Increase In Hydroxyproline Content Research Articles

MMP-3 mediates copper oxide nanoparticle-induced pulmonary inflammation and fibrosis

BackgroundThe increasing production and usage of copper oxide nanoparticles (Nano-CuO) raise human health concerns. Previous studies have demonstrated that exposure to Nano-CuO could induce lung inflammation, injury, and fibrosis. However, the potential underlying mechanisms are still unclear. Here, we proposed that matrix metalloproteinase-3 (MMP-3) might play an important role in Nano-CuO-induced lung inflammation, injury, and fibrosis.ResultsExposure of mice to Nano-CuO caused acute lung inflammation and injury in a dose-dependent manner, which was reflected by increased total cell number, neutrophil count, macrophage count, lactate dehydrogenase (LDH) activity, and CXCL1/KC level in bronchoalveolar lavage fluid (BALF) obtained on day 3 post-exposure. The time-response study showed that Nano-CuO-induced acute lung inflammation and injury appeared as early as day 1 after exposure, peaked on day 3, and ameliorated over time. However, even on day 42 post-exposure, the LDH activity and macrophage count were still higher than those in the control group, suggesting that Nano-CuO caused chronic lung inflammation. The Nano-CuO-induced pulmonary inflammation was further confirmed by H&E staining of lung sections. Trichrome staining showed that Nano-CuO exposure caused pulmonary fibrosis from day 14 to day 42 post-exposure with an increasing tendency over time. Increased hydroxyproline content and expression levels of fibrosis-associated proteins in mouse lungs were also observed. In addition, Nano-CuO exposure induced MMP-3 overexpression and increased MMP-3 secretion in mouse lungs. Knocking down MMP-3 in mouse lungs significantly attenuated Nano-CuO-induced acute and chronic lung inflammation and fibrosis. Moreover, Nano-CuO exposure caused sustained production of cleaved osteopontin (OPN) in mouse lungs, which was also significantly decreased by knocking down MMP-3.ConclusionsOur results demonstrated that short-term Nano-CuO exposure caused acute lung inflammation and injury, while long-term exposure induced chronic pulmonary inflammation and fibrosis. Knocking down MMP-3 significantly ameliorated Nano-CuO-induced pulmonary inflammation, injury, and fibrosis, and also attenuated Nano-CuO-induced cleaved OPN level. Our study suggests that MMP-3 may play important roles in Nano-CuO-induced pulmonary inflammation and fibrosis via cleavage of OPN and may provide a further understanding of the mechanisms underlying Nano-CuO-induced pulmonary toxicity.Graphical

Exploring the wound healing activity of phytosomal gel of Annona squamosa and Cinnamomum tamala leaves ethanolic extracts with antioxidant and antimicrobial activities in S aureus infected excision wound model

Wound healing is a natural process but it is impaired in certain conditions like age, stress, health, immunity status and microbial infection. Particularly in cases of chronic wounds, infection is nearly often the main and unavoidable obstacle to wound healing. For this purpose, leaves of Annona squamosa and Cinnamomum tamala were selected based on their ethnopharmacological uses and reported pharmacological activities. The ethanolic extracts of both plant parts i.e. ethanolic extracts of Annona squamosa (ASEE) and Cinnamomum tamala (CTEE) were evaluated for their antioxidant and antimicrobial activities individually as well as in 1:1 combination as Polyherbal Ethanolic extract (PHEE). In our previous work both these ethanolic extracts were combined and phytosomes were prepared by thin layer hydration method and optimized for vesicle size and entrapment efficiency. The phytosomes were then incorporated into Carbopol gel matrix. In this present study the selected phytosomal gel was tested in two different concentrations (2% and 5%) for in vivo wound healing activity using S. aureus infected excision wound model. The various parameters examined were percentage wound contraction, epithelization period, bacteriological quantification, biochemical parameters like Superoxide dismutase (SOD), Catalase and hydroxyproline. The PHEE exhibited synergistic antioxidant activity. The PHEE also showed enhanced antimicrobial activity against bacteria namely gram-positive S. aureus, gram-negative E. Coli. The phytosomal gel showed increased wound contraction, reduced time of epithelization, increased hydroxyproline content, increased levels of SOD and Catalase enzymes and reduced bacterial load when compared with Povidone iodine ointment as standard in S. aureus infected excision wound model.

