Production and transplantation of bioengineered lung into a large-animal model.

The inability to produce perfusable microvasculature networks capable of supporting tissue survival and of withstanding physiological pressures without leakage is a fundamental problem facing the field of tissue engineering. Microvasculature is critically important for production of bioengineered lung (BEL), which requires systemic circulation to support tissue survival and coordination of circulatory and respiratory systems to ensure proper gas exchange. To advance our understanding of vascularization after bioengineered organ transplantation, we produced and transplanted BEL without creation of a pulmonary artery anastomosis in a porcine model. A single pneumonectomy, performed 1 month before BEL implantation, provided the source of autologous cells used to bioengineer the organ on an acellular lung scaffold. During 30 days of bioreactor culture, we facilitated systemic vessel development using growth factor-loaded microparticles. We evaluated recipient survival, autograft (BEL) vascular and parenchymal tissue development, graft rejection, and microbiome reestablishment in autografted animals 10 hours, 2 weeks, 1 month, and 2 months after transplant. BEL became well vascularized as early as 2 weeks after transplant, and formation of alveolar tissue was observed in all animals (n = 4). There was no indication of transplant rejection. BEL continued to develop after transplant and did not require addition of exogenous growth factors to drive cell proliferation or lung and vascular tissue development. The sterile BEL was seeded and colonized by the bacterial community of the native lung.

Body Composition Changes in Severely Burned Children During ICU Hospitalization.

OBJECTIVES: Prolonged hospitalization due to burn injury results in physical inactivity and muscle weakness. However, how these changes are distributed among body parts is unknown. The aim of this study was to evaluate the degree of body composition changes in different anatomical regions during ICU hospitalization. DESIGN: Retrospective chart review. SETTING: Children's burn hospital. PATIENTS: Twenty-four severely burned children admitted to our institution between 2000 and 2015. INTERVENTIONS: All patients underwent a dual-energy x-ray absorptiometry within 2 weeks after injury and 2 weeks before discharge to determine body composition changes. No subject underwent anabolic intervention. We analyzed changes of bone mineral content, bone mineral density, total fat mass, total mass, and total lean mass of the entire body and specifically analyzed the changes between the upper and lower limbs. MEASUREMENTS AND MAIN RESULTS: In the 24 patients, age was 10 ± 5 years, total body surface area burned was 59% ± 17%, time between dual-energy x-ray absorptiometries was 34 ± 21 days, and length of stay was 39 ± 24 days. We found a significant (p < 0.001) average loss of 3% of lean mass in the whole body; this loss was significantly greater (p < 0.001) in the upper extremities (17%) than in the lower extremities (7%). We also observed a remodeling of the fat compartments, with a significant whole-body increase in fat mass (p < 0.001) that was greater in the truncal region (p < 0.0001) and in the lower limbs (p < 0.05). CONCLUSIONS: ICU hospitalization is associated with greater lean mass loss in the upper limbs of burned children. Mobilization programs should include early mobilization of upper limbs to restore upper extremity function.

Integrity of airway epithelium in pediatric burn autopsies: Association with age and extent of burn injury.

UNLABELLED: This study examines the structural integrity of the airway epithelium in autopsy tissues from pediatric burn subjects. METHODS: A semi-quantitative score for the degree of airway epithelial integrity was made for seventy- two pediatric burn autopsies. Multivariate ordinal logistic regression was performed to identify relationships between epithelial integrity and conditions related to tissue fixation, time of death after injury, age, total body surface area burn (TBSA), extent of 3rd degree burn, presence of inhalation injury, ventilator days and pneumonia. RESULTS: No significant difference in epithelial integrity scores was identified between burn only cases and those with inhalation injury. Significant correlations with bronchiolar epithelial integrity scores were identified for age, p=0.02, and percent 3rd degree burn, p=0.02. There was no significant relationship between epithelial integrity and time between death and autopsy, p>0.44. CONCLUSIONS: Airway epithelial loss seen in autopsy tissue is not simply an artifact of tissue fixation. The degree of compromise correlates most strongly with age and degree of burn. Further studies are needed to identify physiological or critical care factors following burn injury that contribute to compromise in the structural and functional properties of the airway epithelium.

Airway obstruction and bacterial invasion in autopsy tissue of pediatric burn victims.

