Changes in the levels of free sialic acid during ex vivo lung perfusion do not correlate with pulmonary function. Experimental model.

BACKGROUND: Ex vivo lung perfusion (EVLP) constitutes a tool with great research potential due to its advantages over in vivo and in vitro models. Despite its important contribution to lung reconditioning, this technique has the disadvantage of incurring high costs and can induce pulmonary endothelial injury through perfusion and ventilation. The pulmonary endothelium is made up of endothelial glycocalyx (EG), a coating of proteoglycans (PG) on the luminal surface. PGs are glycoproteins linked to terminal sialic acids (Sia) that can affect homeostasis with responses leading to edema formation. This study evaluated the effect of two ex vivo perfusion solutions on lung function and endothelial injury. METHODS: We divided ten landrace swine into two groups and subjected them to EVLP for 120 min: Group I (n = 5) was perfused with Steen® solution, and Group II (n = 5) was perfused with low-potassium dextran-albumin solution. Ventilatory mechanics, histology, gravimetry, and sialic acid concentrations were evaluated. RESULTS: Both groups showed changes in pulmonary vascular resistance and ventilatory mechanics (p < 0.05, Student's t-test). In addition, the lung injury severity score was better in Group I than in Group II (p < 0.05, Mann-Whitney U); and both groups exhibited a significant increase in Sia concentrations in the perfusate (p < 0.05 t-Student) and Sia immunohistochemical expression. CONCLUSIONS: Sia, as a product of EG disruption during EVLP, was found in all samples obtained in the system; however, the changes in its concentration showed no apparent correlation with lung function.

Clinical, Histological, and Profibrotic Extracellular Matrix Protein Changes in a Model of Tracheal Stenosis Induced by Cervical Tracheal Autotransplantation.

BACKGROUND: Tracheal stenosis (TS) is a complication of prolonged intubation, tracheotomy, and tracheal surgery that compromises the vascular supply. Animal models are essential for studying its pathophysiology and the effect of interventions. OBJECTIVE: To establish a TS model in rats secondary to tracheal autotransplantation with a graft submerged in bleomycin (Atx-Bleo). Additionally, to evaluate the clinical and histological changes, as well as the expression of newly formed collagen (NFC), isoforms of transforming growth factor beta (TGFß), fibronectin (FN), elastin (ELN), integrin ß1 (ITGß1), and matrix metalloproteinase 1 (MMP1) in TS. METHODS: Twenty Wistar rats were divided into three groups: group I (n = 20) control; group II (n = 10) end-to-end anastomosis of the trachea (tracheoplasty); and group III (n = 10) Atx-Bleo. The animals were evaluated clinically, tomographically, macroscopically, morphometrically, and microscopically. NFC deposition, and the expression of profibrotic and antifibrotic proteins were evaluated in tracheal scars. RESULTS: All animals survived the surgical procedure and the study period. Compared with the other study groups, the Atx-Bleo group developed TS and fibrosis, exhibited higher expression of NFC, TGFß1, TGFß2, FN, ELN, and ITGß1, and mild expression of TGFß3 and MMP1 (p < 0.005; analysis of variance, Dunnett and Tukey tests). CONCLUSION: Atx-Bleo in TS model rats produces tomographic and histological changes, and induces the upregulation of profibrotic proteins (TGFß1, TGFß2, collagen, FN, ELN, ITGß1) and downregulation of antifibrotic proteins (TGFß3, MMP1). Therefore, this model may be used to test new pharmacological treatments for reversing or preventing TS, and conduct basic studies regarding its pathophysiology.

Endothelin-Converting Enzyme 1 and Vascular Endothelial Growth Factor as Potential Biomarkers during Ex Vivo Lung Perfusion with Prolonged Hypothermic Lung-Sparing.

