Collagen Type Research Articles

Dextromethorphan inhibits collagen and collagen-like cargo secretion to ameliorate lung fibrosis.

Excessive deposition of fibrillar collagen in the interstitial extracellular matrix (ECM) of human lung tissue causes fibrosis, which can ultimately lead to organ failure. Despite our understanding of the molecular mechanisms underlying the disease, no cure for pulmonary fibrosis has yet been found. We screened a drug library and found that dextromethorphan (DXM), a cough expectorant, reduced the amount of excess fibrillar collagen deposited in the ECM in cultured primary human lung fibroblasts, a bleomycin mouse model, and a cultured human precision-cut lung slice model of lung fibrosis. The reduced extracellular fibrillar collagen upon DXM treatment was due to reversible trafficking inhibition of collagen type I (COL1) in the endoplasmic reticulum (ER) in TANGO1- and HSP47-positive structures. Mass spectrometric analysis showed that DXM promoted hyperhydroxylation of proline and lysine residues on various collagens (COL1, COL3, COL4, COL5, COL7, and COL12) and latent transforming growth factor-β-binding protein (LTBP1 and LTBP2) peptides, coinciding with their secretion block. Additionally, proteome profiling of DXM-treated cells showed increased thermal stability of prolyl-hydroxylases P3H2, P3H3, P3H4, P4HA1, and P4HA2, suggesting a change in their activity. Transcriptome analysis of profibrotic stimulated primary human lung fibroblasts and human ex vivo lung slices after DXM treatment showed activation of an antifibrotic program through regulation of multiple pathways, including the MMP-ADAMTS axis, WNT signaling, and fibroblast-to-myofibroblast differentiation. Together, these data obtained from in vitro, in vivo, and ex vivo models of lung fibrogenesis show that DXM has the potential to limit fibrosis through inhibition of COL1 membrane trafficking in the ER.

Collagen type III formation but not degradation is associated with risk of kidney disease progression and mortality after acute kidney injury

Abstract Background Acute kidney injury (AKI), a rapid decrease in kidney function, is associated with increased risk of adverse outcomes including development and progression of CKD. Kidney fibrosis is one of the pathological processes central to this AKI to CKD transition. Here we investigate the association of biomarkers of collagen type III turnover with adverse outcome following AKI. Methods We measured three biomarkers reflecting collagen type III formation (PRO-C3) and degradation (C3M and C3C) in plasma samples collected 1 year after an episode of AKI in 800 patients (392 patients with AKI and 408 non-AKI controls) from the prospective AKI Risk in Derby (ARID) study. Patients were followed until 3 years after the episode of AKI and the following outcomes were assessed: kidney disease progression, mortality, heart failure, CV events, and hospital readmission. Results PRO-C3 levels were elevated in the AKI group compared to the controls (P<0.001), whereas C3M and C3C levels were not different between groups. In multivariate models including common risk factors, PRO-C3 was prognostic for kidney disease progression and mortality in the AKI group and for heart failure in the control group. C3M and C3C were not prognostic for any of the investigated outcomes. Conclusions Circulating PRO-C3, a biomarker of fibroblast activity, was prognostic for kidney disease progression and mortality when measured one year after an episode of AKI. Biomarkers of fibroblast activity may help patient stratification after an episode of AKI by identifying patients at higher risk of kidney disease progression.

The Multifaced Role of Collagen in Cancer Development and Progression

Collagen is a crucial protein in the extracellular matrix (ECM) essential for preserving tissue architecture and supporting crucial cellular functions like proliferation and differentiation. There are twenty-eight identified types of collagen, which are further divided into different subgroups. This protein plays a critical role in regulating tissue homeostasis. However, in solid tumors, the balance can be disrupted, due to an abundance of collagen in the tumor microenvironment, which significantly affects tumor growth, cell invasion, and metastasis. It is important to investigate the specific types of collagens in cancer ECM and their distinct roles in tumor progression to comprehend their unique contribution to tumor behavior. The diverse pathophysiological functions of different collagen types in cancers illustrate collagen’s dual roles, offering potential therapeutic options and serving as prognostic markers.