An analysis combining proteomics and transcriptomics revealed a regulation target of sea cucumber autolysis

Sea cucumber is a valuable seafood product and autolysis is the main concern for the aquaculture industry. This study employed proteomics and transcriptomics to investigate the autolysis mechanism of sea cucumbers. The fresh sea cucumber was exposed to UV light to induce autolysis. The body wall samples were cut off to analyze by proteomics and transcriptomics. The angiotensin-converting enzyme (ACE) inhibitor of teprotide and the activator of imatinib were gastric gavage to live sea cucumbers, respectively, to identify the regulation target. Autolysis occurrence was evaluated by appearance, soluble peptide, and hydroxyproline content. Four gene-protein pairs were ACE, AJAP10923, Heme-binding protein 2-like, and Ficolin-2-like. Only the ACE protein and gene changed synchronously and a significant down-regulation of ACE occurred in the autolysis sea cucumbers. Teprotide led to a 1.58-fold increase in the TCA-soluble protein content and a 1.57-fold increase in hydroxyproline content. No significant differences were observed between imatinib-treated sea cucumbers and fresh ones regarding TCA-soluble protein content or hydroxyproline levels (P > 0.05). ACE inhibitor accelerated the autolysis of sea cucumber, but ACE activator inhibited the autolysis. Therefore, ACE can serve as a regulatory target for autolysis in sea cucumbers.

Remodeled Extracellular Matrix Guides Endothelial Cell Phenotype and Transcriptome to Fibrotic Niche

Impairment of endothelial barrier integrity causes vascular injury leading to acute lung injury (ALI), which, if not resolved, can compromise a patient’s lung functions and mortality. While it is known that both endothelial cells (EC) and their underlying extracellular matrix (ECM) are required for preserving lung homeostasis and functions, much of the efforts to understand mechanisms regulating these functions have focused mainly on EC. A fundamental unresolved question is whether lung-remodeled ECM formed due to leaky endothelium impacts EC functions, and if so, how? Herein, using lung ECM derived from a mouse lacking focal adhesion kinase (FAK), specifically in the endothelium (remodeled ECM), we assessed ECM components and their role in regulating EC functions and their relevance in vivo. Previously, we have shown that conditional deletion of FAK in endothelium increases EC permeability and induces vascular injury at homeostasis. Mass spectrometric analysis showed 805 proteins in control ECM (FAKfl/fl mice lung derived ECM) consisting of glycoproteins, matrisome-associated proteins, and secreted factors. However, the remodeled ECM had 722 proteins and was highly enriched with fibrillin 1, fibrinogen, and collagen I. Moreover, the remodeled ECM was two-fold stiffer than the control ECM. Remodeled ECM with its inherent composition recapitulates the 3D microenvironment, encouraging EC morphological alteration, increased cell stiffness, and enhanced focal adhesion (FA) formation compared to control ECM. In addition, RNA sequencing identified increased expression of genes linked with fibrosis, including integrin b8, TGFb2, GDF15, SMAD7, COL1A1, IL33, ID3, BMPR1A, and ADAMTS4 in EC seeded on remodeled ECM. Interestingly, fibrotic signaling observed in EC plated on remodeled ECM mimicked in vivo in EC-FAK−/− mice as they showed increased hydroxyproline content and collagen deposition in the lungs, indicating loss of EC-FAK as the crucial initiator of fibrotic signaling. In summary, studies for the first time show disrupted EC signaling as the instructor of fibrotic niche in the lungs for targeting pulmonary fibrosis. NIH. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Chemical composition, antioxidant, antimicrobial, and wound healing effects of Trachyspermum roxburghianum (DC.) H. Wolff essential oil: An in vivo and in silico approach

