Anti-inflammatory Environment Research Articles

Immunological role of zinc in preterm neonates

Zinc (Zn), an essential trace element, plays a significant role in fetal development and biological defense during the embryonic and neonatal periods. Therefore, exploring the kinetics of Zn related to immune disturbances in preterm neonates is important. We here performed the measurement of Zn concentration along with immunological analysis of neonates and investigated the role of Zn in the neonatal period. Serum Zn concentrations were measured immediately after birth in neonates (329 cases). Moreover, for 25 cases, the kinetics of various immune cells and cytokines were measured by flow cytometry and electrochemiluminescence. We observed that Zn levels were inversely correlated with gestational weeks. Immune cell and cytokine analysis revealed an inverse correlation between HLA-DR on monocytes and Zn levels and between inflammatory cytokine interleukin-12 and Zn levels. Furthermore, oxidative stress status was inversely correlated with Zn levels. Our results suggested that the Zn dynamics immediately after birth, which show a negative correlation with the gestational week, can provide an anti-inflammatory and anti-oxidative environment for preterm neonates. The increased Zn concentration in the blood of preterm neonates may consequently protect neonates from perinatal stress.

Intestinal microbiome dysbiosis increases Mycobacteria pulmonary colonization in mice by regulating the Nos2-associated pathways

Increasing researches reveal gut microbiota was associated with the development of tuberculosis (TB). How to prevent or reduce Mycobacterium tuberculosis colonization in the lungs is a key measure to prevent TB. However, the data on gut microbiota preventing Mycobacterium colonization in the lungs were scarce. Here, we established the clindamycin-inducing intestinal microbiome dysbiosis and fecal microbial transplantation models in mice to identify gut microbiota’s effect on Mycobacterium’s colonization in the mouse lungs and explore its potential mechanisms. The results showed that clindamycin treatment altered the diversity and composition of the intestinal bacterial and fungal microbiome, weakened the trans-kingdom network interactions between bacteria and fungi, and induced gut microbiome dysbiosis in the mice. Gut microbiota dysbiosis increases intestinal permeability and enhances the susceptibility of Mycobacterium colonization in the lungs of mice. The potential mechanisms were gut microbiota dysbiosis altered the lung transcriptome and increased Nos2 expression through the ‘gut–lung axis’. Nos2 high expression disrupts the intracellular antimicrobial and anti-inflammatory environment by increasing the concentration of nitric oxide, decreasing the levels of reactive oxygen species and Defb1 in the cells, and promoting Mycobacteria colonization in the lungs of mice. The present study raises a potential strategy for reducing the risks of Mycobacteria infections and transmission by regulating the gut microbiome balance.

Immune response to viscerotropic Leishmania: a comprehensive review.

L. donovani and L. infantum infections are associated with a broad clinical spectrum, ranging from asymptomatic cases to visceral leishmaniasis (VL) with high mortality rates. Clinical manifestations such as post-kala-azar dermal leishmaniasis (PKDL) and visceral leishmaniasis-associated hemophagocytic lymphohistiocytosis-mimic (VL-associated HLH-mimic) further contribute to the diversity of clinical manifestations. These clinical variations are intricately influenced by the complex interplay between the host's immune response and the parasite's escape mechanisms. This narrative review aims to elucidate the underlying immunological mechanisms associated with each clinical manifestation, drawing from published literature within the last 5 years. Specific attention is directed toward viscerotropic Leishmania sinfection in patients with inborn errors of immunity and acquired immunodeficiencies. In VL, parasites exploit various immune evasion mechanisms, including immune checkpoints, leading to a predominantly anti-inflammatory environment that favors parasite survival. Conversely, nearly 70% of individuals are capable of mounting an effective pro-inflammatory immune response, forming granulomas that contain the parasites. Despite this, some patients may experience reactivation of the disease upon immunosuppression, challenging current understandings of parasite eradication. Individuals living with HIV and those with inborn errors of immunity present a more severe course of infection, often with higher relapse rates. Therefore, it is crucial to exclude both primary and acquired immune deficiencies in patients presenting disease relapse and VL-associated HLH-mimic. The distinction between VL and HLH can be challenging due to clinical similarities, suggesting that the nosological entity known as VL-associated HLH may represent a severe presentation of symptomatic VL and it should be considered more accurate referring to this condition as VL-associated HLH-mimic. Consequently, excluding VL in patients presenting with HLH is essential, as appropriate antimicrobial therapy can reverse immune dysregulation. A comprehensive understanding of the immune-host interaction underlying Leishmania infection is crucial for formulating effective treatment and preventive strategies to mitigate the disease burden.