Innovative Ink-Based 3D Hydrogel Bioprinted Formulations for Tissue Engineering Applications

Three-dimensional (3D) models with improved biomimicry are essential to reduce animal experimentation and drive innovation in tissue engineering. In this study, we investigate the use of alginate-based materials as polymeric inks for 3D bioprinting of osteogenic models using human bone marrow stem/stromal cells (hBMSCs). A composite bioink incorporating alginate, nano-hydroxyapatite (nHA), type I collagen (Col) and hBMSCs was developed and for extrusion-based printing. Rheological tests performed on crosslinked hydrogels confirm the formation of solid-like structures, consistently indicating a superior storage modulus in relation to the loss modulus. The swelling behavior analysis showed that the addition of Col and nHA into an alginate matrix can enhance the swelling rate of the resulting composite hydrogels, which maximizes cell proliferation within the structure. The LIVE/DEAD assay outcomes demonstrate that the inclusion of nHA and Col did not detrimentally affect the viability of hBMSCs over seven days post-printing. PrestoBlueTM revealed a higher hBMSCs viability in the alginate-nHA-Col hydrogel compared to the remaining groups. Gene expression analysis revealed that alginate-nHA-col bioink favored a higher expression of osteogenic markers, including secreted phosphoprotein-1 (SPP1) and collagen type 1 alpha 2 chain (COL1A2) in hBMSCs after 14 days, indicating the pro-osteogenic differentiation potential of the hydrogel. This study demonstrates that the incorporation of nHA and Col into alginate enhances osteogenic potential and therefore provides a bioprinted model to systematically study osteogenesis and the early stages of tissue maturation in vitro.

A prospective study of the effect of laparoscopic splenectomy and azygoportal disconnection on liver synthetic function and liver cirrhosis

Objective: To investigate the effect of laparoscopic splenectomy and azygoportal disconnection (LSD) on liver synthesis and development of liver cirrhosis. Methods: This is a prospective case series study.The clinical data of liver cirrhotic patients with portal hypertension who received LSD at the Department of Hepatobiliary Surgery of Northern Jiangsu People's Hospital Affiliated to Yangzhou University from September 2014 to January 2016 were included. This study analyzed the diameter of the portal vein, the velocity of portal blood flow, the routine blood parameters, the liver function, the synthetic proteins of liver (antithrombin Ⅲ (AT-Ⅲ), protein S, protein C), and the serum content of liver fibrotic markers(collagen type Ⅳ, procollagen type Ⅲ, laminin, hyaluronidase). Repeated measures ANOVA was used for comparison between multiple groups, and least significance difference was used for post-hoc multiple comparison. Results: A total of 106 patients were included in the study, including 70 males and 36 females, aged (51.8±9.8) years(range: 28 to 75 years).Compared with the preoperative results, the diameter of portal vein and the velocity of portal vein decreased after surgery (F=14.03, 12.15, respectively, both P<0.01). Compared with the preoperative results, the total bilirubin, albumin, prothrombin time, international normalized ratio, Child-Pugh score and classification were improved (F=17.96, 56.01, 66.63, 35.83, 33.49, and 27.50, respectively, all P<0.01), and the AT-Ⅲ, protein S, protein C,collagen type Ⅳ, procollagen type Ⅲ, laminin and hyaluronidase levels were also improved (F=47.87, 36.26, 18.02, 2.79, 14.58, 44.35, and 14.38, respectively, all P<0.01). Compared with the preoperative period, the diameter of portal vein was reduced from the first week to the 24th month after surgery (t=5.45 to 9.39, all P<0.01). Compared with the preoperative period, the velocity of portal vein blood from the first week after surgery to the 24th month after surgery was decreased (t=4.02 to 8.43, all P<0.01). Compared with the preoperative period, routine blood parameters (white blood count, hemoglobin, platelet count), liver function (total bilirubin, albumin, prothrombin time, international normalized ratio, Child-Pugh score), liver synthetic protein (AT-Ⅲ, protein S, protein C) and liver fibrotic markers (collagen type Ⅳ, procollagen type Ⅲ, laminin, hyaluronidase) were improved to varying degrees at the 24th month after surgery (t=-20.46 to 11.93, all P<0.01). Conclusion: Preliminary findings show that LSD can reduce portal vein pressure, restore blood cell number, and improve liver synthesis function and the degree of liver fibrosis in patients with portal hypertension in liver cirrhosis.

