Dynamics of the Activity of MAP-Kinase Cascades in the Healing Process of Postoperative Musculocutaneous Wounds

Background. Management of the reparative process is an urgent task of modern medicine. In our opinion, the development of pathogenetically grounded approaches to optimizing the repair process for managing the interrelations of stromal cells is promising. One of the promising areas in this regard is the impact on the MAPK-cascades.Aim: to study the expression of MAP-kinase mechanisms in the regulation of repair by the example of a musculocutaneous wound.Methods. A linear muscular skin wound was modeled using Wistar rats weighing 220–250 g at the age of 9 months (n = 24). Immunofluorescence staining was performed to detect the activity of p38, JNK, and ERK MAPK cascades from 1 to 30 days.Results. It was established that specific staining in the area of connective tissue formation during staining with p38 MAPK and its phosphorylated form was first observed on the 3rd day, and its maximum severity occurred at the same time. On the 7th and 14th day, small zones in the area of scar formation were minimally stained. The phosphorylated part of the JNK-cascade in the zone of traumatic injury was detected starting from the 1st day after the injury. Bright color persisted on the 3rd day. On the 7th day, the color was minimal, and by the 14th day a second wave of expression was observed. ERK-staining was observed from the 1st to the 14th day with a peak activity on the 3rd day.Conclusion. Thus, we revealed the simultaneous involvement of p38, JNK-, and ERK-cascades in the regulation of the reparative process in the conditions of a musculoskeletal wound. At the same time, it is noteworthy that the peak activity of all cascades coincides and falls on the 3rd day

Innovations for diagnostics and treatment of injuries and disorders of musculoskeletal system in Irkutsk Scientific Center of Surgery and Traumatology (to the 70th anniversary of Scientific Research Institute of Traumatology and Orthopedics)

The article summarizes experience and achievements of Irkutsk Scientific Research Institute of Traumatology and Orthopedics (now - Irkutsk Scientific Center of Surgery and Traumatology) over a 70-year period. The development of new methods of diagnostics and treatment and organization of health care for patients with traumatological and orthopedic pathology are highly influenced by the Institute. The team of Irkutsk Scientific Center of Surgery and Traumatology developed a new promising direction of combined tension-band osteosynthesis, substantiated organ-sparing endoprosthesis replacement, created new technologies of treatment of chronic osteomyelitis, developed concept of treatment of spinal trauma and spinal cord injuries, determined regularities of cross-system organization of physiological functions at the musculoskeletal system pathology, created original technologies and instruments for the treatment of musculoskeletal system traumas and disorders. Innovations created in ISCST include 330 patents. Irkutsk Scientific Center of Surgery and Traumatology was awarded Governor's Award in Science and Technics for the development and introduction in clinical medicine of new diagnostic and treatment methods and of candidate medicines in 2012 and 2013

Using corrugation stitch in submammary fold at the reconstructive breast surgery

The aim of the research was to develop a new technique for suturing the surgical incision in submammary area after subpectoral reconstructive breast surgery with good cosmetic result. Clinical observation group included 10 patients. It was found that the new technique of suturing the surgical incision, in comparison with the method offered by M.B. Calobrace et al. (2014), allows to decrease the duration of suturing from 6 min 10 ± 0.06 sec down to 5 min 29 ± 0.04 sec (р < 0.01), to reduce number of stitches from 24 down to 15, to shorten postoperative stitch from 4.24 ± 0.06 cm down to 3.71 ± 0.07 cm (р < 0.01), and to shorten cicatrix length in 1 year after the operation from 4.78 ± 0.19 cm down to 4.25 ± 0.22 cm (р < 0.05). Therefore, the developed technique of suturing the surgical wound provides the location of postoperative cicatrix in the area of submammary fold which makes it less noticeable. Moreover, it allows to shorten the length of postoperative cicatrix, decrease the duration of the surgery and to secure adequate stability of implant fixation

