Effects of circular myotomy on the healing of esophageal suture anastomosis: an experimental study

For esophageal reconstruction in newborns with esophageal atresia, esophageal reunion with an end-to-end anastomosis is the ideal procedure, although it may result in leaks and strictures due to tension on the suture line, mainly in cases with a wide gap between the ends. Circular myotomy (Livaditis' procedure) is the best method to elongate the proximal esophageal pouch and reduce anastomotic tension. This experimental investigation in dogs was undertaken to attempt to verify that circular myotomy decreases the anastomotic leak rate in newborns with wide gap esophageal atresia, and to analyze whether the technique promotes morphologic changes in the anastomotic scar. A pilot study demonstrated that it is necessary to resect more than 8 cm (40% of the total esophageal length) in order to obtain high leak rates. In the experimental project, such resection was performed in dogs divided into two groups (control group, anastomosis only, and experimental group, anastomosis plus circular myotomy in the proximal esophageal segment). The animals were killed in the 14th postoperative day, submitted to autopsy, and were evaluated as to the presence of leaks and strictures, as well as to the features (macroscopic and microscopic aspects) of the anastomosis. Leak rates were the same in both groups. Morphometric analysis revealed that in animals in the experimental group, the anastomotic scar was thinner than the control animals, and the isolated muscular manchette distal to the site of myotomy was replaced by fibrous tissue. Correspondingly, a decreased number of newly formed small vessels were noted in the experimental animals, compared to control animals. We concluded that circular myotomy does not decrease the incidence of anastomotic leaks, and it also promotes deleterious changes in anastomotic healing.

Stromal vascular fraction cell sheets angiogenic potential for tissue engineering and regenerative medicine applications

One of the biggest concerns in the Tissue Engineering field is the correct vascularization of engineered constructs. Strategies involving the use of endothelial cells are promising but adequate cell sourcing and neo-vessels stability are enduring challenges. In this work, we propose the hypoxic pre-conditioning of the stromal vascular fraction (SVF) of human adipose tissue to obtain highly angiogenic cell sheets (CS). For that, SVF was isolated after enzymatic dissociation of adipose tissue and cultured until CS formation in normoxic (pO2=21%) and hypoxic (pO2=5%) conditions for 5 and 8 days, in basal medium. Immunocytochemistry against CD31 and CD146 revealed the presence of highly branched capillary-like structures, which were far more complex for hypoxia. ELISA quantification showed increased VEGF and TIMP-1 secretion in hypoxia for 8 days of culture. In a Matrigel assay, the formation of capillary-like structures by endothelial cells was more prominent when cultured in conditioned medium recovered from the cultures in hypoxia. The same conditioned medium increased the migration of adipose stromal cells in a scratch assay, when compared with the medium from normoxia. Histological analysis after implantation of 8 days normoxic- and hypoxic-conditioned SVF CS in a hindlimb ischemia murine model showed improved formation of neo-blood vessels. Furthermore, Laser Doppler results demonstrated that the blood perfusion of the injured limb after 30 days was enhanced for the hypoxic CS group. Overall, these results suggest that SVF CS created under hypoxia can be used as functional vascularization units for tissue engineering and regenerative medicine.

Release of insulin from PLGA-alginate dressing stimulates regenerative healing of burn wounds in rats

Burn wound healing involves a complex set of overlapping processes in an environment conducive to ischemia, inflammation, and infection costing $7.5 billion/year in the US alone, in addition to the morbidity and mortality that occur when the burns are extensive. We previously showed that insulin, when topically applied to skin excision wounds, accelerates re-epithelialization, and stimulates angiogenesis. More recently, we developed an alginate sponge dressing (ASD) containing insulin encapsulated in PLGA microparticles that provides a sustained release of bioactive insulin for >20days in a moist and protective environment. We hypothesized that insulin-containing ASD accelerates burn healing and stimulates a more regenerative, less scarring, healing. Using a heat-induced burn injury in rats, we show that burns treated with dressings containing 0.04mg insulin/cm2, every three days for 9 days, have faster closure, faster rate of disintegration of dead tissue, and decreased oxidative stress.In addition, in insulin-treated wounds the pattern of neutrophil inflammatory response suggests faster clearing of the burn dead tissue. We also observe faster resolution of the pro-inflammatory macrophages. We also found that insulin stimulates collagen deposition and maturation with the fibers organized more like a basket weave (normal skin) than aligned and crosslinked (scar tissue). In summary , application of ASD-containing insulin-loaded PLGA particles on burns every three days stimulates faster and more regenerative healing. These results suggest insulin as a potential therapeutic agent in burn healing and, because of its long history of safe use in humans, insulin could become one of the treatments of choice when repair and regeneration are critical for proper tissue function.

