morphological examinations of the uterine walls daily in 5 animals from day 1 to 7 and on day 15. The sections from paraffin blocks were stained with hematoxylin and eosin and Mallory’s trichrome staining. Immunohistochemistry detected FABP4+ adipocytes and CD68+ macrophages. The morphometric study was carried out using the Leica system (Leica, Germany). The results underwent processing in the Statistica 12 software (StatSoft). Results. We noticed the period of the most active interaction of adipose tissue with the damaged horn to last from day 3 to 15 and coincide with the macrophage activation in the healing zone. The intact left uterine horn was not involved in the interaction processes with the mesenteric adipose tissue. Starting from day 3 FABP4+ cells in the uterine wall of the operated horn formed groups, creating rounded nest-like structures. Clusters of FABP4+ cells were localized in the healing zone, near the suture material, and in the perime-trium near the mesentery attachment sites. The changes over time of the indicators of the area of cell nests depended on the localization and duration of healing. There were no FABP4+ cells in the left intact horn. Conclusion. We characterized the morphological interaction of adipose tissue with the damaged uterine wall during the first two weeks after a full-thickness surgical incision of the rat uterine horn. The results of the study indicate that adipocytes take an active part in the healing after a surgical incision of the rat uterine wall at the earliest stages. Keywords: rat uterus, hysterotomy, healing, adipocytes, macrophages
{"title":"The role of adipose tissue in the healing of rat uterine wall after a full-thickness surgical incision","authors":"N. Tikhonova, A. Milovanov, V. Aleksankina, T. V. Fokina, M. N. Boltovskaya, A. Aleksankin","doi":"10.31088/cem2021.10.4.72-80","DOIUrl":"https://doi.org/10.31088/cem2021.10.4.72-80","url":null,"abstract":"Introduction. Data about the role of components of adipose tissue in the repair of damaged uterine walls are limited, although a number of authors claim that cell-based drugs from adipose tissue have a positive effect on the repair of damaged uterine walls in laboratory rodents. Meanwhile, stem cells derived from adipose tissue are considered to be the most promising type of cells in regenerative medicine. The aim of the studywas to evaluate the adipocyte components in the uterine wall of rats in healing after a full-thickness surgi-cal incision. Materials and methods. We conducted the study on 40 female Sprague Dawley rats. The animals were sub-jected to a full-thickness longitudinal incision in the wall of the right uterine horn, with the left one serving as an intact control. We carried out morphological examinations of the uterine walls daily in 5 animals from day 1 to 7 and on day 15. The sections from paraffin blocks were stained with hematoxylin and eosin and Mallory’s trichrome staining. Immunohistochemistry detected FABP4+ adipocytes and CD68+ macrophages. The morphometric study was carried out using the Leica system (Leica, Germany). The results underwent processing in the Statistica 12 software (StatSoft). Results. We noticed the period of the most active interaction of adipose tissue with the damaged horn to last from day 3 to 15 and coincide with the macrophage activation in the healing zone. The intact left uterine horn was not involved in the interaction processes with the mesenteric adipose tissue. Starting from day 3 FABP4+ cells in the uterine wall of the operated horn formed groups, creating rounded nest-like structures. Clusters of FABP4+ cells were localized in the healing zone, near the suture material, and in the perime-trium near the mesentery attachment sites. The changes over time of the indicators of the area of cell nests depended on the localization and duration of healing. There were no FABP4+ cells in the left intact horn. Conclusion. We characterized the morphological interaction of adipose tissue with the damaged uterine wall during the first two weeks after a full-thickness surgical incision of the rat uterine horn. The results of the study indicate that adipocytes take an active part in the healing after a surgical incision of the rat uterine wall at the earliest stages. Keywords: rat uterus, hysterotomy, healing, adipocytes, macrophages","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.2.32-39
O. Kovaleva, A. Gratchev, P. Podlesnaya, M. Rashidova, D. Samoilova, N. Sokolov, Z. Mamedli, D. Kudlay, N. Kushlinskii