P016 METAPROTEOMICS ANALYSIS IN HEALTHY INDIVIDUALS REVEALS A HIGH CARDIOVASCULAR RISK PROFILE ACROSS BLOOD PRESSURE, DIET, HUMAN AND MICROBIAL PROTEINS

Background/Objective: The gut microbiota is a crucial link between diet and cardiovascular health by producing beneficial metabolites, such as short-chain fatty acids (SCFAs), through fibre fermentation. However, most studies have focused on microbial DNA to infer microbiota function. Here, we employed metaproteomics, a novel method to capture gut human and microbiota protein expression profiles concurrently. We aimed to investigate the crosstalk between gut microbiota, diet, host gut health, and blood pressure (BP). Methods: Mass spectrometry-based metaproteomic analysis was performed on 61 faecal samples with paired ambulatory BP monitoring, food intake (food frequency questionnaire), and demographic data. Results: We identified 21,594 microbial and 1,072 human proteins. Traditional risk factors (age, BMI, sex, BP) explained only ∼10% of the proteome variance. However, unsupervised clustering analysis of both human and microbial proteome revealed two distinct clusters (low-risk and high-risk) with significantly different 24-hour systolic BP (mean±SD, low-risk 116±13mmHg vs high-risk: 126±15mmHg, P=0.016), women prevalence (72.4% vs 40.6%, P=0.025), and diet quality (assessed as Athlete Diet Index, 61.6±6.7 vs 57.8±7.6, P=0.044). In the human proteome, the low-risk group had lower expression of the VEGFA-VEGFR2 signalling pathway and tight junction proteins and higher SCFA production (particularly propionate), suggesting an anti-inflammatory gut environment. In the microbial proteome, the low-risk group had higher activity of inorganic ion transport and metabolism-related proteins, with increased expression of acetate kinase that produces both the SCFAs acetate and propionate, particularly in primary fibre-fermenting bacteria, including Phocaeicola dorei and Blautia wexlerae. Finally, via co-expression networks, we identified profilin-1 (PFN1), which contributes to hypertension and atherosclerosis, as a crucial human protein associated with several bacteria present in the high-risk group. Conclusions: Employing an unsupervised clustering based on human and microbial proteomics, we identified two groups of participants with varying traditional and emerging cardiovascular risk factors. This was explained by complex interactions among diet, gut microbiota, and BP. This underscores the need for sophisticated models to understand multifactorial risk factors for cardiovascular disease.

Graded Modulation of Inflammation by Metal Ion-Coordinated Peptide-Based Hydrogel Chemical Regulators Promotes Tendon-Bone Junction Regeneration.

After rotator cuff injuries, uncontrolled inflammation hinders tendon-bone junction regeneration and induces scar formation in situ. Therefore, precisely controlling inflammation could be a solution to accelerate tendon-bone junction regeneration. In this study, we synthesized a peptide-metal ion complex hydrogel with thermosensitive capability that can be used as a hydrogel chemical regulator. By the coordination complex between Mg2+ and BMP-12, the free and coordinated Mg2+ can be programmability released from the hydrogel. The fast release of free Mg2+ can prevent inflammation at the early stage of injuries, according to the results of RT-qPCR and immunofluorescence staining. Then, the coordinated Mg2+ was slowly released from the hydrogel and provided an anti-inflammatory environment for tendon-bone junction regeneration in the long term. Finally, the hydrogel demonstrated enhanced therapeutic effects in a rat rotator cuff tear model. Overall, the Mg2+/BMP-12 peptide-metal ion complex-based hydrogel effectively addresses the regenerative requirements of the tendon-bone junction across various stages by graded modulating inflammation.

The immunomodulatory role of paracrine signalling factor VSIG4 in peritoneal metastases

Peritoneal metastasis (PM), the regional progression of intra-abdominal malignancies, is a common sequelae of colorectal cancer (CRC). Immunotherapy is slated to be effective in generating long-lasting anti-tumour response as it utilizes the specificity and memory of the immune system. In the tumour microenvironment, tumour associated macrophages (TAMs) are posited to create an anti-inflammatory pro-tumorigenic environment. In this paper, we aimed to identify immunomodulatory factors associated with colorectal PM (CPM). A publicly available colorectal single cell database (GSE183916) was analysed to identify possible immunological markers that are associated with the activation of macrophages in cancers. Immunohistochemical analysis for V-set and immunoglobin containing domain 4 (VSIG4) expression was performed on tumour microarrays (TMAs) of tumours of colorectal origin (n = 211). Expression of VSIG4 in cell-free ascites obtained from CPM patients (n = 39) was determined using enzyme-linked immunosorbent assay (ELISA). CD163-positive TAMs cluster expression was extracted from a publicly available single cell database and evaluated for the top 100 genes. From these macrophage-expressed genes, VSIG4, a membrane protein produced by the M2 macrophages, mediates the up-regulation of anti-inflammatory and down-regulation of pro-inflammatory macrophages, contributing to an overall anti-inflammatory state. CRC TMA IHC staining showed that low expression of VSIG4 in stromal tissues of primary CRC are associated with poor prognosis (p = 0.0226). CPM ascites also contained varying concentrations of VSIG4, which points to a possible role of VSIG4 in the ascites. The contribution of VSIG4 to CPM development can be further evaluated for its potential as an immunotherapeutic agent.

