The Influence of Haemostatic Dressing Prototypes for the Emergency Services on the Histopathological Parameters of Porcine Muscle.

BACKGROUND/AIM: Haemostatic dressings for the uniformed and rescue services are an integral part of life-saving equipment for controlling post-traumatic haemorrhage. The aim of this study was to assess the influence of active constituent substances and materials of haemostatic dressings on muscle tissue and muscle regeneration after traumatic injury. MATERIALS AND METHODS: Three hemostatic dressing prototypes were analysed: OBR/G/S sponge: dressing material sponge made of Na-Ca chitosan/algal composite microfibers and nanofibers; OBR/MBT/S: tactic gauze modified with a polymer mixture of Na-Ca chitosan/algal composite microfibers and nanofibers, impregnated with a moderate amount of procoagulants (22.9 g/m2); and OBR/MS/S: seton gauze modified with a polymer mixture of Na-Ca chitosan/algal composite microfibers and nanofibers, impregnated with a moderate amount of procoagulants (18.0 g/m2), with chitosan (ChitoClearhqg 95) and sodium alginate (Protanal LF10/60 FT) as the coagulants. The experiment was conducted on 20 pigs which were euthanised 24 h, 7 or 14 days after wound dressing. Samples of porcine muscle tissue were subjected to qualitative histopathological analysis. RESULTS: Histopathological analysis of muscle tissues from the experimental pigs revealed that the application of modified seton (OBR/MS/S) produced the most satisfactory results. The observed changes were similar on all dates that samples were collected and in all experimental groups, and minor differences in their extent were observed between groups. Regenerative processes were most advanced, and retrograde changes were least apparent in animals treated with OBR/MS/S. CONCLUSION: Modified seton (OBR/MS/S) induced the least tissue reaction and was most effective in promoting tissue regeneration after injury.

The Effect of Haemostatic Dressing Prototypes for the Emergency Services in the Porcine Haemostatic System.

BACKGROUND/AIM: Coagulopathy can develop when hemostatic dressings are used to stop massive bleeding, even in patients without prior history of clotting disorders. The selection of procoagulants, which effectively control bleeding and prevent disseminated intravascular coagulation (DIC) and thrombosis, is a significant challenge. The aim of this study was to evaluate the effect of two prototypes of haemostatic dressing in the porcine haemostatic system. MATERIALS AND METHODS: The total number of animals used in our experiments was 24. Group I: pigs were treated with the developed prototype of sponge dressing, made of Na-Ca chitosan/algal composite of microfibers and nanofibers. Group II: animals were treated with a seton gauze modified with a polymer mixture of Na-Ca chitosan/algal composite of microfibers and nanofibers. Group III: animals were treated with non-hemostatic dressing and this group was the control. Blood was sampled five times to determine changes in the coagulation and fibrinolytic profiles: before injury: i) at 1 h, ii) at 24 h, iii) at 7, and iv) at 14 days following injury. RESULTS: Significant changes were observed in the coagulation parameters, in the total numbers of white blood cells and platelets in groups I and II, compared to controls. CONCLUSION: The modified haemostatic dressings used in this study produced a strong procoagulant effect in pigs. This, together with high fibrinogen concentrations, which can cause DIC, require further studying.

Plasma and cerebrospinal fluid interleukin-1ß during lipopolysaccharide-induced systemic inflammation in ewes implanted or not with slow-release melatonin.

BACKGROUND: Interleukin-1ß (IL-1ß) is important mediator of inflammatory-induced suppression of reproductive axis at the hypothalamic level. At the beginning of inflammation, the main source of cytokines in the cerebrospinal fluid (CSF) is peripheral circulation, while over time, cytokines produced in the brain are more important. Melatonin has been shown to decrease pro-inflammatory cytokines concentration in the brain. In ewes, melatonin is used to advance the onset of a breading season. Little is known about CSF concentration of IL-1ß in ewes and its correlation with plasma during inflammation as well as melatonin action on the concentration of IL-1ß in blood plasma and the CSF, and brain barriers permeability in early stage of lipopolysaccharide (LPS)-induced inflammation. METHODS: Systemic inflammation was induced through LPS administration in melatonin- and sham-implanted ewes. Blood and CSF samples were collected before and after LPS administration and IL-1ß and albumin concentration were measured. To assess the functions of brain barriers albumin quotient (QAlb) was used. Expression of IL-1ß (Il1B) and its receptor type I (Il1r1) and type II (Il1r2) and matrix metalloproteinase (Mmp) 3 and 9 was evaluated in the choroid plexus (CP). RESULTS: Before LPS administration, IL-1ß was on the level of 62.0 ± 29.7 pg/mL and 66.4 ± 32.1 pg/mL in plasma and 26.2 ± 5.4 pg/mL and 21.3 ± 8.7 pg/mL in the CSF in sham- and melatonin-implanted group, respectively. Following LPS it increased to 159.3 ± 53.1 pg/mL and 197.8 ± 42.8 pg/mL in plasma and 129.8 ± 54.2 pg/mL and 139.6 ± 51.5 pg/mL in the CSF. No correlations was found between plasma and CSF IL-1ß concentration after LPS in both groups. The QAlb calculated before LPS and 6 h after was similar in all groups. Melatonin did not affected mRNA expression of Il1B, Il1r1 and Il1r2 in the CP. The mRNA expression of Mmp3 and Mmp9 was not detected. CONCLUSIONS: The lack of correlation between plasma and CSF IL-1ß concentration indicates that at the beginning of inflammation the local synthesis of IL-1ß in the CP is an important source of IL-1ß in the CSF. Melatonin from slow-release implants does not affect IL-1ß concentration in plasma and CSF in early stage of systemic inflammation.

