Peripheral nerve repair is associated with augmented cross-tissue inflammation following vascularized composite allotransplantation.

Introduction: Vascularized composite allotransplantation (VCA), with nerve repair/coaptation (NR) and tacrolimus (TAC) immunosuppressive therapy, is used to repair devastating traumatic injuries but is often complicated by inflammation spanning multiple tissues. We identified the parallel upregulation of transcriptional pathways involving chemokine signaling, T-cell receptor signaling, Th17, Th1, and Th2 pathways in skin and nerve tissue in complete VCA rejection compared to baseline in 7 human hand transplants and defined increasing complexity of protein-level dynamic networks involving chemokine, Th1, and Th17 pathways as a function of rejection severity in 5 of these patients. We next hypothesized that neural mechanisms may regulate the complex spatiotemporal evolution of rejection-associated inflammation post-VCA. Methods: For mechanistic and ethical reasons, protein-level inflammatory mediators in tissues from Lewis rats (8 per group) receiving either syngeneic (Lewis) or allogeneic (Brown-Norway) orthotopic hind limb transplants in combination with TAC, with and without sciatic NR, were compared to human hand transplant samples using computational methods. Results: In cross-correlation analyses of these mediators, VCA tissues from human hand transplants (which included NR) were most similar to those from rats undergoing VCA + NR. Based on dynamic hypergraph analyses, NR following either syngeneic or allogeneic transplantation in rats was associated with greater trans-compartmental localization of early inflammatory mediators vs. no-NR, and impaired downregulation of mediators including IL-17A at later times. Discussion: Thus, NR, while considered necessary for restoring graft function, may also result in dysregulated and mis-compartmentalized inflammation post-VCA and therefore necessitate mitigation strategies. Our novel computational pipeline may also yield translational, spatiotemporal insights in other contexts.

Divergence of acetate uptake in proinflammatory and inflammation-resolving macrophages: implications for imaging atherosclerosis.

BACKGROUND: Metabolic divergence of macrophages polarized into different phenotypes represents a mechanistically relevant target for non-invasive characterization of atherosclerotic plaques using positron emission tomography (PET). Carbon-11 (11C)-labeled acetate is a clinically available tracer which accumulates in atherosclerotic plaques, but its biological and clinical correlates in atherosclerosis are undefined. METHODS AND RESULTS: Histological correlates of 14C-acetate uptake were determined in brachiocephalic arteries of western diet-fed apoE-/- mice. The effect of polarizing stimuli on 14C-acetate uptake was determined by proinflammatory (interferon-γ + lipopolysaccharide) vs inflammation-resolving (interleukin-4) stimulation of murine macrophages and human carotid endarterectomy specimens over 2 days. 14C-acetate accumulated in atherosclerotic regions of arteries. CD68-positive monocytes/macrophages vs smooth muscle actin-positive smooth muscle cells were the dominant cells in regions with high vs low 14C-acetate uptake. 14C-acetate uptake progressively decreased in proinflammatory macrophages to 25.9 ± 4.5% of baseline (P < .001). A delayed increase in 14C-acetate uptake was induced in inflammation-resolving macrophages, reaching to 164.1 ± 21.4% (P < .01) of baseline. Consistently, stimulation of endarterectomy specimens with interferon-γ + lipopolysaccharide decreased 14C-acetate uptake to 66.5 ± 14.5%, while interleukin-4 increased 14C-acetate uptake to 151.5 ± 25.8% compared to non-stimulated plaques (P < .05). CONCLUSIONS: Acetate uptake by macrophages diverges upon proinflammatory and inflammation-resolving stimulation, which may be exploited for immunometabolic characterization of atherosclerosis.

The Effects of Tacrolimus on Tissue-Specific, Protein-Level Inflammatory Networks in Vascularized Composite Allotransplantation.

