Expression Of Collagen Type Research Articles

The Effect of Stichopus hermanii to TLR-4 in Mediating Periodontal Ligament Remodeling During Orthodontic Relapse

Stichopus hermanii contains an active compound that plays apart in the innate immune system. This research was aimed to investigate the effect of S. hermanii on the TLR-4 in mediating periodontal ligament (PDL) remodeling through fibroblast growth factor (FGF-2) and collagen type 1 parameter during orthodontic relapse. The true experimental using a post-test for the control group. Thirty-two male Guinea pig were distributed into four groups: K(−) group was the negative control group, K(+) group was the positive control group applied with orthodontic relapse forces, P1 group was applied with orthodontic relapse forces and 3% S. hermanii 3%, and the P2 group was applied with 3.5%. TLR-4, FGF-2, and collagen type-1 expression at the tension site was examined using immunohistochemistry. Analysis of Variance and Least Significance Difference tests were conducted for all specimens. TLR-4 in P1 and P2 were decreasing significantly (p≤0.05), while FGF-2 and collagen type 1 expression in P1 and P2 increased significantly (p≤0.05). Multiple regression test to correlate between parameters was also significant at 0.00 (p≤0.05). This study concluded that decrease of TLR-4 expression during orthodontic relapse showed a strong tendency to coincide with PDL remodeling along with increasing FGF-2 and collagen type 1 parameter. The optimum result in decreasing TLR-4 expression occurs when administered with S. hermanii 3.5%, which induces decreasing relapse to 29% and increasing periodontal remodeling, demonstrated by FGF-2 and collagen type-1 expression.

Circular RNA circANAPC2 mediates the impairment of endochondral ossification by miR-874-3p/SMAD3 signalling pathway in idiopathic short stature.

Idiopathic short stature (ISS) is a main reason for low height among children. Its exact aetiology remains unclear. Recent findings have suggested that the aberrant expression of circRNAs in peripheral blood samples is associated with many diseases. However, to date, the role of aberrant circRNA expression in mediating ISS pathogenesis remains largely unknown. The up‐regulated circANAPC2 was identified by circRNA microarray analysis and RT‐qPCR. Overexpression of circANAPC2 inhibited the proliferation of human chondrocytes, and cell cycle was arrested in G1 phase. The expressions of collagen type X, RUNX2, OCN and OPN were significantly down‐regulated following circANAPC2 overexpression. Moreover, Von Kossa staining intensity and alkaline phosphatase activity were also decreased. Luciferase reporter assay results showed that circANAPC2 could be targeted by miR‐874‐3p. CircANAPC2 overexpression in human chondrocytes inhibits the expression of miR‐874‐3p. The co‐localization of circANAPC2 and miR‐874‐3p was confirmed in both human chondrocytes and murine femoral growth plates via in situ hybridization. The rescue experiment demonstrated that the high expression of miR‐874‐3p overexpression antagonized the suppression of endochondral ossification, hypertrophy and chondrocyte growth caused by circANAPC2 overexpression. A high‐throughput screening of mRNA expression and RT‐qPCR verified SMAD3 demonstrated the highest different expressions following overcircANAPC2. Luciferase reporter assay results indicated that miR‐874‐3p could be targeted by Smad3, thus down‐regulating the expression of Smad3. Subsequent rescue experiments of SMAD3 further confirmed that circANAPC2 suppresses endochondral ossification, hypertrophy and chondrocyte growth through miR‐874‐3p/Smad3 axis. The present study provides evidence that circANAPC2 can serve as a promising target for ISS treatment.

Pheophorbide a Derivatives Exert Antiwrinkle Effects on UVB-Induced Skin Aging in Human Fibroblasts.

