Wound fluid sampling methods and analysis of cytokine mRNA expression in ulcers from patients with diabetes mellitus.

The development of diabetic foot ulcers is a common and severe complication of diabetes that can significantly affect quality of life. The physiological healing cascade does not progress tissue repair in diabetic foot ulcerations in a timely manner. Serum markers from foot ulcers have been used to characterize the healing process of the diabetic foot using various collection techniques. This study aimed to compare the use of cervical brushes and the Levine technique to collect wound fluid from foot ulcers of people with diabetes in order to determine the presence of cytokines. The collected material was used for gene expression analysis of macrophage/monocyte-associated cytokines IL1-ß, IL-6, TNF-α, regulatory cytokine IL-10 and growth factor TGFß, via quantitative polymerase chain reaction (qPCR). Both collection methods produced sufficient amounts of RNA, but significantly more RNA was collected using a cervical brush (brush 224.82 ng/µL vs. Levine 80.90 ng/µL p = 0.0001). Significantly higher levels of expression of the following cytokine genes were detected in samples collected using a cervical brush: IL1-ß (p = 0.0001), IL-6 (p = 0.0106), IL-10 (p = 0.0277) and TGFß (p = 0.0002). Understanding why some wounds are difficult to heal is important for developing more effective treatments, and biomarkers may be useful for predicting the healing trajectory. These results demonstrate that it is possible to collect material from the wound bed for RT-qPCR analysis, and the cervical brush proved to be a simple and rapid method for monitoring cytokine gene expression.

Impact of oral nutritional supplement composition on healing of different chronic wounds: A systematic review.

Chronic wounds are characterized by prolonged non-healing, significantly affecting patients' quality of life. Oral formulas may enhance the wound healing process and contribute to cost reduction in care. This review aimed to evaluate the effects of oral nutritional supplementation on chronic wound healing and provide insights into formula characteristics. A comprehensive search across Cinahl, Embase, PubMed, and Web of Science databases yielded nine studies from the past decade involving 741 patients ages 52 to 81.7 across various care settings: hospitals, long-term care facilities, and home care. Primary wound types included pressure injuries (58%), diabetic foot ulcers (40%), and venous ulcers (2%). The intervention duration ranged from 2 to 16 wk, with sample sizes varying from 24 to 270 patients. Notably, four studies reported a reduction in wound area and an increased healing rate with a hypercaloric, hyperproteic formula enriched with zinc and vitamins A, C, and E. However, two studies found no significant differences compared with control groups. Two other studies investigated a combination of arginine, glutamine, and ß-hydroxy-ß-methylbutyrate; however, they did not yield significant results, and one study favored a hyperproteic formula instead of a hyperproteic formula with arginine. This review provides evidence supporting the potential of oral nutritional supplementation to enhance the healing process of chronic wounds. Based on our findings, a desirable formula should be characterized by a high calorie and protein content and the inclusion of antioxidant micronutrients, including, but not limited to, vitamins A, E, C, and zinc.

Nanosilver-Functionalized Hybrid Hydrogels of Carboxymethyl Cellulose/Poly(Vinyl Alcohol) with Antibacterial Activity for Prevention and Therapy of Infections of Diabetic Chronic Wounds.

Diabetic foot ulcers (DFUs) are considered one of the most severe chronic complications of diabetes and can lead to amputation in severe cases. In addition, bacterial infections in diabetic chronic wounds aggravate this scenario by threatening human health. Wound dressings made of polymer matrices with embedded metal nanoparticles can inhibit microorganism growth and promote wound healing, although the current clinical treatments for diabetic chronic wounds remain unsatisfactory. In this view, this research reports the synthesis and characterization of innovative hybrid hydrogels made of carboxymethyl cellulose (CMC) and poly(vinyl alcohol) (PVA) chemically crosslinked by citric acid (CA) functionalized with silver nanoparticles (AgNPs) generated in situ using an eco-friendly aqueous process. The results assessed through comprehensive in vitro and in vivo assays demonstrated that these hybrid polymer hydrogels functionalized with AgNPs possess physicochemical properties, cytocompatibility, hemocompatibility, bioadhesion, antibacterial activity, and biocompatibility suitable for wound dressings to support chronic wound healing process as well as preventing and treating bacterial infections. Hence, it can be envisioned that, with further research and development, these polymer-based hybrid nanoplatforms hold great potential as an important tool for creating a new generation of smart dressings for treating chronic diabetic wounds and opportunistic bacterial infections.