Systems-level insights into inflammatory events after vascularized composite allotransplantation (VCA) are critical to the success of immunomodulatory strategies of these complex procedures. To date, the effects of tacrolimus (TAC) immunosuppression on inflammatory networks in VCA, such as in acute rejection (AR), have not been investigated. We used a systems biology approach to elucidate the effects of tacrolimus on dynamic networks and principal drivers of systemic inflammation in the context of dynamic tissue-specific immune responses following VCA. Lewis (LEW) rat recipients received orthotopic hind limb VCA from fully major histocompatibility complex-mismatched Brown Norway (BN) donors or matched LEW donors. Group 1 (syngeneic controls) received LEW limbs without TAC, and Group 2 (treatment group) received BN limbs with TAC. Time-dependent changes in 27 inflammatory mediators were analyzed in skin, muscle, and peripheral blood using Principal Component Analysis (PCA), Dynamic Bayesian Network (DyBN) inference, and Dynamic Network Analysis (DyNA) to define principal characteristics, central nodes, and putative feedback structures of systemic inflammation. Analyses were repeated on skin + muscle data to construct a "Virtual VCA", and in skin + muscle + peripheral blood data to construct a "Virtual Animal." PCA, DyBN, and DyNA results from individual tissues suggested important roles for leptin, VEGF, various chemokines, the NLRP3 inflammasome (IL-1ß, IL-18), and IL-6 after TAC treatment. The chemokines MCP-1, MIP-1α; and IP-10 were associated with AR in controls. Statistical analysis suggested that 24/27 inflammatory mediators were altered significantly between control and TAC-treated rats in peripheral blood, skin, and/or muscle over time. "Virtual VCA" and "Virtual Animal" analyses implicated the skin as a key control point of dynamic inflammatory networks, whose connectivity/complexity over time exhibited a U-shaped trajectory and was mirrored in the systemic circulation. Our study defines the effects of TAC on complex spatiotemporal evolution of dynamic inflammation networks in VCA. We also demonstrate the potential utility of computational analyses to elucidate nonlinear, cross-tissue interactions. These approaches may help define precision medicine approaches to better personalize TAC immunosuppression in VCA recipients.

In-vivo efficacy of biodegradable ultrahigh ductility Mg-Li-Zn alloy tracheal stents for pediatric airway obstruction.

Pediatric laryngotracheal stenosis is a complex congenital or acquired airway injury that may manifest into a potentially life-threatening airway emergency condition. Depending on the severity of obstruction, treatment often requires a combination of endoscopic techniques, open surgical repair, intraluminal stenting, or tracheostomy. A balloon expandable biodegradable airway stent maintaining patency while safely degrading over time may address the complications and morbidity issues of existing treatments providing a less invasive and more effective management technique. Previous studies have focused on implementation of degradable polymeric scaffolds associated with potentially life-threatening pitfalls. The feasibility of an ultra-high ductility magnesium-alloy based biodegradable airway stents was demonstrated for the first time. The stents were highly corrosion resistant under in vitro flow environments, while safely degrading in vivo without affecting growth of the rabbit airway. The metallic matrix and degradation products were well tolerated by the airway tissue without exhibiting any noticeable local or systemic toxicity.

Subglottic Stenosis: Development of a Clinically Relevant Endoscopic Animal Model.

OBJECTIVE: Develop a clinically relevant and reproducible endoscopic animal model for subglottic stenosis amenable to testing of minimally invasive therapeutic modalities. STUDY DESIGN: Cohort study. SETTING: Division of Laboratory Animals Research, University of Pittsburgh. SUBJECTS AND METHODS: Subglottic stenosis was induced endoscopically via microsuspension laryngoscopy in 26 New Zealand white rabbits. A trimmed polypropylene brush connected to a novel electronic stenosis induction apparatus was used to create circumferential trauma to the subglottis. By using open source image analysis software, the cross-sectional areas of the stenotic and native airways were compared to calculate the percentage of stenosis and the Myer-Cotton classification grade. RESULTS: Of the 26 rabbits, 24 (92%) exhibited stenosis after the first attempt. The mean percentage of airway stenosis was 57% (range, 34%-85%; SD, 15%). Five rabbits (19.2%) died on the day of stenosis induction from procedural complications. Of the 21 rabbits, 2 demonstrated no stenosis 7 days after initial injury and so underwent reinduction of airway injury, upon which they developed stenosis. Overall, 14 of the 21 rabbits (67%) exhibited moderate to severe stenosis (grade 2 or 3). CONCLUSION: The stenosis induction apparatus reliably induced stenosis with a low mortality rate as compared with that of other methods in the literature. The device could be improved to generate a predetermined potentially reproducible grade of stenosis as desired by the operator. This method sets the stage for a clinically relevant and reproducible subglottic stenosis disease model that is amenable to testing of minimally invasive treatment modalities.