Wnt5a manipulate the progression of osteoarthritis via MMP-13 dependent signaling pathway.

The object of this study was to propose a Wnt5a-matrix metalloproteinase (MMP)-13 dependent signaling axis for osteoarthritis (OA) progression. To this end, the chondrocytes were isolated from both OA patients and normal controls. The chondrocytes were treated with diverse concentrations of Wnt5a (0, 50, 100, and 200 ng/mL), respectively. The expression levels of Wnt5a, MMP-13, and Collagen type II were examined using reverse transcription-polymerase chain reaction and western blotting. At the same time, the cell proliferation and cell apoptosis of chondrocytes were also observed. Compared with control tissues, the activities of Wnt5a and MMP-13 were significantly enhanced in chondrocytes of OA patients. Treated with different concentrations of Wnt5a (0, 50, 100, and 200 ng/mL), chondrocyte cell proliferation was clearly downregulated. At the same time, the chondrocyte cell apoptosis was obviously accelerated. The expression pattern of Collagen type II was same as cell proliferation manner. Co-treatment of MMP-13 siRNA could significantly compensate the functions of Wnt-5a administration, suggesting MMP-13 was a direct target of Wnt-5a. Collectively, the study speculated a novel Wnt5a-MMP-13 molecular mechanism for OA progression and shed an innovative signaling axis for the disorder.

Multi-omic profiling highlights factors associated with resistance to immuno-chemotherapy in non-small-cell lung cancer.

Although immune checkpoint blockade (ICB) therapies have shifted the treatment paradigm for non-small-cell lung cancer (NSCLC), many patients remain resistant. Here we characterize the tumor cell states and spatial cellular compositions of the NSCLC tumor microenvironment (TME) by analyzing single-cell transcriptomes of 232,080 cells and spatially resolved transcriptomes of tumors from 19 patients before and after ICB-chemotherapy. We find that tumor cells and secreted phosphoprotein 1-positive macrophages interact with collagen type XI alpha 1 chain-positive cancer-associated fibroblasts to stimulate the deposition and entanglement of collagen fibers at tumor boundaries, obstructing T cell infiltration and leading to poor prognosis. We also reveal distinct states of tertiary lymphoid structures (TLSs) in the TME. Activated TLSs are associated with improved prognosis, whereas a hypoxic microenvironment appears to suppress TLS development and is associated with poor prognosis. Our study provides novel insights into different cellular and molecular components corresponding to NSCLC ICB-chemotherapeutic responsiveness, which will benefit future individualized immuno-chemotherapy.

Phytoestrogens as Natural Anti-Aging Solutions for Enhanced Collagen Synthesis in Skin.

The dermal extracellular matrix (ECM) is a dynamic scaffold composed mainly of proteins, with collagen as the key structural component providing resilience and support to the skin. Post-menopause, declining estrogen levels lead to a significant reduction in skin health, notably a 30% decrease in collagen types I and III within 5 years. To discover natural extracts that stimulate collagen production. We utilized PathwayStudio to analyze protein-protein interactions and identify regulators of essential collagen types. Our study assessed Glycyrrhiza uralensis extract's ability to boost collagen production and enhance dermal density both invitro and invivo. PathwayStudio analysis highlighted phytoestrogens including glycyrrhizin, isoliquiritigenin, liquiritigenin, liquiritin, and glabrol, as potential candidates. Liquorice rhizome (G. uralensis), used in traditional Chinese medicine, is rich in phytoestrogens like liquiritigenin. The G. uralensis extract increased collagen I and III gene expression and pro-collagen I protein levels in human dermal fibroblasts and inhibited UVB and pollution-induced matrix metalloproteinase-1 (MMP1) production. In an invivo study, a topical formulation containing the extract significantly improved dermal density after 56 days, measured by the DUB SkinScanner. These findings suggest G. uralensis extract as a promising agent for enhancing collagen production and skin health, particularly in postmenopausal women. Further research is needed to explore its mechanisms and long-term effects.

