Treatment of Critical Limb Ischemia by pIRES/VEGF165/HGF Administration.

BACKGROUND: Prognosis of peripheral artery disease (PAD), especially critical limb ischemia (CLI), is very poor despite the development of endovascular therapy and bypass surgery. Many patients result in having leg amputation. We decided to investigate the safety and efficacy of plasmid of internal ribosome entry site/vascular endothelial growth factor (VEGF) 165/hepatocyte growth factor (HGF) gene therapy (GT) in patients suffered from CLI. METHODS: Administration of plasmid of internal ribosome entry site/VEGF165/HGF was performed in 12 limbs of 12 patients with rest pain and ischemic ulcers due to CLI. Plasmid was injected into the muscles of the ischemic limbs. The levels of VEGF in serum and the ankle-brachial index (ABI) were measured before and after treatment. RESULTS: Mean (±SD) plasma levels of VEGF increased nonsignificantly from 258 ± 81 pg/L to 489 ± 96 pg/L (P > 0.05) 2 weeks after therapy, and the ABI improved significantly from 0.27 ± 0.20 to 0.50 ± 0.22 (P < 0.001) 3 months after therapy. Ischemic ulcers healed in 9 limbs. Amputation was performed in 3 patients because of advanced necrosis and wound infection. However, the level of amputations was lowered below knee in these cases. Complications were limited to transient leg edema in 3 patients and fever in 2 patients. CONCLUSIONS: Intramuscular administration of plasmid of internal ribosome entry site/VEGF165/HGF is safe, feasible, and effective for patients with critical leg ischemia.

Role of vascular endothelial growth factor in inducing production of angiopoetin-1 - in vivo study in Fisher rats.

The aim of the study was to investigate how an intramuscular injection of plasmids with genes coding various pro-angiogenic factors: angiopoetin-1 (ANGPT1), vascular endothelial growth factor (VEGF165) and hepatic growth factor (HGF), influences the production of ANGPT1. 40 Healthy Fisher rats received i.m. injections containing plasmids encoding pro-angiogenic genes in thigh muscles. They were divided into four equal groups. The first group received the plANGPT1 plasmid and the second group- the pIRES/ANGPT1/VEGF165 bicistronic plasmid. The pIRES/VEGF165/HGF bicistronic plasmid was administered to the third group and an empty plasmid (control group) to the fourth group. The animals were euthanized after 12 weeks. In each group, the number of vessels stained with the anti-ANGPT1 antibody was assessed under an optical microscope. The anti-ANGPT1 antibodies stained the vessels in all the groups. There were on average 14.1 ±2.3 vessels in the the plANGPT1 group, 32.5 ±10.5 in the pl/RESANGPT1/VEGF group and 30.8 ±13.3 in the plRES/HGV/VEGF group. There were on average 7.3 ±2.3 stained vessels (p < 0.0001) in the control group . The VEGF plays a role in the induction of the production of ANGPT1. The administration of plasmids only encoding ANGPT1 does not induce its production.

Gene Therapy of Thromboangiitis Obliterans with Growth Factor Plasmid (VEGF165) and Autologous Bone Marrow Cells.

BACKGROUND: We performed gene therapy for critical limb ischemia in thromboangiitis obliterans (TAO) by the intramuscular administration of plasmids of the vascular endothelial growth factor gene (VEGF 165) with or without bone marrow-derived stem cells. METHODS: The 21 patients were randomly assigned to three groups: A-with dual therapy, cells and plasmid; B-plasmid only; and C-control group, where patients received intramuscular injections of saline. Serum VEGF levels, the ankle-brachial index (ABI), transcutaneous oxygen pressure (TcPO2), and the rest pain measured by the visual analog scale (VAS) were determined sequentially before treatment, and then 1 and 3 months after treatment. RESULTS: In the treatment groups, serum VEGF levels increased by 4 weeks and returned to baseline values after 3 months. ABI after 12 weeks increased by an average of 0.18 in group A, and 0.09 in group B and group C. TcPO2 increased by an average of 17.3 mmHg in group A, 14.1 mmHg in group B, and 10.7 mmHg in group C. The largest pain decrease was observed in group A and averaged 5.43 less pain intensity. CONCLUSIONS: Gene therapy using the VEGF plasmid along with or without bone marrow-derived mononuclear cells administered intramuscularly into an ischemic limb in TAO is a safe and effective therapy.