Lung transplantation requires optimization of donor's organ use through ex vivo lung perfusion (EVLP) to avoid primary graft dysfunction. Biomarkers can aid in organ selection by providing early evidence of suboptimal lungs during EVLP and thus avoid high-risk transplantations. However, predictive biomarkers of pulmonary graft function such as endothelin-converting enzyme (ECE-1) and vascular endothelial growth factor (VEGF) have not been described under EVLP with standard prolonged hypothermic preservation, which are relevant in situations where lung procurement is difficult or far from the transplantation site. Therefore, this study is aimed at quantifying ECE-1 and VEGF, as well as determining their association with hemodynamic, gasometric, and mechanical ventilatory parameters in a swine model of EVLP with standard prolonged hypothermic preservation. Using a protocol with either immediate (I-) or delayed (D-) initiation of EVLP, ECE-1 levels over time were found to remain constant in both study groups (p > 0.05 RM-ANOVA), while the VEGF protein was higher after prolonged preservation, but it decreased throughout EVLP (p > 0.05 RM-ANOVA). Likewise, hemodynamic, gasometric, mechanical ventilatory, and histological parameters had a tendency to better results after 12 hours of hypothermic preservation in the delayed infusion group.

Expression of Claudin-4 in Lung Ischemia-Reperfusion Injury in Experimental Lung Transplantation.

OBJECTIVE: To assess the presence of CLDN4 in bronchoalveolar lavage fluid (BALF) and pulmonary tissue as an early indicator of LIRI and its relationship with changes in pulmonary physiology, edema formation and histology in an experimental porcine model of LTx with CIT of 50 min or 6 h. METHODS: In 12 pigs, LIRI was produced by: group I (n = 6) LTx with 50 min of CIT (LTx-50 min-CIT); and group II (n = 6) LTx with 6 h of CIT (LTx-6h-CIT). The lung function, edema formation, macroscopic and microscopic changes were assessed. CLDN4 expression in BALF and pulmonary tissue were determined. RESULTS: Both groups presented similar clinical, edema, and histological damage, as well as similar expression of CLDN4 in BALF and tissue (p > 0.05, RM-ANOVA). CONCLUSION: CLDN4 expressed in BALF and the pulmonary tissue during the first 5 h within 72 h of the PGD window are not associated by the deterioration of lung function, edema and lung histological injury, in LTx with CIT 50 min or 6 h, CLDN4 does not seem to be a valuable indicator of LIRI.

Treatment with Hyaluronic Acid and Collagen-Polyvinylpyrrolidone Improves Extracellular Matrix Assembly for Scarring after Tracheal Resection.

Treatment of tracheal stenosis is occasionally performed in combination with wound healing modulators to manipulate new extracellular matrix (ECM) formation and prevent fibrosis. Hyaluronic acid (HA) and collagen-polyvinylpyrrolidone (collagen-PVP) decrease fibrosis in experimental tracheal healing. However, they have not been used clinically as their effect on ECM components, which modify tracheal scarring, has not been described. Objective. To evaluate the effect of the application of HA, collagen-PVP, a mixture of HA and collagen-PVP (HA+collagen-PVP), and mitomycin C on the expression of decorin, matrix metalloproteinase 1 (MMP1), and MMP9, as well as the type of collagen and deposits formed in the scar after resection and end-to-end anastomosis (REEA) of the cervical trachea using an experimental model. Materials and Methods. Thirty dogs underwent REEA of the cervical trachea and were treated with different wound healing modulators: group I (n = 6), control; group II (n = 6), HA; group III (n = 6), collagen-PVP; group IV (n = 6), HA+collagen-PVP; and group V (n = 6), mitomycin C. The dogs were evaluated clinically and endoscopically for 4 weeks. Subsequently, macroscopic and microscopic changes, expression of ECM proteins, and collagen deposition in tracheal scars were analysed. Results. Groups II, III, and IV showed reduced endoscopic, macroscopic, and microscopic inflammation, improved neovascularization, high decorin expression (p < 0.01, analysis of variance (ANOVA)), and moderate expression of MMP1 (p < 0.003, ANOVA) and type I and III collagen (p < 0.05, Kruskal-Wallis). Groups IV and V developed fewer collagen deposits (p < 0.001, ANOVA). Conclusion. Treatment with HA and collagen-PVP improved post-REEA healing by increasing neovascularization, stimulating the expression of decorin, and regulating the expression of MMP1, as well as type I and III collagen and their deposition.

Effects of Pirfenidone and Collagen-Polyvinylpyrrolidone on Macroscopic and Microscopic Changes, TGF-ß1 Expression, and Collagen Deposition in an Experimental Model of Tracheal Wound Healing.