Pheophorbide a is a chlorophyll metabolic breakdown product. This study investigated the antiwrinkle effect of pheophorbide a (PA) and its derivatives, including pyropheophorbide a (PyroPA) and pyropheophorbide a methyl ester (PyroPA-ME), on ultraviolet (UV) B-stimulated CCD-986sk fibroblasts. PA, PyroPA, and PyroPA-ME effectively suppressed reactive oxygen species accumulation in UVB-exposed CCD-986sk fibroblasts. All three pheophorbides also reduced UVB-induced matrix metalloproteinase (MMP)-1 secretion and mRNA expression of MMP-1, MMP-2, and MMP-9. Treatment with pheophorbides resulted in increased procollagen synthesis, and this required enhancement of procollagen type I C-peptide content and mRNA expression of collagen type I alpha 1 (COL1A1) and COL1A2 in CCD-986sk cells. These antiwrinkle effects were more potent with PA and PyroPA than with PyroPA-ME. Furthermore, PA and PyroPA suppressed UVB-induced phosphorylation of extracellular signal-regulated protein kinase and c-Jun N-terminal kinase but not p38. Moreover, all three pheophorbides inhibited NF-κB p65 phosphorylation. Therefore, these pheophorbides, especially PA and PyroPA, can be used as antiwrinkle agents, and PA- or PyroPA-rich natural resources can be used in functional cosmetics.

Growth factor stimulation for the healing of traumatic ulcers with liquid rice hull smoke

ObjectiveThe healing process of a traumatic ulcer requires growth factors to rebuild the lost tissue after the inflammatory process has been completed. Liquid rice hull smoke (LR-HS) has shown unique anti-inflammatory properties. This study analyses the role of LR-HS in growth factor stimulation for the healing of traumatic ulcers, such as fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and collagen type 1 (COL-1) expression. MethodsWe obtained LR-HS from the pyrolysis of rice hulls. Traumatic ulcers were created in the labial fornix incisive inferior of Wistar rats and treated with LR-HS once a day for 3, 5, and 7 days. The control group was treated with sterile water. Each animal was sacrificed after treatment, and its labial fornix incisive inferior tissues were biopsied and immunohistochemically stained to examine FGF, VEGF, PDGF, and COL-1 expression. ResultThe treatment of traumatic ulcers with LR-HS showed an increase in FGF, VEGF, PDGF, and COL-1 expression. VEGF expression increased under LR-HS treatment compared with the control 7-day treatment groups (p < 0.000). FGF and COL-1 expression increased under LR-HS treatment compared with the control 5- and 7-day treatment groups (p < 0.000). PDGF expression increased after treatment with LR-HS for 3, 5, and 7 days (p < 0.000). ConclusionThis study has demonstrated that LR-HS can induce the expression of growth factors during the healing of a traumatic ulcer using immunohistochemical staining. We suggest that LR-HS can be used as a herbal medicine for oral ulcer therapy.

Nitric Oxide Produced by the Antioxidant Activity of Verapamil Improves the Acute Wound Healing Process.

Verapamil is used in the treatment of hypertension, angina pectoris, cardiac arrhythmia, hypertrophic scars, and keloids to block transmembrane calcium ion flux. Verapamil has antioxidant activity, which enhances the production of nitric oxide (NO). NO promotes the proliferation of fibroblasts, keratinocytes, endothelial cells, and epithelial cells during wound healing. In this study, we investigated the effect of verapamil and its antioxidant properties on the enhancement of acute wound healing via NO. A full-thickness wound healing model was created on the rat dorsal with a silicone ring. The wound closure rate was estimated every 2days for 14days. A histological study was performed to evaluate wound healing. Immunofluorescence staining was analyzed for angiogenesis. The expressions of collagen type I (COL I), collagen type III (COL III), and vascular endothelial growth factor (VEGF) were assessed by Western blot. Real-time polymerase chain reaction (qRT-PCR) was performed to examine the expression of endothelial NO synthase and inducible NO synthase, which are related to antioxidant activity in the process of wound healing. The wound closure rate was faster in the verapamil group compared to the control and silicone groups. Histologic analysis revealed capillaries and stratum basale in the verapamil group. Immunofluorescence staining was shown vessel formation in the verapamil group. Western blot and qRT-PCR analysis revealed high expression levels of COL I, VEGF, eNOS, and FGF in the verapamil. Verapamil's antioxidant activity enhances NO production in acute wound healing. We suggest that verapamil can be used to promote acute wound healing.

In vivo Therapeutic Effects and Mechanisms of Hydroxyasiaticoside Combined With Praziquantel in the Treatment of Schistosomiasis Induced Hepatic Fibrosis.