Near-Infrared Responsive Nanocomposite Hydrogel Dressing with Anti-Inflammation and Pro-Angiogenesis for Wound Healing.

Anti-inflammatory and angiogenesis are two important factors in wound healing. Wound dressings with anti-inflammation and vascularization are essential to address complex interventions, expensive treatments, and uncontrolled release mechanisms. Based on the above considerations, we designed a near-infrared (NIR)-responsive hydrogel dressing, which is composed of mPDA-DFO@LA nanoparticles (mPDA: dopamine hydrochloride nanoparticles, DFO: deferoxamine, LA: lauric acid), valsartan (abbreviated as Va), and dopamine-hyaluronic acid hydrogel. The hydrogel dressing demonstrated injectability, bioadhesive, and photothermal properties. The results indicated the obtained dressing by releasing Va can appropriately regulate macrophage phenotype transformation from M1 to M2, resulting in an anti-inflammatory environment. In addition, DFO encapsulated by LA can be sustainably released into the wound site by NIR irradiation, which further prevents excessive neovascularization. Notably, the results in vivo indicated the mPDA-DFO@LA/Va hydrogel dressing significantly enhanced wound recovery, achieving a healing rate of up to 96% after 11 days of treatment. Therefore, this NIR-responsive hydrogel dressing with anti-inflammation, vascularization, and on-demand programmed drug release will be a promising wound dressing for wound infection.

P-799 Augmenting endometrial thickness and implantation rates: investigating the efficacy of autologous blood cell derivative - endosera in women struggling with thin endometrium and infertility

Abstract Study question Could autologous platelet derived growth factor based strategies promote endometrial growth in women with refractory thin endometrium resulting in sustained implantation and live births? Summary answer Autologous platelet derived growth factor based strategies remarkably promote endometrial thickness and receptivity in women with refractory thin endometrium and pregnancy outcomes What is known already Repeated implantation failure (RIF) attributed to thin endometrium poses significant challenge in reproductive medicine. In cases where conventional treatments prove ineffective, couples often contemplate stressful option of surrogacy. Regenerative strategies, such as platelet-rich plasma (PRP), exhibit promise by capitalizing on anti-inflammatory and pro-regenerative attributes of platelet-derived growth factors. Recognizing the imperative for anti-inflammatory microenvironment conducive to successful implantation, Endosera, a cell-free growth factors concentrate tailored for implantation has been formulated. This study seeks to evaluate the effectiveness of Endosera in augmenting endometrial thickness and implantation rates, offering potential remedies for RIF and thin endometrium. Study design, size, duration In this prospective self-controlled study 62 women (24-47 years) were enrolled spanning May 2021 to December 2023. Women experienced >2 failed IVF cycles and multiple cycle cancellations due to refractory thin endometrium with poor endometrial receptivity defined as endometrial thickness of < 7mm despite standard medical therapy, abnormal endometrial pattern and suboptimal endometrialvascularity, negative hysteroscopic screening for endometrial pathology, and negative bacteriologicscreening and despite multiple failed conventional therapies were selected to undergo Endosera treatment. Participants/materials, setting, methods After obtaining informed consent, patients underwent three intrauterine infusions of Endosera on menstrual cycle Day 7, Day 12, and 2 days before frozen embryo transfer (FET). Alongside standard hormone replacement therapy (HRT), 0.8 ml of Endosera was introduced into the uterine cavity. Pregnancy confirmation relied on a positive serum β-HCG test two weeks post-ET, while clinical pregnancy was established by detecting fetal heartbeat through transvaginal ultrasound four weeks post-ET. Main results and the role of chance In this study involving 62 patients facing multiple cancelled IVF cycles due to refractory thin endometrium, the administration of three doses of Endosera demonstrated significant positive outcomes. Average endometrial thickness notably increased from 6.2 ± 1.2 mm to 8.4 ± 1.2 mm (P < 0.0001) after the 2nd Endosera dose, reaching an optimal thickness of ≥ 8mm. Among the participants, 43 achieved positive pregnancy result (69.4%), with 26 (41.9%) delivering healthy babies and 7 (11.3%) uneventful ongoing pregnancies. Poor outcomes included for 2 (3.2%) patients cycle was cancelled due to no endometrial improvement 5(8.1%) miscarriages before 10 weeks, while 12 (19.4%) women didn’t conceive and for 4.8%(3) patients results awaited post ET. No adverse effects were reported, indicating safety of Endosera.Cumulative positive pregnancy outcomes recorded to be 53.2% (33). The formulation, standardized to contain 6-9 times higher growth factor levels than peripheral blood, emphasizes implantation-friendly and anti-inflammatory cytokines. This optimized composition aligns with cyclical requirements of the endometrial phases (proliferation, secretion, and implantation), creating an angiogenic and anti-inflammatory environment crucial for successful implantation. Endosera’s safety, reproducibility, and efficacy in mimicking natural tissue repair and regeneration processes position it as promising solution for refractory thin endometrium-related challenges. Limitations, reasons for caution This study, though demonstrating improved outcomes in refractory thin endometrium cases with Endosera treatment, acknowledges limitations such as a small sample size and absence of a randomized control group. To establish Endosera as a routine adjuvant in IVF, larger study with stringent inclusion criteria for refractory thin endometrium is recommended. Wider implications of the findings Endosera demonstrated notable success with significant improvement in implantation, pregnancy, and live birth rates for refractory thin endometrium patients prompts rigorous randomized controlled studies. These will include diverse populations, validating results and offering patients hope to reduce financial and psychological stress and challenges associated with cycle cancellations and surrogacy. Trial registration number Not Applicable