[Stem and progenitor cells in biostructure of blood vessel walls]. / Komórki macierzyste i progenitorowe w biostrukturze scian naczyn krwionosnych.

Development of vascular and hematopoietic systems during organogenesis occurs at the same time. During vasculogenesis, a small part of cells does not undergo complete differentiation but stays on this level, "anchored" in tissue structures described as stem cell niches. The presence of blood vessels within tissue stem cell niches is typical and led to identification of niches and ensures that they are functioning. The three-layer biostructure of vessel walls for artery and vein, tunica: intima, media and adventitia, for a long time was defined as a mechanical barrier between vessel light and the local tissue environment. Recent findings from vascular biology studies indicate that vessel walls are dynamic biostructures, which are equipped with stem and progenitor cells, described as vascular wall-resident stem cells/progenitor cells (VW-SC/PC). Distinct zones for vessel wall harbor heterogeneous subpopulations of VW-SC/PC, which are described as "subendothelial or vasculogenic zones". Recent evidence from in vitro and in vivo studies show that prenatal activity of stem and progenitor cells is not only limited to organogenesis but also exists in postnatal life, where it is responsible for vessel wall homeostasis, remodeling and regeneration. It is believed that VW-SC/PC could be engaged in progression of vascular disorders and development of neointima. We would like to summarize current knowledge about mesenchymal and progenitor stem cell phenotype with special attention to distribution and biological properties of VW-SC/PC in biostructures of intima, media and adventitia niches. It is postulated that in the near future, niches for VW-SC/PC could be a good source of stem and progenitor cells, especially in the context of vessel tissue bioengineering as a new alternative to traditional revascularization therapies.

Complementary Gene Therapy after Revascularization with the Saphenous Vein in Diabetic Foot Syndrome.

Diabetic foot syndrome (DFS) is one of the most serious macroangiopathic complications of diabetes. The primary treatment option is revascularization, but complementary therapies are still being sought. The study group consisted of 18 patients diagnosed with ischemic ulcerative and necrotic lesions in DFS. Patients underwent revascularization procedures and, due to unsatisfactory healing of the lesions, were randomly allocated to two groups: a group in which bicistronic VEGF165/HGF plasmid was administered and a control group in which saline placebo was administered. Before gene therapy administration and after 7, 30, 90, and 180 days, color duplex ultrasonography (CDU) was performed, the ankle-brachial index (ABI) and transcutaneous oxygen pressure (TcPO2) were measured, and DFS changes were described and documented photographically. In the gene therapy group, four out of eight patients (50%) healed their DFS lesions before 12 weeks. During this time, the ABI increased by an average of 0.25 and TcPO2 by 30.4 mmHg. In the control group, healing of the lesions by week 12 occurred in six out of nine patients (66.67%), and the ABI increased by an average of 0.14 and TcPO2 by 27.1 mmHg. One major amputation occurred in each group. Gene therapy may be an attractive option for complementary treatment in DFS.

Evaluation of the humoral and cellular immune responses after implantation of a PTFE vascular prosthesis.