Schistosomiasis has been a fatal obstinate disease that threatens global human health, resulting in the granulomatous inflammation and liver fibrosis.Objective:The aim of this study was to evaluate the therapeutic effects and mechanisms of hydroxyasiaticoside combined with praziquantel in the treatment of schistosomiasis-induced liver fibrosis.Methods:Mice were randomly distributed into four experimental groups: normal control group, model group, praziquantel group, praziquantel + hydroxyasiaticoside group. Except for the normal control group, they were infected with Schistosomia cercariae through the abdominal skin to induce liver fibrosis. In the intervention group, mice were administered with the respective drugs by gavage after 8 weeks of infection. At the end of the treatment, mice were sacrificed to collect blood for the determination of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) serum levels. Moreover, the liver was excised, weighed, and liver indices were calculated. Histopathological examination was performed to assess liver morphology. Besides, the expression of collagen type I and III in liver was determined; the mRNA expression levels of IL-6 and TNF-α in liver tissues were measured using Real-time PCR while ELISA and western blotting were performed on liver tissue homogenate to determine the protein expression of IL-6 and TNF-α.Results:The combination of praziquantel and hydroxyasiaticoside lowered the pathological scores of schistosomiasis-induced hepatic fibrosis, the liver indice, serum AST and ALT levels, improved liver morphology, downregulated the expression levels of hepatic type I and III collagen, inhibited the mRNA expression levels of pro-inflammatory factors (IL-6 and TNF-α) in the liver of mice relative to the praziquantel alone.Conclusion:The combination of hydroxyasiaticoside and praziquantel is a potential therapeutic option for schistosomiasis-induced hepatic fibrosis. Notably, this combination noticeably suppresses the protein and mRNA expression levels of pro-inflammatory factors (TNF-α and IL-6) in the liver.

The NF-kB and Collagen Type 1 Expression in Dental Pulp after Treated Calcium Hydroxide Combined with Propolis.

Objective To analyze the expression of nuclear factor kappa B (NF-kB) and collagen type 1 on dental pulp after a treated combination of calcium hydroxide and propolis. Materials and Methods The first maxillary molars of 30 rats were mechanically perforated. Teeth were divided into three groups of 10 for two separate extraction time frames, giving a total of 60 rats. The control groups were treated with Cention, the second treatment groups were treated with calcium hydroxide, and the third treatment groups were treated with a combination of calcium hydroxide and propolis. Final restoration was done with Cention. The teeth were extracted on days 7 and 14, and the expression of NF-kB and collagen type I was analyzed using immunohistochemistry. Results There is lowest NF-kB expression and highest collagen type 1 expression on dental pulp after treated with a combination of calcium hydroxide and propolis on days 7 and 14 ( p < 0.05). Conclusion The combination of calcium hydroxide and propolis inhibits pulp inflammation and stimulates regeneration through decreasing the NF-kB expression and increasing collagen type 1.

Which Culture System Is better for Chondrogenesis of Adipose-Derived Stem Cells ; Pellet or Micromass?

Background and Aims: The current study was conducted to compare the expression levels of collagen type Π and X during chondrogenesis of human adipose-derived mesenchymal stem cells (hADMSCs) pellet and micromass cultures. Materials and Methods: Extracted hADMSCs were cultured until three passages and then transferred to pellet and micromass cultures in the experimental groups of day 7 and day14. For pellet and micromass cultures, aliquots of 5×105 cells/ml were centrifuged and respectively cultured in the conical tubes and droplets (12.5 µl) of the 24-well plates containing chondrogenic medium. Realtime-polymerase chain reaction technique was performed for gene expression levels. Results: Increased expression of collagen type Π was shown in micromass day14 compared to micromass day 7, pellet day 14 (p&lt;0.01) and pellet day 7 (p&lt;0.001). Also, an increased expression of collagen type Π was seen in micromass day 7 and pellet day 14 compared to pellet day 7 (p&lt; 0.05). Expression of collagen type X increased in pellet day 14 compared to micromass on days 7 and 14 (p&lt;0.001, p&lt;0.01) and pellet day14 compared to pellet day7 (p&lt; 0.05). An increased expression of collagen type X was shown in pellet day 7 compared to micromass on days 7 and 14 (p&lt;0.05). Conclusions: According to the results, higher expression of collagen type Π and lower expression of collagen type X in micromass cultures that are prepared by cell suspension play a better role during cellular condensation that leads to the formation of large nodules exhibiting cartilage-like morphology, suggests a higher efficiency for micromass cultures.