Chondrogenic commitment of human umbilical cord blood and umbilical cord-derived mesenchymal stem cells induced by the supernatant of chondrocytes: A comparison study.

Native cartilage has low capacity for regeneration because it has very few progenitor cells. Human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) and human umbilical cord-derived MSCs (hUC-MSCs) have been employed as promising sources of stem cells for cartilage injury repair. Reproduction of hyaline cartilage from MSCs remains a challenging endeavor. The paracrine factors secreted by chondrocytes possess the capability to induce chondrogenesis from MSCs. The conditioned medium derived from chondrocytes was utilized to induce chondrogenic differentiation of hUCB-MSCs and hUC-MSCs. The expression levels of collagen type I alpha 1 chain (Col1a1), collagen type II alpha 1 chain (Col2a1), and SRY-box transcription factor 9 (SOX9) were assessed through quantitative real-time polymerase chain reaction (qRT-PCR), Western blot (WB), and immunofluorescence (IF) assays. To elucidate the mechanism of differentiation, the concentration of transforming growth factor-β1 (TGF-β1) in the conditioned medium of chondrocytes was quantified using enzyme-linked immunosorbent assay (ELISA). Meanwhile, the viability of cells was assessed using Cell Counting Kit-8 (CCK-8) assays. The expression levels of Col2a1 and SOX9 were found to be higher in induced hUC-MSCs compared to those in induced hUCB-MSCs. The conditioned medium of chondrocytes contained TGF-β1. The CCK-8 assays revealed that the proliferation rate of hUC-MSCs was significantly higher compared to that of hUCB-MSCs. The chondrogenic potential and proliferation capacity of hUC-MSCs surpass those of hUCB-MSCs, thereby establishing hUC-MSCs as a superior source of seed cells for cartilage tissue engineering.

Effect of Dendrobium nobile powder combined with conventional therapy on mild to moderate fatty liver.

Nonalcoholic fatty liver disease (NAFLD) encompasses a variety of liver conditions impacting individuals who consume minimal or no alcohol. Recently, traditional Chinese medicine has been gradually used to treat mild to moderate fatty liver, among which Dendrobium nobile Lindl. powder has been affirmed by many doctors and patients to be effective. However, there is limited research on combining this treatment with standard therapies for mild to moderate NAFLD. To survey the effect of combining Dendrobium nobile Lindl. powder with standard treatment on liver function and lipid metabolism disorder in patients with mild to moderate NAFLD. Eighty patients with mild to moderate NAFLD participated in this retrospective study, classified into two groups: The observation group (n = 40) and the control group (n = 40). In November 2020 and November 2022, the study was conducted at People's Hospital of Chongqing Liang Jiang New Area. The control group received standard treatment, while the observation group received Dendrobium nobile Lindl. powder based on the control group. The study compared differences in traditional Chinese medicine clinical syndrome scores, liver fibrosis treatment, liver function indicators, lipid levels, and serum inflammatory factor levels before and after treatment, and we calculated the incidence of adverse reactions for both groups. The total effective rate was 97.50% in the observation group and 72.5% in the control group. After 8 weeks of treatment, the main and secondary symptom scores remarkably decreased, especially in the observation group (P < 0.05), and there was a significant reduction in the serum levels of hyaluronic acid (HA), laminin (LN), human rocollagen III (PC III), and collagen type IV (CIV). The levels of HA, LN, PC III, and CIV were significantly lower in the observation group (P < 0.05). After 8 weeks, both groups indicated remarkable improvements in liver function and blood lipid levels, with the observation group having even lower levels (P < 0.05). Serum levels of interleukin-1β, tumor necrosis factor-α, and interleukin-8 also dropped significantly. The observation group had a lower rate of adverse reactions (5.00%) compared to the control group (22.50%). Adding Dendrobium nobile Lindl. powder to standard treatment has been found to remarkably improve symptoms and reduce inflammation in patients with mild to moderate fatty liver disease. It also enhances hepatic function and lipid profile, ameliorates liver fibrosis indices, and lowers the risk of side effects. Consequently, this therapeutic protocol shows promise for clinical implementation and dissemination.