INTRODUCTION: The experiment was designed in order to determine the immunological processes that occur during the healing in synthetic vascular grafts, especially to establish the differences in the location of the complement system proteins between the proximal and distal anastomosis and the differences in the arrangement of inflammatory cells in those anastomoses. The understanding of those processes will provide a true basis for determining risk factors for complications after arterial repair procedures. MATERIAL/METHODS: The experiment was carried out on 16 dogs that underwent implantation of unilateral aorto-femoral bypass with expanded polytetrafluoroethylene (ePTFE). After 6 months all animals were euthanized to dissect the vascular grafts. Immunohistochemical assays and electron microscopic examinations were performed. RESULTS: Immunohistochemical findings in the structure of neointima between anastomoses of vascular prostheses demonstrated significant differences between humoral and cellular responses. The area of proximal anastomosis revealed the presence of fibroblasts, but no macrophages were detected. The histological structure of the proximal anastomosis indicates that inflammatory processes were ended during the prosthesis healing. The immunological response obtained in the distal anastomosis corresponded to the chronic inflammatory reaction with the presence of macrophages, myofibroblasts and deposits of complement C3. DISCUSSION: The identification of differences in the presence of macrophages and myofibroblasts and the presence of the C3 component between the anastomoses is the original achievement of the present study. In the available literature, no such significant differences have been shown so far in the humoral and cellular immune response caused by the presence of an artificial vessel in the arterial system.

Two-Stage Gene Therapy (VEGF, HGF and ANG1 Plasmids) as Adjunctive Therapy in the Treatment of Critical Lower Limb Ischemia in Diabetic Foot Syndrome.

One of the most serious problems in people with diabetes is diabetic foot syndrome. Due to the peripheral location of atherosclerotic lesions in the arterial system of the lower extremities, endovascular treatment plays a dominant role. However, carrying out these procedures is not always possible and does not always bring the expected results. Gene therapy, which stimulates angiogenesis, improves not only the inflow from the proximal limb but also the blood redistribution in individual angiosomes. Due to the encouraging results of sequential treatment consisting of intramuscular injections of VEGF/HGF bicistronic plasmids followed by a month of ANG1 plasmids, we decided to use the described method for the treatment of critical ischemia of the lower limbs in the course of diabetes and, more specifically, in diabetic foot syndrome. Twenty-four patients meeting the inclusion criteria were enrolled in the study. They were randomly divided into two equal groups. The first group of patients was subjected to gene therapy, where the patients received intramuscular injections of pIRES/VEGF165/HGF plasmids and 1 month of ANG-1 plasmids. The remaining patients constituted the control group. Gene therapy was well tolerated by most patients. The wounds healed significantly better in Group 1. The minimal value of ABI increased significantly in Group 1 from 0.44 ± 0.14 (± standard deviation) to 0.47 ± 0.12 (with p = 0.028) at the end of the study. There were no significant differences in the control group. In the gene treatment group, PtcO2 increased significantly (from 28.71 ± 10.89 mmHg to 33.9 ± 6.33 mmHg with p = 0.001), while in Group 2, no statistically significant changes were found. The observed resting pain decreased significantly in both groups (Group 1 decreased from 6.80 ± 1.48 to 2.10 ± 1.10; p < 0.001; the control group decreased from 7.44 ± 1.42 to 3.78 ± 1.64 with p < 0.001). In our study, we evaluated the effectiveness of gene therapy with the growth factors described above in patients with CLI in the course of complicated DM. The therapy was shown to be effective with minimal side effects. No serious complications were observed.

Local intramuscular administration of ANG1 and VEGF genes using plasmid vectors mobilizes CD34+ cells to peripheral tissues and promotes angiogenesis in an animal model.

INTRODUCTION: Patients with peripheral artery disease have poor prognosis despite advances in vascular surgery. Therefore, attempts have been made at using gene and cell therapy to stimulate angiogenesis in the lower limbs in patients with critical lower limb ischemia (CLI). METHODS: The study included 30 rats divided into 3 groups. An intramuscular injection of a therapeutic gene or cells in the right hind limb was administered in each group: angiopoietin-1 (ANG1) plasmid in group 1, ANG1/vascular endothelial growth factor (ANG1/VEGF) bicistronic construct in group 2, and naked plasmid in group 3 (control). After 3 months of follow-up, tissue samples were harvested, and vessels that stained positively for CD34 cells were quantified. RESULTS: The highest CD34+ cell count was noted in the ANG1/VEGF group (98.26 cells), followed by the ANG1 group (80.31) and control group (47.93). The CD34+ cell count was significantly higher in the ANG1/VEGF and ANG1 groups than in the control group. There was no significant difference in the CD34+ cell count between the ANG1/VEGF and ANG1 groups. CONCLUSION: Our study confirmed that therapy with ANG1 plasmid alone or ANG1/VEGF bicistronic construct is safe and effective in a rat model. The therapy resulted in the recruitment of more CD34+ vascular endothelial cells than in the control group receiving naked plasmid.