Ethnopharmacological relevanceTrachyspermum roxburghianum (DC.) H. Wolff, commonly known as ‘Ajamoda,’ is a neglected Indian spice highly used in Ayurveda and folklore remedies as an antimicrobial for chronic wounds and discharges, along with many other disease conditions. Aim of the studyThe objective of the study was to explore chemical composition and to investigate the antioxidant, antimicrobial, analgesic, and wound healing activities of T. roxburghianum fruit essential oil from India. Materials and methodsThe phytochemical characterization of the oil was determined through standard qualitative procedures and the gas chromatography-mass spectrometry (GC-MS) technique. The in vitro antioxidant aptitude was assessed by scavenging DPPH and ABTS radicals. The antimicrobial potential of the oil was investigated using the disc diffusion method, followed by the determination of minimum inhibitory concentration against Gram-positive and Gram-negative bacterial and fungal strains. The analgesic potential was evaluated using thermal and chemically induced pain models in Swiss albino mice. Wound healing was assessed in vivo, including determining wound contraction rates, histopathology, and hydroxyproline estimation, using the excision wound model in Swiss albino mice. ResultsGC-MS analysis identified 55 compounds with major terpenoids, including thymol (13.8%), limonene (11.5%), and others. Substantial radical-scavenging activity was exhibited by T. roxburghianum fruit essential oil (TREO) (IC50 94.41 ± 2.00 μg/mL in DPPH assay and 91.28 ± 1.94 μg/mL in ABTS assay). Microorganisms were inhibited with low MIC (2 μL/mL for the inhibition of Staphylococcus aureus and Bacillus subtilis; 4 μL/mL against Salmonella typhi and 16 μL/mL against Candida albicans). In the cytotoxicity study, no cytotoxicity was observed on the Monkey Normal Kidney Cell line (Vero). Significant antinociceptive effects were observed (25.47 ± 1.10 % of inhibition at 100 mg/kg and 44.31 ± 1.69 % at 200 mg/kg). A remarkable rate of wound closure and epithelization, along with a marked increase in hydroxyproline content, were observed for the oil during wound healing in mice. ConclusionsThe results suggested that oil could be utilized as a potential source of wound healing therapeutics.

Bisphenol A exposure stimulates prostatic fibrosis via exosome-triggered epithelium changes

Fibrosis is the pathological basis for the clinical progression of benign prostatic hyperplasia (BPH). Prostatic fibrosis is an important risk factor in patients with BPH who experience lower urinary tract symptoms. Bisphenol A (BPA) is an environmental endocrine disruptor (EED) that causes prostate defects. The effects of BPA on the prostate were investigated in this study using mouse and human prostate cell models. BPA-induced mouse prostatic fibrosis is characterized by collagen deposition and an increase in hydroxyproline concentration. Furthermore, BPA-exposed prostatic stromal fibroblasts exosomes promote the epithelial-mesenchymal transition of epithelial cells. High-throughput RNA sequencing and functional enrichment analyses show that substantially altered mRNAs, lncRNAs and circRNAs play roles in cellular interactions and the hypoxia-inducible factor-1 signaling pathway. The results showed that exosomes participated in the pro-fibrogenic effects of BPA on the prostate by mediating communication between stromal and epithelial cells and triggering epithelial changes.