This study measured airway obstruction and bacterial invasion in systematically sampled lung tissue of burn victims at autopsy. Lung tissue from victims of combined smoke inhalation and burn injury (n = 5) and burn injury alone (n = 9) was examined histologically and the degree of bronchial and bronchiolar obstruction was measured. The walls of both bronchi and bronchioles were examined for bacterial invasion. Correlation analysis was performed for the association of airway obstruction with TBSA burn, number of ventilatory days, maximum inspiratory pressure, and days after injury. There was no significant difference in the mean degree of airway obstruction in smoke inhalation and burn victims compared with victims of burn-only injury (P > .05). Increased bronchiolar obstruction scores were detected in victims with pneumonia (55.3 ± 24.2%) compared with victims without pneumonia (9.3 ± 0.2%; P = .03). Bacterial invasion of the bronchial wall was present in one case, and invasion into the walls of bronchioles was seen in five cases. Burned children who died had extensive bronchiolar obstruction whether or not they had smoke inhalation injury. There was bacterial invasion into the airway wall in six of 14 cases (43%). Improved understanding of the mechanisms of airway obstruction is important for improved care of burned children.

Intensive insulin therapy in severely burned pediatric patients: a prospective randomized trial.

RATIONALE: Hyperglycemia and insulin resistance have been shown to increase morbidity and mortality in severely burned patients, and glycemic control appears essential to improve clinical outcomes. However, to date no prospective randomized study exists that determines whether intensive insulin therapy is associated with improved post-burn morbidity and mortality. OBJECTIVES: To determine whether intensive insulin therapy is associated with improved post-burn morbidity. METHODS: A total of 239 severely burned pediatric patients with burns over greater than 30% of their total body surface area were randomized (block randomization 1:3) to intensive insulin treatment (n = 60) or control (n = 179). MEASUREMENTS AND MAIN RESULTS: Demographics, clinical outcomes, sepsis, glucose metabolism, organ function, and inflammatory, acute-phase, and hypermetabolic responses were determined. Demographics were similar in both groups. Intensive insulin treatment significantly decreased the incidence of infections and sepsis compared with controls (P < 0.05). Furthermore, intensive insulin therapy improved organ function as indicated by improved serum markers, DENVER2 scores, and ultrasound (P < 0.05). Intensive insulin therapy alleviated post-burn insulin resistance and the vast catabolic response of the body (P < 0.05). Intensive insulin treatment dampened inflammatory and acute-phase responses by deceasing IL-6 and acute-phase proteins compared with controls (P < 0.05). Mortality was 4% in the intensive insulin therapy group and 11% in the control group (P = 0.14). CONCLUSIONS: In this prospective randomized clinical trial, we showed that intensive insulin therapy improves post-burn morbidity. Clinical trial registered with www.clinicaltrials.gov (NCT00673309).

In vitro analog of human bone marrow from 3D scaffolds with biomimetic inverted colloidal crystal geometry.

In vitro replicas of bone marrow can potentially provide a continuous source of blood cells for transplantation and serve as a laboratory model to examine human immune system dysfunctions and drug toxicology. Here we report the development of an in vitro artificial bone marrow based on a 3D scaffold with inverted colloidal crystal (ICC) geometry mimicking the structural topology of actual bone marrow matrix. To facilitate adhesion of cells, scaffolds were coated with a layer of transparent nanocomposite. After seeding with hematopoietic stem cells (HSCs), ICC scaffolds were capable of supporting expansion of CD34+ HSCs with B-lymphocyte differentiation. Three-dimensional organization was shown to be critical for production of B cells and antigen-specific antibodies. Functionality of bone marrow constructs was confirmed by implantation of matrices containing human CD34+ cells onto the backs of severe combined immunodeficiency (SCID) mice with subsequent generation of human immune cells.

Upper airway mucus deposition in lung tissue of burn trauma victims.