O-280 Vaginal microbiota transplant (VMT) without antibiotic pretreatment reveals changes in vaginal microbiota and immune profile in women with asymptomatic dysbiosis – a randomized, placebo-controlled trial

Abstract Study question Investigate the safety of VMT and explore its effects on the vaginal microbiomes and local inflammation in healthy, asymptomatic volunteer women screened for vaginal dysbiosis Summary answer We demonstrate safety and engraftment of donor bacteria in 50% of recipients, and a distinct shift in local inflammatory markers following resolution of vaginal dysbiosis What is known already Dominance by a subset of vaginal Lactobacillus species is considered healthy. Dysbiosis characterized by high abundance of anaerobic species is observed in around one third of women, and affects fertility, pregnancy outcomes, risk of STIs and UTIs, and potentially cancer treatment. Current use of antibiotics is often insufficient with high recurrence rates and potentially antibiotic resistance. There is a clear need for alternatives and recently it was shown that VMT following antibiotic treatment resulted in long-lasting improvements in symptoms of bacterial vaginosis in four out of five patients, but RCTs without the use of antibiotics remain to be reported. Study design, size, duration We performed a randomized, placebo-controlled double-blinded trial in 34 recipients randomized in an active:placebo 3:1 ratio. Subjects in the active arm received VMTs from donor cervicovaginal secretions (CVS) while placebo participants received saline. All recipients were dosed on 3 consecutive days with active arm recipients receiving all 3 VMTs originating from the same donor. All recipients were followed for safety and efficacy assessments for 6 menstrual cycles after dosing. Participants/materials, setting, methods Donors: CVS was obtained from healthy subjects screened to be negative for a broad panel of infections and having lactobacilli-dominant vaginal microbiomes. RCT: Healthy, asymptomatic women aged 18 to 45 whose vaginal microbiome had combined lactobacilli relative abundance <10% and combined relative abundance of dysbiosis-associated bacteria >20% were included. VMTs were applied vaginally on three consecutive days. Follow-up visits were performed 1 week after the last VMT and then once every cycle for 6 cycles. Main results and the role of chance Here, we report the successful implementation of a donor program to obtain screened, frozen, and banked donor CVS to be used in the first VMT intervention RCT. In the ITT population, the primary efficacy analysis showed statistically significant increase from baseline in relative abundance of vaginal lactobacilli to follow-up timepoints 1 week and 3 cycles post intervention in the active arm, while no statistical significant change was observed in the placebo arm at any follow-up timepoint. Also, time-resolved metagenomic analysis showed that 50% of women treated with VMT displayed a shift of the vaginal microbiome toward the Lactobacillus-dominated donor CVS sample, confirmed with strain-level engraftment analysis. We observed no serious treatment-associated adverse events and the intervention was well tolerated. Further, the successful “twinning” of the recipient to match the donor microbiome post-VMT also led to a dramatic shift toward an anti-inflammatory environment based on transcriptomic analysis of CVS. Limitations, reasons for caution Recipients were asymptomatic and mainly Caucasian, which limits generalizability to other ethnicities and the symptomatic spectrum of vaginal dysbiosis. Due to COVID-19-related recruitment issues, the cohort size ended below the targeted number of n = 40. Future studies will aim to identify factors driving engraftment and its impact on clinical outcomes. Wider implications of the findings Vaginal microbial interventions including VMT have great potential as the first therapeutic intervention that provides effective and long-term vaginal microbiota restoration, positively impacting a wide variety of diseases and conditions affecting the female reproductive tract. Trial registration number NCT05114031

P-305 Enhancing endometrial thickness and implantation rates: a study on the effectiveness of autologous blood cell derivative - endosera in infertile women with thin endometrium