Effect of Resistance Training Along with Curcumin Supplementation on Expression of Some Regulator Genes Associated with Cardiac Muscle Structure in Obese Rats

Background: Reduced physical activity can cause obesity and metabolic syndrome, leading to fibrosis in cardiac muscles and premature cardiac aging. Physical activity, along with herbal supplements, can have a synergistic effect on preventing cardiac muscle proteolysis. Objectives: In this study, the effects of curcumin and resistance training were assessed on cardiac muscle atrophy in obese rats. Methods: Twenty-four male Sprague rats were categorized into four groups, including the placebo, resistance training, curcumin, and resistance training + curcumin. Resistance training was performed three times a week with three sets in each session, repeated five times for eight weeks. During this time, 150 mg/kg curcumin was administered through gavage. Twenty-four hours after finishing resistance training, surgery was performed on the cardiac muscle, and gene expressions of PGC1-α, FOXO1, Murf-1, Atrogin, Collagen1, and Collagen 3 were assessed with real-time PCR. Results: The expression of PGC1-α and FOXO1 genes in both resistance training and resistance training+curcumin groups significantly increased and decreased, respectively, compared to the control group (P = 0.001). The MuRF1 expression in the curcumin+resistance training group decreased significantly (P = 0.013) compared to the placebo and curcumin groups. The expression of collagen type 1 and type 2 in all the three treatment groups had significant decreases compared to the placebo group (P &lt; 0.05). Conclusions: Considering the results of this study, resistance training and curcumin supplement each alone can prevent cardiac muscle atrophy. However, the simultaneous use of curcumin supplement and resistance training can lead to synergistic effects.

UTERINE SCAR FORMATION AND EXPRESSION OF SPECIFIC CONNECTIVE TISSUE PROTEINS

The aim of the study – to evaluate the role of expression of specific connective tissue proteins in the formation of the scar on the uterus.&#x0D; Materials and Methods. The study was performed on the basis of the regional perinatal center and KU PB No. 5 during 2017–2019. 426 women with a scar on the uterus were examined, including 115 (27.0 %) with two or more scars. The mean age of the subjects was (33.4±1.1) years. The results of sonographic studies were analyzed. For pathomorphological examination, three cases of intranatal rupture of the uterine wall along the “old” scar after cesarean section from middle-aged women with a gestational age of 33 to 40 weeks are presented. The expression of collagen types 1 and 3 and the protein of the intermediate filaments of the muscle tissue of desmin was determined using a semi-quantitative method with the calculation of H-score.&#x0D; Results and Discussion. Analysis of ultrasound data showed that the average assessment of the degree of insolvency of the scar on the uterus was in the examined women X=(1.3±0.2) points, a high level of insolvency was found in 114 (26.8 %) pregnant women. At the same time, 225 (52.8 %) pregnant women showed no signs of scar failure at all. In 87 (20.4 %) one sign of uterine scar failure was found, in 76 (17.8 %) – two signs, in 20 (4.7 %) – three signs, and in 18 (4.2 %) – four signs. Subsequently, 44 (10.3 %) women were born operatively, the rest gave birth per via naturales. Local myometrial defect after ultrasound delivery was detected in 25 of 182 (13.7 %) women who gave birth independently.&#x0D; Conclusions. A high level of scar failure risk is found in 26.8 % of pregnant women. H-score valuesfor collagen types 1 and 3 averaged (212±24) and (188±22), for desmin – (193±17). Thus, reparative processes at the site of previous surgery on the uterine wall are by incomplete regeneration (substitution) and compensatory hyperplasia of structural tissue elements.

