Wound healing with "spray-on" autologous skin grafting (ReCell) compared with standard care in patients with large diabetes-related foot wounds: an open-label randomised controlled trial.

There is an urgent need for interventions that improve healing time, prevent amputations and recurrent ulceration in patients with diabetes-related foot wounds. In this randomised, open-label trial, participants were randomised to receive an application of non-cultured autologous skin cells ("spray-on" skin; ReCell) or standard care interventions for large (>6 cm2 ), adequately vascularised wounds. The primary outcome was complete healing at 6 months, determined by assessors blinded to the intervention. Forty-nine eligible foot wounds in 45 participants were randomised. An evaluable primary outcome was available for all wounds. The median (interquartile range) wound area at baseline was 11.4 (8.8-17.6) cm2 . A total of 32 (65.3%) index wounds were completely healed at 6 months, including 16 of 24 (66.7%) in the spray-on skin group and 16 of 25 (64.0%) in the standard care group (unadjusted OR [95% CI]: 1.13 (0.35-3.65), P = .845). Lower body mass index (P = .002) and non-plantar wounds (P = .009) were the only patient- or wound-related factors associated with complete healing at 6 months. Spray-on skin resulted in high rates of complete healing at 6 months in patients with large diabetes-related foot wounds, but was not significantly better than standard care (Australian New Zealand Clinical Trials Registry: ACTRN12618000511235).

Experimental hookworm infection and gluten microchallenge promote tolerance in celiac disease.

BACKGROUND: Celiac disease (CeD) is a common gluten-sensitive autoimmune enteropathy. A gluten-free diet is an effective treatment, but compliance is demanding; hence, new treatment strategies for CeD are required. OBJECTIVE: Parasitic helminths hold promise for treating inflammatory disorders, so we examined the influence of experimental hookworm infection on the predicted outcomes of escalating gluten challenges in CeD subjects. METHODS: A 52-week study was conducted involving 12 adults with diet-managed CeD. Subjects were inoculated with 20 Necator americanus larvae, and escalating gluten challenges consumed as pasta were subsequently administered: (1) 10 to 50 mg for 12 weeks (microchallenge); (2) 25 mg daily + 1 g twice weekly for 12 weeks (GC-1g); and (3) 3 g daily (60-75 straws of spaghetti) for 2 weeks (GC-3g). Symptomatic, serologic, and histological outcomes evaluated gluten toxicity. Regulatory and inflammatory T cell populations in blood and mucosa were examined. RESULTS: Two gluten-intolerant subjects were withdrawn after microchallenge. Ten completed GC-1g, 8 of whom enrolled in and completed GC-3g. PRIMARY OUTCOMES: median villous height-to-crypt depth ratios (2.60-2.63; P = .98) did not decrease as predicted after GC-1g, and the mean IgA-tissue transglutaminase titers declined, contrary to the predicted rise after GC-3g. SECONDARY OUTCOMES: quality of life scores improved (46.3-40.6; P = .05); celiac symptom indices (24.3-24.3; P = .53), intra-epithelial lymphocyte percentages (32.5-35.0; P = .47), and Marsh scores were unchanged by gluten challenge. Intestinal T cells expressing IFNγ were reduced following hookworm infection (23.9%-11.5%; P = .04), with corresponding increases in CD4(+) Foxp3(+) regulatory T cells (0.19%-1.12%; P = .001). CONCLUSIONS: Necator americanus and gluten microchallenge promoted tolerance and stabilized or improved all tested indices of gluten toxicity in CeD subjects.

Study of the degradation in an ultisol of alginate-chitosan complex and its stability and applicability as a soil conditioner.

The present work describes the process of degradation of a polyelectrolytic complex (PEC) based on sodium alginate (ALG) and chitosan (CHI), buried for different time intervals, in a clayey soil (ultisol) collected from the municipality of Campos dos Goytacazes, in the northern region of the state of Rio de Janeiro, Brazil. The influence of PEC on soil moisture was also investigated. The results showed that soil moisture increased with the presence of PEC after 7 days of testing, and remained high until the end of the study. FTIR and Raman spectra showed that the breaking of the glycosidic bond (C-O-C) was responsible for the PEC degradation. Thermogravimetry results revealed that alginate was possibly degraded faster than chitosan. Microscopic analysis of the PEC revealed a fragile and fragmented surface of the samples that were buried, in comparison with those not buried. The microbiological assays of the soil confirmed the biodegradation of the polysaccharides. Chemical analysis of soil indicated that PEC did not significantly influence soil fertility. Therefore, we conclude that the PEC (ALG: CHI), formed only by electrostatic interaction, buried in clayey soil, even being biodegraded, can be a promising soil conditioner for agricultural applications.