Intra-aneurysm sac pressure measurement using a thin pressure wire during endovascular aneurysm repair.

BACKGROUND: An endoleak is a typical complication of endovascular aneurysm repair (EVAR). It is characterized by persistent blood flow between a stent graft and the aneurysm sac. Usually, it can be visualized during primary EVAR, but in many cases, this remains impossible. Therefore, other methods of endoleak assessment are urgently needed. The measurement of aneurysm sac pressure (ASP) seems to be a promising direction of research in this area. OBJECTIVES: We aimed to evaluate the safety and efficacy of a new method for invasive pressure measurement inside the abdominal aortic aneurysm (AAA) during EVAR. We also assessed a correlation between pressure values and early angiographic occurrence of an endoleak after the procedure. MATERIAL AND METHODS: A total of 20 patients with AAA were included in this experimental prospective study. During EVAR, systolic, diastolic and mean pressure values were recorded both for ASP and aortic pressure (AP) before procedure, after stent graft opening and after final stent graft ballooning. RESULTS: The measurements were successfully obtained in all participants without any complications. There were no significant differences between all ASP and AP before procedure. After the procedure, blood pressure significantly decreased in the aneurysm sac but not in the aorta. Systolic ASP was significantly lower than systolic AP both after stent graft opening (80.4 ±20.9 mm Hg compared to 110.7 ±21.6 mm Hg, p < 0.01) and after its balloon post-dilatation (65.6 ±26.1 mm Hg compared to 107.4 ±22.1 mm Hg, p < 0.001). Diastolic ASP decreased significantly in comparison to diastolic AP only after stent graft ballooning (48.0 ±14.6 mm Hg compared to 56.4 ±13.6 mm Hg, p < 0.05). CONCLUSIONS: Our study confirmed that the novel method for the measurement of ASP during EVAR, using a thin pressure wire, is feasible and safe.

Double VEGF/HGF Gene Therapy in Critical Limb Ischemia Complicated by Diabetes Mellitus.

Critical leg ischemia (CLI) complicated by diabetes mellitus (DM), which is a very common and dangerous disease, represents the ultimate stage of peripheral arterial disease. Patients are treated with antiplatelet drugs, statins and limb revascularization, but a significant number of patients are not candidate for revascularization. Literature shows that in such cases, gene therapy could be a perfect therapeutic option. The aim of our study was to evaluate efficacy of double vascular endothelial growth factor/hepatocyte growth factor (VEGF/HGF) gene therapy in patients with CLI complicated by DM. We observed that 90 days after administration, serum level of VEGF and ankle-brachial index increased significantly (p < 0.001) and rest pain decreased significantly compared with the control group (p < 0.002). Moreover considerable improvement in vascularization was observed in computed tomography angiography (P = 0.04). Based on the results of this study, we suggest that the therapy with pIRES/VEGF165/HGF bicistronic plasmid administration is a safe and effective method of treatment of patients with both CLI and DM. Graphical abstract.

LC-QTOF-MS and 1H NMR Metabolomics Verifies Potential Use of Greater Omentum for Klebsiella pneumoniae Biofilm Eradication in Rats.