Use of Cultivated Epidermal Allograft in Chronic Wounds: A Novel Therapeutic Alternative.

Introduction: Chronic wounds represent a frequent cause of consultation for plastic and reconstructive surgeons. The use of epidermal culture stands out because they provide complete epithelialization, adequate aesthetic-functional results, and no morbidity for the patient. Epifast® is a pre-manufactured cultured epidermal allograft derived from the amplification in vitro of human keratinocytes. Materials and Methods: A prospective longitudinal multicenter study was carried out in four chronic wound reference centers, which were in charge of plastic and reconstructive surgery services. For a standardized wound bed preparation, the protocol synthesized by the acronym "TIME" was used. At the end of the "TIME" protocol, the pre-fabricated allograft was applied and removed 7 days after its application. Results: A total of 133 patients with diagnosis of chronic wound were included in the study. The median age was 69.3 ± 13.6 years. The most common comorbidity found was diabetes mellitus type 2 in 71.4% of the patients (n = 95) and systemic arterial hypertension in 60.2% of the patients (n = 80). The most frequent location of chronic wounds was seen in the lower extremity with 45.1% (n = 60). The mean duration for it to close was 46 ± 14 days, in which they closed within the first 3 months in 93% (n = 125) of the cases. About 91.7% (n = 122) of the wounds achieved total closure. Conclusion: Cultured epidermal allograft, combined with a meticulous technique and an adequate selection of patients, represents a safe and effective tool for chronic wounds.

Role of copper nanoparticles in wound healing for chronic wounds: literature review.

Chronic wounds are defined as wounds that fail to proceed through the normal phases of wound healing in an orderly and timely manner. The most common and inevitable impairment to wound healing is the installation of an infection, usually in the case of chronic wounds. Therefore, the objective of the present review was to identify the importance of copper nanoparticles in dressings for wound healing. Nanoparticles such as silver, gold and copper combat infectious processes through the inhibition of protein synthesis, peroxidation of the cell membrane and destroying the nucleic acids of bacteria and viruses. Among bioactive nanoparticles, copper plays a complex role in various cells, it modulates several cytokines and growth factor mechanisms of action and is essentially involved in all stages of the wound healing process. More importantly, copper plays a key role in skin regeneration and angiogenesis and accelerates the healing process through induction of vascular endothelial growth factor (VEGF) and angiogenesis by hypoxia-induced factor-1-alpha (HIF-1α) action where copper enhances HIF-1α expression and HIF-1α binding to the critical motifs in the promoter and putative enhancer regions of HIF-1-regulated genes.

Hard-to-heal wounds: a randomised trial of an oral proline-containing supplement to aid repair.

OBJECTIVE: We evaluated the effects of a specialised oral nutritional supplement (ONS) containing arginine and proline, with high vitamin A, C and E, zinc and selenium content, on the repair of hard-to-heal wounds. METHOD: Patients with hard-to-heal wounds were evaluated at five timepoints (S0-S4) over four consecutive weeks. At S0 patients were randomised to the specialised ONS (n=15; 25 wounds) or control (n=15; 25 wounds) groups. Posology was 200ml twice daily over the research period. Wound surface area and perimeter were monitored. In addition to the metric data, it was also possible to calculate the rate of wound contraction and the linear growth of the wound edges, looking for wound-healing predictive factors. RESULTS: A total of 30 patients took part in the study. Mean age was 65 years and 50% of patients had diabetes. Of the total evaluated wounds, 78% were <50cm2, 14% were 50-150cm2 and 8% were >250cm2. In 96% of cases, the wounds were in the lower limbs. A statistically significant reduction (p=0.004) in surface area of the wounds due to the specialised ONS, with a performance peak between S1 and S2, was observed. This specialised ONS did not induce changes in blood pressure, blood glucose level or renal function. A mean weekly wound edge growth of 1.85mm in patients with diabetes and 3.0mm in those without diabetes was observed. These results were 2.9 and 4.6 times, respectively, higher than expected, according to the literature. CONCLUSION: Specialised ONS can be a therapeutic option for hard-to-heal wounds.