Modulation of Biomaterial-Associated Fibrosis by Means of Combined Physicochemical Material Properties.

Biomaterial-associated fibrosis remains a significant challenge in medical implants. To optimize implant design, understanding the interplay between biomaterials and host cells during the foreign body response (FBR) is crucial. Material properties are known to influence cellular behavior and can be used to manipulate cell responses, but predicting the right combination for the desired outcomes is challenging. This study exploreshow combined physicochemical material properties impact early myofibroblast differentiation using the Biomaterial Advanced Cell Screening (BiomACS) technology, which assesseshundreds of combinations of surface topography, stiffness, and wettability in a single experiment. Normal human dermal fibroblasts (NHDFs) are screened for cell density, area, and myofibroblast markers α-smooth muscle actin (α-SMA) and Collagen type I (COL1) after 24hand 7 days of culture, with or without transforming growth factor-beta (TGF-β). Results demonstrated that material properties influence fibroblast behavior after 7 days with TGF-β stimulation, with wettability emerging as the predominant factor, followed by stiffness. The study identified regions with increased cell adhesion while minimizing myofibroblast differentiation, offering the potential for implant surface optimization to prevent fibrosis. This research provides a powerful tool for cell-material studies andrepresents a critical step toward enhancing implant properties and reducing complications, ultimately improving patient outcomes.

Self-Assembled DNA-Collagen Bioactive Scaffolds Promote Cellular Uptake and Neuronal Differentiation.

Different modalities of DNA/collagen complexes have been utilized primarily for gene delivery studies. However, very few studies have investigated the potential of these complexes as bioactive scaffolds. Further, no studies have characterized the DNA/collagen complex formed from the interaction of the self-assembled DNA macrostructure and collagen. Toward this investigation, we report herein the fabrication of novel bioactive scaffolds formed from the interaction of sequence-specific, self-assembled DNA macrostructure and collagen type I. Varying molar ratios of DNA and collagen resulted in highly intertwined fibrous scaffolds with different fibrillar thicknesses. The formed scaffolds were biocompatible and presented as a soft matrix for cell growth and proliferation. Cells cultured on DNA/collagen scaffolds promoted the enhanced cellular uptake of transferrin, and the potential of DNA/collagen scaffolds to induce neuronal cell differentiation was further investigated. The DNA/collagen scaffolds promoted neuronal differentiation of precursor cells with extensive neurite growth in comparison to the control groups. These novel, self-assembled DNA/collagen scaffolds could serve as a platform for the development of various bioactive scaffolds with potential applications in neuroscience, drug delivery, tissue engineering, and in vitro cell culture.

ULK1 methylation promotes TGF-β1-induced endometrial fibrosis via the FOXP1/DNMT1 axis.

Intrauterine adhesion (IUA) is the second most common cause of secondary infertility in women and can also lead to menstrual abnormalities and multiple adverse pregnancy outcomes. Therefore, elucidating the mechanism of its development is crucial for the prevention and treatment of IUA. This study will investigate the function and mechanism of forkhead box P1 (FOXP1)/DNA methyltransferase 1 (DNMT1)/unc-51-like autophagy activating kinase 1 (ULK1) in IUA. Expression levels of key genes were detected using western blot and quantitative - real time reverse transcription polymerase chain reaction. Cell proliferation was detected by CCK-8 and EdU staining. Transcriptional regulation relationships were detected by dual luciferase reporter gene and chromatin immunoprecipitation (ChIP) assay. Methylation station of ULK1 was detected by methylmion specific PCR (MSP). Fibrosis and pathological changes in the uterine cavity tissues were detected by Masson and hematoxylin and eosin staining. It was observed that the expression of FOXP1 and DNMT1 increased in transforming growth factor (TGF)-β1-induced cells, while ULK1 expression decreased. Downregulation of FOXP1 could inhibit human endometrial stromal cells proliferation and autophagy, as well as decrease the expression of fibrogenic factors (collagen type I alpha 1 chain [COL1A1], fibronectin [FN], and alpha-smooth muscle actin [α-SMA]). The results of MSP and ChIP experiments showed that DNMT1 promotes methylation of the ULK1 promoter region and inhibits its transcription. In an animal model, knockdown of FOXP1 alleviated pathological fibrosis and uterine adhesions. Knockdown of FOXP1 can inhibit endometrial fibrosis in IUA rats; FOXP1 could be a potential target for the treatment of IUA.