Bacterial wound infections are a common problem associated with surgical interventions. In particular, biofilm-forming bacteria are hard to eradicate, and alternative methods of treatment based on covering wounds with vascularized flaps of tissue are being developed. The greater omentum is a complex organ covering the intestines in the abdomen, which support wound recovery following surgical procedures and exhibit natural antimicrobial activity that could improve biofilm eradication. We investigated changes in rats' metabolome following Klebsiella pneumoniae infections, as well as the greater omentum's ability for Klebsiella pneumoniae biofilm eradication. Rats received either sterile implants or implants covered with Klebsiella pneumoniae biofilm (placed in the peritoneum or greater omentum). Metabolic profiles were monitored at days 0, 2, and 5 after surgery using combined proton nuclear magnetic resonance (1H NMR) and high performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF­MS) measurements of urine samples followed by chemometric analysis. Obtained results indicated that grafting of the sterile implant to the greater omentum did not cause major disturbances in rats' metabolism, whereas the sterile implant located in the peritoneum triggered metabolic perturbations related to tricarboxylic acid (TCA) cycle, as well as choline, tryptophan, and hippurate metabolism. Presence of implants colonized with Klebsiella pneumoniae biofilm resulted in similar levels of metabolic perturbations in both locations. Our findings confirmed that surgical procedures utilizing the greater omentum may have a practical use in wound healing and tissue regeneration in the future.

[The use of the synthetic silver coated prosthesis and penduculated omentum flap in the treatment of prostheses infection]. / Zastosowanie syntetycznej protezy impregnowanej solami srebra i plata sieci wiekszej w leczeniu zakazen protez naczyniowych.

There is a methods of surgical treatment of secondary enteroaortal fistula after infection of vascular prostheses with the use silver prosthesis and great omentum. Secondary enteroaortal fistula is the most serious complication followed infection of vascular prosthesis, with high risk of death rate regardless of different operative methods. Patients is operated and the dracon prosthesis was exchanged to arterial homograft, which is more resistant to infections or subclavio-femoral bypasses. Removing of intestinal fistula included resection of duodenum and gastroenterotomy or duodenorrhaphy. The modification of duodenal fistula provision relies on duodenorraphy and protection of that area and prosthesis with pediculated flap of gastrocolic omentum conducted retrocolicaly in retroperitoneum space. The retrocolical and retroperitoneal omentoplasty seems to be a practical modification of aorto-duodenal fistula treatment with useful protection of duodenal plastic.

[Complex gene-cell therapy in treatment of critical lower limbs ischemia]. / Kompleksowa terapia komórkowo-genowa w leczeniu krytycznego niedokrwienia konczyn dolnych.

THE AIM: The paper presents the results of our hybrid gene-cell therapy in case of critical lower limb ischemia. Eighteen patients with critical limb ischemia were enrolled to the study. The bone marrow from each patient was harvested. It had been incubated with VEGF165 gene plasmid for two hours before it was administrated intramuscularly into patient's lower limb. METHODS: In the study we evaluated: safety, clinical outcomes of the therapy, venous blood VEGF protein concentration before and after (at: 7th, 14th, 28th and 90th day) administration of the stem cells. We obtained samples of muscles from the patients who underwent the limb amputation, which were examined histological and by PCR (Polymerase Chain Reaction) for detection of plasmid genes. RESULTS: We observed very good clinical outcomes of the therapy. In ten of eighteen patients (-55%) critical lower limb ischemia symptoms subsided--they saved their limbs. The serum VEGF concentration was higher than in healthy controls (p < 0.05). We observed the increase of the concentration of the cytokine at the 14th day an decrease at the 90th day after administration of cells with plasmid. We found expression of plasmid VEGF in transfected stem cells and in tissues taken from amputated limbs. However histological examination did not reveal any sings of new blood vessels formation in the samples taken from ischemic limbs. CONCLUSIONS: The therapy is safe and effective. We observed significant improvement in patient's clinical state. The therapy needs further investigations.

[Molecular basics of angiogenesis]. / Molekularne podstawy angiogenezy.

