Collagen Content Research Articles

Insulin-like growth factor-1 infusion in preterm piglets does not affect growth parameters of skeletal muscle or tendon tissue.

Prematurity has physical consequences, such as lower birth weight, decreased muscle mass and increased risk of adult-onset metabolic disease. Insulin-like growth factor 1 (IGF-1) has therapeutic potential to improve the growth and quality of muscle and tendon in premature births, and thus attenuate some of these sequalae. We investigated the effect of IGF-1 on extensor carpi radialis muscle and biceps brachii tendon of preterm piglets. The preterm group consisted of 19-day-old preterm (10 days early) piglets, treated with either IGF-1 or vehicle. Term controls consisted of groups of 9-day-old piglets (D9) and 19-day-old piglets (D19). Muscle samples were analysed by immunofluorescence to determine the cross-sectional area (CSA) of muscle fibres, fibre type composition, satellite cell content and central nuclei-containing fibres in the muscle. Tendon samples were analysed for CSA, collagen content and maturation, and vascularization. Gene expression of the tendon was measured by RT-qPCR. Across all endpoints, we found no significant effect of IGF-1 treatment on preterm piglets. Preterm piglets had smaller muscle fibre CSA compared to D9 and D19 control group. Satellite cell content was similar across all groups. For tendon, we found an effect of age on tendon CSA, and mRNA levels of COL1A1, tenomodulin and scleraxis. Immunoreactivity for elastin and CD31, and several markers of tendon maturation, were increased in D9 compared to the preterm piglets. Collagen content was similar across groups. IGF-1 treatment of preterm-born piglets does not influence the growth and maturation of skeletal muscle and tendon.

L-Wnk1 Deletion in Smooth Muscle Cells Causes Aortitis and Inflammatory Shift.

The long isoform of the Wnk1 (with-no-lysine [K] kinase 1) is a ubiquitous serine/threonine kinase, but its role in vascular smooth muscle cells (VSMCs) pathophysiology remains unknown. AngII (angiotensin II) was infused in Apoe-/- to induce experimental aortic aneurysm. Mice carrying an Sm22-Cre allele were cross-bred with mice carrying a floxed Wnk1 allele to specifically investigate the functional role of Wnk1 in VSMCs. Single-cell RNA-sequencing of the aneurysmal abdominal aorta from AngII-infused Apoe-/- mice revealed that VSMCs that did not express Wnk1 showed lower expression of contractile phenotype markers and increased inflammatory activity. Interestingly, WNK1 gene expression in VSMCs was decreased in human abdominal aortic aneurysm. Wnk1-deficient VSMCs lost their contractile function and exhibited a proinflammatory phenotype, characterized by the production of matrix metalloproteases, as well as cytokines and chemokines, which contributed to local accumulation of inflammatory macrophages, Ly6Chi monocytes, and γδ T cells. Sm22Cre+Wnk1lox/lox mice spontaneously developed aortitis in the infrarenal abdominal aorta, which extended to the thoracic area over time without any negative effect on long-term survival. AngII infusion in Sm22Cre+Wnk1lox/lox mice aggravated the aortic disease, with the formation of lethal abdominal aortic aneurysms. Pharmacological blockade of γδ T-cell recruitment using neutralizing anti-CXCL9 antibody treatment, or of monocyte/macrophage using Ki20227, a selective inhibitor of CSF1 receptor, attenuated aortitis. Wnk1 deletion in VSMCs led to aortic wall remodeling with destruction of elastin layers, increased collagen content, and enhanced local TGF-β (transforming growth factor-beta) 1 expression. Finally, in vivo TGF-β blockade using neutralizing anti-TGF-β antibody promoted saccular aneurysm formation and aorta rupture in Sm22 Cre+ Wnk1lox/lox mice but not in control animals. Wnk1 is a key regulator of VSMC function. Wnk1 deletion promotes VSMC phenotype switch toward a pathogenic proinflammatory phenotype, orchestrating deleterious vascular remodeling and spontaneous severe aortitis in mice.

Peritoneal Dialysis Aggravates and Accelerates Atherosclerosis in Uremic ApoE-/- Mice.