CHANGES IN TRYPTOPHAN METABOLISM AND CYTOKINE CONCENTRATION IN SYNOVIAL JOINT TISSUE IN RHEUMATOID ARTHRITIS

The aim of the research. To identify changes in tryptophan and cytokine metabolism in synovial joint tissue in rheumatoid arthritis. Research materials and methods. The experiments were performed on 40 WISTAR rats. At the beginning of the study, each animal of the experimental groups received an intraperitoneal injection of collagen type 2 solution (Chondrex, Inc., USA) in Freund's incomplete adjuvant. The content of tryptophan, kynurenine, 3-hydroxykinurinine, L-5-hydrotryptophan by HPLC with fluorimetric and spectrophotometric detection, and the concentration of cytokines: IL1b, IL6, IL10, IL17, IL18, TNFa were determined in the tissues of the synovial membrane of the knee joint. They were determined using Rat Inflammation Panel V02 multiplex assay kits (Biolegend, USA) on a Cytomics FC500 flow cytofluorimeter (Beckman Coulter, USA). Statistical analysis was performed using the Jamovi program version 2.3.Results. The concentration of cytokines: IL1b, IL6, IL10, IL17, IL18, TNFa was found to be high during the acute development of rheumatoid arthritis. With experimental rheumatoid arthritis, the content of tryptophan metabolites along the kynurenine pathway increases and the concentration of metabolites along the serotonin pathway decreases. Direct positive correlations of IL1b, IL6, IL10, IL17, IL18, TNFa with the content of tryptophan metabolites along the kynurenine pathway have been established.Conclusions. Tryptophan metabolism and IL1b, IL6, IL10, IL17, IL18, and TNFa content are altered in rheumotoid arthritis.

FRESH 3D Bioprinting of Collagen Types I, II, and III.

Collagens play a vital role in the mechanical integrity of tissues as well as in physical and chemical signaling throughout the body. As such, collagens are widely used biomaterials in tissue engineering; however, most 3D fabrication methods use only collagen type I and are restricted to simple cast or molded geometries that are not representative of native tissue. Freeform reversible embedding of suspended hydrogel (FRESH) 3D bioprinting has emerged as a method to fabricate complex 3D scaffolds from collagen I but has yet to be leveraged for other collagen isoforms. Here, we developed collagen type II, collagen type III, and combination bioinks for FRESH 3D bioprinting of millimeter-sized scaffolds with micrometer scale features with fidelity comparable to scaffolds fabricated with the established collagen I bioink. At the microscale, single filament extrusions were similar across all collagen bioinks with a nominal diameter of ∼100 μm using a 34-gauge needle. Scaffolds as large as 10 × 10 × 2 mm were also fabricated and showed similar overall resolution and fidelity across collagen bioinks. Finally, cell adhesion and growth on the different collagen bioinks as either cast or FRESH 3D bioprinted scaffolds were compared and found to support similar growth behaviors. In total, our results expand the range of collagen isoform bioinks that can be 3D bioprinted and demonstrate that collagen types I, II, III, and combinations thereof can all be FRESH printed with high fidelity and comparable biological response. This serves to expand the toolkit for the fabrication of tailored collagen scaffolds that can better recapitulate the extracellular matrix properties of specific tissue types.