Introduction. Tumor-associated macrophages (TAMs) are traditionally considered to be a pro-tumor fac-tor that promotes the growth of various tumors; however, for colorectal carcinomas (CRC), the prognostic significance of TAMs has not been fully determined, which may be due to the lack of macrophage markers suitable for this tumor type. The aim of this work was to study the expression of the nuclear marker of stromal cells PU.1 in colorectal tumors and its association with the clinical and morphological tumor characteristics. Materials and methods. We performed an immunohistochemical analysis to assess the expression of PU.1, CD68, and CD20 in 85 primary CRCs. The Mann-Whitney test was used to determine statistically significant differences in independent groups. Correlation analysis of the expression of the studied protein was carried out by determining the Spearman’s rank correlation coefficient. Differences were considered statistically significant at p <0.05. Results. We analyzed the expression of PU.1, CD68, and CD20 in CRC and detected positive PU.1 and CD68 expressions in tumor stromal cells in all of the studied samples. Expression of CD20 was observed in 87% of cases. We showed that in colorectal tumors all CD68+ or CD20+ cells express PU.1 and that PU.1 and CD20 were significantly associated with the disease stage (p=0.036 and p=0.002) and the presence or absence of regional metastases (p=0.022 and p=0.007). In addition, PU.1 showed a significant correlation with the distant metastases’ presence and tumor localization (p=0.031 and p=0.022). Higher content of PU.1 was typical for colon tumors without metastases. CD20 also showed a significant association with tumor size (p=0.025). No significant correlations with clinical and morphological features were found for CD68. We also demonstrated that the number of PU.1+ cells in tumors significantly positively correlates with CD68 (r=0.231, p=0.036) and CD20 (r=0.267, p=0.015). Conclusion. The results of this study indicate that PU.1 can be considered as an independent marker of a favorable prognosis in CRC patients. Keywords: colorectal cancer, expression, CD20, CD68, PU.1, macrophages, B-cells
{"title":"PU.1 is a nuclear factor of immunocompetent cells of tumor stroma in colorectal cancer","authors":"O. Kovaleva, A. Gratchev, P. Podlesnaya, M. Rashidova, D. Samoilova, N. Sokolov, Z. Mamedli, D. Kudlay, N. Kushlinskii","doi":"10.31088/cem2021.10.2.32-39","DOIUrl":"https://doi.org/10.31088/cem2021.10.2.32-39","url":null,"abstract":"Introduction. Tumor-associated macrophages (TAMs) are traditionally considered to be a pro-tumor fac-tor that promotes the growth of various tumors; however, for colorectal carcinomas (CRC), the prognostic significance of TAMs has not been fully determined, which may be due to the lack of macrophage markers suitable for this tumor type. The aim of this work was to study the expression of the nuclear marker of stromal cells PU.1 in colorectal tumors and its association with the clinical and morphological tumor characteristics. Materials and methods. We performed an immunohistochemical analysis to assess the expression of PU.1, CD68, and CD20 in 85 primary CRCs. The Mann-Whitney test was used to determine statistically significant differences in independent groups. Correlation analysis of the expression of the studied protein was carried out by determining the Spearman’s rank correlation coefficient. Differences were considered statistically significant at p <0.05. Results. We analyzed the expression of PU.1, CD68, and CD20 in CRC and detected positive PU.1 and CD68 expressions in tumor stromal cells in all of the studied samples. Expression of CD20 was observed in 87% of cases. We showed that in colorectal tumors all CD68+ or CD20+ cells express PU.1 and that PU.1 and CD20 were significantly associated with the disease stage (p=0.036 and p=0.002) and the presence or absence of regional metastases (p=0.022 and p=0.007). In addition, PU.1 showed a significant correlation with the distant metastases’ presence and tumor localization (p=0.031 and p=0.022). Higher content of PU.1 was typical for colon tumors without metastases. CD20 also showed a significant association with tumor size (p=0.025). No significant correlations with clinical and morphological features were found for CD68. We also demonstrated that the number of PU.1+ cells in tumors significantly positively correlates with CD68 (r=0.231, p=0.036) and CD20 (r=0.267, p=0.015). Conclusion. The results of this study indicate that PU.1 can be considered as an independent marker of a favorable prognosis in CRC patients. Keywords: colorectal cancer, expression, CD20, CD68, PU.1, macrophages, B-cells","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279202","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.2.6-12
I. Arutyunyan, T. Dubovaya
The transplantation of artificial tissues and organs is gradually becoming a part of our reality. At the same time, researchers are facing a problem common to all transplantologists, i.e. the need for a long-term storage of a biomedical product (transplant) without losing its properties. The possibility to cryopreserve cells adhered to various scaffolds' surface was first presented about 20 years ago. However, the data on the technology as a whole remains unsystematized and controversial. This review aimed to analyze the literature on tissue-engineered constructs (TEC) cryopreservation of different scientific groups to create a unified approach in assessing the technique's efficacy necessary for further regenerative medicine development. The comparison of studies on TEC cryopreservation conducted by various research groups is hampered not only by the lack of standardized protocols but also by different approaches to assessing the result. As experimental data were accumulated, the cryopreservation efficacy was reassessed from meeting the basic requirements for the structure preservation (thawed TEC retains its integrity, cells are partially alive and attached to the matrix) to focusing on the final result (thawed TEC retains its functional properties and is ready to be transplanted). Many of the currently used in vitro research methods presented in the review allow one to look for new ways of increasing the TEC cryopreservation efficacy; however, in our opinion, the next step on the way to introducing the technology into clinical practice should be research on experimental animals. Keywords: tissue engineered construction, cryopreservation, efficacy estimation