Cultivo mediante hisopado superficial versus cultivo de la biopsia de tejidos profundos en la infección del pie diabético / Culture through superficial swab versus deep tissue biopsy in the diabetic foot infection

RESUMEN Introducción: para lograr el adecuado y precoz diagnóstico de la infección en pie diabético, es necesario la obtención de una muestra bacteriológica de calidad para la identificación del germen causal. Objetivo: identificar posibles relaciones entre los resultados obtenidos, en el cultivo realizado mediante hisopado superficial versus el obtenido mediante biopsia de los tejidos profundos en la infección del pie diabético. Materiales y métodos: se realizó un estudio explicativo observacional, longitudinal, prospectivo en el Servicio Provincial de Angiología y Cirugía Vascular del Hospital Provincial Clínico Quirúrgico Universitario "Comandante Faustino Pérez", durante un periodo de 3 años desde enero del 2016 hasta diciembre del 2018. Una selección muestral no probabilística determinó una muestra constituida por 138 extremidades en 132 pacientes con diagnóstico clínico de pie diabético infectado, que requirieron cirugía para desbridamiento de la lesión. Aceptaron ser incluidos en la investigación y para el aislamiento del germen causal fueron empleados ambos métodos de cultivo: hisopado superficial y biopsia de los tejidos profundos. Resultados: el promedio de microorganismos aislados se incrementó en relación con la severidad de la infección del pie diabético, con mayor incremento en el aislamiento hecho por el hisopado superficial. El hisopado superficial posee pobre correlación con los gérmenes aislados mediante el cultivo de la biopsia de los tejidos profundos. Conclusiones: las muestras deben ser obtenidas preferentemente por curetaje. En el diagnóstico de la infección del pie diabético es de gran utilidad, por su rapidez y concordancia con los resultados del cultivo, efectuar siempre una tinción de Gram a partir del mismo sitio (AU).

ABSTRACT Introduction: to arrive to an adequate and precocious diagnosis of the diabetic foot infection, it is necessary to obtain a qualitative bacteriological sample to identify the causing germ. Objective: to identify possible relationships between the results obtained both, in the culture made through superficial swab and the culture obtained from deep tissues biopsy in the diabetic foot infection. Materials and methods: a prospective, longitudinal, observational, explicative study was carried out in the Provincial Service of Angiology and Vascular Surgery of Provincial University Clinical Surgical Hospital "Comandante Faustino Pérez", in a period of three years, from January 2016 to December 2018. A non-probabilistic sampling choose a sample of 138 lower limbs in 132 patients with clinical diagnosis of infected diabetic foot, who required surgery for lesion debridement. They gave their consent to be included in the research; for the isolation of the casual germ were used both culture methods, superficial swab and deep tissues biopsy. Results: the average of isolated microorganism increased in relation to the severity of the diabetic food infection, with higher increase in the isolation obtained by superficial swab. The superficial swab shows poor correlation with the germ isolates by the culture the deep tissue biopsy. Conclusions: the samples should be gathered preferably by curettage. In the diagnosis of the diabetic foot infection, it is very useful, due to its speed and concordance with the culture results, to make always a Gram staining beginning from the same place (AU).

Relationship between Wound Severity, Discomfort, and Psychological Problems in Patients with a Diabetic Foot Ulcer in Indonesia: A Cross-sectional Study / Relação entre gravidade da ferida, desconforto e problemas psicológicos em pacientes com úlcera diabética do pé na Indonésia: um estudo transversal / Relación entre la gravedad de la herida, la molestia y los problemas psicológicos en pacientes con úlcera del pie diabético en Indonesia: estudio transversal

ABSTRACT Objectives: The purpose of this study is to identify the relationship between wound severity, discomfort, and psychological problems in patients with a diabetic foot ulcer in Indonesia. Methods: A cross-sectional study is conducted in three general hospitals and one clinic in Indonesia. The Bates-Jensen wound assessment tool (BWAT), the discomfort evaluation of wound instrument (DEWI), and the depression, anxiety, and stress scale (DASS) are used to measure the variables of interest. Path analysis is performed to evaluate the association between wound severity, discomfort, and psychological problems. Results: Of 140 patients with diabetic foot ulcers who joined this study, the majority experienced immobilization (74.3 %), pain (69.3 %), and sleep disturbance (63.6 %). The means were as follows: discomfort (2.35 ± 0.33), depression (1.34 ± 0.41), stress (1.49 ± 0.48), anxiety (1.43 ± 0.40), and wound severity (31.35 ± 9.96). Discomfort partially mediated the relationship between wound severity and psychological problems, which indirect effect was 0.11. Conclusion: High prevalence of discomfort, both physical and psychological, was found in patients with a diabetic foot ulcer. Discomfort mediates the relationship between wound severity and psychological problems. Integrating comfort into wound care management may help to reduce the psychological burden.