In the article we present the latest knowledge about angiogenesis. We have divided the paper into three main parts, in which the involvement of the extracellular matrix, cells, and cytokines/growth factors in the growth of new blood vessels is described. In brief, the extracellular compartment plays a crucial role in the formation of new vasculature. Degradation of matrix is a very important and precisely controlled process performed mostly by a family of proteins called matrix metallproteinases (MMPs). The extracellular compartment, through the special transmembrane proteins integrins, transmit a wide variety of signals into the cells and thus influence such cell behavior as proliferation, invasion, shape, migration, and maturation. Many products of matrix degradation are potent (mostly negative) regulators of angiogenesis; this self-limiting system prevents excessive proteolysis of the matrix components. The cells involved in the process are endothelial progenitor cells (EPCs), which are derived from bone marrow. The major surface antigens of the cells are CD34+, CD133+, and VEGFR2+. It has been demonstrated that EPCs are responsible for maintaining the functional integrity of endothelium. The number of EPCs in peripheral blood samples inversely correlates with cardiovascular risk factors. In the last section of the article the role of cytokines/growth factors is described. VEGF, as a key regulator of the initial steps of angiogenesis, controls the mobilization and incorporation of EPCs into the site of ischemia. The most important cytokine that facilitates the mobilization of EPCs from bone marrow is SDF-1, which is the strongest chemoattractant for EPCs. Ang-1, on the other hand, controls new blood vessel maturation and stabilization.

[Application of VEGF165 plasmid in treatment of critical lower limb ischemia]. / Zastosowanie plazmidu VEGF165 w leczeniu krytycznego niedokrwienia konczyn dolnych.

UNLABELLED: In the study we assessed the clinical and histological benefits of gene therapy in no-option patients with critical lower limb ischemia. MATERIAL AND METHODS: We administrated VEGF165 (Vascular Endothelial Growth Factor) plasmid to 10 patients. The plasmid was injected intramuscularly into lower limb. In the study we evaluated: clinical outcomes, venous blood VEGF protein concentration before and after (7th, 14th, 28th and 90th day) administration of plasmid. From patients who required amputation we obtained samples of muscles, which were examined histological and by PCR (Polymerase Chain Reaction) for detection of plasmid genes. RESULTS: Only two patients from the group did not require amputation due to the therapy. In eight patients the operation was essential. Serum VEGF concentration was higher than in healthy controls (p < 0.05). We observed increase of the concentration of the cytokine on the 14th day an decrease on the 90th day after administration of plasmid (p < 0.05). Histological analysis did not reveal any sings of new blood vessels formation in the samples taken from amputated limbs. There was also no signs of expression of plasmid primers in PCR. CONCLUSIONS: The therapy is safe. We did not observe significant improvement in patient's clinical state. The therapy needs further investigations.

The Formation of Blood Vessel After the Administration of the Plasmid Encoding Ang-1 Gene in Fischer Rats.

BACKGROUND: Chronic limb ischemia is a serious clinical problem. Patients who do not qualify for standard treatment may benefit from novel gene therapies. OBJECTIVES: This study evaluated angiogenesis following intramuscular injections of angiogenic plasmid Ang-1 in Fisher rats. MATERIAL AND METHODS: Twenty rats had plasmids injected intramuscularly in their hind limbs. The study group consisted of 10 animals which received the Ang-1 plasmid, while the control group consisted of 10 rats that received an empty plasmid. All the animals were euthanized after 12 weeks and tissue samples from the hind limb thigh muscles and internal organs were harvested for histological and immunohistochemical examinations. To assess the angiogenesis the number of vessels in the hind limb muscles visualized by the SMA and FVIII markers was counted for each animal in five separate microscopic fields. RESULTS: There were no pathological lesions or any signs of neoplastic angiogenesis in any of the 20 rats. The number of vessels visualized by the FVIII marker in the study group was two times higher than in the control group (median: 12, range: 7-25 vs. median: 6, range: 2-15; p < 0.0001). The median estimated that the number of vessels visualized by the SMA marker is 63% higher in the study group compared to the control group (median: 6.5, range: 1-12 vs. median: 4, range: 0-10; p = 0.0008). CONCLUSIONS: Intramuscular injections of Ang-1 plasmids induced angiogenesis in the rat hind limb muscles.

Combined autologous bone marrow mononuclear cell and gene therapy as the last resort for patients with critical limb ischemia.