Previous study in an ovine model of smoke inhalation and burn (S + B) injury has shown distal migration of upper airway mucus. This study examines the localization of an upper airway gland specific mucus, mucin 5B (MUC5B) in lung autopsy tissues of burn-only injury and in victims of S + B injury. We hypothesize that victims with S + B injury would exhibit increased distal migration of MUC5B than that seen in victims of burn-only injury. Autopsy lung tissue from victims of burn injury alone (n = 38) and combined S + B injury (n = 22) were immunostained for MUC5B. No normal lung tissues were included in the study. Semiquantitative analysis of the extent of MUC5B in bronchioles and parenchyma was performed on masked slides. Irrespective of injury conditions, all victims showed MUC5B in bronchioles. Mucin 5B was seen in the parenchyma except in two burn victims. No statistically significant difference was seen in the mean bronchiolar and parenchyma MUC5B scores between S + B and burn-only victims (P > 0.05). No strong statistical correlation of MUC5B scores with days postinjury or to the number of ventilatory days was evident. The percentage of pneumonia, identified histologically, was also similar between study groups. This study did not confirm our results in an ovine model of S + B injury. In contrast, virtually all pediatric burn victims, regardless of concomitant inhalation injury, showed MUC5B in their bronchioles and parenchyma. Increased mucus synthesis and/or impaired mucociliary function may contribute to the pulmonary pathophysiology associated with burn injury.

Sex differences in the long-term outcome after a severe thermal injury.

We have recently shown that during the acute phase, postburn female pediatric burn patients had significantly increased levels of anabolic hormones with an associated decreased hypermetabolism leading to a significant shorter intensive care unit stay compared with male patients. The aim of the present study was to determine possible differences between girls and boys in body composition, hypermetabolism, and hormone pattern in the long term. Sixty-two children (1-16 years old) who sustained a severe thermal injury (>or=40% total body surface area) were included into the study. Patients were further divided into girls (n = 22) and boys (n = 40). Patient demographics, nutritional support, and mortality were noted. Resting energy expenditure (REE) was measured by indirect calorimetry, body composition by dual-energy x-ray absorptiometry (Hologic Inc, Waltham, Mass) at discharge, 3, 6, 9, 12, 18, and 24 months after burn. In addition, blood was drawn at the same time points, and serum hormones were measured. There were no significant differences between girls and boys for demographics, nutritional intake, or concomitant injuries. Predicted REE was significantly decreased in girls at discharge, 6, 12, and 18 months postburn (P < 0.05). Dual-energy x-ray absorptiometry scan showed that girls had improved change in bone mineral content and percent fat compared with boys (P < 0.05). There were no differences in changes in height, body weight, lean body mass, and total fat between groups. Girls had significantly higher levels of insulinlike growth factor 1, insulinlike growth factor binding protein 3, free thyroxine index, T4, and insulin when compared with boys (P < 0.05). No differences were found for T3 uptake, osteocalcin, cortisol, growth hormone, and parathyroid hormone (PTH) between groups. Data indicate that girls have a reduced REE associated with changes in bone content and endogenous anabolic hormones.

Endogenous anabolic hormones and hypermetabolism: effect of trauma and gender differences.

OBJECTIVE: Protein degradation, negative nitrogen balance and compromised structure of essential organs have been associated with resistance and decreased production of anabolic hormones. In turn, increased levels of anabolic hormones are associated with improved survival. The aims of the present study were to determine the pattern of anabolic hormones, resting energy expenditure and cytokines in severely thermally injured pediatric patients and to compare these parameters in female and male patients. METHODS: Sixty-five children (1 to 16 years of age) sustaining a severe thermal injury (> or =40% TBSA) were included into the study. Patients were further divided into females (n = 22) and males (n = 43). Patient demographics, nutritional support, incidence of sepsis, inhalation injury, and mortality were noted. Resting energy expenditure was measured during hospital course by indirect calorimetry. Blood was drawn 0, 10, 20, and 40 days postburn and serum insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-1 and -3 (IGFBP-1, and -3), growth hormone, insulin, and cytokines were measured. RESULTS: There were no significant differences between females and males for demographics, nutritional intake, or concomitant injuries. In both groups, endogenous anabolic agents were drastically decreased by 3- to 5-fold up to 40 days posttrauma. Females had significantly higher levels of IGF-I, IGFBP-3, growth hormone, and insulin when compared with males, P < 0.05. Increased levels of anabolic hormones were associated with decreased stay on the ICU (females 36 +/- 22 days versus males 53+/- 39 days), decreased serum IL-1beta and TNF-alpha as well as resting energy expenditure, P < 0.05. CONCLUSION: Data indicate that despite adequate nutritional support, severe thermal injury leads to decreased anabolic hormones over a prolonged period of time. Female patients had significantly increased levels of anabolic hormones, which are associated with decreased proinflammatory mediators and hypermetabolism, leading to a significant shorter ICU stay compared with male patients.