Atherosclerosis is highly prevalent in people with chronic kidney disease (CKD), including those receiving peritoneal dialysis (PD). Although it is lifesaving, PD induces profound systemic inflammation, which may aggravate atherosclerosis. Therefore, the hypothesis is that this PD-induced inflammation aggravates atherosclerosis via immune cell activation. ApoE-/- mice were subjected to a 5/6 nephrectomy to induce CKD. Three weeks later, mice were fed a high-cholesterol diet. Half of the nephrectomized mice then received daily peritoneal infusions of 3.86% Physioneal for 67 further days (CKD+PD) until the end of the experiment, and were compared with mice without CKD. Sham operated and PD-only mice were additional controls. CKD+PD mice displayed more severe atherosclerotic disease than control mice. Plaque area increased, and plaques were more advanced with a vulnerable phenotype typified by decreased collagen content and decreased fibrous cap thickness. Increased CD3+ T-cell numbers were present in plaques and perivascular adipose tissue of CKD and CKD+PD mice. Plaques of CKD+PD mice contained more iNOS+ immune cells. Spleens of CKD+PD mice showed more CD4+ central memory, terminally differentiated type 1 T-helper (Th1), Th17, and CX3C motif chemokine receptor 1+ (CX3CR1) CD4+ T-cells with less regulatory and effector T-cells. PD-fluid exposure in uremic mice potentiates systemic and vascular T-cell-driven inflammation and aggravates atherosclerosis. PD polarized CD4+ T-cells toward an inflammatory Th1/Th17 phenotype, and increased CX3CR1+ CD4+ T-cells, which are associated with vascular homing in CKD-associated atherosclerosis. Targeting CD4+ T-cell activation and CX3CR1+ polarization has the potential to attenuate atherosclerosis in PD patients.

Promoting Biological Similarity by Collagen Microfibers in 3D Colorectal Cancer-Stromal Tissue: Replicating Mechanical Properties and Cancer Stem Cell Markers

The extracellular matrix (ECM) of cancer tissues is rich in dense collagen, contributing to the stiffening of these tissues. Increased stiffness has been reported to promote cancer cell proliferation, invasion, metastasis, and prevent drug delivery. Replicating the structure and mechanical properties of cancer tissue in vitro is essential for developing cancer treatment drugs that target these properties. In this study, we recreated specific characteristics of cancer tissue, such as collagen density and high elastic modulus, using a colorectal cancer cell line as a model. Using our original material, collagen microfibers (CMFs), and a constructed three-dimensional (3D) cancer-stromal tissue model, we successfully reproduced an ECM highly similar to in vivo conditions. Furthermore, our research demonstrated that cancer stem cell markers expressed in the 3D cancer-stromal tissue model more closely mimic in vivo conditions than traditional two-dimensional cell cultures. We also found that CMFs might affect an impact on how cancer cells express these markers. Our 3D CMF-based model holds promise for enhancing our understanding of colorectal cancer and advancing therapeutic approaches. Statement of SignificanceReproducing the collagen content and stiffness of cancer tissue is crucial in comprehending the properties of cancer and advancing anticancer drug development. Nonetheless, the use of collagen as a scaffold material has posed challenges due to its poor solubility, hindering the replication of a cancer microenvironment. In this study, we have successfully recreated cancer tissue-specific characteristics such as collagen density, stiffness, and the expression of cancer stem cell markers in three-dimensional (3D) colorectal cancer stromal tissue, utilizing a proprietary material known as collagen microfiber (CMF). CMF proves to be an ideal scaffold material for replicating cancer stromal tissue, and these 3D tissues constructed with CMFs hold promise in contributing to our understanding of cancer and the development of therapeutic drugs.

Collagen concentration regulates neutrophil extravasation and migration in response to infection in an endothelium dependent manner

IntroductionAs the body’s first line of defense against disease and infection, neutrophils must efficiently navigate to sites of inflammation; however, neutrophil dysregulation contributes to the pathogenesis of numerous diseases that leave people susceptible to infections. Many of these diseases are also associated with changes to the protein composition of the extracellular matrix. While it is known that neutrophils and endothelial cells, which play a key role in neutrophil activation, are sensitive to the mechanical and structural properties of the extracellular matrix, our understanding of how protein composition in the matrix affects the neutrophil response to infection is incomplete.MethodsTo investigate the effects of extracellular matrix composition on the neutrophil response to infection, we used an infection-on-a-chip microfluidic device that replicates a portion of a blood vessel endothelium surrounded by a model extracellular matrix. Model blood vessels were fabricated by seeding human umbilical vein endothelial cells on 2, 4, or 6 mg/mL type I collagen hydrogels. Primary human neutrophils were loaded into the endothelial lumens and stimulated by adding the bacterial pathogen Pseudomonas aeruginosa to the surrounding matrix.ResultsCollagen concentration did not affect the cell density or barrier function of the endothelial lumens. Upon infectious challenge, we found greater neutrophil extravasation into the 4 mg/mL collagen gels compared to the 6 mg/mL collagen gels. We further found that extravasated neutrophils had the highest migration speed and distance in 2mg/mL gels and that these values decreased with increasing collagen concentration. However, these phenomena were not observed in the absence of an endothelial lumen. Lastly, no differences in the percent of extravasated neutrophils producing reactive oxygen species were observed across the various collagen concentrations.DiscussionOur study suggests that neutrophil extravasation and migration in response to an infectious challenge are regulated by collagen concentration in an endothelial cell-dependent manner. The results demonstrate how the mechanical and structural aspects of the tissue microenvironment affect the neutrophil response to infection. Additionally, these findings underscore the importance of developing and using microphysiological systems for studying the regulatory factors that govern the neutrophil response.

