Wound Fluid Research Articles

Inflammatory Breast Cancer: The Cytokinome of Post-Mastectomy Wound Fluid Augments Proliferation, Invasion, and Stem Cell Markers.

Inflammatory breast cancer (IBC) is an aggressive phenotype with a high recurrence and low survival rate. Approximately 90% of local breast cancer recurrences occur adjacent to the same quadrant as the initial cancer, implying that tumor recurrence may be caused by residual cancer cells and/or quiescent cancer stem cells (CSCs) in the tumor. We hypothesized that wound fluid (WF) collected after modified radical mastectomy (MRM) may activate cancer cells and CSCs, promoting epithelial mesenchymal transition (EMT) and invasion. Therefore, we characterized the cytokinome of WF drained from post-MRM cavities of non-IBC and IBC patients. The WF of IBC patients showed a significantly higher expression of various cytokines than in non-IBC patients. In vitro cell culture models of non-IBC and IBC cell lines were grown in media conditioned with and/without WF for 48 h. Afterwards, we assessed cell viability, the expression of CSCs and EMT-specific genes, and tumor invasion. Genes associated with CSCs properties and EMT markers were regulated in cells seeded in media conditioned by WF. IBC-WF exhibited a greater potential for inducing IBC cell invasion than non-IBC cells. The present study demonstrates the role of the post-surgical tumor cavity in IBC recurrence and metastasis.

A novel orchiectomy surgical procedure in donkeys ( Equus asinus africanus) with parascrotal access.

Donkeys are a public health concern in the Northeast region of Brazil, with thousands of stray animals. Orchiectomy is an important population control measure; however, the long postoperative period with daily treatment of open wounds in the scrotum makes it difficult to perform a large number of castrations in sheltering centers. We evaluate a novel surgical procedure for orchiectomy in donkeys using parascrotal access. Twelve donkeys were used, divided into two groups: I - submitted to orchiectomy through parascrotal surgical access (novel procedure), and II - submitted to orchiectomy through scrotal access (conventional). Postoperative evaluations consisted of a macroscopic evaluation of the surgical wound (bleeding and intensity of edema), hematological parameters, and peritoneal fluid, which occurred in both groups at the moments (M): M0 - before the surgical procedure. The others moments occurred after surgery: M12 (twelve hours); M24 (twenty-four hours); M48 (forty-eight hours); M72 (seventy-two hours); M8D (eight days); and M16D (sixteen days). The surgical techniques did not generate an important systemic inflammatory response to the point detected by the leukogram, fibrinogen dosage, and peritoneal fluid. The parascrotal technique required long surgery but promoted less bleeding, less edema, and faster healing. The techniques used did not promote sufficient systemic inflammation to alter the number of leukocytes and the fibrinogen concentration; however, evaluation of the peritoneal fluid proved to be important for evaluating inflammatory processes involving the scrotum and inguinal canal. We describe a novel surgical procedure for orchiectomy in Donkeys using a parascrotal access that promoted less risk of bleeding, shorter period of edema, and healing time, but required longer surgery time.

Cytokines and Venous Leg Ulcer Healing-A Systematic Review.

Venous leg ulcers (VLUs) are the most common type of leg ulcers with a significant socioeconomic burden due to slow healing. Cytokines may be involved in the pathogenesis of VLUs. In this systematic review, our objective was to investigate the association between cytokine levels, including growth factors, with the healing of VLUs. PubMed, Embase, Web of Science and Cochrane Library were searched from their inception to August 2021. We retrieved 28 articles investigating 38 different cytokines in 790 patients. Cytokines were most commonly investigated in wound fluid and less frequently in biopsies and serum. The studies were judged as having a moderate to high risk of bias, and the results were often inconsistent and sometimes conflicting. A meta-analysis was not performed due to clinical and methodological heterogeneities. We found weak evidence for elevated IL-1α, IL-6, IL-8, TNF-α and VEGF levels in non-healing VLUs, an elevation that declined with healing. TGF-β1 levels tended to increase with VLU healing. Other cytokines warranting further investigations include EGF, FGF-2, GM-CSF, IL-1β, IL-1Ra and PDGF-AA/PDGF-BB. We conclude that non-healing VLUs may be associated with an elevation of a palette of pro-inflammatory cytokines, possibly reflecting activated innate immunity in these wounds. There is a paucity of reliable longitudinal studies monitoring the dynamic changes in cytokine levels during wound healing.