Tracheal stenosis (TS) is a fibrosis originated by prolonged inflammation and increased transforming growth factor beta 1 (TGF-ß1) expression and collagen deposition (CD) in the tracheal wound. Several wound-healing modulators (WHMs) have been used to modulate the tracheal healing process and prevent TS, but they have failed, justifying the need to evaluate alternative WHM. The pirfenidone (PFD) and collagen-polyvinylpyrrolidone (Collagen-PVP) decrease inflammation and fibrosis. This study assessed the effect of PFD administration and Collagen-PVP topical application on macroscopic and microscopic changes, TGF-ß1 expression, and CD in an experimental model of tracheal wound healing. Forty Wistar rats underwent cervical tracheoplasty, were divided into 4 groups (n = 10), and were treated with different WHM: group I, saline solution (SS); group II, Collagen-PVP; group III, mitomycin C (MMC); and group IV, 40 mg/kg PFD. Four weeks after surgery, the macroscopic and microscopic changes, in situ TGF-ß1 expression, and CD in posttracheoplasty scars were evaluated. The animals treated with Collagen-PVP and PFD developed less inflammation and fibrosis than animals in the other study groups (p < 0.05, Kruskal-Wallis) and, moreover, showed lower TGF-ß1 expression and CD than animals in group I (p < 0.05, ANOVA and Tukey's test). In conclusion, PFD and Collagen-PVP decrease inflammation, fibrosis, TGFß-1 expression, and CD in the posttracheoplasty rats' scar.

Comparison of lyophilized glutaraldehyde-preserved bovine pericardium with different vascular prostheses for use as vocal cords implants: experimental study.

This study compared the use of lyophilized glutaraldehyde-preserved bovine pericardium (LGPBP), polytetrafluoroethylene (PTFE), polyethylene terephthalate (PET), and Teflon felt (TF) as implants for vocal cords (VC) medialization and aimed to assess the endoscopic, macroscopic, and microscopic VC changes after medialization in a canine model. In 18 mongrel dogs, the right VC were medialized with LGPBP and the left were implanted as follows: Group I (n = 6): LGPBP and PTFE; Group II (n = 6): LGPBP and PET; Group III (n = 6): LGPBP and TF. Surgical handling of the implants was compared. Three months after surgery, macroscopic and microscopic changes of VC and implants were evaluated. LGPBP offered the best surgical handling (p = 0.005, Kruskal-Wallis). TF implants showed extrusion (p = 0.005, Kruskal-Wallis) and severe inflammation. All VC formed fibrous capsules around the implants; the ones developed by LGPBP implants were thinner (p = 0.001, ANOVA, Tukey). VC implanted with synthetic materials showed eosinophilic infiltration (p = 0.01, Kruskal-Wallis). We concluded that the LGPBP could be used as an implant for VC medialization because it is biocompatible, easy to handle and remove during surgical procedures, and nonabsorbable or extrudable and produces an inflammatory reaction similar to PTFE and PET.

Antigen-induced airway hyperresponsiveness and obstruction is related to caveolin-1 expression in airway smooth muscle in a guinea pig asthma model.

BACKGROUND: Caveolin-1 is a fundamental signalling scaffold protein involved in contraction; however, the role of caveolin-1 in airway responsiveness remains unclear. We evaluated the relationship between caveolin-1 expression in airway smooth muscle (ASM) and antigen-induced airway responsiveness and obstruction in a guinea pig asthma model. METHODS: Airway obstruction in sensitised guinea pigs, induced by antigenic (ovalbumin) challenges administered every 10 days, was measured. Antigen-induced responsiveness to histamine and the expression of caveolin-1 and cavin 1, 2 and 3 were evaluated at the third ovalbumin challenge. The control group received saline solution instead of ovalbumin. RESULTS: After the first challenge, antigen exposure induced a transient airway obstruction and airway hyperresponsiveness, high levels of IL-4 and IL-5 in lung and airway globet cells proliferation at the third antigenic challenge. Caveolin-1 mRNA levels in total lung decreased in the experimental group compared with controls. Flow cytometric analysis of ASM from the experimental group showed a high number of cells expressing caveolin-1 compared with controls. This increase was confirmed by western blot. Airway obstruction and hyperresponsiveness correlated with the degree of increased caveolin-1 expression in ASM cells (P < 0.05; r = 0.69 and -0.52, respectively). The expression of cavins 1, 2 and 3 in ASM also increased in the experimental group compared to controls. Immunohistochemical findings reveal that differences in ASM caveolin-1 were not evident between groups. Nevertheless, a marked decrease in caveolin-1 and caspase 3 was observed in the pulmonary vascular smooth muscle of asthma model compared with controls. Histological analysis did not reveal differences in smooth muscles mass or subepithelial fibrosis levels in airways between groups. However, an enlargement of smooth muscle mass was observed in the pulmonary microvessels of experimental animals. This enlargement did not induce changes in pulmonary or systemic arterial pressures. CONCLUSIONS: Our data suggest that caveolin-1 expression in ASM has a crucial role in the development of antigen-induced airway obstruction and hyperresponsiveness in a guinea pig asthma model. In addition, the asthma model in guinea pigs appears to induce a contractile smooth muscle phenotype in the airways and a proliferative smooth muscle phenotype in pulmonary vessels.