Hookworm excretory/secretory products induce interleukin-4 (IL-4)+ IL-10+ CD4+ T cell responses and suppress pathology in a mouse model of colitis.

Evidence from human studies and mouse models shows that infection with parasitic helminths has a suppressive effect on the pathogenesis of some inflammatory diseases. Recently, we and others have shown that some of the suppressive effects of hookworms reside in their excretory/secretory (ES) products. Here, we demonstrate that ES products of the hookworm Ancylostoma caninum (AcES) suppress intestinal pathology in a model of chemically induced colitis. This suppression was associated with potent induction of a type 2 cytokine response characterized by coexpression of interleukin-4 (IL-4) and IL-10 by CD4(+) T cells, downregulation of proinflammatory cytokine expression in the draining lymph nodes and the colon, and recruitment of alternatively activated (M2) macrophages and eosinophils to the site of ES administration. Protease digestion and heat denaturation of AcES resulted in impaired induction of CD4(+) IL-4(+) IL-10(+) cell responses and diminished ability to suppress colitis, indicating that protein component(s) are responsible for some of the immunosuppressive effects of AcES. Identification of the specific parasite-derived molecules responsible for reducing pathology during chemically induced colitis could lead to the development of novel therapeutics for the treatment of human inflammatory bowel disease.

Effect of pH variation and crosslinker absence on the gelling mechanism of high acyl gellan: Morphological, thermal and mechanical approaches.

This article describes the preparation of superabsorbent hydrogels made of high acyl gellan without the addition of a crosslinker . Variations of the pH of the medium and different gellan solution concentrations were evaluated. The samples produced were investigated regarding their morphology by scanning electron microscopy, thermal resistance by thermogravimetry, and compressive strength by mechanical testing. Rheological tests were also performed and the swelling decree of the hydrogels was calculated. The results showed that the morphology of the samples could be modified by varying the preparation conditions, ranging from a foliaceous structure to a brittle structure, containing voids with different sizes. The hydrogels prepared with acidic pH presented less compressive strength than those prepared with basic pH. Although all the samples had swelling degree greater than 1000 %, the highest values were obtained with the samples prepared at pH 4, regardless of the concentration of the gellan solution.

Spray on skin for diabetic foot ulcers: an open label randomised controlled trial.

BACKGROUND: One Australian loses a limb every 3 h as a result of infected diabetic foot ulcers (DFU). This common condition accounts for substantial morbidity and mortality for affected individuals and heavy economic costs for the health sector and the community. There is an urgent need to test interventions that improve wound healing time, prevent amputations and recurrent ulceration in patients presenting with DFU whilst improving quality of life and reducing health care costs. METHODS: One hundred and fifty eligible participants will be randomised to receive an autologous skin cell suspension, also termed 'spray-on' skin (ReCell®) or standard care interventions for their DFU. The primary outcome is complete wound healing at 6 months, but participants will be followed up for a total of 12 months to enable secondary outcomes including total overall costs, ulcer free days at 12 months and quality of life to be assessed. DISCUSSION: Outpatient costs for dressings, home nursing visits and outpatient appointments are key cost drivers for DFU. If spray-on skin is effective, large cost savings to WA Health will be realised immediately through a shortened time to healing, and through a higher proportion of patients achieving complete healing. Shortened healing times may enable participants to return to work earlier. Any economic benefits are likely to be amplified across Australia and other similar demographic settings where aging populations with increased diabetes rates are considered major future challenges. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry ACTRN12618000511235. Registered on 9 April 2018.

Interaction of polyelectrolyte complex between sodium alginate and chitosan dimers with a single glyphosate molecule: A DFT and NBO study.

The formation of a polyelectrolyte complex through dimers of alginate and chitosan in the presence of sodium cations (SA/CS), and its interaction with the glyphosate herbicide, has been investigated at the DFT level (B3LYP/6‒311+G(d,p)). The lowest energy structure for SA/CS presents one Na+ cation coordinated to both dimers and formation of two H-bonds involving COO- and NH3+. The coordination energy of Na+ contributes with about 40% of the total complex stabilization energy. LMOEDA method indicates important contribution of covalent nature to stabilization of SA/CS. This result is corroborated by NBO analysis which shows high contribution of lp(O)→σ*(NH) overlapping, with average energy of 30 kcal mol-1 for the formed H-bonds. Two water molecules neighboring the complex increases its stability and promotes an octahedral coordination arrangement around Na+. The glyphosate interacts with SA/CS coordinating to Na+ and bonding to the chitosan dimer by H-bond, in agreement to performed fluorescence microscopy measurements.