Extraction and physicochemical characterization of native and broiler chicken feet gelatin.

Poultry processing generates a large amount of industrial waste, which is rich in collagen content. This waste can be utilized for the extraction of valuable components such as gelatin, which can be used as an alternative to mammalian gelatin (porcine and bovine). Gelatins were analyzed for their yield, proximate analysis, pH, color, viscosity, bloom strength, and texture profile analysis. The yield of broiler chicken feet gelatin (BCFG) was slightly higher (7.93%) as compared to native chicken feet gelatin (NCFG) (7.06%). The protein content was 85.92% and 82.53% for BCFG and NCFG. Both gelatin had moisture content in the standard range (< 15) as given by Gelatin Manufacturers of Europe (GME). Both gelatins showed higher bloom strength (326 g for NCFG and 203 g for BCFG) at 6.67% gelatin concentration, classified as high bloom. Fourier-transform infrared (FTIR) analysis showed amide I, amide A, amide B at 1636 cm-1, 3302 cm-1, 2945 cm-1 for NCFG and 1738 cm-1, 3292 cm-1, 2920 cm-1 for BCFG. At 6.67% gelatin concentration, hardness and cohesiveness values were also higher than commercial gelatin previously studied. The pH values for NCFG were 5.43 and BCFG was 5.31. Both NCFG and BCFG viscosities (4.43 and 3.85 cP) were in the optimum range of commercial gelatins (2-7 cP). Hence, the present study concluded that both NCFG and BCFG have a huge potential to replace commercial mammalian gelatins (porcine and bovine) in the food industries. However further studies should be done to optimize the extraction process. © 2024 Society of Chemical Industry.

P-589 Effects of phthalates on the 3D spheroid cell culture as a model for the human ovarian extracellular matrix

Abstract Study question Do phthalates affect the ovary by impacting its extracellular matrix (ECM)? Summary answer Phthalate mixture (PM) negatively affects the components of ovarian ECM, as demonstrated by spheroids as a model. What is known already The widespread production and pervasive use of phthalates as plasticizers in everyday products significantly increases human exposure to these substances. Their documented toxic effects on reproductive and endocrine functions raise substantial public health concerns. Yet, a full understanding of how phthalates affect ovarian functions and especially the ECM structure in females—more exposed to these products than men—remains incomplete. Moreover, there has been limited investigation in identifying a suitable model capable of mimicking the 3D ovarian ECM structure in vitro for studying phthalate impacts. Study design, size, duration Stromal cells were isolated from the ovarian cortex of post-menopausal organ donors. Cells were seeded onto agarose micro-well molds to form the spheroids. After 6 days, spheroids were treated with a PM (0 and 100 µg/ml) for 4 days. Cell proliferation, spheroids diameter and protein levels of ECM components were measured for both groups. Participants/materials, setting, methods Ovaries from four deceased post-menopausal multi-organ donors (IRB reference 2018/19DEC/475) were used for cell isolation. After seeding of cells, their diameter was measured on days 6 (start of treatment) and 10 (end of treatment) using an inverted microscope to assess their development. The study measured the protein levels of collagen VI, Emilin, and Fibrilin using immunohistochemistry and Ki67 using immunofluorescence in ovarian spheroids. The picrosirius red staining (PSR) was used to measure the collagen content. Main results and the role of chance The evaluation of spheroid diameter indicated comparable sizes on day 6 for both groups (0 µg/ml PM: 109±1.3 µm; 100 µg/ml PM: 112.5±1.1 µm). By day 10, there were no notable differences in spheroid size for the group exposed to PM in comparison to the control group (0 µg/ml PM: 104±1.2 µm; 100 µg/ml PM: 107.2±1.2 µm). The assessment of proliferation revealed no significant difference in the expression of ki67-positive cells between the groups (0 µg/ml PM: 3.211±0.64 %; 100 µg/ml PM: 3.379±0.61%; Fig 1). On the other hand, immunostaining results displayed a decrease in EMILIN-1 (171.6±5.31 vs 201.9±4.71 in control, P &amp;lt; 0.0001) and fibrillin levels (53.1±3.34 vs. 88.3±3.68 in control, P &amp;lt; 0.0001) post-PM treatment. Conversely, collagen VI levels were significantly higher in spheroids treated with PM compared to the control group (122.8±4.99 vs 104.8±4.04, P = 0.005). Although PSR revealed an increase following treatment (9.62±0.75 vs 8.64±0.70 in control), this change did not reach statistical significance (P = 0.344). Limitations, reasons for caution This study exclusively focused on ovarian stromal cells from post-menopausal women. Further research is needed to explore the impacts of PM on ECM of prepubertal and reproductive-age women. Wider implications of the findings Our study highlights the detrimental impact of phthalates on the ECM of postmenopausal women. Findings suggest ovarian stiffness by decreased elastogenesis-associated proteins (EMILIN-1, fibrillin-1) and increased collagen VI (not I, III) post-phthalate exposure.This pioneering study prompts further exploration of phthalate-induced ECM changes and their potential influence on steroidogenesis and folliculogenesis. Trial registration number not applicable