PPARγ modulates refractive development and form deprivation myopia in Guinea pigs

Form deprivation myopia (FDM) is characterized by loss of choroidal thickness (ChT), reduced choroidal blood perfusion (ChBP), and consequently scleral hypoxia. In some tissues, changes in levels of peroxisome proliferator-activated receptor γ (PPARγ) expression modulate hypoxia-induced pathological responses. We determined if PPARγ modulates FDM through changes in ChT, ChBP, scleral hypoxia-inducible transcription factor (HIF-1α) that in turn regulate scleral collagen type 1 (COL1) expression levels in guinea pigs. Myopia was induced by occluding one eye, while the fellow eye served as control. They received daily peribulbar injections of either the PPARγ antagonist GW9662, or the GW1929 agonist, with or without ocular occlusion for 4 weeks. Ocular refraction and biometric parameters were estimated at baseline, 2 and 4 weeks post-treatment. ChT and ChBP were measured at the 2- and 4-week time points. Western blot analysis determined the expression levels of scleral HIF-1α and COL1. GW9662 induced a myopic shift in unoccluded eyes. Conversely, GW1929 inhibited FDM progression without affecting the refraction in unoccluded eyes. GW9662 reduced both ChT and ChBP in unoccluded eyes, while GW1929 inhibited their declines in occluded eyes. Scleral HIF-1α expression rose in GW9662-treated unoccluded eyes whereas GW1929 reduced HIF-1α upregulation in occluded eyes. GW9662 downregulated scleral COL1 expression in unoccluded eyes, while GW1929 reduced their decreases in occluded eyes. Therefore, PPARγ modulates collagen expression levels and FDM through an inverse relationship between changes in PPARγ and HIF-1α expression levels.

The Inhibition of Cathepsin G on Endometrial Explants With Endometrosis in the Mare

Although proteases found in neutrophil extracellular traps (NETs) have antimicrobial properties, they also stimulate collagen type 1 (COL1) production by the mare endometrium, contributing for the development of endometrosis. Cathepsin G (CAT), a protease present in NETs, is inhibited by specific inhibitors, such as cathepsin G inhibitor I (INH; β-keto-phosphonic acid). Matrix metallopeptidases (MMPs) are proteases involved in the equilibrium of the extracellular matrix. The objective of this study was to investigate the effect of CAT and INH (a selective CAT inhibitor) on the expression of MMP-2 and MMP-9 and on gelatinolytic activity. In addition, the putative inhibitory effect of INH on CAT-induced COL1 production in mare endometrium was assessed. Endometrial explants retrieved from mares in follicular phase or midluteal phase were treated for 24 or 48 h with CAT, inhibitor alone, or both treatments. In explants, transcripts (quantitative polymerase chain reaction) of COL1A2, MMP2, and MMP9, as well as the relative abundance of COL1 protein (Western blot), and activity of MMP-2 and MMP-9 (zymography) were evaluated. The protease CAT induced COL1 expression in explants, at both estrous cycle phases and treatment times. The inhibitory effect of INH was observed on COL1A2 transcripts in follicular phase at 24-h treatment, and in midluteal phase at 48 h (P < 0.05), and on the relative abundance of COL protein in follicular phase and midluteal phase explants, at 48 h (P < 0.001). Our study suggests that MMP-2 might also be involved in an earlier response to CAT, and MMP-9 in a later response, mainly in the follicular phase. While the use of INH reduced CAT-induced COL1 endometrial expression, MMPs might be involved in the fibrogenic response to CAT. Therefore, in mare endometrium, the use of INH may be a future potential therapeutic means to reduce CAT-induced COL1 formation and to hamper endometrosis establishment.

Expression of collagen type 1 alpha 1 indicates lymph node metastasis and poor outcomes in squamous cell carcinomas of the lung.