The impact of intraoperative radiotherapy on breast cancer: focus on the levels of angiogenic factors

ObjectiveAngiogenesis is one of the hallmarks of cancers that is involved in tumor progression. Angiogenic factors induce the formation of new blood vessels and tumor extension, and finally reduce the survival of patients. Intraoperative radiotherapy (IORT), in which radiation is delivered to the tumor bed can kill cells and change tumor microenvironment. Here, we compared the impact of IORT on the levels of angiogenic factors in the blood and surgical wound fluids (SWF) of the breast cancer patients.Patients and methodsThree hundred sixty patients, who had undergone breast-conserving surgery between 2013 and 2018, were enrolled in IORT and non-IORT groups non-randomly. Blood and drained wound fluid (WF) samples were collected from the patients before and after surgery, followed by quantification of the amounts of TGF-β, EGF, FGF, VEGF, and DLL4 in the patients using ELISA.ResultsOur results were indicative of significant differences between the pre-surgery and post-surgery serum levels of EGF, DLL4, and VEGF. Furthermore, ROC analyses showed that TGF-β and DLL4 can differentiate of the early-stage from late-stage of the disease. Interestingly, the rate of the death and recurrence was reduced in IORT group.ConclusionsIn summary, IORT is a safe and effective treatment that can affect angiogenic factors and improve the overall- and recurrence-free survival of breast cancer patients.

Influence of Wound Fluid on the Transdifferentiation of Human Mesenchymal Bone Marrow Stem Cells into Cancer-Associated Fibroblasts

Cancer-associated fibroblasts (CAF) in the tumor microenvironment have a decisive influence on tumor growth and metastatic behavior. The cellular origins as well as the stimuli leading to CAF formation are heterogenous, impeding a precise characterization. Aim of this study was to analyze the influence of cytokines secreted in the process of wound healing, tumor cell-associated paracrine-secreted factors, and direct cell-cell contact on the expression of the CAF-associated markers fibroblast activation protein (FAP), α-smooth muscle actin (α-SMA), thrombospondin-1 (THBS1), and tenascin-c (TNC) by RT-PCR in mesenchymal stem cells (MSC). Cells developed different morphological characteristics after incubation with wound fluid (WF). Moreover, expression of FAP and α-SMA in MSC was significantly reduced after WF compared to tumor-conditioned medium and in co-culture with tumor cells; THBS1 and TNC were not significantly altered after any of the different incubation methods. There were no alterations of expression patterns of FAP and α-SMA in the immunohistochemical analysis. Differ­ences in the cytokine composition of the media were found in the dot blot. The heterogeneity of the results emphasizes the complexity of the interactions of tumor cells and cells of the microenvironment, particularly through the addition of human-derived WF.

Characteristics of cytokine profile of wound fluid in breast cancer patients after mammoplasty with textured implants.