Acinar cells of the pancreas as an independent predictor of the development of postoperative pancreatic fistula: A review

Analysis of data published in the modern literature on the role of the number of functioning acinar cells of the pancreas as a prognostic marker of the development of pancreatic fistula. The search for sources was carried out in the systems Clinicaltrials.gov, PubMed, Medline, NCCN, Scopus, Elibrary. In writing the literature review, 52 sources published from 2004 to 2022 were used. Included are studies reflecting the importance of functioning acinar cells at the edge of pancreatic resection, as well as other factors associated with the development of pancreatic fistula and other post-resection complications. A large number of acinar cells (40%) and a low collagen content (15%) in the edge of pancreatic resection significantly correlates with the development of pancreatic fistula. The content of adipose tissue in the edge of pancreatic resection showed no connection with the development of pancreatic fistula and other complications after surgery. Palpatory assessment of the structure of the pancreatic parenchyma is subjective and can lead to erroneous interpretation and adoption of inadequate tactics of preventive measures. Intraoperative counting of acinuses at the edge of pancreatic resection is easy to use, while not yielding to more complex methods for assessing the risk of post-resection complications and can be recommended as a routine method for predicting the occurrence of pancreatic fistula

Mechanical Stress-induced Connective Tissue Growth Factor Plays a Critical Role in Intestinal Fibrosis in Crohn's-like Colitis.

Crohn's disease (CD) is an inflammatory bowel disease characterized by transmural inflammation and intestinal fibrosis. Mechanisms of fibrosis in CD are not well understood. Transmural inflammation is associated with inflammatory cell infiltration, stenosis, and distention, which present mechanical stress (MS) to the bowel wall. We hypothesize that MS induces gene expression of pro-fibrotic mediators such as connective tissue growth factor (CTGF), which may contribute to fibrosis in CD. A rodent model of CD was induced by intracolonic instillation of TNBS to the distal colon. TNBS instillation induced a localized transmural inflammation (site I), with a distended colon segment (site P) proximal to site I. We detected significant fibrosis and collagen content not only in site I, but also in site P in CD rats by day 7. CTGF expression increased significantly in sites P and I, but not in the segment distal to the inflammation site. Increased CTGF expression was detected mainly in the smooth muscle cells (SMC). When rats were fed exclusively with clear liquid diet to prevent mechanical distention in colitis, expression of CTGF in sites P and I was blocked. Direct stretch led to robust expression of CTGF in colonic SMC. Treatment of CD rats with anti-CTGF antibody FG-3149 reduced fibrosis and collagen content in both sites P and I and exhibited consistent trends towards normalizing expression of collagen mRNAs. In conclusion, our studies suggest that mechanical stress, by up-regulating pro-fibrotic mediators i.e. CTGF, may play a critical role in fibrosis in CD.

Effects of Bulleyaconitine A on Extracellular Matrix Secretion and Expression of Related Proteins in Acetaldehyde-Activated Hepatic Stellate Cells.