BackgroundSquamous cell carcinomas of the lung are an extremely common and deadly form of non-small cell lung cancers. Clinical management of the disease is dependent on staging and metastatic status. Metastasis to the lymph node is especially crucial to diagnose as it occurs at an earlier stage. However, lymphadenectomies are invasive and tumor cells may be overlooked during evaluation.There are limited approved biomarkers for predicting lymph node metastasis with squamous cell carcinomas of the lung (LSCC).MethodsGenome data of 60 tumor-adjacent samples were downloaded from Genome Expression Omnibus. We identified over-expressed HUB genes using Cytoscape as key prognostic markers. The selected markers were further evaluated based on gene ontology and overall expression levels compared to normal tissue using The Cancer Genome Atlas. We further validated these results using clinical biopsy tissue taken from squamous cell carcinoma patients.ResultsAnalysis of the genome expression data resulted in 13 relevant hub genes that were differentially expressed in cancerous samples. All of these genes are associated with collagen biosynthesis within the tumor microenvironment. We chose Collagen Type 1 Alpha 1 (COL1A1) as the most relevant prognostic marker due to its high number of pathway connections and over expression in the tumor microenvironment compared to the other 12 genes. Additionally, based on analysis of The Cancer Genome Atlas, tumors with higher levels of COL1A1 expression are associated with poorer overall survival. Finally, evaluation of clinical biopsy samples suggests that overexpression of COL1A1 in the LSCC microenvironment highly correlates with lymph node metastasis. These results suggest COL1A1 is a clinically relevant marker that should be used to justify lymphadenectomies.

The Impact of a Polyphenol-Rich Extract from the Berries of Aronia melanocarpa L. on Collagen Metabolism in the Liver: A Study in an In Vivo Model of Human Environmental Exposure to Cadmium.

This study examined whether a polyphenol-rich extract from the berries of Aronia melanocarpa L. (AE; chokeberries) may protect from the impact of cadmium (Cd) on the metabolism of collagen in the liver. The study was conducted in an experimental model (rats that were fed a diet containing 1 or 5 mg Cd/kg for 3–24 months) of human exposure to this xenobiotic during a lifetime. The concentration of total collagen and the expression of collagen types I and III at the mRNA and protein levels, as well as the concentrations of matrix metalloproteinases (MMP-1 and MMP-2) and their tissue inhibitors (TIMP-1 and TIMP-2), were assayed. The administration of Cd and/or AE had only a slight and temporary impact on the concentration of total collagen in the liver. The supplementation with AE significantly prevented Cd-mediated changes in the expression of collagen types I and III at the mRNA and protein levels and their ratio (collagen III/collagen I), as well as a rise in the concentrations of MMPs and TIMPs in this organ. The results allow the conclusion that the intake of chokeberry products in the case of Cd intoxication may be effective in prevention from this xenobiotic-induced disturbance in collagen homeostasis in the liver.

Hepatocyte Growth Factor Secreted from Human Adipose-Derived Stem Cells Inhibits Fibrosis in Hypertrophic Scar Fibroblasts.

To study the effect of Adipose-derived stem cells (ADSCs) on fibrosis of hypertrophic scar-derived fibroblasts (HSFs) and its concrete mechanism. ADSCs have been reported to reduce collagen production and fibroblast proliferation in co-culture experiments. Conditioned medium from adipose-derived stem cells (ADSCs-CM) has successfully inhibited fibrosis by decreasing the expression of collagen type І (Col1) and α-smooth muscle actin (α-SMA) in rabbit ear scar models. Hepatocyte growth factor (HGF), the primary growth factor in ADSCs-CM, has been shown to reverse fibrosis in various fibrotic diseases. To test the hypothesis that ADSCs inhibit fibrosis of HSFs through the secretion of HGF. HSFs were treated with DMEM containing 0%, 10%, 50% and 100% concentration of ADSCs-CM. The effect of ADSCs-CM on the viability was determined by cell viability assay, and the collagen production in HSFs was examined by Sirius red staining. Expression and secretion of fibrosis and degradation proteins were detected separately. After measuring the concentration of HGF in ADSCs-CM, the same number of HSFs were treated with 50% ADSCs-CM or HGF. HGF activity in ADSCs-CM was neutralized with a goat anti-human HGF antibody. The results demonstrated that ADSCs-CM dose-dependently decreased cell viability, expression of fibrosis molecules, and tissue inhibitor of metalloproteinases-1 (TIMP-1), and significantly increased matrix metalloproteinase-1 (MMP-1) expression in HSFs. Collagen production and the ratio of collagen type І and type III (Col1/Col3) were also suppressed by ADSCs-CM in a dose-dependent manner. When HSFs were cultured with either 50% ADSCs-CM or HGF (1 ng/ml), a similar trend was observed in gene expression and protein secretion. Adding an HGF antibody to both groups returned protein expression and secretion to basal levels but did not significantly affect the fibrosis factors in the control group. Our findings revealed that adipose-derived stem cell-secreted HGF effectively inhibits fibrosis-related factors and regulates extracellular matrix (ECM) remodeling in hypertrophic scar fibroblasts.