Changes in protein expression after treatment with Ancylostoma caninum excretory/secretory products in a mouse model of colitis.

Different reports have highlighted the potential use of helminths and their secretions in the treatment of inflammatory bowel disease (IBD) conditions; however, no reports have investigated their effects at a proteome level. Herein, we characterise the protein expression changes that occur in lamina propria (LP) and the intestinal epithelial cells (IEC) of mice with dextran sulfate sodium (DSS)-induced colitis treated with Ancylostoma caninum excretory/secretory (ES) products using a quantitative proteomic approach. We have shown how parasite products can significantly alter the expression of proteins involved in immune responses, cell death and with an antioxidant activity. Interestingly, significant changes in the expression levels of different mucins were observed in this study. MUC13, a mucin implicated in gastrointestinal homeostasis, was upregulated in the LP of mice with DSS-induced colitis treated with ES, while MUC2, a major component of mucus, was upregulated in the IEC. In addition, A. caninum proteins have an important effect on proteins with antioxidant functions and proteins involved in intestinal homeostasis and tissue integrity and regeneration. Understanding how parasites can ameliorate IBD pathogenesis can help us design novel treatments for autoimmune diseases.

Suppression of inflammation and tissue damage by a hookworm recombinant protein in experimental colitis.

Gastrointestinal parasites, hookworms in particular, have evolved to cause minimal harm to their hosts when present in small numbers, allowing them to establish chronic infections for decades. They do so by creating an immunoregulatory environment that promotes their own survival, but paradoxically also benefits the host by protecting against the onset of many inflammatory diseases. To harness the therapeutic value of hookworms without using live parasites, we have examined the protective properties of the recombinant protein anti-inflammatory protein (AIP)-1, secreted in abundance by hookworms within the intestinal mucosa, in experimental colitis. Colitic inflammation assessed by weight loss, colon atrophy, oedema, ulceration and necrosis, as well as abdominal adhesion was significantly suppressed in mice treated with a single intraperitoneal dose of AIP-1 at 1 mg kg-1. Local infiltration of inflammatory cells was also significantly reduced, with minimal goblet cell loss and preserved mucosal architecture. Treatment with AIP-1 promoted the production of colon interleukin (IL)-10, transforming growth factor (TGF)-ß and thymic stromal lymphopoietin (TSLP), resulting in the suppression of tumour necrosis factor (TNF)-α, IL-13 and IL-17 A cytokines and granulocyte macrophage colony-stimulating factor (GM-CSF), CX motif chemokine (CXCL)-11 and cyclooxygenase synthase (COX)-2 mRNA transcripts. AIP-1 promoted the accumulation of regulatory T cells in the colon likely allowing rapid healing of the colon mucosa. Hookworm recombinant AIP-1 is a novel therapeutic candidate for the treatment of inflammatory bowel diseases that can be explored for the prevention of acute inflammatory relapses, an important cause of colorectal cancer.

New polyelectrolyte complex from pectin/chitosan and montmorillonite clay.

A new nanocomposite hydrogel was prepared by forming a crosslinked hybrid polymer network based on chitosan and pectin in the presence of montmorillonite clay. The influence of clay concentration (0.5 and 2% wt) as well as polymer ratios (1:1, 1:2 and 2:1) was investigated carefully. The samples were characterized by different techniques: transmission and scanning electron microscopy, X-ray diffraction, thermogravimetric analysis, infrared spectroscopy, swelling degree and compression test. Most samples presented swelling degree above 1000%, which permits characterizing them as superabsorbent material. Images obtained by transmission electron microscopy showed the presence of clay nanoparticles into hydrogel. The hydrogels' morphological properties were evaluated by scanning electron microscope in high and low-vacuum. The micrographs showed that the samples presented porous. The incorporation of clay produced hydrogels with differentiated morphology. Thermogravimetric analysis results revealed that the incorporation of clay in the samples provided greater thermal stability to the hydrogels. The compression resistance also increased with addition of clay.

Evaluation of different methods to prepare superabsorbent hydrogels based on deacetylated gellan.