Respiratory mechanics and plasma levels of tumor necrosis factor alpha and interleukin 6 are affected by gas humidification during mechanical ventilation in dogs.

The use of dry gases during mechanical ventilation has been associated with the risk of serious airway complications. The goal of the present study was to quantify the plasma levels of TNF-alpha and IL-6 and to determine the radiological, hemodynamic, gasometric, and microscopic changes in lung mechanics in dogs subjected to short-term mechanical ventilation with and without humidification of the inhaled gas. The experiment was conducted for 24 hours in 10 dogs divided into two groups: Group I (n = 5), mechanical ventilation with dry oxygen dispensation, and Group II (n = 5), mechanical ventilation with oxygen dispensation using a moisture chamber. Variance analysis was used. No changes in physiological, hemodynamic, or gasometric, and radiographic constants were observed. Plasma TNF-alpha levels increased in group I, reaching a maximum 24 hours after mechanical ventilation was initiated (ANOVA p = 0.77). This increase was correlated to changes in mechanical ventilation. Plasma IL-6 levels decreased at 12 hours and increased again towards the end of the study (ANOVA p>0.05). Both groups exhibited a decrease in lung compliance and functional residual capacity values, but this was more pronounced in group I. Pplat increased in group I (ANOVA p = 0.02). Inhalation of dry gas caused histological lesions in the entire respiratory tract, including pulmonary parenchyma, to a greater extent than humidified gas. Humidification of inspired gases can attenuate damage associated with mechanical ventilation.

Pericardio bovino tratado con glutaraldehído y liofilizado como materiales para la medialización de cuerdas vocales en modelo animal / Glutaraldehyde-preserved and lyophilized bovine pericardium as materials for medialization of the vocal folds in an animal model

Introducción: El pericardio bovino tratado con glutaraldehído (PBTG) y el pericardio bovino tratado con glutaraldehído liofilizado (PBTGL)ha sido utilizado exitosamente en la reparación de varios defectos anatómicos, pero su eficacia y seguridad como implantes de cuerdas vocales (CV) no se ha descrito. Objetivo: Evaluar la utilidad del PBTG y PBTGL como material para la medialización tiroplástica y valorar los cambios endoscópicos, macroscópicos y microscópicos de las CV posmedialización en un modelo experimental canino. Material y métodos: En 12 perros mestizos, se medializó la CV derecha con pericardio y la izquierda con politetrafluoroetileno (PTFE). Grupo i (n=6): PBTG, y Grupo ii (n=6): PBTGL. Se comparó el manejo quirúrgico de los implantes. Los animales se valoraron clínica y endoscópicamente. Tres meses poscirugía se evaluaron macroscópica y microscópicamente las laringes. Resultados: El PBTG y PBTGL mostraron mejor manejo quirúrgico (Kruskal-Wallis, p=0,005). No se presentaron granulomas, absorción o extrusión del implante en ningún caso endoscópica ni macroscópicamente. Al final del estudio las CV medializadas con PTFE se observaron más engrosadas. Microscópicamente todas las CV formaron una cápsula fibrosa alrededor del implante y una reacción inflamatoria crónica similar, pero las implantadas con PTFE mostraron infiltrado eosinofílico (Kruskal-Wallis, p<0,05). Conclusión: El PBTG y PBTGL pueden ser utilizados para la medialización de las CV debido a que son biocompatibles, de fácil manejo quirúrgico, no se absorben, no migran, ni extruyen y producen una reacción inflamatoria similar a la del PTFE (AU)