Activated hepatic stellate cells differentiate into myofibroblasts, which synthesize and secrete extracellular matrix (ECM) leading to liver fibrosis. It was previously demonstrated that bulleyaconitine A (BLA), an alkaloid from Aconitum bulleyanum, inhibits proliferation and promotes apoptosis of human hepatic Lieming Xu-2 (LX-2) cells. In this study, we analyzed the effect of BLA on the production of ECM and related proteins by LX-2 cells activated with acetaldehyde (AA). The cells were randomized into the control group, AA group (cells activated with 400 μM AA), and BLA+AA group (cells cultured in the presence of 400 μM AA and 18.75 μg/ml BLA). In the BLA+AA group, the contents of collagens I and III and the expression of α-smooth muscle actin and transforming growth factor-β1 (TGF-β1) were statistically significantly higher than in the control, but lower than in the AA group. Expression of MMP-1 in the BLA+AA group was also significantly higher than in the AA group, but lower than in the control. Expression of TIMP-1 in the BLA+AA group was significantly higher than in the control, but lower than in the AA group. Thus, BLA suppressed activation and proliferation of LX-2 cells by inhibiting TGF-β1 signaling pathway and decreasing the content of collagens I and III by reducing the MMP-1/TIMP-1 ratio.

Alterations in the Structure, Composition, and Organization of Galactosaminoglycan-Containing Proteoglycans and Collagen Correspond to the Progressive Stages of Dupuytren’s Disease

Dupuytren’s disease (DD) is a prevalent fibroproliferative disorder of the hand, shaped by genetic, epigenetic, and environmental influences. The extracellular matrix (ECM) is a complex assembly of diverse macromolecules. Alterations in the ECM’s content, structure and organization can impact both normal physiological functions and pathological conditions. This study explored the content and organization of glycosaminoglycans, proteoglycans, and collagen in the ECM of patients at various stages of DD, assessing their potential as prognostic indicators. This research reveals, for the first time, relevant changes in the complexity of chondroitin/dermatan sulfate structures, specifically an increase of disaccharides containing iduronic acid residues covalently linked to either N-acetylgalactosamine 6-O-sulfated or N-acetylgalactosamine 4-O-sulfated, correlating with the disease’s severity. Additionally, we noted an increase in versican expression, a high molecular weight proteoglycan, across stages I to IV, while decorin, a small leucine-rich proteoglycan, significantly diminishes as DD progresses, both confirmed by mRNA analysis and protein detection via confocal microscopy. Coherent anti-Stokes Raman scattering (CARS) microscopy further demonstrated that collagen fibril architecture in DD varies importantly with disease stages. Moreover, the urinary excretion of both hyaluronic and sulfated glycosaminoglycans markedly decreased among DD patients.Our findings indicate that specific proteoglycans with galactosaminoglycan chains and collagen arrangements could serve as biomarkers for DD progression. The reduction in glycosaminoglycan excretion suggests a systemic manifestation of the disease.

Exosomes from umbilical cord mesenchymal stromal cells promote the collagen production of fibroblasts from pelvic organ prolapse

BACKGROUND Pelvic organ prolapse (POP) involves pelvic organ herniation into the vagina due to pelvic floor tissue laxity, and vaginal structure is an essential factor. In POP, the vaginal walls exhibit abnormal collagen distribution and decreased fibroblast levels and functions. The intricate etiology of POP and the prohibition of transvaginal meshes in pelvic reconstruction surgery present challenges in targeted therapy development. Human umbilical cord mesenchymal stromal cells (hucMSCs) present limitations, but their exosomes (hucMSC-Exo) are promising therapeutic tools for promoting fibroblast proliferation and extracellular matrix remodeling. AIM To investigate the effects of hucMSC-Exo on the functions of primary vaginal fibroblasts and to elucidate the underlying mechanism involved. METHODS Human vaginal wall collagen content was assessed by Masson’s trichrome and Sirius blue staining. Gene expression differences in fibroblasts from patients with and without POP were assessed via RNA sequencing (RNA-seq). The effects of hucMSC-Exo on fibroblasts were determined via functional experiments in vitro . RNA-seq data from fibroblasts exposed to hucMSC-Exo and microRNA (miRNA) sequencing data from hucMSC-Exo were jointly analyzed to identify effective molecules. RESULTS In POP, the vaginal wall exhibited abnormal collagen distribution and reduced fibroblast 1 quality and quantity. Treatment with 4 or 6 μg/mL hucMSC-Exo suppressed inflammation in POP group fibroblasts, stimulated primary fibroblast growth, and elevated collagen I (Col1) production in vitro . High-throughput RNA-seq of fibroblasts treated with hucMSC-Exo and miRNA sequencing of hucMSC-Exo revealed that abundant exosomal miRNAs downregulated matrix metalloproteinase 11 (MMP11) expression. CONCLUSION HucMSC-Exo normalized the growth and function of primary fibroblasts from patients with POP by promoting cell growth and Col1 expression in vitro . Abundant miRNAs in hucMSC-Exo targeted and downregulated MMP11 expression. HucMSC-Exo-based therapy may be ideal for safely and effectively treating POP.