RESUMEN Objetivos: el presente estudio tiene como objetivo identificar la relación entre la gravedad de la herida, la molestia y los problemas psicológicos en pacientes con úlcera del pie diabético en Indonesia. Métodos: se realizó un estudio transversal en tres hospitales generales y una clínica de Indonesia. La herramienta de evaluación de heridas Bates-Jensen (BWAT, por sus siglas en inglés), el instrumento de evaluación de molestia de la herida (DEWI, por sus siglas en inglés) y la escala de depresión, ansiedad y estrés (DASS, por sus siglas en inglés) se utilizaron para medir las variables de interés. Se realizó un análisis de ruta para evaluar la asociación entre la gravedad de la herida, la molestia y los problemas psicológicos. Resultados: de 140 pacientes con úlcera del pie diabético que participaron en este estudio, la mayoría experimentó inmovilización (74,3 %), dolor (69,3 %) y trastornos del sueño (63,6 %). Las medias fueron las siguientes: molestia (2,35 ± 0,33), depresión (1,34 ± 0,41), estrés (1,49 ± 0,48), ansiedad (1,43 ± 0,40) y gravedad de la herida (31,35 ± 9,96). La molestia medió en parte la relación entre la gravedad de la herida y los problemas psicológicos, cuyo efecto indirecto fue de 0,11. Conclusión: se encontró una alta prevalencia de molestias, tanto físicas como psicológicas, en pacientes con úlcera del pie diabético. Además, la molestia media la relación entre la gravedad de la herida y los problemas psicológicos. La integración de la comodidad en el manejo del cuidado de las heridas puede ayudar a reducir la carga psicológica.

RESUMO Objetivos: o objetivo deste estudo é identificar a relação entre a gravidade da ferida, o desconforto e os problemas psicológicos en pacientes com úlcera do pé diabético na Indonésia. Métodos: foi realizado um estudo transversal em três hospitais gerais e uma clínica da Indonésia. A ferramenta da avaliação de feridas Bates-Jensen (BWAT, por sua sigla em inglês), o instrumento de avaliação do desconforto da ferida (DEWI, por sua sigla em inglês) e a escala de depressão, ansiedade e estresse (DASS, por sua sigla em inglês) foram utilizados para medir as variáveis de interesse. Uma análise de rota foi realizada para avaliar a associação entre a gravidade da ferida, o desconforto e os problemas psicológicos. Resultados: de 140 pacientes com úlcera diabética do pé que participaram deste estudo, a maioria experimentou imobilização (74,3 %), dor (69,3 %) e transtornos do sono (63,6 %). As médias foram: desconforto (2,35 ± 0,33), depressão (1,34 ± 0,41), estresse (1,49 ± 0,48), ansiedade (1,43 ± 0,40) e gravidade da ferida (31,35 ± 9,96). O desconforto foi o mediador em parte da relação entre a gravidade da ferida e os problemas psicológicos, cujo efeito indireto foi de 0,11. Conclusões: verificou-se alta prevalência de desconforto, tanto físico quanto psicológico, em pacientes com úlcera diabética do pé. Além disso, o desconforto intermedeia a relação entre a gravidade da ferida e os problemas psicológicos. A integração do conforto na gestão do cuidado das feridas pode ajudar a reduzir a carga psicológica.

[Marjolin's ulcer. A 10 year experience in a diabetic foot unit]. / Úlcera de Marjolin: experiencia de 10 años en una unidad de pie diabético.

BACKGROUND: Marjolin's ulcer is defined as the appearance of a neoplasm within a chronic wound. The most common histological type is squamous. A total of 2 cases treated in our hospital are presented. CLINICAL CASE: Case 1. A 71 year old man who presented with redness and suppuration from the wounds he had in his right foot after an electric shock 40 years earlier. The radiology showed involvement of the 4° and 5° metatarsal. Supracondylar amputation was performed, showing a well-differentiated invasive squamous cell carcinoma. CASE 2: A 56 year old male, paraplegic for 20 years. He was treated due to an infected right heel ulcer, with partial improvement, but the ulcers persisted. Biopsy was performed, reporting as epidermoid carcinoma. Infracondylar amputation was performed. The diagnosis was a well-differentiated squamous cell carcinoma infiltrating the dermis. CONCLUSION: The prevalence of Marjolin's ulcer is 1.3-2.2% of all ulcers. Diagnosis is difficult, so biopsy is recommended on any suspicious lesion or ulcer that has received conservative treatment for one month without improvement, although this time limit is not clear. The treatment is the surgery. Local excision with a margin of an inch is enough. If the ulcer is extensive, amputation is required. Survival is estimated between 66 and 80% at 2 years, with recurrence rates of 23%. Unfavourable factors are poor tumour differentiation and metástasis, appearing in 20% of cases.