Introduction: Glutaraldehyde-preserved bovine pericardium (GBP) and lyophilized GBP (LGBP) have been used successfully in repairing several anatomical defects, but their effectiveness and safety as implants to vocal cords (VC) have not been reported. Objective: The aim of this study was to evaluate the use of GBP and LGBP as materials for medialization thyroplasty, as well as to assess the endoscopic, macroscopic and microscopic VC changes after medialization in an experimental canine model. Material and methods: In 12 healthy mongrel dogs, the right VC were medialized using pericardium and the left with polytetrafluoroethylene (PTFE). Group 1 (n=6): GBP and Group 2 (n=6): LGBP. The surgical manoeuvrability of the implants was compared. The animals were evaluated clinically and endoscopically. Three months after surgery, the larynges were assessed macro- and microscopically. Results: Both GBP and LGBP implants showed better surgical manoeuvrability (Kruskal-Wallis, P=0.005). Endoscopic and macroscopic studies showed no evidence of granulomas, absorption or extrusion of the implant. At the end of the study, greater thickness was observed in VC implanted with PTFE. Microscopically, all the VC developed fibrous capsules surrounding the implants and similar chronic inflammation reaction. The VC implanted with PTFE presented eosinophilic infiltration (Kruskal-Wallis, P<0.05). Conclusion: Both GBP and LGBP can be used as implants for VC medialization because they are biocompatible, have easy surgical manoeuvrability, do not suffer absorption, migration or extrusion and produce inflammation reactions similar to those of PTFE (AU)

[Glutaraldehyde-preserved and lyophilized bovine pericardium as materials for medialization of the vocal folds in an animal model]. / Pericardio bovino tratado con glutaraldehído y liofilizado como materiales para la medialización de cuerdas vocales en modelo animal.

INTRODUCTION: Glutaraldehyde-preserved bovine pericardium (GBP) and lyophilized GBP (LGBP) have been used successfully in repairing several anatomical defects, but their effectiveness and safety as implants to vocal cords (VC) have not been reported. OBJECTIVE: The aim of this study was to evaluate the use of GBP and LGBP as materials for medialization thyroplasty, as well as to assess the endoscopic, macroscopic and microscopic VC changes after medialization in an experimental canine model. MATERIAL AND METHODS: In 12 healthy mongrel dogs, the right VC were medialized using pericardium and the left with polytetrafluoroethylene (PTFE). Group 1 (n=6): GBP and Group 2 (n=6): LGBP. The surgical manoeuvrability of the implants was compared. The animals were evaluated clinically and endoscopically. Three months after surgery, the larynges were assessed macro- and microscopically. RESULTS: Both GBP and LGBP implants showed better surgical manoeuvrability (Kruskal-Wallis, P=.005). Endoscopic and macroscopic studies showed no evidence of granulomas, absorption or extrusion of the implant. At the end of the study, greater thickness was observed in VC implanted with PTFE. Microscopically, all the VC developed fibrous capsules surrounding the implants and similar chronic inflammation reaction. The VC implanted with PTFE presented eosinophilic infiltration (Kruskal-Wallis, P<.05). CONCLUSION: Both GBP and LGBP can be used as implants for VC medialization because they are biocompatible, have easy surgical manoeuvrability, do not suffer absorption, migration or extrusion and produce inflammation reactions similar to those of PTFE.

[Effects of propofol pretreatment on endothelin in oleic acid-induced acute lung injury]. / Efectos del pretratamiento con propofol sobre la endotelina en daño pulmonar agudo inducido por ácido oleico.

INTRODUCTION: Acute lung injury (ALI) is a pathological condition characterized by injury in the alveolar-capillary membrane that triggers local and systemic inflammation. Endothelin (ET) is a protein that regulates immune response and constricts blood vessels; when it is over-expressed, it may contribute to high blood pressure and lung injury. This work tries to determine if propofol may decrease hemodynamic, gasometric, microscopic, ET-1 plasmatic concentration, and immuno-histochemical alterations in an experimental model of oleic acid-induced acute lung injury. MATERIALS AND METHODS. Animals were classified into three groups (n = 6): group I was the control group; in group II, there was oleic acid-induced ALI with no treatment, and group III with propofol pre-treatment and oleic acid-induced ALI. RESULTS: All animals survived until the end of the study, and 100% of group II and group III developed ALI, with hemodynamic, gasometric and gravimetric alterations. However, group III showed less inflammatory infiltration and lower ET-1 expression in lung tissue. CONCLUSIONS: Pretreatment with propofol in a canine model of OA-induced ALI indicates that the drug has anti-inflammatory action, with a potential therapeutic role against progression of anti-inflammation and lung damage.