This study stands out for analyzing distinct ways of preparing hydrogels from deacetylated gellan gum that have high swelling capacity and good thermal resistance. We carried out a thorough investigation, applying various combinations of different experimental parameters. Two preparation methods were evaluated, in which the pH was adjusted before or after thermal treatment of the gellan solution, with subsequent addition of the crosslinking agent, to assess the influence of preparation method on the conformation of the gellan chains regarding formation of double helices. The pH range tested varied from acid (2, 3 or 4) to basic (8, 9 or 10). Gellan solution was prepared in different concentrations. Both pure gellan and hydrogel samples were characterized by Fourier-transform infrared spectroscopy and thermogravimetry. Pure gellan was also characterized by atomic absorption spectroscopy. The swelling degree of the hydrogels was analyzed. The results showed that all the hydrogels had high swelling capacity (>400%), so they can be considered superabsorbent materials. Hydrogels prepared with acid pH in general had lower thermal resistance than samples prepared in alkaline pH, regardless of the preparation method. Samples prepared with alkaline pH tended to have initial decomposition temperature similar to that of pure gellan.

Hookworm recombinant protein promotes regulatory T cell responses that suppress experimental asthma.

In the developed world, declining prevalence of some parasitic infections correlates with increased incidence of allergic and autoimmune disorders. Moreover, experimental human infection with some parasitic worms confers protection against inflammatory diseases in phase 2 clinical trials. Parasitic worms manipulate the immune system by secreting immunoregulatory molecules that offer promise as a novel therapeutic modality for inflammatory diseases. We identify a protein secreted by hookworms, anti-inflammatory protein-2 (AIP-2), that suppressed airway inflammation in a mouse model of asthma, reduced expression of costimulatory markers on human dendritic cells (DCs), and suppressed proliferation ex vivo of T cells from human subjects with house dust mite allergy. In mice, AIP-2 was primarily captured by mesenteric CD103+ DCs and suppression of airway inflammation was dependent on both DCs and Foxp3+ regulatory T cells (Tregs) that originated in the mesenteric lymph nodes (MLNs) and accumulated in distant mucosal sites. Transplantation of MLNs from AIP-2-treated mice into naïve hosts revealed a lymphoid tissue conditioning that promoted Treg induction and long-term maintenance. Our findings indicate that recombinant AIP-2 could serve as a novel curative therapeutic for allergic asthma and potentially other inflammatory diseases.

Severe trauma in young permanent tooth: a case report / Trauma severo em dente permanente jovem: relato de caso

ABSTRACT Dentoalveolar trauma is a severe traumatic injury involving alveolar bone and dental structures. Like any trauma, it can leave irreparable sequelae and even cause tooth loss. The aim of this study is to report on the case of a dentoalveolar trauma in a 10-year-old male patient and the treatment performed with 12-month follow-up. The child had lateral luxation and displacement of teeth 11 and 21 in labial direction after a fall from his own height. The patient was examined at a hospital, received medication and was referred to a dental clinic. At the dental office, the teeth were repositioned, as well as received endodontic treatment and dental reanatomization. Two months after the trauma, root resorption was observed externally, which remained stable for 12 months. It was concluded that post-trauma treatment should be immediate and that follow-up sessions should be done carefully to minimize sequelae and to receive better prognosis.

RESUMO Traumatismo dentoalveolar é uma lesão traumática grave envolvendo osso alveolar e estruturas dentárias. Como qualquer trauma, pode deixar sequelas irreparáveis e até causar perda dentária. O objetivo deste estudo é relatar o caso de um trauma dentoalveolar em um paciente de 10 anos e o tratamento realizado com 12 meses de acompanhamento. A criança apresentou luxação lateral e deslocamento dos dentes 11 e 21 na direção labial após uma queda da própria altura. O paciente foi examinado em um hospital, recebeu medicação e foi encaminhado a uma clínica odontológica. No consultório odontológico, os dentes foram reposicionados, posteriormente, tratados endodônticamente e realizado a reanatomização dentária. Dois meses após o trauma, a reabsorção radicular foi observada externamente, permanecendo estável por 12 meses. Concluiu-se que o tratamento pós-trauma deve ser imediato e que as sessões de acompanhamento devem ser realizadas com cuidado, a fim de minimizar sequelas e obter melhor prognóstico.