The effect of hemostatic dressing prototypes for the uniformed services on selected blood coagulation parameters in pigs.

BACKGROUND: Serious injuries accompanied by severe bleeding are life-threatening. Post-traumatic hemorrhage involves the risk of developing coagulopathy. Hemostatic dressings are widely used to minimize bleeding. The application of procoagulants in control of hemorrhage may lead to thrombosis or disseminated intravascular coagulation. The aim of this study was to evaluate the effect of hemostatic dressing prototypes on the porcine coagulation system. RESULTS: Fibrinogen and D-dimer concentrations were significantly higher in the experimental groups where hemostatic dressings were used in comparison with the control group. Considerable differences in antithrombin III activity and thrombin-antithrombin complex concentrations were also observed between groups. CONCLUSIONS: The hemostatic dressing comprising modified seton impregnated with 18.0 g/m2 of procoagulant was most effective in preserving the physiological equilibrium between fibrinogenesis and fibrinolysis.

Assessment of adhesion formation after laparoscopic intraperitoneal implantation of Dynamesh IPOM mesh.

INTRODUCTION: Formation of adhesions after laparoscopic hernia repair using the intra-peritoneal onlay mesh (IPOM) procedure can lead to intestinal obstruction or mesh erosion into intestinal lumen. The aims of this study included: measurement of adhesion formation with Dynamesh IPOM after laparoscopic intraperitoneal implantation, and assessment of the occurrence of isolated adhesions at the fastening sites of slowly absorbable sutures. MATERIAL AND METHODS: Twelve healthy pigs underwent laparoscopic implantation of 2 Dynamesh IPOM mesh fragments each, one was fastened with PDSII, and the other with Maxon sutures. An assessment of adhesion formation was carried out after 6 weeks and included an evaluation of surface area, hardness according to the Zhulke scale, and index values. The occurrence of isolated adhesions at slowly absorbable suture fixation points was also analyzed. RESULTS: Adhesions were noted in 83.3% of Dynamesh IPOM meshes. Adhesions covered on average 37.7% of the mesh surface with mean hardness 1.46 and index value 78.8. In groups fixed with PDS in comparison to Maxon sutures adhesions covered mean 31.6% vs. 42.5% (p = 0.62) of the mesh surface, mean hardness was 1.67 vs.1.25 (p = 0.34) and index 85.42 vs. 72.02 (p = 0.95). CONCLUSIONS: The Dynamesh IPOM mesh, in spite of its anti-adhesive layer of PVDF, does not prevent the formation of adhesions. Adhesion hardness, surface area, and index values of the Dynamesh IPOM mesh are close to the mean values of these parameters for other commercially available 2-layer meshes. Slowly absorbable sutures used for fastening did not increase the risk of adhesion formation.

Neurochemical characterization of zinc transporter 3-like immunoreactive (ZnT3(+)) neurons in the intramural ganglia of the porcine duodenum.