Advances in thermoplastic pultruded composites

Pultrusion is a versatile continuous high speed production technology allowing the production of fibre reinforced complex profiles. Thermosetting resins are normally used as matrices in the production of structural constant cross section profiles. Although only recently thermoplastic matrices have been used in long and continuous fibre reinforced composites replacing with success thermosetting matrices, the number of their applications is increasing due to their better ecological and mechanical performance. Composites with thermoplastic matrices offers increased fracture toughness, higher impact tolerance, short processing cycle time and excellent environmental stability. They are recyclable, post-formable and can be joined by welding. The use of long/continuous fibre reinforced thermoplastic matrix composites involves, however, great technological and scientific challenges since thermoplastics present much higher viscosity than thermosettings, which makes much difficult and complex the impregnation of reinforcements and consolidation tasks. In this work continuous fibres reinforced thermoplastic matrix towpregs were produced using equipment developed by the Institute for Polymers and Composites (IPC). The processing of the towpregs was made by pultrusion, in a developed prototype equipment existing in the Engineering School of the Polytechnic Institute of Porto (ISEP). Different thermoplastic matrices and fibres raw-materials were used in this study to manufacture pultruded composites for commercial applications (glass and carbon fibre/ polypropylene) and for advanced markets (carbon fibre/Primospire®). To improve the temperature distribution profile in heating die, different modifications were performed. In order to optimize both processes, towpregs production and pultruded composites profiles were analysed to determine the influence of the most relevant processing arameters in the final properties. The final pultruded composite profiles were submitted to mechanical tests to obtain the relevant properties.

A new empirical system for rock slope stability analysis in exploitation stage

An empirical system was developed to obtain a quality index for rock slopes in road infrastructures, named Slope Quality Index (SQI), and it was applied to a set of real slopes.The SQI is supported in nine factors affecting slope stability that contemplate the evaluation of different parameters. Consequently, each factor is classified by the degree of importance and influence by assigned weights. These weights were established through a statistical analysis of replies to a survey that was distributed to several experienced professionals in the field. The proposed SQI varies between1 and 5, corresponding to slopes in very good and very bad condition state, respectively. Besides the advantage linked to a quantitative and qualitative evaluation of slopes, theSQI also allows identifying the most critical factors on the slope stability, which is a fundamental issue for an efficient management of the slope network in the road infrastructure, namely in the planning of conservation and maintenance operations.

Rényi entropy and the thermodynamic stability of black holes

Thermodynamic stability of black holes, described by the Rényi formula as equilibrium compatible entropy function, is investigated. It is shown that within this approach, asymptotically flat, Schwarzschild black holes can be in stable equilibrium with thermal radiation at a fixed temperature. This implies that the canonical ensemble exists just like in anti-de Sitter space, and nonextensive effects can stabilize the black holes in a very similar way as it is done by the gravitational potential of an anti-de Sitter space. Furthermore, it is also shown that a Hawking–Page-like black hole phase transition occurs at a critical temperature which depends on the q-parameter of the Rényi formula.

Spontaneous healing of Mycobacterium ulcerans lesions in the guinea pig model

Buruli Ulcer (BU) is a necrotizing skin disease caused by Mycobacterium ulcerans infection. BU is characterized by a wide range of clinical forms, including non-ulcerative cutaneous lesions that can evolve into severe ulcers if left untreated. Nevertheless, spontaneous healing has been reported to occur, although knowledge on this process is scarce both in naturally infected humans and experimental models of infection. Animal models are useful since they mimic different spectrums of human BU disease and have the potential to elucidate the pathogenic/protective pathway(s) involved in disease/healing. In this time-lapsed study, we characterized the guinea pig, an animal model of resistance to M. ulcerans, focusing on the macroscopic, microbiological and histological evolution throughout the entire experimental infectious process. Subcutaneous infection of guinea pigs with a virulent strain of M. ulcerans led to early localized swelling, which evolved into small well defined ulcers. These macroscopic observations correlated with the presence of necrosis, acute inflammatory infiltrate and an abundant bacterial load. By the end of the infectious process when ulcerative lesions healed, M. ulcerans viability decreased and the subcutaneous tissue organization returned to its normal state after a process of continuous healing characterized by tissue granulation and reepethelialization. In conclusion, we show that the experimental M. ulcerans infection of the guinea pig mimics the process of spontaneous healing described in BU patients, displaying the potential to uncover correlates of protection against BU, which can ultimately contribute to the development of new prophylactic and therapeutic strategies.