Treg-inducing microparticles promote donor-specific tolerance in experimental vascularized composite allotransplantation.

For individuals who sustain devastating composite tissue loss, vascularized composite allotransplantation (VCA; e.g., hand and face transplantation) has the potential to restore appearance and function of the damaged tissues. As with solid organ transplantation, however, rejection must be controlled by multidrug systemic immunosuppression with substantial side effects. As an alternative therapeutic approach inspired by natural mechanisms the body uses to control inflammation, we developed a system to enrich regulatory T cells (Tregs) in an allograft. Microparticles were engineered to sustainably release TGF-ß1, IL-2, and rapamycin, to induce Treg differentiation from naïve T cells. In a rat hindlimb VCA model, local administration of this Treg-inducing system, referred to as TRI-MP, prolonged allograft survival indefinitely without long-term systemic immunosuppression. TRI-MP treatment reduced expression of inflammatory mediators and enhanced expression of Treg-associated cytokines in allograft tissue. TRI-MP also enriched Treg and reduced inflammatory Th1 populations in allograft draining lymph nodes. This local immunotherapy imparted systemic donor-specific tolerance in otherwise immunocompetent rats, as evidenced by acceptance of secondary skin grafts from the hindlimb donor strain and rejection of skin grafts from a third-party donor strain. Ultimately, this therapeutic approach may reduce, or even eliminate, the need for systemic immunosuppression in VCA or solid organ transplantation.

Demonstration of technical feasibility and viability of whole eye transplantation in a rodent model.

INTRODUCTION: Whole eye transplantation (WET) holds promise for vision restoration in devastating/disabling visual loss (congenital or traumatic) not amenable to surgical or neuroprosthetic treatment options. The eye includes multiple tissues with distinct embryonic lineage and differential antigenicity. Anatomically and immunologically, the eye is unique due to its avascular (cornea) and highly vascular (retina) components. Our goal was to establish technical feasibility, demonstrate graft viability, and evaluate histologic changes in ocular tissues/adnexae in a novel experimental model of WET that included globe, adnexal, optic nerve (ON), and periorbital soft tissues. METHODS: Outbred Sprague-Dawley rats (n = 5) received heterotopic vascularized WET from donors. Each WET included the entire globe, adnexa, ON, and periorbital soft tissues supplied by the common carotid artery and external jugular vein. Viability and perfusion were confirmed by clinical examination, angiography and magnetic resonance imaging (MRI). Globe, adnexal, and periorbital tissues were analyzed for histopathologic changes, and the ON was examined for neuro-regeneration at study endpoint (30 days) or Banff Grade 3 rejection in the periorbital skin (whichever was earlier). RESULTS: Gross examination confirmed transplant viability and corneal transparency. Average operative duration was 64.0 ±â€¯5.8 min. Average ischemia time was 26.0 ±â€¯4.2 min. MRI revealed loss of globe volume by 36.0 ±â€¯2.8% after transplantation. Histopathology of globe and adnexal tissues showed unique and differential patterns of inflammatory cell infiltration. The ON revealed a neurodegeneration pattern. CONCLUSION: The present study is the first in the literature to establish an experimental model of WET. This model holds significant potential in investigating mechanistic pathways, monitoring strategies or developing management approaches involving ocular viability, immune rejection, and ON regeneration after WET.