Abstract Study question Could personalized regenerative medicine based strategies promote endometrial growth and receptivity in women with refractory thin endometrium resulting in successful implantation and live births? Summary answer Personalized regenerative medicine based strategies remarkably promote endometrial growth and receptivity in women with refractory thin endometrium resulting in successful implantation and live births. What is known already Repeated implantation failure (RIF) due to thin endometrium is a formidable challenge in reproductive medicine, often steering couples toward the stressful option of surrogacy. Regenerative approaches like platelet-rich plasma (PRP) have shown promise, leveraging the anti-inflammatory and pro-regenerative properties of platelet-derived growth factors. Recognizing the need for an anti-inflammatory milieu for successful implantation, Endosera, a cell-free implantation-friendly growth factors concentrate, has been developed. This study at Bloom IVF Centre-Mumbai, India, aims to assess Endosera’s efficacy in enhancing endometrial thickness and implantation rates, providing potential solutions for RIF and thin endometrium. Study design, size, duration In this prospective self-controlled study 72 women (27-49 years) were enrolled spanning December 2022 to December 2023. Women experienced two or more failed IVF cycles and multiple cycle cancellations due to refractory thin endometrium, negative hysteroscopic screening for endometrial pathology, and negative bacteriologic screening and despite multiple failed conventional therapies were selected to undergo Endosera treatment. Participants/materials, setting, methods After obtaining informed consent, patients underwent three intrauterine infusions of Endosera on menstrual cycle Day 7, Day 12, and 2 days before frozen embryo transfer (FET). Alongside standard hormone replacement therapy (HRT), Endosera was introduced into the uterine cavity. Pregnancy confirmation relied on a positive serum β-HCG test two weeks post-ET, while clinical pregnancy was established by detecting fetal heartbeat through transvaginal ultrasound four weeks post-ET. Main results and the role of chance In this study involving 72 patients facing recurrent implantation failure due to refractory thin endometrium, the administration of three doses of Endosera demonstrated significant positive outcomes. The average endometrial thickness notably increased from 6.4 ± 1.1 mm to 8.7 ± 1.0 mm (P < 0.0001) after the 2nd Endosera dose, reaching an optimal thickness of ≥ 7mm. Among the participants, 34 achieved a positive pregnancy result (47.2%), with 6 (8.3%) delivering healthy babies and 23 (31.9%) experiencing ongoing clinical pregnancies. Adverse outcomes included 2 (2.8%) ectopic pregnancies and 3 (4.2%) miscarriages before 10 weeks, while 36 (50%) women returned negative HCG results. No adverse effects were reported, indicating the safety of Endosera. The formulation, standardized to contain 6-9 times higher growth factor levels than peripheral blood, emphasizes implantation-friendly and anti-inflammatory cytokines. This optimized composition aligns with the cyclical requirements of the endometrial phases (proliferation, secretion, and implantation), creating an angiogenic and anti-inflammatory environment crucial for successful implantation. Endosera’s safety, reproducibility, and efficacy in mimicking natural tissue repair and regeneration processes position it as a promising solution for refractory thin endometrium-related challenges. Limitations, reasons for caution This study, though demonstrating improved outcomes in refractory thin endometrium cases with Endosera treatment, acknowledges limitations such as small sample size and absence of a randomized control group. To establish Endosera as a routine adjuvant in IVF, a larger study with stringent inclusion criteria for refractory thin endometrium is recommended. Wider implications of the findings Endosera demonstrated notable success with significant improvement in implantation, pregnancy, and live birth rates for refractory thin endometrium patients prompts rigorous randomized controlled studies. These will include diverse populations, validating results and offering patients hope to reduce financial and psychological stress and challenges associated with cycle cancellations and surrogacy. Trial registration number Not applicable

Antiinflammatory and antibacterial polytetrahydropyrimidine loaded on degradable thermosensitive hydrogels for treatment of open wounds

Open wound is one of the concerns of modern medicine. The early wound has not been closed, there will be easy to enter the bacteria leading to bacterial infection. When the wound deteriorates further, the inflammation at the wound will continue to strengthen, and eventually become a chronic wound or non-healing wound. Therefore, an antibacterial anti-inflammatory tissue recovery environment will greatly promote the wound healing process. In this work, we prepared a hyaluronic acid/gelatin hydrogel that provides a comfortable recovery environment for wound healing. We also synthesized an anti-bacterial and anti-inflammatory polytetrahydropyrimidine (PTHP) polymer and loaded it into the hydrogel. The PTHP can be released in the hydrogel at a human body temperature of 37 °C, thereby achieving bactericidal and anti-inflammatory effects on the wound. Moreover, the hyaluronic acid/gelatin hydrogel can be degraded automatically at 37 °C. Therefore, PTHP-loaded hyaluronic acid/gelatin hydrogel system is an open wound care material with antibacterial, anti-inflammatory and microenvironment-responsive tissue recovery environment. It is an ideal candidate for the treatment of open wounds.