e12589 Background: Breast reconstruction after mastectomy for breast cancer (BC) is an important rehabilitation stage. Breast prosthesis implantation is sometimes accompanied by different complications which can be influenced by the immune system status. The aim of this study was to identify characteristics of the local cytokine profile in patients with breast cancer in the early postoperative period after reconstructive surgery with textured implants (TI). Methods: The study included 30 patients aged 32-68 years, mean age 42.9±1.98 years, with stage I-IIb BC (monocentric nodular BC, T1N0M0 - T2N1M0). All patients underwent skin-sparing mastectomy with level II axillary lymph node dissection and immediate implant reconstruction in 2017-2019. Levels of cytokines IL-1β, IL-2, IL-4, IL-6, IL-8, IFN-γ, and TNF-α were determined by ELISA in wound fluid from the cavity with TI on days 1, 3-4 and 7 after the surgery. Results: Levels of IL-6 in wound fluid of patients with TI statistically significantly decreased by 43% (p=0.036) and 41% (p=0.039) on days 3-4 and 7 after the surgery, and IL-2 decreased by 27% (p=0.045) with its further increase by 44% (p=0.037) compared to day 1. A pronounced increase of the IFN-γ levels by 118% (p=0.018) and 158% (p=0.012) was registered respectively on days 3-4 and 7 after surgery. Conclusions: Reconstructive surgery with TI is followed by a local increase in levels of IL-2 and IFN-γ which may indicate increasing antitumor potential of the microenvironment. The revealed decrease in IL-6 levels after TI implantation indicates certain features of the inflammatory reaction development in the intervention site.

Lymphocyte populations in wound fluid in breast cancer patients after mammoplasty with textured implants.

e12588 Background: Breast reconstruction after skin-sparing mastectomy for breast cancer (BC) is an important rehabilitation stage. Its results depend on many factors, with the immune system status playing the main role. The purpose of this study was to identify characteristics of the local lymphocyte populations in patients with breast cancer in the early postoperative period after reconstructive surgery with textured implants (TI). Methods: The study included 30 patients aged 32-68 years, mean age 42.9±1.98 years, with stage I-IIb BC (monocentric nodular BC, T1N0M0 - T2N1M0). All patients underwent skin-sparing mastectomy with level II axillary lymph node dissection and immediate implant reconstruction in 2017-2019. Populations and subpopulations of lymphocytes were determined in wound fluid from the cavity with TI on days 1, 3-4 and 7 after the surgery using the FacsCanto II flow cytometer (Becton Dickinson, USA) with markers: CD3 FITC /CD15+56 PE /CD45 PerCP /CD4 PE-Cy7/CD19 APC/ CD8 APC-Cy7; CD45RA FITC /CD45RO PE /CD3 PerCP /CD8 APC; CD45RA FITC /CD62L PE /CD3 PerCP /CD4 APC; CD4 FITC /CD127 PE /CD3 PerCP / CD25 APC-Cy7; CD4 FITC /CD38PE /CD3 PerCP / HLADR APC. At least 50,000 cells were accumulated in each sample for the analysis. Results: The relative number of total lymphocytes exceeded the initial values by 3.3 (p=0.025) and 10.9 (p=0.012) times, respectively, on days 3-4 and 7 after surgery. Levels of CD3+CD4+ cells increased gradually and were 29% (p=0.042) higher by day 7, while levels of CD3+CD8+ decreased during the whole observation period. Levels of Tregs did not change, while B lymphocytes decreased by 36% (p=0.035) and 67% (p=0.026), respectively, on days 3-4 and 7. The levels of activated T lymphocytes increased by 33% (p=0.038) on days 3-4, compared with the initial values, probably due to the elevation of CD3+CD8+ levels (by 32%, p=0.037). The number of activated T lymphocytes with the CD3+CD4+ phenotype increased by 44% (p=0.024) on days 6-7. While the content of CD3+CD4+ cells with early activation markers (CD38+) significantly decreased (by 40%, p=0.031) and remained the same on days 3-4 and 7, the number of CD3+CD8+ with similar markers (CD38+) significantly increased by 28% (p=0.044) and 43%. Conclusions: TI implantation was accompanied by the activation of the cytotoxic T unit during the observation period, together with a decrease in immunosuppressive populations of lymphocytes, which may indicate a favorable development of the patient's body reaction.