Anti-biofilm activity of chitosan gels formulated with silver nanoparticles and their cytotoxic effect on human fibroblasts.

The development of multi-species biofilms in chronic wounds is a serious health problem that primarily generates strong resistance mechanisms to antimicrobial therapy. The use of silver nanoparticles (AgNPs) as a broad-spectrum antimicrobial agent has been studied previously. However, their cytotoxic effects limit its use within the medical area. The purpose of this study was to evaluate the anti-biofilm capacity of chitosan gel formulations loaded with AgNPs, using silver sulfadiazine (SSD) as a standard treatment, on strains of clinical isolates, as well as their cytotoxic effect on human primary fibroblasts. Multi-species biofilm of Staphylococcus aureus oxacillin resistant (MRSA) and Pseudomonas aeruginosa obtained from a patient with chronic wound infection were carried out using a standard Drip Flow Reactor (DFR) under conditions that mimic the flow of nutrients in the human skin. Anti-biofilm activity of chitosan gels and SSD showed a log-reduction of 6.0 for MRSA when chitosan gel with AgNPs at a concentration of 100 ppm was used, however it was necessary to increase the concentration of the chitosan gel with AgNPs to 1000 ppm to get a log-reduction of 3.3, while the SSD showed a total reduction of both bacteria in comparison with the negative control. The biocompatibility evaluation on primary fibroblasts showed better results when the chitosan gels with AgNPs were tested even in the high concentration, in contrast with SSD, which killed all the primary fibroblasts. In conclusion, chitosan gel formulations loaded with AgNPs effectively prevent the formation of biofilm and kill bacteria in established biofilm, which suggest that chitosan gels with AgNPs could be used for prevention and treatment of infections in chronic wounds. The statistic significance of the biocompatibility of chitosan gel formulations loaded with AgNPs represents an advance; however further research and development are necessary to translate this technology into therapeutic and preventive strategies.

Anti-biofilm and cytotoxicity activity of impregnated dressings with silver nanoparticles.

Infections arising from bacterial adhesion and colonization on chronic wounds are a significant healthcare problem. Silver nanoparticles (AgNPs) impregnated in dressing have attracted a great deal of attention as a potential solution. The goal of the present study was to evaluate the anti-biofilm activities of AgNPs impregnated in commercial dressings against Pseudomonas aeruginosa, bacteria isolated of chronic wounds from a hospital patient. The antimicrobial activity of AgNPs was tested within biofilms generated under slow fluid shear conditions using a standard bioreactor. A 2-log reduction in the number of colony-forming units of P. aeruginosa was recorded in the reactor on exposure to dressing impregnated with 250ppm of AgNPs, diameter 9.3±1.1nm, and also showed compatibility to mammalian cells (human fibroblasts). Our study suggests that the use of dressings with AgNPs may either prevent or reduce microbial growth in the wound environment, and reducing wound bioburden may improve wound-healing outcomes.

Heridas agudas y crónicas causadas por picaduras y mordeduras de animales / Acute and chronic wounds caused by bites and stings of animals

Se presenta la metodología de trabajo en el servicio, para el manejo de heridas agudas y crónicas en pacientes con mordeduras, las características de este tipo de lesiones y una revisión bibliográfica sobre el tema.(AU)

We present the methodology of work in the service, for the management of acute and chronic wounds in patients with bites, the characteristics of this type of injury and a literature review on the subject.(AU)

Heridas agudas y crónicas causadas por picaduras y mordeduras de animales / Acute and chronic wounds caused by bites and stings of animals

Se presenta la metodología de trabajo en el servicio, para el manejo de heridas agudas y crónicas en pacientes con mordeduras, las características de este tipo de lesiones y una revisión bibliográfica sobre el tema.

We present the methodology of work in the service, for the management of acute and chronic wounds in patients with bites, the characteristics of this type of injury and a literature review on the subject.