A biodegradable synthetic graft for small arteries matches the performance of autologous vein in rat carotid arteries.

Autologous veins are the most widely used grafts for bypassing small arteries in coronary and peripheral arterial occlusive diseases. However, they have limited availability and cause donor-site morbidity. Here, we report a direct comparison of acellular biodegradable synthetic grafts and autologous veins as interposition grafts of rat carotid arteries, which is a good model for clinically relevant small arteries. Notably, extensive but transient infiltration of circulating monocytes at day 14 in synthetic grafts leads to a quickly-resolved inflammation and arterial-like tissue remodeling. The vein graft exhibits a similar inflammation phase except the prolonged presence of inflammatory monocytes. The walls of the remodeled synthetic graft contain many circumferentially aligned contractile non-proliferative smooth muscle cells (SMCs), collagen and elastin. In contrast, the walls of the vein grafts contain disorganized proliferating SMCs and thicken over time, suggesting the onset of stenosis. At 3 months, both grafts have a similar patency, extracellular matrix composition, and mechanical properties. Furthermore, synthetic grafts exhibit recruitment and re-orientation of newly synthesized collagen fibers upon mechanical loading. To our knowledge, this is the first demonstration of a biodegradable synthetic vascular graft with a performance similar to an autologous vein in small artery grafting.

Comparison of Resorbable Mesh (Poly L-Lactide/Glycolic Acid) and Porous Polyethylene in Orbital Floor Fractures in an Experimental Model.

BACKGROUND: Resorbable mesh and porous polyethylene are frequently used alloplastic materials for the treatment of the orbital blowout fractures. The literature lacks reports comparing their long-term effects on experimental models. OBJECTIVE: Our aim was to radiologically and histologically evaluate the effectiveness and safety of porous polyethylene and resorbable mesh in a rabbit orbital blowout fracture model. METHODS: Twelve New Zealand white rabbits (24 orbits) were randomized to 4 groups. In group 1, only orbital floor dissection was done. In group 2, following orbital floor dissection, a 10-mm defect was created without any extra procedure. In group 3, following a 10-mm defect creation, a 12-mm-round cut porous polyethylene was placed on the defect. In group 4, following a 10-mm defect creation, a 12-mm-round cut resorbable mesh was placed on the defect. Computed tomographic analysis was performed during follow-up period. Orbital floors were evaluated histologically at month 6. RESULTS: No clinical complications were observed during follow-up period. In radiological evaluation, there was no statistically significant difference between groups regarding bone formation. In histological evaluation, the connective tissue was denser, and organized and better bone formation was observed in group 3 and 4 when compared with other groups. CONCLUSION: Although no significant radiological changes were present, porous polyethylene and resorbable mesh performed better histologically. They were effective and well tolerated for reconstruction of the isolated orbital floor defects.

HISTORIQUE: Le treillis résorbable et le polyéthylène poreux sont des matériaux alloplastiques souvent utilisés pour traiter les fractures isolées du plancher de l'orbite. Les publications ne contiennent pas de rapports sur les effets à long terme de ces matériaux dans des modèles expérimentaux. OBJECTIF: Les chercheurs visaient à évaluer l'efficacité et la sécurité du polyéthylène poreux et du treillis résorbable sur le plan radiologique et histologique dans un modèle de fracture isolée du plancher de l'orbite chez un lapin. MÉTHODOLOGIE: Douze lapins blancs néo-zélandais (24 orbites) ont été répartis au hasard en quatre groupes. Le groupe 1 a seulement subi la dissection du plancher de l'orbite. Dans le groupe 2, après cette dissection, une anomalie de 10 mm a été créée sans intervention supplémentaire. Dans le groupe 3, après la création d'une anomalie de 10 mm, une coupe ronde de polyéthylène poreux de 12 mm a été placée sur l'anomalie. Dans le groupe 4, après la création d'une anomalie de 10 mm, une coupe ronde de treillis résorbable de 12 mm a été placée sur l'anomalie. Les chercheurs ont effectué une analyse tomodensitométrique pendant la période de suivi. Au sixième mois, ils ont évalué les planchers orbitaux à l'histologie. RÉSULTATS: Les chercheurs n'ont observé aucune complication clinique pendant la période de suivi. À l'évaluation radiologique, la formation osseuse ne présentait aucune différence statistiquement significative entre les groupes. À l'évaluation histologique, les tissus conjonctifs étaient plus denses et la formation osseuse était organisée et de meilleure qualité dans les groupes 3 et 4 que dans les autres groupes. CONCLUSION: Malgré l'absence de modification significative à la radiologie, le polyéthylène poreux et le treillis résorbable donnaient de meilleurs résultats sur le plan histologique. Ces matériaux étaient efficaces et bien tolérés pour la reconstruction des anomalies isolées du plancher orbital.