Supplemental Protein and a Multinutrient Beverage Speed Wound Healing after Acute Sleep Restriction in Healthy Adults

BackgroundPhysiologic and psychologic stress slow healing from experimental wounds by impairing immune function. ObjectivesWe aimed to determine whether supplemental protein and multinutrient supplementation improved wound healing markers after acute stress induced by acute sleep restriction. MethodsIn this single-blind, crossover study in generally healthy young adults (18 males/2 females; mean ± SD age: 19.7 ± 2.30 y), experimental wounds were created by removing the top layer of forearm blisters induced via suction after 48 h of 72-h sleep restriction (2-h nightly sleep), a protocol previously shown to delay wound healing. Skin barrier restoration (measured by transepidermal water loss) assessed wound healing ≤10 d postblistering, and local immune responses were evaluated by serial measurement of cytokine concentrations in fluid collected at wound sites for 48 h postblistering. Participants consumed controlled, isocaloric diets with either 0.900 g · kg−1 · d−1 protein plus placebo (PLA) or 1.50 g · kg−1 · d−1 protein plus multinutrient beverage [l-arginine: 20.0 g/d; l-glutamine: 30.0 g/d; omega-3 (n–3) fatty acids: 1.00 g/d; zinc sulfate: 24.0 mg/d; cholecalciferol: 800 IU/d; and vitamin C: 400 mg/d] (NUT) during sleep restriction and for 4 d afterwards. ResultsSkin barrier restoration (primary outcome) was shorter for NUT (median: 3.98 d; IQR: 1.17 d) than for PLA (median: 5.25 d; IQR: 1.05 d) (P = 0.001). Cytokines from wound fluid (secondary outcome) increased over time (main effect of time P ≤ 0.001), except IL-13 (P = 0.07); however, no effects of treatment were observed. ConclusionsSupplemental nutrition may promote wound healing after sleep restriction in healthy adults including military personnel, the latter of which also have a high incidence of wounds and infection.This trial was registered at clinicaltrials.gov as NCT03525184.

Bio‐Inspired Self‐Hydrophobized Sericin Adhesive with Tough Underwater Adhesion Enables Wound Healing and Fluid Leakage Sealing

AbstractGiven that adhesion‐resistance of water severely weakens the bonding strength of tissue adhesives, instant adhesion to wet biological tissue surfaces remains challenging. Inspired by the robust underwater adhesion of natural creatures (such as mussels and barnacles whose underwater adhesion derives from the synergy of hydrophobic and adhesive matrix), a self‐hydrophobized adhesive is developed by co‐assembly of disulfide‐bond hydrolyzed hydrophobic natural sericin protein (a major component of silkworm silk fibers) and tannic acid. Once exposed to water, the self‐aggregation of hydrophobic chains within the adhesive repels water and enhances interfacial hydrogen bonding or electrostatic interactions, mechanistically leading to a robust (>0.5 MPa for solid plates and >0.1 MPa for tissues) and durable (still maintained at 0.4 Mpa even after five cycles) underwater adhesion. Owing to its robust underwater adhesion property, this adhesive possesses multiple advantages outperforming commercial adhesives, such as in vivo wound healing‐promoting effects, effective fluid leakage sealing, and rapid hemostasis activity. This study not only offers a novel strategy for designing and fabricating an underwater adhesive with natural protein but also provides a new adhesive for various potential applications, including promoting wound healing and hemostasis.

Role of Nitric Oxide-Releasing Glycosaminoglycans in Wound Healing.

Two glycosaminoglycan (GAG) biopolymers, hyaluronic acid (HA) and chondroitin sulfate (CS), were chemically modified via carbodiimide chemistry to facilitate the loading and release of nitric oxide (NO) to develop a multi-action wound healing agent. The resulting NO-releasing GAGs released 0.2-0.9 μmol NO mg-1 GAG into simulated wound fluid with NO-release half-lives ranging from 20 to 110 min. GAGs containing alkylamines with terminal primary amines and displaying intermediate NO-release kinetics exhibited potent, broad spectrum bactericidal action against three strains each of Pseudomonas aeruginosa and Staphylococcus aureus ranging in antibiotic resistance profile. NO loading of the GAGs was also found to decrease murine TLR4 activation, suggesting that the therapeutic exhibits anti-inflammatory mechanisms. In vitro adhesion and proliferation assays utilizing human dermal fibroblasts and human epidermal keratinocytes displayed differences as a function of the GAG backbone, alkylamine identity, and NO-release properties. In combination with antibacterial properties, the adhesion and proliferation profiles of the GAG derivatives enabled the selection of the most promising wound healing candidates for subsequent in vivo studies. A P. aeruginosa-infected murine wound model revealed the benefits of CS over HA as a pro-wound healing NO donor scaffold, with benefits of accelerated wound closure and decreased bacterial burden attributable to both active NO release and the biopolymer backbone.