Identification of potential biomarkers for predicting the early onset of diabetic cardiomyopathy in a mouse model

Type 2 diabetes (T2D) is characterized by metabolic derangements that cause a shift in substrate preference, inducing cardiac interstitial fibrosis. Interstitial fibrosis plays a key role in aggravating left ventricular diastolic dysfunction (LVDD), which has previously been associated with the asymptomatic onset of heart failure. The latter is responsible for 80% of deaths among diabetic patients and has been termed diabetic cardiomyopathy (DCM). Through in silico prediction and subsequent detection in a leptin receptor-deficient db/db mice model (db/db), we confirmed the presence of previously identified potential biomarkers to detect the early onset of DCM. Differential expression of Lysyl Oxidase Like 2 (LOXL2) and Electron Transfer Flavoprotein Beta Subunit (ETFβ), in both serum and heart tissue of 6–16-week-old db/db mice, correlated with a reduced left-ventricular diastolic dysfunction as assessed by high-resolution Doppler echocardiography. Principal component analysis of the combined biomarkers, LOXL2 and ETFβ, further displayed a significant difference between wild type and db/db mice from as early as 9 weeks of age. Knockdown in H9c2 cells, utilising siRNA of either LOXL2 or ETFβ, revealed a decrease in the expression of Collagen Type I Alpha1 (COL1A1), a marker known to contribute to enhanced myocardial fibrosis. Additionally, receiver-operating curve (ROC) analysis of the proposed diagnostic profile showed that the combination of LOXL2 and ETFβ resulted in an area under the curve (AUC) of 0.813, with a cut-off point of 0.824, thus suggesting the favorable positive predictive power of the model and further supporting the use of LOXL2 and ETFβ as possible early predictive DCM biomarkers.

Resveratrol counteracts IL-1β-mediated impairment of extracellular matrix deposition in 3D articular chondrocyte constructs.

When aiming at cell-based therapies in osteoarthritis (OA), proinflammatory conditions mediated by cytokines such as IL-1β need to be considered. In recent studies, the phytoalexin resveratrol (RSV) has exhibited potent anti-inflammatory properties. However, long-term effects on 3D cartilaginous constructs under inflammatory conditions with regard to tissue quality, especially extracellular matrix (ECM) composition, have remained unexplored. Therefore, we employed long-term model cultures for cell-based therapies in an in vitro OA environment and evaluated effects of RSV. Pellet constructs made from expanded porcine articular chondrocytes were cultured with either IL-1β (1-10 ng/ml) or RSV (50 μM) alone, or a cotreatment with both agents. Treatments were applied for 14 days, either directly after pellet formation or after a preculture period of 7 days. Culture with IL-1β (10 ng/ml) decreased pellet size and DNA amount and severely compromised glycosaminoglycan (GAG) and collagen content. Cotreatment with RSV distinctly counteracted the proinflammatory catabolism and led to partial rescue of the ECM composition in both culture systems, with especially strong effects on GAG. Marked MMP13 expression was detected in IL-1β-treated pellets, but none upon RSV cotreatment. Expression of collagen type I was increased upon IL-1β treatment and still observed when adding RSV, whereas collagen type X, indicating hypertrophy, was detected exclusively in pellets treated with RSV alone. In conclusion, RSV can counteract IL-1β-mediated degradation and distinctly improve cartilaginous ECM deposition in 3D long-term inflammatory cultures. Nevertheless, potential hypertrophic effects should be taken into account when considering RSV as cotreatment for articular cartilage repair techniques.