Tropoelastin modulates systemic and local tissue responses to enhance wound healing

Wound healing is facilitated by biomaterials-based grafts and substantially impacted by orchestrated inflammatory responses that are essential to the normal repair process. Tropoelastin (TE) based materials are known to shorten the period for wound repair but the mechanism of anti-inflammatory performance is not known. To explore this, we compared the performance of the gold standard Integra Dermal Regeneration Template (Integra), polyglycerol sebacate (PGS), and TE blended with PGS, in a murine full-thickness cutaneous wound healing study. Systemically, blending with TE favorably increased the F4/80+ macrophage population by day 7 in the spleen and contemporaneously induced elevated plasma levels of anti-inflammatory IL-10. In contrast, the PGS graft without TE prompted prolonged inflammation, as evidenced by splenomegaly and greater splenic granulocyte and monocyte fractions at day 14. Locally, the inclusion of TE in the graft led to increased anti-inflammatory M2 macrophages and CD4+T cells at the wound site, and a rise in Foxp3+ regulatory T cells in the wound bed by day 7. We conclude that the TE-incorporated skin graft delivers a pro-healing environment by modulating systemic and local tissue responses. Statement of significanceTropoelastin (TE) has shown significant benefits in promoting the repair and regeneration of damaged human tissues. In this study, we show that TE promotes an anti-inflammatory environment that facilitates cutaneous wound healing. In a mouse model, we find that inserting a TE-containing material into a full-thickness wound results in defined, pro-healing local and systemic tissue responses. These findings advance our understanding of TE's restorative value in tissue engineering and regenerative medicine, and pave the way for clinical applications.

Decoding the complexity of delayed wound healing following Enterococcus faecalis infection

Wound infections are highly prevalent and can lead to delayed or failed healing, causing significant morbidity and adverse economic impacts. These infections occur in various contexts, including diabetic foot ulcers, burns, and surgical sites. Enterococcus faecalis is often found in persistent non-healing wounds, but its contribution to chronic wounds remains understudied. To address this, we employed single-cell RNA sequencing (scRNA-seq) on infected wounds in comparison to uninfected wounds in a mouse model. Examining over 23,000 cells, we created a comprehensive single-cell atlas that captures the cellular and transcriptomic landscape of these wounds. Our analysis revealed unique transcriptional and metabolic alterations in infected wounds, elucidating the distinct molecular changes associated with bacterial infection compared to the normal wound healing process. We identified dysregulated keratinocyte and fibroblast transcriptomes in response to infection, jointly contributing to an anti-inflammatory environment. Notably, E. faecalis infection prompted a premature, incomplete epithelial-mesenchymal transition in keratinocytes. Additionally, E. faecalis infection modulated M2-like macrophage polarization by inhibiting pro-inflammatory resolution in vitro, in vivo, and in our scRNA-seq atlas. Furthermore, we discovered macrophage crosstalk with neutrophils, which regulates chemokine signaling pathways, while promoting anti-inflammatory interactions with endothelial cells. Overall, our findings offer new insights into the immunosuppressive role of E. faecalis in wound infections.

Decoding the complexity of delayed wound healing following Enterococcus faecalis infection.

Wound infections are highly prevalent and can lead to delayed or failed healing, causing significant morbidity and adverse economic impacts. These infections occur in various contexts, including diabetic foot ulcers, burns, and surgical sites. Enterococcus faecalis is often found in persistent non-healing wounds, but its contribution to chronic wounds remains understudied. To address this, we employed single-cell RNA sequencing (scRNA-seq) on infected wounds in comparison to uninfected wounds in a mouse model. Examining over 23,000 cells, we created a comprehensive single-cell atlas that captures the cellular and transcriptomic landscape of these wounds. Our analysis revealed unique transcriptional and metabolic alterations in infected wounds, elucidating the distinct molecular changes associated with bacterial infection compared to the normal wound healing process. We identified dysregulated keratinocyte and fibroblast transcriptomes in response to infection, jointly contributing to an anti-inflammatory environment. Notably, E. faecalis infection prompted a premature, incomplete epithelial-mesenchymal transition in keratinocytes. Additionally, E. faecalis infection modulated M2-like macrophage polarization by inhibiting pro-inflammatory resolution in vitro, in vivo, and in our scRNA-seq atlas. Furthermore, we discovered macrophage crosstalk with neutrophils, which regulates chemokine signaling pathways, while promoting anti-inflammatory interactions with endothelial cells. Overall, our findings offer new insights into the immunosuppressive role of E. faecalis in wound infections.

The role of the microscopic world: Exploring the role and potential of intratumoral microbiota in cancer immunotherapy.