[Experimental model of canine oleic acid-induced acute lung injury: establishment of a working model]. / Modelo experimental canino de daño pulmonar agudo producido con ácido oleico: estableciendo un modelo de trabajo.

INTRODUCTION: There are several experimental model of acute lung injury induced by oleic acid (OA); however, there are few studies that show how this injury develops. OBJECTIVE: This study seeks to detail the x-ray, hemodynamic, gasometrical, gravimetrical, macroscopic and microscopic alterations developed in an experimental model of canine OA-induced acute lung injury (ALI). MATERIAL AND METHODS: Twelve dogs were divided in 2 study groups: Group I (n=6): Control group without ALI. Group II (n=6); OA-induced ALI. All dogs were submitted to X-ray, hemodynamic and gasometric evaluation before ALI induction, and later every 15 minutes during 150 minutes. At the end of the study, the animals were euthanatized and were evaluated the changes gravimetric, macroscopic and microscopic in injured lungs. RESULTS: All the animals survived through the study. In group II, 100% of the animals developed x-ray (p < 0.003 Wilcoxon), hemodynamic, gasometrical and gravimetric (p < 0.5 ANOVA, Tukey), macroscopically and microscopically (p < 0.001 Wilcoxon) ALI. CONCLUSIONS: The OA-induced ALI is a model in which dogs develop X-ray, hemodynamic, gasometrical, gravimetrical, macroscopically and microscopically injuries of the exudative phase that lung with ALI injury presents.

Lyophilized glutaraldehyde-preserved bovine pericardium for experimental atrial septal defect closure.

A variety of patch materials has been used to close large atrial septal defects (ASD). Autologous pericardium and glutaraldehyde-preserved bovine pericardium are the most used. Lyophilized bovine pericardium has not been tested inside the cardiovascular system. The aim of this work was to study the behaviour and effectiveness of lyophilized glutaraldehyde-preserved bovine pericardium in ASD closure. Sixteen mongrel dogs were operated on. A 3 cm diameter atrial septal defect was created, and closed with: Group I (n=8): Lyophilized glutaraldehyde preserved bovine pericardium (LGPBP). Group II (n=8): Vascular Dacron patch. The animals were evaluated clinically, by echocardiography, macroscopically, and microscopically. Statistical analysis was done with analysis of variance (ANOVA) and Student's t-test. All the animals survived the surgical procedure and study time (6 months). Clinically all the animals displayed normal physical activity, with normal cardiac sounds. Echocardiography showed that both groups had a normal heart without intracardiac shunts, no thrombus formation, and no vegetations. Macroscopically all the animals showed good integration of the lyophilized bioprosthesis and Dacron patch. All group I animals presented a decrease of the area of the ASD in the left atrium (p<0.001 by ANOVA and Student's t-test). Microscopically, group I presented dense and well-organized collagenous tissue, areas of cartilaginous metaplasia and remnants of the lyophilized bioprosthesis (p<0.001 by ANOVA and Student's t-test). Group II showed encapsulated Dacron patch covered with collagenous tissue and cartilaginous metaplasia. In conclusion, the new lyophilized bioprosthesis is well integrated into the atrial septum, without complications and is effective for ASD closure.

[Middle ear packing with collagen-polyvinylpyrrolidone sponge. Experimental study in guinea pigs]. / Empaquetamiento del oído medio con esponja de colágena polivinil pirrolidona. Estudio experimental en cobayos.

INTRODUCTION: Packing material is mandatory in middle ear (ME) surgery in order to avoid inflammation, adhesions and fibrotic healing. Collagen polivynil-pirrolidone (CPVP) is a healing modulator, which reduces inflammation and fibrosis. Hence we can hypothesize that packing of the ME with CPVP sponge will lead a good ME healing. OBJECTIVE: The aim of this study was to evaluate the otoscopic and microscopic changes induced on the healthy mucosa of the ME and Tympanic membrane (TM) after packing with CPVP sponge in guinea pigs. MATERIAL AND METHODS: Twelve guinea pigs were operated on of right myringotomy. The ME was packed with: Group I (n = 6): Absorbable gelatin sponge (AGS) in SS; Group II (n = 6): CPVP soaked in SS. TM and ME integrity was evaluated otoscopically, as well as residual packing material. Euthanasia was performed on the 4th post-operative week. ME mucosa histologic examination was done. RESULTS: Group I in all the cases showed residual packing material (p < 0.007 Student's test p < 0.001 ANOVA). Histologically both groups presented inflammation with polymorphonuclears, in addition group I showed severe lymphocytosis (p < 0.003 Student's, test p < 0.001, ANOVA). CONCLUSION: The CPVP sponge when it is used as material of packing in the OM of guinea pigs produces less chronic inflammatory changes, but more studies with the injured mucosa are required to validate their utility in the otologic surgery.