Research Note: Basic chemical composition, physicochemical, and textural characteristics of male and female guinea fowl meat

The purpose of this study was to compare analyze the quality of meat depending on the type of muscle: breast muscles (m. Pectoralis superficialis and m. Pectoralis profundus) and leg muscles (m. Sartorius and m. Femorotibialis) in relation to the sex of the common guinea fowl (Numida meleagris). For the study, pectoral muscles and leg muscles from 10 females and 10 males at 20 wk of age were used to determine their chemical composition, physicochemical properties, and texture characteristics of the pectoral muscle were determined. Overall, pectoral muscles were characterized by higher protein content, lightness (L*), and electrical conductivity (EC24), and lower redness (a*), and pH24 compared to leg muscles. Leg muscles, on the other hand, were characterized by higher, collagen, intramuscular fat and salt content. In terms of pectoral muscle texture characteristics, males were characterized by lower chewiness value and were less gumminess. In conclusion, it can be said that the sex of the birds affected some of the pectoral muscle texture traits, but did not affect the remaining analyzed features of the pectoral and leg muscles. However, from the consumer's point of view, breast muscles contained more protein and less fat, so they are more dietary compared to leg muscles.

Blackcurrant extract promotes differentiation of MC3T3‑E1 pre‑osteoblasts.

Osteoporosis risk increases in menopausal individuals owing to the decrease in estrogen secretion. Blackcurrant extract (BCE) ameliorates osteoporosis; however, the underlying mechanisms are unclear. Furthermore, although BCE has phytoestrogenic activity, its effects on osteoblasts are unknown. In the present study, we investigated BCE-mediated attenuation of osteoporosis using mouse MC3T3-E1 pre-osteoblasts, with a focus on osteogenesis. After treating MC3T3-E1 cells with BCE for 48 h, cell proliferation was assessed using Cell Counting Kit-8. Levels of osteoblast differentiation markers, namely alkaline phosphatase activity and total collagen content in the cells, were evaluated after 3 and 14 days of BCE treatment, respectively. The expression of genes encoding osteoblast differentiation markers, including collagen type I (Col-I), alkaline phosphatase (Alp), bone γ-carboxyglutamate protein (Bglap), and runt-related transcription factor 2 (Runx2), was evaluated using reverse transcription-quantitative polymerase chain reaction. Mineralization of the cells was evaluated using Alizarin Red staining. Femoral tissues of ovariectomized (OVX) rats with or without 3% BCE were stained using ALP to evaluate osteogenic differentiation in femoral tissue. After treating MC3T3-E1 cells with BCE, cell proliferation had increased. BCE treatment increased Alp activity and total collagen content. Moreover, the expression of Col-I, Alp, Bglap, and Runx2 increased in BCE-treated cells. Furthermore, when MC3T3-E1 cells were treated with BCE for 21 days, the levels of calcified nodules increased. Alp staining intensity was stronger in the epiphyses on femoral tissue of OVX rats treated with 3% BCE than in those of untreated OVX rats. The results suggest that BCE may promote osteogenesis by inducing osteoblast differentiation.

3D-Printed SERS Chips for Highly Specific Detection of Denatured Type I and IV Collagens in Blood for Early Hepatic Fibrosis Diagnosis.