Sphingosine 1-phosphate acts as proliferative and fibrotic cue in leiomyoma cells

ObjectiveTo determine whether the bioactive sphingolipid sphingosine 1-phosphate (S1P) modulates cellular proliferation and synthesis of fibrotic proteins in leiomyoma differently than myometrial cells. DesignA basic science study using human leiomyoma and myometrial cells. SettingAcademic laboratory. ExposureLeiomyoma and myometrial cells were treated with S1P, as well as with selective antagonists for S1P specific G-protein coupled receptors (S1PR) and secondarily with inhibitors of extracellular-signal regulated kinase 1/2 (ERK1/2) and ezrin. Main Outcome Measure(s)Cellular proliferation and fibrogenesis. Bromodeoxyuridine (BrdU) cell proliferation assay was employed to measure DNA synthesis and proliferation, while Western Blot analysis was used to assess the expression of the fibrotic markers N-cadherin, alpha Smooth Muscle Actin (αSMA), transgelin and Collagen type I alpha 1. Result(s)S1P stimulates cellular proliferation of leiomyoma but not myometrial cells. The mitogenic effect elicited by S1P relies on the specific engagement of its specific receptor S1P2 and is mediated by ERK1/2 and ezrin activation. Furthermore, S1P exerts a pro-fibrotic effect in a S1PR-dependent manner in leiomyoma but not myometrial cells. Conclusion(s)These results, beside extending the knowledge on the molecular mechanism underlying uterine leiomyoma development and fibrosis, demonstrate the pathogenetic role of S1P in leiomyoma and support the rationale for targeting S1P signaling pathway as innovative potential treatment.

Identification of SNPs and INDELS associated with duck egg quality traits through a genome-wide association analysis

Egg quality traits are economically important in the poultry industry. To explore the genetic architecture and identify potential candidate genes, a genome-wide association study (GWAS) was performed for 13 egg quality traits using data from whole-genome sequencing of 299 Longyan Shan-ma female ducks, including 12 quantitative traits and one qualitative trait, eggshell color (ESC; white, light green, green). From estimation of pedigree genetic parameters, heritability (h2) ranged from 0.022 to 0.996 for the 12 quantitative traits, with the highest h2 (0.996) for eggshell color a* value (ESCA) and the lowest h2 (0.022) for egg yolk percentage relative to EW. A total of 8,874 single nucleotide polymorphism (SNP)-based significant associations (1.0 × 10-6) and 247 insertion-deletion (indel)-based significant associations (1.00 × 10-5) were identified, including 5,980 SNPs and 159 indel markers. From 5,924 SNPs and 143 indels associated with ESC traits, 181 potential candidate genes were identified, and most significant SNPs and indels (P < 1.0 × 10-20) were located at 1.86 Mb (44.29-46.15 Mb) on chromosome 4. The top SNP (chr4:45325309:C>A; P = 7.97 × 10-43) and the top indel (chr4:45299595:delTTCCACTCCAC; P = 4.20 × 10-36) for the ESC a* value were within two known ESC candidate genes; ATP-binding cassette subfamily G member 2 (ABCG2) and protein kinase cGMP-dependent 2 (PRKG2). Of 56 SNPs and 16 indels associated with other egg quality traits, 46 potential candidate genes were identified including synapse differentiation-inducing 1-like (SYNDIG1L) for EW, and core histone macro-H2A.1 (LOC101795967) and neurogenin 1 (NEUROG1) for egg shape index; and four genes including collagen type VI alpha 3 chain (COL6A3), lysine demethylase 7A (KDM7A), LOC101802169, and sperm-associated antigen 16 (SPAG16) for egg yolk weight and the percentage of yolk to total egg weight. Of the 46 genes, the molecular functions of 22 are related to protein binding, indicating important roles in the formation of egg quality traits. Our findings provide new insight into the genetic basis of egg quality traits in ducks.

OSCC-derived EVs educate fibroblasts and remodel collagen landscape

Cancer-associated myofibroblasts (mCAFs) represent a significant component of the tumor microenvironment due to their contributions to extracellular matrix (ECM) remodeling. The pro-tumor mechanisms of extracellular vesicles (EVs) by regulating mCAFs and related collagens remain poorly understood in oral squamous cell carcinoma (OSCC). In this study, through analysis of single-cell sequencing data and immunofluorescence staining, we confirmed the increased presence of mCAFs and enrichment of specific collagen types in OSCC tissues. Furthermore, we demonstrated that OSCC-derived EVs promote the transformation of fibroblasts into mCAFs, leading to tumor invasion. Proteomic analysis identified the presence of TGF-β1 in EVs and revealed its role in inducing mCAFs via the TGF-β1/Smad3 signaling pathway. Experiments in vivo confirmed that EVs, particularly those carrying TGF-β1, trigger COL18high COL5high matrix deposition, thereby forming the pro-tumor ECM in OSCC. In summary, our investigation unveils the significant involvement of OSCC-derived EVs in orchestrating the differentiation of fibroblasts into mCAFs and modulating specific collagen types within the ECM. Therefore, this study provides a theoretical basis for targeting the EV-mediated TGF-β1 signaling pathway as a potential therapeutic strategy for OSCC.