Wound healing modulators in a tracheoplasty canine model.

Postsurgical tracheal stenosis results from fibrosis formation due to ischemia. There are healing modulators, hyaluronic acid (HA) and collagen polyvinylpyrrolidone (CPVP), which reduce collagen fibers formation. Thus we can hypothesize that the topical application of one of these modulators can diminish postsurgical tracheal scarring and stenosis. The aim of this work was to evaluate the macroscopic, microscopic, and biochemical changes of tracheal healing after the application of HA or CPVP in a canine tracheoplasty model. The study design was prospective experimental investigation in a canine model. Eighteen mongrel dogs underwent three cervical tracheal rings resection and end-to-end anastomosis. They were randomized into three groups according to treatment: group I (control group) (n = 6), topical application of saline solution on tracheal anastomosis; group II (n = 6), topical application of 15 microg HA on tracheal anastomosis; and group III (n = 6), topical application of 2.5 mg CPVP on tracheal anastomosis. They were evaluated clinical, radiological and tracheoscopically during 4 weeks. They were euthanized at the end of the study time. Macroscopic, microscopic, and biochemical changes of tracheal anastomosis healing were analyzed. Collagen formation was quantified by the Woessner method. All the animals survived the surgical procedure and study period. Macroscopic, radiologic, and endoscopic studies showed that animals in group I developed tracheal stenosis, inflammation, and firm fibrous tissue formation, and histological studies also showed severe inflammatory reaction and fibrosis formation. Groups II (HA) and III (CPVP) showed well-organized thin collagen fibers with minimal inflammatory response. Biochemical evaluation revealed a higher collagen concentration in group I animals (analysis of variance [ANOVA] p < .05 and Tukey p < .01). Thus, hyaluronic acid or collagen polyvinylpyrrolidone administered after tracheal anastomosis diminished the degree of stenosis and inflammatory reaction. Both modulators improved tracheal healing.

Trasplante experimental de tráquea cervical combinado con la aplicación de factor de crecimiento de fibroblastos / Experimental cervical trachea transplant combined with application of basic fibroblast growth factor

Antecedentes: El trasplante traqueal de longitudes mayores a 6 cm de longitud ha fallado por complicaciones isquémicas del injerto. Experimenta/mente se han utilizado factores de crecimiento y diferentes técnicas quirúrgicas, como la de trasplante traqueal dividido para favorecer la neoformación de vasos sanguíneos. Objetivo: Evaluar la viabilidad, cambios tráquea cervical, macroscópicos y microscópicos del trasplante de tráquea cervical en perros al utilizar la técnica quirúrgica de trasplante traqueal dividido, combinando la aplicación del factor básico de crecimiento para fibroblastos en los sitios de las anastomosis. Material y métodos: Se operaron 24 perros que fueron divididos en 4 grupos de estudio: Grupo I (n = 6): Trasplante de 9 anillos de tráquea cervical con la técnica convencional (TTCC); Grupo II (n = 6): TTCC e instilación tópica de factor básico de crecimiento de fibroblastos (bFGF) en los sitios de anastomosis; Grupo III (n = 6): Trasplante con la técnica de trasplante dividido (TTCD) y Grupo IV (n = 6): TTCD y aplicación de bFGF. Los animales recibieron triple inmunosupresión (azatioprina, metilprednisolona, ciclos-porína). Se planearon evaluaciones clínica, radiológica y traqueoscópica durante 4 semanas. Al final del estudio los injertos trasplantados se evaluaron macroscópica y microscópicamente. Resultados: Ningún animal concluyó su tiempo de estudio por disnea grave durante la primera semana del estudio. Macroscópicamente todos los injertos desarrollaron fístulas y necrosis. Microscópicamente mostraron necrosis, inflamación, vasculitis, hemorragia y destrucción del cartílago. Conclusión: En este estudio encontramos que el trasplante de tráquea cervical mayor a 6 cm, tiene malos resultados con la técnica quirúrgica convencional o dividida e inmunosupresión, así como con y sin la aplicación de bFGF en los sitios de anastomosis.