Development of a textile based protein sensor for monitoring the healing progress of a wound

This article focuses on the design and fabrication of flexible textile-based protein sensors to be embedded in wound dressings. Chronic wounds require continuous monitoring to prevent further complications and to determine the best course of treatment in the case of infection. As proteins are essential for the progression of wound healing, they can be used as an indicator of wound status. Through measuring protein concentrations, the sensor can assess and monitor the wound condition continuously as a function of time. The protein sensor consists of electrodes that are directly screen printed using both silver and carbon composite inks on polyester nonwoven fabric which was deliberately selected as this is one of the common backing fabric types currently used in wound dressings. These sensors were experimentally evaluated and compared to each other by using albumin protein solution of pH 7. A comprehensive set of cyclic voltammetry measurements was used to determine the optimal sensor design the measurement of protein in solution. As a result, the best sensor design is comprised of silver conductive tracks but a carbon layer as the working and counter electrodes at the interface zone. This design prevents the formation of silver dioxide and protects the sensor from rapid decay, which allows for the recording of consecutive measurements using the same sensor. The chosen printed protein sensor was able to detect bovine serum albumin at concentrations ranging from 30 to 0.3 mg/mL with a sensitivity of 0.0026 mu A/M. Further testing was performed to assess the sensor’s ability to identify BSA from other interferential substances usually present in wound fluids and the results show that it can be distinguishable.

Radiobiological effects of wound fluid on breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic culture

Intraoperative radiotherapy (IORT) could abrogate cancer recurrences, but the underlying mechanisms are unclear. To clarify the effects of IORT-induced wound fluid on tumor progression, we treated breast cancer cell lines and human-derived tumor spheroids in 2D and microfluidic cell culture systems, respectively. The viability, migration, and invasion of the cells under treatment of IORT-induced wound fluid (WF-RT) and the cells under surgery-induced wound fluid (WF) were compared. Our findings showed that cell viability was increased in spheroids under both WF treatments, whereas viability of the cell lines depended on the type of cells and incubation times. Both WFs significantly increased sub-G1 and arrested the cells in G0/G1 phases associated with increased P16 and P21 expression levels. The expression level of Caspase 3 in both cell culture systems and for both WF-treated groups was significantly increased. Furthermore, our results revealed that although the migration was increased in both systems of WF-treated cells compared to cell culture media-treated cells, E-cadherin expression was significantly increased only in the WF-RT group. In conclusion, WF-RT could not effectively inhibit tumor progression in an ex vivo tumor-on-chip model. Moreover, our data suggest that a microfluidic system could be a suitable 3D system to mimic in vivo tumor conditions than 2D cell culture.

High-throughput and high-sensitivity biomarker monitoring in body fluid by FAIMS-enhanced fast LC SureQuant™ IS targeted quantitation