Hepatic fibrosis, the insidious progression of chronic liver scarring leading to life-threatening cirrhosis and hepatocellular carcinoma, necessitates the urgent development of noninvasive and precise diagnostic methodologies. Denatured collagen emerges as a critical biomarker in the pathogenesis of hepatic fibrosis. Herein, we have for the first time developed 3D-printed collagen capture chips for highly specific surface-enhanced Raman scattering (SERS) detection of denatured type I and type IV collagen in blood, facilitating the early diagnosis of hepatic fibrosis. Employing a novel blend of denatured collagen-targeting peptide-modified silver nanoparticle probes (Ag@DCTP) and polyethylene glycol diacrylate (PEGDA), we engineered a robust ink for the 3D fabrication of these collagen capture chips. The chips are further equipped with specialized SERS peptide probes, Ag@ICTP@R1 (S-I) and Ag@IVCTP@R2 (S-IV), tailored for the targeted detection of type I and IV collagen, respectively. The SERS chip platform demonstrated exceptional specificity and sensitivity in capturing and detecting denatured type I and IV collagen, achieving detection limits of 3.5 ng/mL for type I and 3.2 ng/mL for type IV collagen within a 10-400 ng/mL range. When tested on serum samples from hepatic fibrosis mouse models across a spectrum of fibrosis stages (S0-S4), the chips consistently measured denatured type I collagen and detected a progressive increase in type IV collagen concentration, which correlated with the severity of fibrosis. This novel strategy establishes a benchmark for the multiplexed detection of collagen biomarkers, enhancing our capacity to assess the stages of hepatic fibrosis.

Evaluation of Aloe Vera Gel Combined with Cinnamon Oil with Antibacterial Activity for Wound Healing Application.

Treatment of chronic wounds has been shifted to traditional approaches due to surge in antibiotic resistance. Wounds that fail to heal satisfactorily may result in the amputation of the organ. In this research work, cinnamon oil (CO) and aloe vera (AV) that have been traditionally used as antibacterial agents are combined in a unique gel (COVA) and its antibacterial activity has been evaluated through in vitro and in vivo studies. Antibacterial activity was measured through disk diffusion and agar dilution method against Pseudomonas aeruginosa and Staphylococcus aureus. To check antibacterial and wound healing activity, diabetic excision wound healing rat model was used. Wound closure, wound contraction, tissue hydroxyproline content, antioxidant capacity (TAC), and malondialdehyde (MDA) level were monitored. The minimum inhibitory concentrations of CO + AV for bacterium P. aeruginosa and S. aureus were 100 and 200 µg/ml, respectively. After 14 days, the wounds covered with COVA therapy reached to nearly full wound closure (79% wound contraction) compared to control. The collagen content and level of TAC increased significantly (P < 0.05) in treated groups; therefore, 25% fast healing was observed in wounds treated with CO and AV gel combined. Reduced levels of tissue MDA were observed in all treated groups and specially wound covered with COVA (0.43 mM/mg in control vs 0.25 mM/mg in COVA). Histopathological examination also supported the outcomes. Significantly elevated increase in the level of hydroxyproline was found in rats of COVA treatment group (37.1 ± 0.44). Combination of CO and AV can be potentially used to prevent infection in wound; as these herbal agents not only inhibit the growth of pathogenic bacteria but also accelerate tissue repair.

Mapping the unicellular transcriptome of the ascending thoracic aorta to changes in mechanosensing and mechanoadaptation during aging.

Aortic stiffening is an inevitable manifestation of chronological aging, yet the mechano-molecular programs that orchestrate region- and layer-specific adaptations along the length and through the wall of the aorta are incompletely defined. Here, we show that the decline in passive cyclic distensibility is more pronounced in the ascending thoracic aorta (ATA) compared to distal segments of the aorta and that collagen content increases in both the medial and adventitial compartments of the ATA during aging. The single-cell RNA sequencing of aged ATA tissues reveals altered cellular senescence, remodeling, and inflammatory responses accompanied by enrichment of T-lymphocytes and rarefaction of vascular smooth muscle cells, compared to young samples. T lymphocyte clusters accumulate in the adventitia, while the activation of mechanosensitive Piezo-1 enhances vasoconstriction and contributes to the overall functional decline of ATA tissues. These results portray the immuno-mechanical aging of the ATA as a process that culminates in a stiffer conduit permissive to the accrual of multi-gerogenic signals priming to disease development.