Microorganisms, including bacteria, viruses, and fungi, coexist in the human body, forming a symbiotic microbiota that plays a vital role in human health and disease. Intratumoral microbial components have been discovered in various tumor tissues and are closely linked to the occurrence, progression, and treatment results of cancer. The intratumoral microbiota can enhance antitumor immunity through mechanisms such as activating the stimulator of interferon genes signaling pathway, stimulating T and NK cells, promoting the formation of TLS, and facilitating antigen presentation. Conversely, the intratumoral microbiota might suppress antitumor immune responses by increasing reactive oxygen species levels, creating an anti-inflammatory environment, inducing T cell inactivation, and enhancing immune suppression, thereby promoting cancer progression. The impact of intratumoral microbiota on antitumor immunity varies based on microbial composition, interactions with cancer cells, and the cancer's current state. A deep understanding of the complex interactions between intratumoral microbiota and antitumor immunity holds the potential to bring new therapeutic strategies and targets to cancer immunotherapy.

Leishmania exploits host cAMP/EPAC/calcineurin signaling to induce an IL-33–mediated anti-inflammatory environment for the establishment of infection

Host anti-inflammatory responses are critical for the progression of visceral leishmaniasis and the pleomorphic cytokine IL-33 was found to be upregulated in infection. The underlying mechanism is not yet known. Here, we documented that IL-33 induction is a consequence of elevated cAMP-mediated EPAC/calcineurin-dependent signaling and is essential for the sustenance of infection. L. donovani-infected RAW and bone marrow-derived macrophages showed significant up-regulation of IL-33 and its neutralization by anti-IL-33 antibody resulted in decreased parasite survival and increased inflammatory responses. Infection-induced elevated cAMP was involved in IL-33 production and of its two downstream effectors PKA and EPAC, only the latter was responsible for elevated IL-33 levels. EPAC initiated Rap-dependent phospholipase C activation, which triggered the release of intracellular calcium followed by calcium/calmodulin complex formation. Screening of calmodulin-dependent enzymes affirmed the involvement of the phosphatase calcineurin in cAMP/EPAC/calcium/calmodulin signaling-induced IL-33 production and parasite survival. Activated calcineurin thereby ensured nuclear localization of the transcription factors NFATc1 and HIF-1α for IL-33 transcription and we further ascertained their role in IL-33 transcription using a ChIP assay. Administering specific inhibitors of NFATc1 and HIF-1α in a BALB/c mouse model of visceral leishmaniasis led to decreased liver and spleen parasite burden with concomitant decrease in IL-33 levels. Splenocyte supernatants of inhibitor-treated infected mice further documented an increase in TNF-α and IL-12 with simultaneous decrease of IL-10, thereby indicating an overall disease-escalating effect of IL-33. Thus, this study demonstrates that cAMP/EPAC/calcineurin signaling is crucial for the activation of IL-33 and in effect creates anti-inflammatory responses, essential for infection.

Acellular spinal cord scaffold containing quercetin-encapsulated nanoparticles plays an anti-inflammatory role in functional recovery from spinal cord injury in rats.

Inflammatory responses play a crucial role in the pathophysiology of spinal cord injury (SCI) and developing new approaches to establish an anti-inflammatory environment for the promotion of neuroregeneration holds promise as a potential approach. In this study, our aim was to investigate the potential of combining an acellular spinal cord scaffold (ASCS) with quercetin-loaded bovine serum albumin (Qu/BSA) nanoparticles (NPs) for the treatment of SCI. The ASCS was prepared using physical and chemical methods, while the Qu/BSA NPs were prepared through a desolvation technique. The NPs exhibited favorable characteristics, including a mean size of 203nm, a zeta potential of -38, and an encapsulation efficiency of 96%. Microscopic evaluation confirmed the successful distribution of NPs on the walls of ASCS. Animal studies revealed that Qu/BSA NPs group exhibited a significant decrease in NLRP3, ASC, and Casp1 gene expression compared to the SCI group (p < 0.0001). The findings indicated a significant decrease in the NLRP3, ASC, and Casp1 protein level between the Qu/BSA/ASCS group and the SCI group (p < 0.0001). Moreover, treatment with ASCS containing either blank BSA (B/BSA) NPs or Qu/BSA NPs effectively promoted functional recovery via increasing the amount of nestin- and glial fibrillary acidic protein (GFAP)-positive cells in the site of injury. Notably, Qu/BSA/ASCS exhibited superior outcomes compared to B/BSA/ASCS. Overall, the combination of ASCS with the Qu delivery system presents a promising therapeutic approach for SCI by inhibiting inflammatory responses and promoting neuroregeneration, leading to the restoration of motor function in animals. This study demonstrates the potential of utilizing biomaterials and NPs to enhance the effectiveness of SCI treatment.

Melasma: A Step-by-Step Approach Towards a Multimodal Combination Therapy.