The level of VEGF-A in the lacrimal fluid of diabetic retinopathy

The lacrimal fluid was investigated in 56 people (95 eyes) with type II diabetes, of which 11 (11 eyes) with no signs of diabetic retinopathy and 45 patients (84 eyes) suffering from diabetic retinopathy of varying severity. Investigation of VEGF-A (vascular endothelial growth factor A) in stimulated lacrimal fluid revealed its level in all patients. It was significantly higher in patients with diabetic retinopathy compared with data that have been obtained in patients with diabetes, but without signs of diabetic retinopathy. Statistically significant differences in the level of VEGF-A in patients with non-proliferative and proliferative retinopathy of 1-3 stages have not been identified. At the same time in patients with far-advanced retinopathy with marked variations of fatal retina and vitreous body (detachment expressed gliosis), but without evidence of neovascularization of the anterior eye, a significant decrease in the level of vasoproliferative factors has revealed

Angiogenesis as an adaptive mechanism in ischemia

The article is devoted to angiogenesis as one of the ways to increase the stability of the ischemic tissue. It is shown that the reduction in ischemic tissue damage is based on four basic processes - vasculogenesis of endothelial progenitor cells, angiogenesis and arteriogenesis with the growth and subsequent stabilization of the outgrowths of mural cells and the growth of collaterals (expansive growth of pre-existing vessels). However, despite the great progress in the understanding of ischemia-induced angiogenesis, and a lot of positive results from its stimulation using the disclosed arrangements natural response to the reduction of tissue perfusion is still an unsolved problem is the practical use of this knowledge to improve the metabolic processes of organs and tissues with compromised vascular bed. The effect of a variety of recombinant growth factors is not enough long-term, and besides, is not without serious side effects. From this perspective, the greatest opportunities is the development of techniques using natural pool of pluripotent cells with high angiogenic potential

EXPRESSION OF COLLAGENS IN THE DAMAGE AREA AT ABDOMINAL ADHESIONS

Background. Postoperative adhesions are a serious problem in surgery. However, at the present time molecular mechanisms of the adhesion process are insufficiently studied. Aim. To study the dynamics of expression of genes encoding the synthesis of collagen in case of damage to the serosa on the example of the peritoneum in conditions of aseptic inflammation. Materials and methods. Aseptic inflammatory process in the abdominal cavity was modeled (Wistar rats, n = 40). A micro- and macroscopic picture of the damage area was studied. Immunofluorescent staining for Type I collagen (Col 1A1) was performed. The expression of genes encoding collagen of different types was evaluated using the RT2-Profiler PCR kit Array Rat Wound Healing. Results. It has been established that the adhesion process with peritoneal damage in aseptic conditions reaches its maximum by the 30th day of observation. The same period coincides with the maximum of collagen synthesis in fibroblasts in the repair area, revealed by immunofluorescence study. The interrelation of synthesis of type I and III collagens went as expected - the onset of expression of type III collagen (from day 3) is ahead of the expression of collagen type I (from day 7). Peak gene expression of collagens type I, Alpha-1 and -2; type III Alpha-1, type IV Alpha-1 and -3, type V Alpha-1, -2 and -3; type XIV Alpha-1 (Col14a1) falls on the 14th day. For the first time, active involvement of type V alpha-3 collagen in the adhesion process was noted - we detected both early (from day 1) and maximum intensive (up to 166.96 times increase in comparison with intact animals). Conclusion. Perhaps, the hyperexpression of collagen V alpha-3 that we revealed is an important link in the pathogenesis of adhesion in the abdominal cavity

Involvement of the ERK MAPK Cascade in the Formation of Adhesions in the Abdominal Cavity

Background. ERK1 and ERK2 (extracellular signal-regulated kinase) are related protein serine / threonine kinases that are involved in the regulation of the cell cycle, cell migration and survival, differentiation, metabolism, and proliferation. Evaluation of this cascade is promising to determine the possibility of influencing  pathological conditions through the regulation of ERK1/2.Aim of the study. To assess the involvement of the ERK MAPK cascade in the development of adhesions in the abdominal cavity.Methods. The experiments were carried out on male Wistar rats. The adhesion process in the abdominal cavity was simulated (n = 40). The research time is from 2 hours to 30 days. Tissue samples from 5 intact animals were examined as a control. Histological sections were prepared and stained histochemically for ERK1. The expression of ERK1 and ERK2 was assessed by PCR.Results. Using the PCR method, the involvement of both the ERK1 and ERK2 cascades in the process was established, and, judging by the change in gene expression, the latter is more actively involved in the process. Histochemical study confirmed the role of ERK cascades in the management of repair in serous membrane injury. Histochemically revealed an increase in ERK1 expression with two peaks of activity – at 1 and 30 days.Conclusion. Our data establish multichannel control of the reparative process in the area of damage to the serous membrane with the involvement of both the p38 and ERK MAPK cascades, which is important to consider when developing methods of influencing the wound healing process in the abdominal cavity

