In vivo performance of decellularized tracheal grafts in the reconstruction of long length tracheal defects: Experimental study.

BACKGROUND: The repair of long-segment tracheal lesions remains an important challenge. Nowdays no predictable and dependable substitute has been found. Decellularized tracheal scaffolds have shown to be a promising graft for tracheal transplantation, since it is non-immunogenic. OBJECTIVE: Evaluate in vivo decellularized tracheal allografts performance to replace long tracheal segment. METHODS: Forty-five swines underwent surgery as follows: Fifteen trachea donors and 30 receptors of decellularized trachea allografts. The receptors were randomly divided in five groups (n = 6). In groups I and II, donor trachea segment was decellularized by 15 cycles with sodium deoxycholate and deoxyribonuclease, in group II, the allograft was reinforced with external surgical steel wire. Groups, III, IV, and V decellularization was reduced to seven cycles, supplemented with cryopreservation in group IV and with glutaraldehyde in group V. A 10 rings segment was excised from the receptor swine and the decellularized trachea graft was implanted to re-establish trachea continuity. RESULTS: Both decellularization cycles caused decreased stiffness. All trachea receptors underwent euthanasia before the third post-implant week due to severe dyspnea and trachea graft stenosis, necrosis, edema, inflammation, hemorrhage, and granulation tissue formation in anastomotic sites. Histologically all showed total loss of epithelium, separation of collagen fibers, and alterations in staining. CONCLUSIONS: Both decellularization techniques severely damaged the structure of the trachea and the extracellular matrix of the cartilage, resulting in a no functional graft, in spite of the use of surgical wire, cryopreservation or glutaraldehyde treatment. An important drawback was the formation of fibrotic stenosis in both anastomosis.

Experimental Tracheal Replacement: Angiogenesis and Null Apoptosis Promote Stenosis.

BACKGROUND: Tracheal replacement is a challenge for thoracic surgeons due to stenosis in the trachea-prosthesis anastomosis. We propose that stenosis occurs due to fibrosis as a result of an abnormal healing process, characterized by an increased expression of wound healing growth factors (vascular endothelial growth factor [VEGF], survivin, and CD31), which promote angiogenesis and decrease apoptosis. We analyzed the immunoreactivity of VEGF, survivin, CD31, and caspase-3 in the development of fibrotic stenosis in prosthetic tracheal replacement. METHODS: Fourteen dogs were operated on: group I (n=7) received a 6-ring cervical tracheal segment autograft, while in group II (n=7), a 6-ring segment of the cervical trachea was resected and tracheal continuity was restored with a Dacron prosthesis. The follow-up was 3 months. Immunoreactivity studies for VEGF, survivin, CD31, and caspase-3 were performed. A statistical analysis was done using the Wilcoxon signed rank test. RESULTS: Four animals in group I were euthanized on the 10th postoperative day due to autograft necrosis. Three animals completed the study without anastomotic stenosis. Moderate expression of VEGF (p=0.038), survivin (p=0.038), and CD31 (p=0.038) was found. All group II animals developed stenosis in the trachea-prosthesis anastomotic sites. Microscopy showed abundant collagen and neovascularization vessels. Statistically significant immunoreactive expression of VEGF (p=0.015), survivin (p=0.017), and CD31 (p=0.011) was observed. No expression of caspase-3 was found. CONCLUSION: We found a strong correlation between fibrosis in trachea-prosthesis anastomoses and excessive angiogenesis, moderate to intense VEGF, CD31, and survivin expression, and null apoptotic activity. These factors led to uncontrolled collagen production.

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.

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.

[Evaluation of the use of bovine pericardium in non-anatomical lung resections in dogs]. / Evaluación de la utilidad del pericardio bovino en resecciones pulmonares no anatómicas en perros.

UNLABELLED: In this study we assessed the usefulness, healing, as well as the integration to lung tissue of glutaraldehyde preserved at 0.5% bovine pericardium GPBP and lyophilized (GPBPL), after reinforced resection of lung tissue in dogs by thoracotomy or thoracoscopy. MATERIAL AND METHODS: GPBP and GPBPL were prepared and used to reinforce the suture line of lung resection in 30 mongrel dogs: Group I (n = 6): The GPBP were fixed on the lung with 4-0 polypropylene by thoracotomy. Group II (n = 6): The resection and fixed of the GPBP were performed with an linear stapler by thoracotomy. Group III (n = 6): The resection and fixed of the GPBPL were performed with an linear stapler by thoracotomy. Group IV (n = 6): The resection and fixed of the GPBP strips were performed with a linear stapler by thoracoscopy. Group V: The resection and fixed of the GPBPL strips were performed with a linear stapler by thoracoscopy. Clinico-radiological evaluation was done until euthanasia of all animals at week 8 postop. Progressive insufflation up to 40 cm H2O of airway pressure was done to evaluated resistance of the heal in the suture line reinforced. Macroscopic, and microscopic examination of the GPBP, GPBPL and lung were evaluated. RESULTS: All animals survived the surgical procedure and study time (8 weeks). No airleaks were evident at any time during the study including the insufflation test. Macroscopic examination of the GPBP and GPBPL showed good adaptation to the lung tissue. Microscopically all animals presented good healing with deposition of fibrotic tissue layer on the GPBP and GPBPL. CONCLUSION: GPBP and GPBPL are an adequate materials to reinforce lung staple line, when resection of lung tissue was performed in dogs by thoracotomy or thoracoscopy.

Evaluación de la utilidad del pericardio bovino en resecciones pulmonares no anatómicas en perros / Evaluation of the use of bovine pericardium in non-anatomical lung resections in dogs

In this study we assessed the usefulness, healing, as well as the integration to lung tissue of glutaraldehyde preserved at 0.5 bovine pericardium GPBP and lyophilized (GPBPL), after reinforced resection of lung tissue in dogs by thoracotomy or thoracoscopy. MATERIAL AND METHODS: GPBP and GPBPL were prepared and used to reinforce the suture line of lung resection in 30 mongrel dogs: Group I (n = 6): The GPBP were fixed on the lung with 4-0 polypropylene by thoracotomy. Group II (n = 6): The resection and fixed of the GPBP were performed with an linear stapler by thoracotomy. Group III (n = 6): The resection and fixed of the GPBPL were performed with an linear stapler by thoracotomy. Group IV (n = 6): The resection and fixed of the GPBP strips were performed with a linear stapler by thoracoscopy. Group V: The resection and fixed of the GPBPL strips were performed with a linear stapler by thoracoscopy. Clinico-radiological evaluation was done until euthanasia of all animals at week 8 postop. Progressive insufflation up to 40 cm H2O of airway pressure was done to evaluated resistance of the heal in the suture line reinforced. Macroscopic, and microscopic examination of the GPBP, GPBPL and lung were evaluated. RESULTS: All animals survived the surgical procedure and study time (8 weeks). No airleaks were evident at any time during the study including the insufflation test. Macroscopic examination of the GPBP and GPBPL showed good adaptation to the lung tissue. Microscopically all animals presented good healing with deposition of fibrotic tissue layer on the GPBP and GPBPL. CONCLUSION: GPBP and GPBPL are an adequate materials to reinforce lung staple line, when resection of lung tissue was performed in dogs by thoracotomy or thoracoscopy.