The SLC30 family of divalent cation transporters is thought to be involved in the transport of zinc in a variety of cellular pathways. Zinc transporter 3 (ZnT3) is involved in the transport of zinc into synaptic vesicles or intracellular organelles. As the presence of ZnT3 immunoreactive neurons has recently been reported in both the central and peripheral nervous systems of the rat, the present study was aimed at disclosing the presence of a zinc-enriched neuron enteric population in the porcine duodenum to establish a preliminary insight into their neurochemical coding. Double- and triple-immunofluorescence labeling of the porcine duodenum for ZnT3 with the pan-neuronal marker (PGP 9.5), substance P, somatostatin, vasoactive intestinal peptide (VIP), nitric oxide synthase (NOS), leu-enkephalin, vesicular acetylcholine transporter (VAChT), neuropeptide Y, galanin (GAL), and calcitonin gene-related peptide were performed. Immunohistochemistry revealed that approximately 35, 43, and 48 % of all PGP9.5-postive neurons in the myenteric (MP), outer submucous (OSP), and inner submucous (ISP) plexuses, respectively, of the porcine duodenum were simultaneously ZnT3(+). In the present study, ZnT3(+) neurons coexpressed a broad spectrum of active substances, but co-localization patterns unique to the plexus were studied. In the ISP, all ZnT3(+) neurons were VAChT positive, and the largest populations among these cells formed ZnT3(+)/VAChT(+)/GAL(+) and ZnT3(+)/VAChT(+)/VIP(+) cells. In the OSP and MP, the numbers of ZnT3(+)/VAChT(+) neurons were two times smaller, and substantial subpopulations of ZnT3(+) neurons in both these plexuses formed ZnT3(+)/NOS(+) cells. The large population of ZnT3(+) neurons in the porcine duodenum and a broad spectrum of active substances which co-localize with this peptide suggest that ZnT3 takes part in the regulation of various processes in the gut both in normal physiology and during pathological processes.

[The assessment of modification of prosthesis fixation method on visceral interposition during laparoscopic abdominal hernia reconstruction]. / Ocena wplywu modyfikacji techniki mocowania protez na ryzyko interpozycji trzewi podczas laparoskopowej naprawy przepuklin brzusznych.

UNLABELLED: During laparoscopic abdominal hernia repair (LAHR) there is a discrepancy in relative position of mesh and fixation points during the procedure (while abdominal cavity is distended with gas) and afterwards. Therefore, after the surgery the prosthesis becomes corrugated, and tension develops in suture anchoring points. One of the proposed solutions of this problem is to place stabilizing sutures outside the mesh edges and to tie them after emptying the abdominal cavity of CO2. However, due to lack of visualization, viscera entrapment between the mesh and integuments may occur during this stage of surgery. The aim of the study was to assess the effect of fixation technique modification on the risk of visceral interposition between the mesh and integuments and deformation of prostheses fixated this way. MATERIAL AND METHODS: The study was performed in an experimental animal model (12 pigs) by implanting laparoscopically two 6.6x6 cm mesh fragments per animals and tying fixation sutures after emptying the abdominal cavity of CO2. After 6 weeks, visceral interposition between mesh and integuments and such fixated fragments corrugation were assessed. RESULTS: Visceral dislocation between parietal and visceral surface was absent in all 24 assessed meshes, despite fixation under no visual guidance. There were no mesh deformities between anchoring points. CONCLUSIONS: The analysed modification of laparoscopic abdominal hernia repair does not create risk of internal organ interposition between the prosthesis and integuments and prevents mesh from being corrugated.

Sources of porcine longissimus dorsi muscle (LDM) innervation as revealed by retrograde neuronal tract-tracing.

The aim of the present study was to establish the origin of the motor, autonomic and sensory innervation of the L1-L2 segment of the porcine longissimus dorsi muscle (LDM), in order to provide morphological basis for further studies focusing on this neural pathway under experimental conditions, e.g. phototerapy and/or lateral electrical surface stimulation. To reach the goal of the study, multiple injections of the fluorescent neuronal tracer Fast Blue (FB) were made into the LDM region between the spinal processes of the vertebrae L1 and L2. The spinal cord (Th13-S1 segments) as well as the sensory and autonomic ganglia of interest, i.e., dorsal root (DRG) and sympathetic chain ganglia from corresponding spinal cord levels were collected three weeks later. FB-positive (FB+) motoneurons were observed exclusively within the nucleus ventromedialis at L1 and L2 spinal cord level, forming the most ventro-medially arranged cell column within this nucleus. Primary sensory and sympathetic chain neurons were found in appropriate ipsilateral ganglia at Th15-L3 levels. The vast majority of retrogradely traced neurons (virtually all motoneurons, approximately 76% of sensory and 99.4% of sympathetic chain ganglia neurons) was found at the L1 and L2 levels. The morphometric evaluation of FB-labeled DRG neurons showed that the majority of them (approximately 66%) belonged to the class of small-diameter perikarya (10-30 microm in diameter), whereas those of medium size (30-80 microm in diameter) and of large diameter (more than 80 microm) constituted 22.6% and 11.5% of all DRG neurons, respectively. The results of the present study demonstrated that the nerve terminals supplying porcine LDM originated from different levels of the spinal cord, dorsal root and sympathetic chain ganglia. Thus, the study has revealed sources and morphological characteristic of somatic, autonomic and spinal afferent neurons supplying porcine LDM, simultaneously pointing out the characteristic features of their distribution pattern.