Matrix metalloprotease 9 and remodeling after myocardial infarction

The objective of the work was to study the distribution of metalloprotease 9 (MMP9) in tissues after myocardial infarction. We studied anatomopathological material from 30 patients, the primary victims of acute transmural myocardial infarction. Average age - 63,7 years. There were 18 men and 12 women. 16 patients died in the period up to 3 days, 14 patients - in a period from 3 days to 1 month. Samples were fixed in formalin and immunohistochemical staining for MMP9 was conducted. We used rabbit IgG monoclonal antibody to MMP9 (Epitomics, Clone 1D: EP1254, Cat. N 2551-1, Lot YG 113001P) in the working dilution of 1: 100 - 1: 250. At fatal outcomes that happened within a few hours after the onset of a heart attack a large number of MMP9 was detected in neutrophils in the blood vessels at near infarction zone. In the period from 1 to 2 hours MMP9 was detected in the extracellular matrix in the area of infarction. In the period from 3 to 30 days MMP9 was detected only in fibroclasts at forming cardiosclerosis zone. Thus, the use of staining for MMP9 is convenient for determination of the time elapsed after myocardial infarction in post mortem examination

Expression of endothelin in experimental myocardial infarction in a changed concentration of FGF and VEGF

The objective of the work was to study the expression of endothelin in experimental myocardial infarction, and. to assess the impact on its altered concentrations of growth factors. The study was performed on a model of myocardial infarction. The experiment was conducted, on 150 female Wistar rats. Five groups of animals were studied: control group, the increased concentration FGF2 and. VEGF, the decreased concentration of FGF or VEGF. Excretion of animal experiments were conducted in the period from 1 to 30 days. The heart of the experimental animal was fixed in a solution of 10% neutral formalin. His-tochemical staining was used. Primary antibodies to endothelin ET-1/2/3 (H-38) rabbit polyclonal IgG (Santa Cruz, Cat. N Sc-98727, Lot # G2209). We determined that in control group there was a stain on endothelin in the area of myocardial infarction from the 1st to the 7th day with a peak on day 3. The use of growth factors increased the staining of the border and. infarct zones on the 1st to the 14th days with a maximum of day 3. The use of antibodies to growth factors reduced the stain of the in farcted area early after myocardial infarction (days 1 to 3), to the 7th day of the stain intensity increases sharply, reaching a maximum, and. to the 14th day again declined. Thus, we established the dynamics of expression of endothelin in the myocardium when the ischemic injury and. heart attack, as well as the significant stimulatory effect of growth factors FGF2 and. VEGF on the expression of endothelin in the area of ischemic damage at experimental myocardial infarction

An examination of hepatotoxicity and nephrotoxicity of a new antiadhesive preparation (experimental study)

Adhesive process in the abdominal cavity was simulated in Wistar rats. The animals were divided into two groups: the main one - simulation of adhesive process in the abdominal cavity and introduction of 3 ml of saline into the abdominal cavity; and the controls - simulation of adhesive process in the abdominal cavity and introduction of 3 ml of new antiadhesive preparation. We evaluated biochemical parameters at eight time points during the period from 2 hours to 28 days. The introduced drug for adhesion prevention did not affect the protein-synthetic function of the liver: the blood level of whole protein was the same in both groups and remained within the normal range throughout the follow-up period. The blood level of creatinine, which is the end product of protein metabolism reflecting the renal excretory function, was within normal values in both groups throughout the observation period. Significant differences between the groups were noted only on the 28th day of the experiment: in the main group creatinine level was higher as compared to the control group (p = 0.014), but remained within the normal range. The same pattern was observed when evaluating the blood urea level, which reflects the renal excretory function - the blood urea level remained normal in both groups throughout the experiment. Hence, the new antiadhesive drug does not produce toxic effect on liver and kidneys in a single intraperitoneal introduction in experiment. Biochemical indices are the same as in single intraperitoneal infusion of saline