The effect of skin expansion on acellular dermis in an experimental model.

OBJECTIVE: Although acellular dermis is widely used in prosthesis-based breast reconstruction, there is no study evaluating its expandability and histological, ultra-structural related changes. The purpose of this study is to determine whether acellular dermis could be expanded and whether the histological changes occurring in the skin with tissue expansion is different from normal when acellular dermis is used. METHODS: Twenty-two male Wistar rats were used in this study. In group 1 (n = 6), a rectangular-shaped tissue expander with final volume of 35 ml was placed subcutaneously. In group 2 (n = 6) tissue expander was placed submuscularly. In group 3 (n = 6), a 4 × 5 cm, acellular dermis was inserted and sutured in place, tissue expander was placed underneath acellular dermis. Group 4 (n = 4) was control, no procedure was done after creation of a pocket. After day 30, tissue expanders were inflated with 2 ml every 2 days with saline solution. On day 66, midvertical, midhorizontal surface expansion measurements were recorded. Samples were examined macroscopically. Biopsies were taken from mid-point of the expanded full-thickness flap. Histological evaluation, total skin thickness, epidermal thickness, and the amount of vascularisation were measured. RESULTS: There was no expansion noted on the acellular dermis; however, acellular dermis increased vascularity and decreased the thinning of the overlying skin and thickening of the epidermis when compared to other groups. CONCLUSION: Acellular dermis use can be beneficial in implant-based breast reconstruction in terms of morbidity and its tissue support.

Humanitarian Activities of Interplast Turkiye: 6 Years of Experience in Uzbekistan for Surgical Treatment of Cleft Patients and Related Secondary Deformities.

Cleft lip and palate (CL/P) is one of the leading congenital deformities among the world. Children born with CL/P experience problems with feeding, speech, hearing, and dentition. In developed countries, CL/P patients are receiving optimal health care involving multidisciplinary team approach and staged surgical operations, whereas in developing countries, there is severe shortage of both medical and financial sources. To overcome these limitations, humanitarian surgical missions are essential. The aim of this article is to share our experience of humanitarian surgical mission in Uzbekistan consisting of 6 consecutive visits between 2009 and 2014. The series of these humanitarian activities consisting of 6 consecutive visits was organized by the cooperation of Interplast Turkiye and governmental Turkish Coordination and Cooperation Agency. After initial evaluation, triage at the initial setting and prompt anesthesia evaluation among many more of them, 529 patients mostly with cleft, craniofacial, or congenital deformities were operated. The success of this type of mission is not solely based on the expertise of the team members, but also meticulous planning, patient selection, good coordination with the local colleagues and communication. At this point, caregivers attending from a culturally close and similar language-spoken countries will certainly have more advantages in achieving a mission. Volunteer surgical missions for congenital deformities can be an important relief for this burden in developing countries. Nevertheless, training the native surgeons and supporting the plastic surgery foundations in these countries are as important as providing the necessary health care by such humanitarian missions.