Widespread Vascularization and Correlation of Glycosaminoglycan Accumulation to Tendon Pain in Human Plantar Fascia Tendinopathy

Background: Plantar fasciitis is a painful tendinous condition (tendinopathy) with a high prevalence in athletes. While a healthy tendon has limited blood flow, ultrasound has indicated elevated blood flow in tendinopathy, but it is unknown if this is related to a de facto increase in the tendon vasculature. Likewise, an accumulation of glycosaminoglycans (GAGs) is observed in tendinopathy, but its relationship to clinical pain is unknown. Purpose: To explore to what extent vascularization, inflammation, and fat infiltration were present in patients with plantar fasciitis and if they were related to clinical symptoms. Study Design: Descriptive laboratory study. Methods: Biopsy specimens from tendinopathic plantar fascia tissue were obtained per-operatively from both the primary site of tendon pain and tissue swelling (“proximal”) and a region that appeared macroscopically healthy at 1 to 2 cm away from the primary site (“distal”) in 22 patients. Biopsy specimens were examined with immunofluorescence for markers of blood vessels, tissue cell density, fat infiltration, and macrophage level. In addition, pain during the first step in the morning (registered during an earlier study) was correlated with the content of collagen and GAGs in tissue. Results: High vascularization (and cellularity) was present in both the proximal (0.89%) and the distal (0.96%) plantar fascia samples, whereas inconsistent but not significantly different fat infiltration and macrophage levels were observed. The collagen content was similar in the 2 plantar fascia regions, whereas the GAG content was higher in the proximal region (3.2% in proximal and 2.8% in distal; P = .027). The GAG content in the proximal region was positively correlated with the subjective morning pain score in the patients with tendinopathy (n = 17). Conclusion: In patients with plantar fasciitis, marked tissue vascularization was present in both the painful focal region and a neighboring nonsymptomatic area. In contrast, the accumulation of hydrophilic GAGs was greater in the symptomatic region and was positively correlated with increased clinical pain levels in daily life. Clinical Relevance: The accumulation of GAGs in tissue rather than the extent of vascularization appears to be linked with the clinical degree of pain symptoms of the disease.

Physicochemical Analysis of Spent Hen Chicken Sausages with Jelly Mushroom (Auricularia auricula) Powder Inclusion as a Substitute for Sodium Tripolyphosphate

Abstract This study aims to evaluate the substitution effect of the Sodium Tripolyphosphate (STPP) with jelly mushroom (Auricularia auricula) powder (JMP)at various concentration on quality attributes of the spent hen chicken sausages. Using a completely randomized design (CRD), this experiment consisting of five treatments and three replications. The treatments including group with 0,3% STPP (P0; w/w), and inclusion of jelly mushroom at 1%, 2%, and 3%, labelled with T1, T2, T3; w/w, respectively. The data obtained were statistically analyzed using analysis of variance (ANOVA) and continued with Duncan’s multiple range test (DMRT) at 5% level. The results showed that the increase in (JMP)substitution up to 3% levels had a significant effect on the tenderness, WHC, and pH value. The addition of (JMP)had no significant effect on the cooking loss, ranging from 4,74 to 6,33%. Substitutions of STPP with JMP in spent hen sausages even at the 1% level generated a significant effect on collagen, fat, and moisture contents and decreased protein content. The addition of jelly mushroom powder up to 1% levels will reduce fat, collagen, and moisture contents. This study inferred that the JMP can be used as an alternative substitution for STPP at the level of 1%, while the use of JMP in spent hen sausages at more than 1% would negatively affect the quality attributes of spent hen chicken sausage.

Fabrication of a decellularized liver matrix-based hepatic patch for the repair of CCl4-induced liver injury.

This article primarily introduces a new treatment for liver fibrosis/cirrhosis. We developed a hepatic patch by combining decellularized liver matrix (DLM) with the hepatocyte growth factor (HGF)/heparin-complex and evaluated its restorative efficacy. In vitro prophylactic results, the HGF/heparin-DLM patches effectively mitigated CCl4-induced hepatocyte toxicity and restored the cytotoxicity levels to the baseline levels by day 5. Furthermore, these patches restored albumin synthesis of injured hepatocytes to more than 70% of the normal levels within 5 days. In vitro therapeutic results, the urea synthesis of the injured hepatocytes reached 91% of the normal levels after 10 days of culture, indicating successful restoration of hepatic function by the HGF/heparin-DLM patches in both prophylactic and therapeutic models. In vivo results, HGF/heparin-DLM patches attached to the liver and gut exhibited a significant decrease in collagen content (4.44 times and 2.77 times, respectively) and an increase in glycogen content (1.19 times and 1.12 times, respectively) compared to the fibrosis group after 1week, separately. In summary, liver function was restored and inflammation was inhibited through the combined effects of DLM and the HGF/heparin-complex in fibrotic liver. The newly designed hepatic patch holds promise for both in vitro and in vivo regeneration therapy and preventive health care for liver tissue engineering.