Evaluating balance, stability and gait symmetry of stroke patients using kinematic and dynamic gait lab data

Dissertação de mestrado integrado em Engenharia Biomédica

Influence of casting condition on the anisotropy of the fracture properties of Steel Fibre Reinforced Self-Compacting Concrete (SFRSCC)

Identification of the tensile constitutive behaviour of Fibre Reinforced Concrete (FRC) represents an important aspect of the design of structural elements using this material. Although an important step has been made with the introduction of guidance for the design with regular FRC in the recently published fib Model Code 2010, a better understanding of the behaviour of this material is still necessary, mainly for that with self-compacting properties. This work presents an experimental investigation employing Steel Fibre Self-Compacting Concrete (SFRSCC) to cast thin structural elements. A new test method is proposed for assessing the post-cracking behaviour and the results obtained with the proposed test method are compared with the ones resulted from the standard three-point bending tests (3PBT). Specimens extracted from a sandwich panel consisting of SFRSCC layers are also tested. The mechanical properties of SFRSCC are correlated to the fibre distribution by analysing the results obtained with the different tests. Finally, the stress-crack width constitutive law proposed by the fib Model Code 2010 is analysed in light of the experimental results.

Investigation on the homogeneity and stability of aqueous nano cellulose suspensions prepared using pluronic F-127

This paper presents the results of experimental investigation on the aqueous dispersion behaviour of micro crystalline cellulose (MCC) prepared using Pluronic F-127. For this purpose, different concentrations (0.5-3.0 wt.%) of MCC were dispersed in water with the help of ultrasonication technique using various concentrations of Pluronic F-127. The homogeneity of the suspensions and agglomerations were characterized by optical and transmission electron microscopy and the concentration of well dispersed MCC was measured using UV-Vis spectroscopy. Also, the suspensions were subjected to high speed ultracentrifugation at 3000 rpm and observed visually for sedimentation and subsequently, concentration was calculated using UV-Vis, in order to assess the long term stability of the suspensions. Based on these experiments, optimum concentration of Pluronic to disperse different MCC concentrations has been suggested.

Efeito da adição de Zr no endurecimento de ligas Al-Sc

Dissertação de mestrado integrado em Engenharia de Materiais

Improved burn wound healing using a bioactive peptide

[Excerpt] Antimicrobial peptides (AMPs) are good candidates to treat burn wounds, a major cause of morbidity, impaired life quality and resources consumption in developed countries. We took advantage of a commercially available hydrogel, Carbopol, a vehicle for topical administration that maintains a moist environment within the wound site. We hypothesized that the incorporation of LLKKK18 conjugated to dextrin would improve the healing process in rat burns. Whereas the hydrogel improves healing, LLKKK18 released from the dextrin conjugates further accelerates wound closure, and simultaneously improving the quality of healing. Indeed, the release of LLKKK18 reduces oxidative stress and inflammation (low neutrophil and macrophage infiltration and pro-inflammatory cytokines levels). Importantly, it induced a faster resolution of the inflammatory stage through early M2 macrophage recruitment. In addition, LLKKK18 stimulates angiogenesis (increased VEGF and microvessel development in vivo), potentially contributing to more effective transport of nutrients and cytokines. Moreover, collagen staining evaluated by Masson’s Trichrome was visually much more intense after treatment with LLKKK18, suggesting higher collagen deposition. (...)

Synthesis of new peptide derivatives that self-assemble into nanostructured hydrogels for biomedical applications

Tese de Doutoramento em Ciências (área de especialização em Química)

PillCam COLON 2© in Crohn's disease: A new concept of pan-enteric mucosal healing assessment

AIM To evaluate mucosal healing in patients with small bowel plus colonic Crohn's disease (CD) with a single non-invasive examination, by using PillCam COLON 2 (PCC2). METHODS Patients with non-stricturing nonpenetrating small bowel plus colonic CD in sustained corticosteroid-free remission were included. At diagnosis, patients had undergone ileocolonoscopy to identify active CD lesions, such as ulcers and erosions, and small bowel capsule endoscopy to assess the Lewis Score (LS). After = 1 year of follow-up, patients underwent entire gastrointestinal tract evaluation with PCC2. The primary endpoint was assessment of CD mucosal healing, defined as no active colonic CD lesions and LS < 135. RESULTS Twelve patients were included (7 male; mean age: 32 years), and mean follow-up was 38 mo. The majority of patients (83.3%) received immunosuppressive therapy. Three patients (25%) achieved mucosal healing in both the small bowel and the colon, while disease activity was limited to either the small bowel or the colon in 5 patients (42%). It was possible to observe the entire gastrointestinal tract in 10 of the 12 patients (83%) who underwent PCC2. CONCLUSION Only three patients in sustained corticosteroid-free clinical remission achieved mucosal healing in both the small bowel and the colon, highlighting the limitations of clinical assessment when stratifying disease activity, and the need for pan-enteric endoscopy to guide therapeutic modification.

Gellan gum-hyaluronate spongy-like hydrogels promote angiogenesis in hindlimb ischemia

"Tissue engineering: part A", vol. 21, suppl. 1 (2015)