Application of miR-29a-Exosome and Multifunctional Scaffold for Full-Thickness Cartilage Defects

BackgroundFull-thickness cartilage defect is a common refractory disease in orthopedics. In this study, we designed a novel composite scaffold composed of silk fibroin-chitosan for the cartilage layer and porous tantalum for the subchondral bone layer, loaded with engineered bone mesenchymal stem cell exosomes, to evaluate its efficacy in repairing full-thickness cartilage defect. MethodsPorous tantalum was 3D printed and combined with silk fibroin-chitosan to form a composite scaffold. Chondrocytes were cultured on the scaffold, and their growth was assessed using the CCK-8 method. Toluidine blue staining confirmed cell morphology, while immunofluorescence revealed collagen type Ⅱ expression. Engineered exosomes loaded with miR-29a were created and characterized using various techniques. Co-culturing with chondrocytes demonstrated their proliferation over 10 days. Immunofluorescence revealed staining for the nucleus, collagen type II, and Aggrecan. In vivo experiments were performed on rats to assess cartilage defect repair, utilizing histological staining and micro-CT scanning at 4 and 8 weeks post-operation. ResultsThe silk fibroin-chitosan scaffold demonstrated good biocompatibility, supporting chondrocyte adhesion, growth, and cartilage tissue formation. Engineered exosomes exhibited promising biological activity, conducive to bone and cartilage regeneration. The implantation of the silk fibroin-chitosan/porous tantalum composite scaffold loaded with engineered exosomes promoted integration with the surrounding bone and cartilage tissues, facilitating repair and regeneration. ConclusionsThe silk fibroin-chitosan combined with porous tantalum scaffold carrying engineered exosomes loaded with miR-29a has good potential for full-thickness cartilage defects regeneration.

Effect of metabolic disorders on reactive gliosis and glial scarring at the early subacute phase of stroke in a mouse model of diabetes and obesity

It is well recognized that type II Diabetes (T2D) and overweight/obesity are established risk factors for stroke, worsening also their consequences. However, the underlying mechanisms by which these disorders aggravate outcomes are not yet clear limiting the therapeutic opportunities. To fill this gap, we characterized, for the first time, the effects of T2D and obesity on the brain repair mechanisms occurring 7 days after stroke, notably glial scarring. In the present study, by performing a 30-minute middle cerebral artery occlusion (MCAO) on db/db (obese diabetics mice) and db/+ (controls) mice, we demonstrated that obese and diabetic mice displayed larger lesions (i.e. increased infarct volume, ischemic core, apoptotic cell number) and worsened neurological outcomes compared to their control littermates. We then investigated the formation of the glial scar in control and db/db mice 7 days post-stroke. Our observations argue in favor of a stronger and more persistent activation of astrocytes and microglia in db/db mice. Furthermore, an increased deposition of extracellular matrix (ECM) was observed in db/db vs control mice (i.e. chondroitin sulfate proteoglycan and collagen type IV). Consequently, we demonstrated for the first time that the db/db status is associated with increased astrocytic and microglial activation 7 days after stroke and resulted in higher deposition of ECM within the damaged area. Interestingly, the injury-induced neurogenesis appeared stronger in db/db as shown by the labeling of migrating neuroblast. This increase appeared correlated to the larger size of lesion. It nevertheless raises the question of the functional integration of the new neurons in db/db mice given the observed dense ECM, known to be repulsive for neuronal migration. Carefully limiting glial scar formation after stroke represents a promising area of research for reducing neuronal loss and limiting disability in diabetic/obese patients.