Targeted proteomics methods have been greatly improved and refined over the last decade and are becoming increasingly the method of choice in protein and peptide quantitative assays. Despite the tremendous progress, targeted proteomics assays still suffer from inadequate sensitivity for lower abundant proteins and throughput, especially in complex biological samples. These attributes are essential for establishing targeted proteomics methods at the forefront of clinical use. Here, we report an assay utilizing the SureQuantTM internal standard triggered targeted method on a newest generation mass spectrometer coupled with a FAIMS (high-field asymmetric waveform ion mobility spectrometry) interface ion mobility device and an EvoSep One liquid chromatography platform, which displays markedly enhanced sensitivity and a high throughput of 100 samples per day. We demonstrate the robustness of this method by quantifying proteins ranging six orders of magnitude in human wound fluid exudates, a biological fluid that exhibits sample complexity and composition similar to plasma. Among the targets quantified were low-abundance proteins such at TNFA and IL1B, highlighting the value of this method in the quantification of trace amounts of invaluable biomarkers that were until recently hardly accessible by targeted proteomics methods. Taken together, this method extends the toolkit of targeted proteomics assays and will help to drive forward mass spectrometry-based proteomics biomarker quantification.

Typing of Leishmania Species Causing Cutaneous Leishmaniasis by Sybr Green Based ITS-1 Real Time Polymerase Chain Reaction Method

Cutaneous leishmaniasis (CL) is an important public health problem, most frequently seen in Şanlıurfa in Turkey. It is important to determine the species in regions where infection occurs with different Leishmania species, as in our province. In this study, it was aimed to genotype 136 samples with suspected Leishmania from Şanlıurfa using the Sybr Green-based ITS-1 real time polymerase chain reaction (Rt-PCR) method and then to compare them with ITS-1 PCR RFLP and direct microscopy methods. Wound fluid samples from patient lesions suspected of leishmaniasis were mounted on a slide, fixed, and stained with Giemsa dye. The preparations were examined under the microscope and evaluated for the presence of amastigote. After the extraction of DNA from Giemsa stained preparations by using the QIAmp DNA Mini Kit (Qiagen, Germany), the samples were studied with the Sybr Green based ITS-1 Rt-PCR method using LITSR and L5.8S primers. As a result of the PCR study, melting curve analysis was determined and the melting curves were compared with the reference strains. Then, PCR was performed in 136 samples for ITS1 region amplification using primers LITSR and L5.8S. PCR products were digested with Hae III restriction enzyme and RFLP process was performed. The products were run on metaphor agarose gel than the gels were stained with ethidium bromide for 15 min and visualized in a UV transilluminator In our study, the results of Sybr Green-based ITS-1 Rt-PCR, ITS-1 PCR-RFLP and direct microscopy methods were compared. The highest positivity rate was determined as 97% (136/132) in ITS-1 Rt-PCR method. With ITS-1 PCR-RFLP method 95.5% (136/130) positivity and with direct microscopy 94.1% (136/128) positivity were obtained, respectively. Of 132 samples, which were studied with the Sybr Green-based ITS-1 Rt-PCR method and found as positive, 121 were genotyped as L.tropica and 11 were genotyped as L.major by melting curve analysis. It was determined that, of 130 samples studied with ITS-1 PCR RFLP method 119 (91.5%) were detected as L.tropica and 11 (8.5%) were detected as L.major. The ITS-1 Rt-PCR method we used in our study was the method that detected the most positivity rate. With this method, Leishmania specimens were typed as L.tropica and L.major. It is thought that this method may be useful for the detection of the presence of Leishmania parasite and in the rapid identification of Leishmania species, as it does not require extra processes such as cutting and staining after PCR and results in a short time, but new studies are needed to observe its effectiveness in detecting other species other than L.tropica and L.major.

Wound fluid sampling methods for proteomic studies: A scoping review.

Understanding why some wounds are hard to heal is important for improving care and developing more effective treatments. The method of sample collection used is an integral step in the research process and thus may affect the results obtained. The primary objective of this study was to summarise and map the methods currently used to sample wound fluid for protein profiling and analysis. Eligible studies were those that used a sampling method to collect wound fluid from any human wound for analysis of proteins. A search for eligible studies was performed using MEDLINE, Embase and CINAHL Plus in May 2020. All references were screened for eligibility by one reviewer, followed by discussion and consensus with a second reviewer. Quantitative data were mapped and visualised using appropriate software and summarised via a narrative summary. After screening, 280 studies were included in this review. The most commonly used group of wound fluid collection methods were vacuum, drainage or use of other external devices, with surgical wounds being the most common sample source. Other frequently used collection methods were extraction from absorbent materials, collection beneath an occlusive dressing and direct collection of wound fluid. This scoping review highlights the variety of methods used for wound fluid collection. Many studies had small sample sizes and short sample collection periods; these weaknesses have hampered the discovery and validation of novel biomarkers. Future research should aim to assess the reproducibility and feasibility of sampling and analytical methods for use in larger longitudinal studies.