Background: Tracheal transplantation of lesions larger than 6 cm fails due to ischemic complications. Growth factors and different surgical techniques, including the divided tracheal graft technique, have been used experimentally to stimulate the neoformation of blood vessels. Objective: Assessment of the viability and macroscopic and microscopic changes of the cervical transplanted trachea in dogs, using the divided tracheal graft technique, combined with the application of basic fibroblast growth factor (bFGF) on the anastomotic site. Material and methods: Twenty four mongrel dogs were divided in 4 study groups: Group I (n = 6): Transplantation of 9 cervical tracheal rings with the conventional surgical technique (TCTC). Group II (n = 6): TCTC combined with topical instillation of bFGF at on the anastomotic line. Group III (n = 6): Transplantation of cervical trachea using the divided tracheal graft technique (TCTD), and Group IV (n = 6): TCTD with topical application of bFGF. The animals received triple immunotherapy (azathioprine, methylprednisolone, cyclosporine) and were to have clinical, radiological and endoscopical evaluation during 4 weeks. At the end of the study, macroscopic and microscopic evaluations of the transplanted grafts were done. Results: No animal completed the study time due to severe dyspnea during the first postoperative week. Macroscopically all grafts showed necrosis and fistulae formation. Microscopically we observed necrosis, inflammation, vasculitis, hemorrhage and destruction of cartilage in all the grafts. Conclusion: In this study, tracheal allotransplanta-tion longer than 6 cm with either surgical technique, with immunosuppression, with or without bFGF, is unsuccessful.

Cryopreserved tracheal grafts: a review of the literature.

Cryopreserved tracheal grafts have been used in several experimental models of long segment replacement. The clinical application of the procedure has been limited due to the fact that contradictory results have been reported. The purpose of this article is to present a review of the literature on tracheal cryopreservation. Despite the fact that most authors indicate that cryopreserved tracheal allografts retain viability and have a low immunological response, though they continue to function after transplantation with good epithelialization and patency, cryopreservation leads to significant damage to cartilage, the degree of which is based on the freezing-storage methods that affect the function and durability of a graft. The long-term storage of cartilage must therefore be investigated in more detail in basic research models of cartilage viability: the evaluation of chondrocyte apoptosis, and the use of different solutions for tracheal cryopreservation other than RPMI-1640, Dulbecco's modified Eagle's, Eurocollins, and TC-199. Furthermore, problems that involve improving the blood supply to the graft after extensive resection and immunosuppression must be resolved before tracheal cryopreservation can become a clinically established method for tracheal grafts.

Uso del pericardio bovino tratado con glutaraldehído / Use of glutaraldehide-treated bovine pericardium

El uso de las prótesis mecánicas y biológicas surgió de la necesidad de reemplazar quirúrgicamente la falta o la falla de un órgano. Las bioprótesis están hechas a partir de tejido autólogo, homólogo y heterólogo. La bioprótesis de pericardio bovino tratada con glutaraldehído es la más estudiada y se utiliza principalmente como bioprótesis cardiaca. Debido a su fácil preparación y uso, este material se ha empleado para reparar defectos hemiarios de la pared abdominal, la pared torácica, de diafragma y para sustituir vasos y tráquea; sin embargo, causa reacción de rechazo y tipo cuerpo extraño, los que han sido estudiados ampliamente para evitarlos. Presentamos una revisión de la literatura sobre el uso y estudio de la bioprótesis de pericardio bovino tratado con glutaraldehído.

The use of biological and mechanical prostheses arose from the need to replace a failing organ or function. Bioprostheses can be manufactured from autologous, homologus or heterologus tissue; glutaraldehyde treated bovine pericardium bioprostheses have been used extensively to repair diaphragmatic, abdominal and chest wall defects and to replace heart valves, blood vessels and tracheal segments, but the implanted tissue can elicit rejection or foreign body reaction; these have been extensively studied in order to avoid them. We review the literature regarding glutaraldehyde treated bovine pericardium bioprostheses.