{"title":"Cryopreservation of tissue-engineered constructs in regenerative medicine","authors":"I. Arutyunyan, T. Dubovaya","doi":"10.31088/cem2021.10.2.6-12","DOIUrl":"https://doi.org/10.31088/cem2021.10.2.6-12","url":null,"abstract":"The transplantation of artificial tissues and organs is gradually becoming a part of our reality. At the same time, researchers are facing a problem common to all transplantologists, i.e. the need for a long-term storage of a biomedical product (transplant) without losing its properties. The possibility to cryopreserve cells adhered to various scaffolds' surface was first presented about 20 years ago. However, the data on the technology as a whole remains unsystematized and controversial. This review aimed to analyze the literature on tissue-engineered constructs (TEC) cryopreservation of different scientific groups to create a unified approach in assessing the technique's efficacy necessary for further regenerative medicine development. The comparison of studies on TEC cryopreservation conducted by various research groups is hampered not only by the lack of standardized protocols but also by different approaches to assessing the result. As experimental data were accumulated, the cryopreservation efficacy was reassessed from meeting the basic requirements for the structure preservation (thawed TEC retains its integrity, cells are partially alive and attached to the matrix) to focusing on the final result (thawed TEC retains its functional properties and is ready to be transplanted). Many of the currently used in vitro research methods presented in the review allow one to look for new ways of increasing the TEC cryopreservation efficacy; however, in our opinion, the next step on the way to introducing the technology into clinical practice should be research on experimental animals. Keywords: tissue engineered construction, cryopreservation, efficacy estimation","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-01-01DOI: 10.31088/cem2021.10.3.14-20
D. S. Shvorob, T. I. Shevchenko, R.B. Kondratyk
Colorectal cancer ranks third in the morbidity structure among all malignant tumors and includes sporadic and hereditary neoplasms. Cancer genome sequencing has revealed numerous mutation variants that determine the ways colorectal carcinoma progresses. The course, prognosis, and management strategy of the disease vary greatly depending on the subtype of a molecular tumor. This literature review discusses the latest data on the variants of colorectal cancer oncogenesis and presents the phenotypic model classification based on them. Immunohistochemistry (IHC) is suggested for determining the individual tumor characteristics. The article also clarifies the Bethesda panel used to detect microsatellite instability, markers for Lynch syndrome, and a list of IHC markers for determining the phenotypic model of colorectal carcinoma. Keywords: colorectal cancer, phenotypic models, consensus molecular subtypes (CMS), immunohisto-chemistry, Bethesda panel, Lynch syndrome
{"title":"Diagnosis of molecular subtypes of colorectal cancer using immunohistochemistry","authors":"D. S. Shvorob, T. I. Shevchenko, R.B. Kondratyk","doi":"10.31088/cem2021.10.3.14-20","DOIUrl":"https://doi.org/10.31088/cem2021.10.3.14-20","url":null,"abstract":"Colorectal cancer ranks third in the morbidity structure among all malignant tumors and includes sporadic and hereditary neoplasms. Cancer genome sequencing has revealed numerous mutation variants that determine the ways colorectal carcinoma progresses. The course, prognosis, and management strategy of the disease vary greatly depending on the subtype of a molecular tumor. This literature review discusses the latest data on the variants of colorectal cancer oncogenesis and presents the phenotypic model classification based on them. Immunohistochemistry (IHC) is suggested for determining the individual tumor characteristics. The article also clarifies the Bethesda panel used to detect microsatellite instability, markers for Lynch syndrome, and a list of IHC markers for determining the phenotypic model of colorectal carcinoma. Keywords: colorectal cancer, phenotypic models, consensus molecular subtypes (CMS), immunohisto-chemistry, Bethesda panel, Lynch syndrome","PeriodicalId":36062,"journal":{"name":"Clinical and Experimental Morphology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"69279311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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