HMGB1 Aggravates Pressure Overload-Induced Left Ventricular Dysfunction by Promoting Myocardial Fibrosis

Aim. Fibrosis had important effects on pressure overload-induced left ventricular (LV) dysfunction. High-mobility group box 1 (HMGB1), which was closely associated with fibrosis, was involved in the pressure overload-induced cardiac injury. This study determines the role of HMGB1 in LV dysfunction under pressure overload. Methods. Transverse aortic constriction (TAC) operation was performed on male C57BL/6J mice to build the model of pressure overload, while HMGB1 or PBS was injected into the LV wall. Cardiac function, collagen volume, and relevant genes were detected. Results. Echocardiography demonstrated that the levels of LV ejection fraction (LVEF) were markedly decreased on day 28 after TAC, which was consistent with raised collagen in the myocardium. Moreover, we found that the exposure of mice to TAC + HMGB1 is associated with higher mortality, BNP, and collagen volume in the myocardium and lower LVEF. In addition, real-time PCR showed that the expression of collagen type I, TGF-β, and MMP2 markedly increased in the myocardium after TAC, while HMGB1 overexpression further raised the TGF-β expression but not collagen type I and MMP2 expressions. Conclusion. This study indicated that exogenous HMGB1 overexpression in the myocardium aggravated the pressure overload-induced LV dysfunction by promoting cardiac fibrosis, which may be mediated by increasing the TGF-β expression.

The Effect of Aerobic Exercise on Collagen Type I and IV Gene Expression and Collagen Type I Protein Changes in the Sciatic Nerve of Diabetic Rats

Background: Neuropathy is one of the complications of diabetes, probably due to the destruction of the extracellular matrix and the thickening of the peripheral nerve basement membrane. However, its mechanisms and the impact of exercise on these disorders has not fully understood. The purpose of the present study was to investigate the effect of aerobic exercise on collagen levels of type I and IV and collagen type I protein changes in the sciatic nerve of diabetic rats. Materials and Methods: Eighteen 10-week-old Wistar male rats weighing 250 ± 20 g were randomly divided into three groups of healthy control (n=6), diabetic (n=6) and diabetic + aerobic exercise (n=6). For this purpose, after introduction and adaptation of rats to new environment, diabetes was induced by single dose injection of dissolved streptozotocin in sodium citrate buffer at pH=4.5 at 45 mg/kg intraperitoneal. After confirming neuropathic conditions (with behavioral tests), diabetic+exercise rats underwent moderate-intensity aerobic exercise on the treadmill for 8 week. At the beginning and at the end of the period, blood glucose of all rats was measured by glucometer and the mean of each group was measured separately. Changes in collagen type I and IV gene expression, and collagen type I protein levels in sciatic nerve of rats were evaluated by real-time PCR technique and immunohistochemistry, respectively. Results: Diabetes increased collagen type I and IV gene expression and collagen type I protein levels in the sciatic nerve samples of rats. However, exercise reduced blood sugar levels and expression of collagen type I and IV genes (p=0.05) and collagen type I protein significantly reduced in sciatic nerve (p=0.001). Conclusion: The results of the present study showed that aerobic exercise as a non-pharmacological strategy by negative regulating type I and IV collagen factors at the gene and protein level, was able to control and inhibit the effects of diabetes on extracellular matrix components in the sciatic nerve.

Decellularized Porcine Epiphyseal Plate-Derived Extracellular Matrix Powder: Synthesis and Characterization

The aim of this study was to develop a porcine epiphyseal plate-derived extracellular matrix powder (PEPEP) for epiphyseal plate regeneration. PEPEP was characterized by chemical assay to determine the contents of DNA and epiphyseal plate complex chemical components (glycosaminoglycan and hydroxyproline). The effects of PEPEP on the viability, proliferation, and differentiation of human bone marrow mesenchymal stem cells (hBMSCs) were also evaluated. hBMSCs cultured in PEPEP exhibited a good distribution with excellent viability after 72 h, demonstrating the ability of PEPEP to support hBMSC proliferation. At week 4 and 6 in vitro, the PEPEP + hBMSCs structure showed chondrogenic ability and an increase in expression of collagen type I, type II, and type X. PEPEP showed a promising ability to enhance cartilage formation and promote chondrocyte differentiation, maturation, and hypertrophy. The results provide insights into the feasibility of PEPEP as a potential material for tissue engineering applications.