Use of intrarradicular pins in primary teeth: case report of one year of accompanying / Uso de pinos intrarradiculares em dentes decíduos: relato de caso de um ano de acompanhamento

ABSTRACT Primary teeth have an important role in the growth and development of the child, besides being important for the good performance of masticatory function, phonation, occlusion and aesthetics, are the best space maintainers and eruption guide for the permanent successor. For anterior region, there are different alternatives of rehabilitation treatment, however, it is up to the professional to choose the best option. Objective To present a therapeutic management of upper incisors affected by carious lesion with intrarradicular pin in early childhood. Case report: male patient, with 2 years and 7 months of age and significant loss of dental structure at an early stage. In the impossibility of performing direct restoration, we opted for the rehabilitation with the use of glass fiber pins for intraradicular reinforcement and composite resin restorations in order to restore shape, function and aesthetics. Conclusion The use of fiberglass intra-radicular pins associated with direct restoration of composite resin is a recommended technique for the reconstruction of anterior teeth with great destruction.

RESUMO Os dentes decíduos possuem um papel importante no crescimento e desenvolvimento da criança, além de serem importantes para o bom desempenho da função mastigatória, fonação, oclusão e estética, são os melhores mantenedores de espaço e guia de erupção para o sucessor permanente. Para região anterior, existem diferentes alternativas de tratamento reabilitador, porém, cabe ao profissional escolher a melhor opção Objetivo Apresentar uma conduta terapêutica dos incisivos superiores acometidos por lesão de cárie com pino intrarradicular na primeira infância. Relato do caso: paciente do sexo masculino, com 2 anos e 7 meses de idade e perda significante de estrutura dentária de forma precoce. Na impossibilidade de ser realizado restauração direta, optou-se pela reabilitação com o uso de pinos de fibra de vidro para reforço intrarradicular e restaurações em resina composta em para restabelecimento da forma, função e estética. Conclusão A utilização de pinos intraradiculares de fibra de vidro associada a restauração direta de resina composta é uma técnica recomendada para a reconstrução de dentes decíduos anteriores com grande destruição.

Do Different Proportions of Antibiotics in the CTZ Paste Interfere with the Antimicrobial Action? In Vitro Study

Abstract Objective: To evaluate the antimicrobial action of the CTZ paste in three different proportions by diffusion in agar with the microorganisms: Enterococcus faecalis,Escherichia coli, and Candida albicans. Material and Methods: Three different proportions of antibiotics were tested: GROUP A - CTZ paste in the ratio of 33.33% chloramphenicol + 33.33% tetracycline + 33.33% zinc oxide, mixed with 2 drops of eugenol (1:1:1 ratio); GROUP B - CTZ paste in the proportion of 25% chloramphenicol + 25% tetracycline + 50% zinc oxide, mixed with 2 drops of eugenol (1: 1: 2 ratio); GROUP C - CTZ paste with 13% chloramphenicol + 13% tetracycline + 74% Zinc Oxide, mixed with 2 drops of eugenol (1:1:6 ratio); PC GROUP - Positive Control (0.12% Chlorhexidine); and NC GROUP - Negative Control (0.9% Saline solution). Data were analyzed through descriptive statistics (means and standard deviation). The one-way ANOVA and Tukey's test were used, with a significance level of 5% Results: No statistical differences for Enterococcus faecalis between groups A, B, and C (p = 0.1986) were found. There were statistical differences for Escherichia coli between groups B and C (p = 0.029), and for Candida albicans between groups A and C (p = 0.006). Groups A, B, and C had significant differences with both Positive and Negative Controls for all the microorganisms Conclusion: The three different ratios of CTZ paste showed antimicrobial efficacy against Enterococcus faecalis,Escherichia coli, and Candida albicans microorganisms.

The hookworm pharmacopoeia for inflammatory diseases.

In the developed world, declining prevalence of parasitic infections correlates with increased incidence of allergic and autoimmune disorders. Current treatments for these chronic inflammatory conditions have little to no effect on their prevalence and are referred to as "controllers" rather than cures. There has been limited success in therapeutically targeting allergic and autoimmune pathways, leaving an unmet need for development of effective anti-inflammatories. We discuss the benefit of hookworm infections and the parasite's ability to condition the immune system to prevent allergic asthma and inflammatory bowel diseases. We then examine the immunomodulatory properties of selected hookworm-derived proteins in these two models of inflammation. While hookworm protein therapy has yet to be fully exploited, the identification of these proteins and the mechanisms by which they skew the immune system will provide new avenues for controlling and optimally reversing key pathological processes important in allergic and inflammatory bowel diseases.