In Vitro-In Vivo Correlation for the Antibacterial Effect of Lactiplantibacillus plantarum as a Topical Healer for Infected Burn Wound.

Difficulties in delivering antimicrobial agents to wound areas and emersion of multiple drug resistant organisms (MDROs) have converted managing burn infections into a complicated task in medicine. Probiotics emerged not only as a probable solution for burn infections but also as an accelerator in the healing process. The probability of in vitro-in vivo correlation (IVIVC) in probiotic activity leads to lower costs in finding new therapeutic options. Simulated wound fluid (SWF) was used to evaluate the antibacterial function of Lactiplantibacillus plantarum in wounds. The growth parameters in SWF were evaluated using a logistic model to predict growth behavior in the wound area. In addition, probiotic antimicrobial activity and secretion of antibacterial substances in SWF were also studied. Data were used to select the initial dose and apply frequency for in vivo study. The wound models were infected by two main pathogens (Pseudomonas aeruginosa or Staphylococcus aureus). In vitro results showed less lag time associated with considerable acid production in SWF. In the following, secretion of antimicrobial substances and co-aggregation with pathogens became more important. The susceptibility of pathogens to these factors was different, and culture medium affected the yield of each factor involved in eliminating pathogens. Histological analysis and macroscopic examination of wounds revealed probiotics as effective as positive control or more. There were some differences in the antibacterial functions of probiotics in simulated and real wound environments. The in vitro effect of probiotics on removal of pathogens was not the same as the trend seen in vivo.

Fabrication and characterization of a myrrh hydrocolloid dressing for dermal wound healing

Common occurrence of dermal injuries has made wound dressing a critical component in wound healing. Hydrocolloid wound dressings have been developed as fanciful and effective wound dressing with ability for wound fluid control, adherence to wound site, and infection prevention. Effectiveness of topical application of myrrh resin to dermal wound has been well documented. We fabricated Mirderm hydrocolloid wound dressings with myrrh resin to enhance wound healing and improve on the mechanical characteristics of the hydrocolloid. The biocompatibility of Mirderm was assayed with CCK-8 while the wound healing efficacy was tested in vivo. The physical and mechanical properties were checked to affirm the suitability of Mirderm as hydrocolloid wound dressing. We observed Mirderm showed superior wound healing and mechanical capability suitable for hydrocolloid polymer relative to investigated commercial hydrocolloid and medical Gauze. These are indications that Mirderm may be an improvement on standard dressings such as Gauze and some commercial hydrocolloids.

Synergism inhibition and eradication activity of silver nitrate/potassium tellurite combination against Pseudomonas aeruginosa biofilm.

Antibiotic resistance, biofilm and persistent infection of Pseudomonas aeruginosa is a perilous challenge in the healthcare system. Hence, a vast number of novel antipseudomonas approaches are currently being pursued. Our group focuses on exploring the efficacy of metal(loid)-based antimicrobials (MBAs) towards novel infection control solutions. Initially, nine MBAs were tested for biofilm prevention and eradication efficacy. Synergistic potentials were then screened systematically in a total of 1920 combinatorial MBA concentrations, in laboratory media [CAMHB and LB] and infection-related simulated wound fluid (SWF). The antibiofilm efficacy of the silver nitrate (AgNO3; 'Ag') with potassium tellurite (K2TeO3; 'Te') combination was examined against clinical antibiotic-resistant isolates and compared with the most used antibiotics. The in vitro resistance acquisition test, for exploring the chance of getting future resistance, and meta-analysis, for estimating Ag/Te human cell cytotoxicity, were carried out. The Ag/Te combination was identified as the most effective agent against P. aeruginosa biofilm. The application of the Ag/Te combination was quite effective against all clinical isolates. Comparison of clinical isolates with indicator strains showed clinical isolates are gaining resistance against the antibiotics (especially gentamicin) and Ag, while they are susceptible to Te and particularly the Ag/Te combination. The chance of getting future resistance against Ag/Te as a mixture was remarkably lower than the individual application of each metal. Te has significantly lower human cell cytotoxicity in comparison with Ag. Te could be an appropriate alternative against P. aeruginosa biofilms (existing or prevention thereof), especially in combination with Ag.