Melasma is a common challenge in the field of pigmentary skin disorders, exerting a significant emotional and psychosocial burden on patients. The persistent and recurring nature of melasma complicates its management in routine clinical practice. This comprehensive review outlines a stepwise, practical approach encompassing diagnostic, preventive and therapeutic strategies for the management of melasma. A thorough exploration of aggravating and exacerbating factors, including sun exposure, hormonal imbalances, photosensitizing medication and cosmetics, is essential for a holistic assessment of the disease. With an emphasis on consistent and effective photoprotection, initial topical treatment modalities target the melanin production and/or the transfer of melanosomes to keratinocytes. Topical tyrosine inhibitors emerge as the first choice for reducing and preventing hyperpigmentation, with compounds such as thiamidol or tranexamic acid (TXA) being preferred for their safety profile over hydroquinone (HQ), kojic acid and arbutin. Combination with chemical peels can further enhance the therapeutic efficacy, even in cases with resistant melasma. In more severe cases, laser- and light-based interventions may be considered, but with the caveat of the likelihood of recurrence within 3-6 months. Assisted TXA delivery, via either fractional non-ablative laser or microneedling techniques, can further improve clinical outcomes. In conclusion, an optimal melasma management strategy is a multimodal approach, which includes effective photoprotection and a mix of different topical treatments targeting melanin synthesis, the anti-inflammatory environment, senescence and vascularity. Complementary procedures, such as chemical peels, and laser, light-based or microneedling procedures, with or without TXA, can further expedite melanin clearance in more severely affected instances. Individual discussions with patients regarding treatment expectations, recurrence likelihood and potential side effects are paramount to a comprehensive and successful therapeutic journey.

Effects of SNPs on TNF-α and IL-10 cytokine expression in TB and HIV patients in the Capricorn district, Limpopo Province, South Africa.

The impact of Tuberculosis (TB) places an immense burden on the health care system. Infection with Human Immunodeficiency Virus (HIV) is a significant risk factor in the development and progression of TB disease. Single Nucleotide Polymorphisms (SNPs) in the promoter region of Interleukin-10 (IL-10) and Tumour Necrotic Factor-Alpha (TNF-α) may play a major role in the disease mechanism and understanding these mechanisms might prove to be a useful diagnostic tool in evaluating the immune regulation and progression of the disease. This study aimed to determine the relationship between cytokine levels and gene variants of Interleukin-10 and Tumour Necrotic Factor Alpha in TB and HIV-infected participants. Cytokine levels were determined by ELISA, and SNPs were determined by MassArray®. The levels of TNF-α were higher in the TB group than the HIV (p < 0.001) and TB-HIV (p = 0.011) groups, but similar to the TNF-α levels in the control group. In the HIV group, IL-10 levels were higher than those of the TB (p < 0.001) and control groups (p = 0.039), whereas there was no difference between the IL-10 levels in the HIV and the TB-HIV infection groups. The ratio was determined and there were no differences between the four infection groups. In this study, no associations were detected between the circulating plasma levels of TNF-α and IL-10 and their genotypes. Our data showed that the gene variants were not associated with circulating plasma levels of TNF-α and IL-10 in our study population. A pro-inflammatory environment was found in the TB and TB-HIV groups, which is suggesting of bacterial clearance, while an anti-inflammatory environment was found in the HIV group, which suggests the suppression of viral replication.

Extracellular Purine Metabolism-Potential Target in Multiple Sclerosis.

The purinergic signaling system comprises a complex network of extracellular purines and purine-metabolizing ectoenzymes, nucleotide and nucleoside receptors, ATP release channels, and nucleoside transporters. Because of its immunomodulatory function, this system is critically involved in the pathogenesis of multiple sclerosis (MS) and its best-characterized animal model, experimental autoimmune encephalomyelitis (EAE). MS is a chronic neuroinflammatory demyelinating and neurodegenerative disease with autoimmune etiology and great heterogeneity, mostly affecting young adults and leading to permanent disability. In MS/EAE, alterations were detected in almost all components of the purinergic signaling system in both peripheral immune cells and central nervous system (CNS) glial cells, which play an important role in the pathogenesis of the disease. A decrease in extracellular ATP levels and an increase in its downstream metabolites, particularly adenosine and inosine, were frequently observed at MS, indicating a shift in metabolism toward an anti-inflammatory environment. Accordingly, upregulation of the major ectonucleotidase tandem CD39/CD73 was detected in the blood cells and CNS of relapsing-remitting MS patients. Based on the postulated role of A2A receptors in the transition from acute to chronic neuroinflammation, the association of variants of the adenosine deaminase gene with the severity of MS, and the beneficial effects of inosine treatment in EAE, the adenosinergic system emerged as a promising target in neuroinflammation. More recently, several publications have identified ADP-dependent P2Y12 receptors and the major extracellular ADP producing enzyme nucleoside triphosphate diphosphohydrolase 2 (NTPDase2) as novel potential targets in MS.