INTRODUCTION: Our study was designed to investigate the safety and efficacy of combined autologous bone marrow mononuclear cell (MNC) and gene therapy in comparison to conventional drug therapy in patients with critical limb ischemia (CLI). MATERIAL AND METHODS: Thirty-two patients with CLI persisting for 12-48 months (average time 27.5 months) were randomized into 2 groups, each group consisting of 16 patients. In the first group, administration of autologous bone marrow MNC and vascular endothelial growth factor (VEGF) plasmid was performed. The patients from the second group were treated pharmacologically with pentoxifylline. Ankle-brachial index (ABI) was measured and angiography was performed before and finally 3 months after treatment. The pain was evaluated using the Visual Analog Scale (VAS) before and after 3 months. RESULTS: Ankle-brachial index improved significantly from 0.29 ±0.21 to 0.52 ±0.23 (p < 0.001) in 12 patients (75.0%) 3 months after the experimental therapy in group 1. In this group angiography showed the development of collateral vessels. Ischemic ulcers healed completely in 11 patients (68.75%). In group 2 the ABI did not improve in any patient; moreover the complete healing of skin ulcers was not found in any of the patients of this group. Amputation was performed in 4 (25.0%) patients in group 1, and in 8 patients (50%) from group 2. CONCLUSIONS: These data after 3-month follow-up indicate that intramuscular injection of MNC combined with gene therapy in patients with chronic CLI is safe, and a more feasible and effective method of treatment than the conventional therapy. However, both therapies are limited by the degree of microcirculation damage.

The morphological evaluation of the homograft wall in an electron microscopic study.

Cases of massive purulent infection of vascular prosthesis are demonstrated in this study. Infected prosthesis was substituted by arterial homograft, harvested during multi-organ procurement, and stored by the cold ischaemia method. In the follow-up period, the patients were divided into two groups, those treated with immunosuppression (n = 16) and those treated without immunosuppressive drugs (n = 13). The patients underwent resurgery, during which a fragment of arterial wall was taken for electron microscopic examination. In the group with immunosuppression, the presence of the following structures was observed: endothelial cells, the intima, with a great number of elastic and collagen fibrils with fibrinogen inclusions, and active phagocyting myoblasts and myofibroblasts. In the group without immunosuppression electron microscopic examination showed the total destruction of the wall of the ruptured arterial homograft - absence of endothelium and sparse, damaged fibroblasts of the media or their degraded fragments, making a picture of cellular death. Morphological analysis of the arterial wall and the clinical state of the patient suggest the necessity of immunosuppressive treatment after fresh arterial homograft transplantation.

Use of mesh fibrous dressing covered with fibrin glue (TachoComb) in hemostasis after vascular anastomoses in the groin.

We present in this paper application of haemostatic device TachoComb onto bleeding after vascular anastomosis of dacron vascular prosthesis (branch of aortobifemoral or bypass aortofemoral) with common femoral artery in the groin. Hemorrhagic complications have influence onto clinical status of operated patients. Haemostatic TachoComb dressing was applied at 30 cases and results were compared to control group consist of 25 cases, in which gas compresses were applied. Mean loss blood in group I with the usage of TachoComb was statistical characteristic (p < 0.003) smallest than in group II. Also mean time of hemostasis was shortest than in group II (p < 0.01). We proved that use of TachoComb limits bleeding from suture line connecting artery with vascular prosthesis.

Intraoperative hemostasis during kidney transplantation and the use collagen mesh dressing covered by fibrin glue (TachoComb).

In this paper we present influence of use of haemostatic dressing TachoComb, onto bleeding from surface of transplanted kidney. Kidney transplantation (KTX) seems to be main method of treatment of terminal renal failure. Enlarging number of KTX results in growing frequency of intra and postoperative complications. Hemorrhagic complications can impact clinical status of recipient and graft function. Haemostatic dressing was applied at 29 cases. Control group in which only gas compresses were used consisted of 25 patients. It was proved, that use of dressing from collagen mesh covered by fibrin glue TachoComb, after kidney transplantation diminished parenchymal bleeding and time necessary to get complete hemostasis.