Does exudate viscosity affect its rate of absorption into wound dressings?

Exudate is a vital component of healing wounds. There are differences between acute and chronic exudate, with the latter seen as highly toxic to the healing environment. Wound exudate assessment is not easy for clinicians. The viscosity of wound fluid/exudate is as important as its quantity. Wound fluid viscosity increases when it contains more protein. As wound dressings exhibit a variety of fluid-handling mechanisms, it is important to understand how they interact with the different exudate types, which may alter the wound/dressing interface. This knowledge will ensure a healing wound environment that is beneficial, not one that leaves harmful exudate on the wound surface. This study aimed to evaluate if the viscosity of exudate affected absorption time in four wound dressing types. This study evaluated the viscosities of two solutions and their effect on the absorption times of four dressing types. The viscosities of the solutions were calculated using Ostwald viscometers, then 2 ml of each of these liquids was applied to each of the four dressing types and repeated 10 times for each dressing, as per British Standards Institution requirements. The parameters for temperature and frequency were applied, and the absorption times recorded. A two-way repeated analysis of variance (ANOVA) was conducted to examine if exudate viscosity and dressing type, or their interaction, affected the absorption time. The results indicated that the viscosity of the solution alone had a significant effect on absorption time (p<0.01), and that the combined effect of dressing type and viscosity also affected absorption (p<0.01). The type of dressing alone was found to have a significant effect on absorption time (p<0.01). When looking at the between-subject effects of the dressings (between the four types) the only non-significant finding was between two dressings-the superabsorbent and the moderate (foam) absorbing dressing (p=0.097). All other dressings had a between-subject effect of (p<0.01). A significant difference in absorption times was found in this investigation of dressing types and viscosity of test solutions. The greater the viscosity of the fluid, the longer it took for it to be absorbed into all dressing types tested. This delay was not determined simply by the dressing or by the viscosity of the fluid, but likely to be a combination of the mode of action of the dressing, its pore size and particulate composition of the exudate, all of which require further investigation.

Myo-Inositol in Fermented Sugar Matrix Improves Human Macrophage Function.

Reactive oxygen species production by innate immune cells plays a central role in host defense against invading pathogens at wound-site. A weakened host-defense results in persistent infection leading to wound chronicity. Fermented Papaya Preparation (FPP), a complex sugar matrix, bolsters respiratory burst activity and improves wound healing outcomes in chronic wound patients. The objective of the current study was to identify underlying molecular factor/s responsible for augmenting macrophage host defense mechanisms following FPP supplementation. In depth LC-MS/MS analysis of cells supplemented with FPP led to identification of myo-inositol as a key determinant of FPP activity towards improving macrophage function. Myo-inositol, in quantities that is present in FPP, significantly improved macrophage respiratory burst and phagocytosis via de novo synthesis pathway of ISYNA1. In addition, myo-inositol transporters, HMIT and SMIT1, played a significant role in such activity. Blocking these pathways using siRNA attenuated FPP-induced improved macrophage host defense activities. FPP supplementation emerged as a novel approach to increase intracellular myo-inositol levels. Such supplementation also modified wound microenvironment in chronic wound patients to augment myo-inositol levels in wound fluid. These observations indicate that myo-inositol in FPP influences multiple aspects of macrophage function critical for host defense against invading pathogens.