The anti-liver fibrosis effect of Tibetan medicine (Qiwei Tiexie capsule) is related to the inhibition of NLRP3 inflammasome activation in vivo and in vitro

Ethnopharmacological relevanceQiwei Tiexie capsule (QWTX) is an improved form of a classical prescription of Tibetan medicine—Qiwei Tiexie pill. It has been employed in the treatment of a variety of chronic liver disorders, including liver fibrosis. Uncertainty still exists regarding the mechanism of QWTX action in liver fibrosis. Aim of the studyConfirm the anti-liver fibrosis effect of QWTX and reveal its mechanism from the perspective of NOD-like receptor protein 3 (NLRP3) inflammasome activation. Materials and methodsIn vivo experiment: A rat model of carbon tetrachloride -induced liver fibrosis was constructed. All rats were randomly divided into six groups: a control group, a model group, a group receiving the positive drug (Biejia Ruangan tablet), and three groups receiving QWTX at high, medium, and low doses. The contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TBil) were detected in serum. Hematoxylin and eosin staining and Masson's staining were used to assess the histomorphological alteration of the liver. The levels of glutathione peroxidase, hydroxyproline, tumor necrosis factor alpha (TNF-α), and interleukin 1 beta (IL-1β) in the liver were determined using the corresponding detection kits. Real-time polymerase chain reaction, immunofluorescence, and western blotting were used to determine the expression levels of NLRP3, adaptor protein (ASC), caspase-1, and alpha-smooth muscle actin (α-SMA). In vitro experiment: Four groups of rat hepatic stellate cell line (HSC-T6) cells were created: the control group, the low-dose QWTX group (0.05 mg/mL), the medium-dose QWTX group (0.1 mg/mL), and the high-dose QWTX group (0.2 mg/mL). Cell viability was assessed using a cell counting kit, and the amounts of collagen type I (Col I) and IL-1β in the cell lysate were measured using an enzyme-linked immunosorbent assay kit. The mRNA and protein expression of NLRP3, ASC, caspase-1, and α-SMA were also estimated. ResultsQWTX had an inhibitory effect on liver fibrosis and a negative effect on HSC activation, while it improved liver histopathological injury and abnormal liver function and increased hydroxyproline content and glutathione peroxidase activity in vivo. QWTX decreased the expression of α-SMA, NLRP3, caspase-1, ASC, and IL-1β both in vitro and in vivo. ConclusionsTibetan medicine QWTX had a significant anti-liver fibrosis effect that was related to the inhibition of NLRP3 inflammasome activation in vivo and in vitro.

Alogliptin exhibits multifaceted effects in thioacetamide-insulted rats: A novel approach to combating hepatic inflammation and fibrogenesis

Hepatic fibrosis arising from chronic liver injury is characterized by dysregulated healing, including hepatic stellate cell activation and excessive deposition of extracellular matrix proteins. Administration of the hepatotoxin thioacetamide (TAA) induces liver injury coupled to fibrogenesis in rodents, mimicking aspects of human disease. Alogliptin is a highly selective inhibitor of dipeptidyl peptidase-4 with purported antifibrotic actions. We investigated the protective effects of alogliptin against TAA-mediated hepatic fibrosis in rats. Adult male Sprague-Dawley rats received intraperitoneal injections of TAA (150 mg/kg) twice weekly for 6 weeks to induce liver fibrosis. A subset of rats also received daily oral alogliptin (20 mg/kg). At 6 weeks, liver injury and fibrosis were assessed by histology, hydroxyproline content, serum liver enzymes, inflammatory cytokines, oxidative stress markers, and genes related to inflammation, apoptosis, and fibrosis. TAA elicited necroinflammation, oxidative stress, upregulation of pro-fibrogenic mediators, increased hydroxyproline content, and excessive collagen deposition, indicating hepatic fibrosis. The administration of Alogliptin led to notable enhancements in liver histology, an extension in survival time, a decrease in hydroxyproline levels and the expression of fibrogenic genes, a reduction in inflammatory cytokines and oxidative stress, and mitigation of hepatocellular apoptosis in rats subjected to TAA treatment. Alogliptin displayed potent antifibrotic, antioxidant, and hepatoprotective properties in this model of toxic liver damage, likely by impeding NFκB while enhanced Nrf2 DNA binding activity which together modulate oxidative stress, inflammation, myofibroblast activation, and apoptosis. These results highlight the potential therapeutic value of alogliptin offering hope for improved treatment of hepatic fibrosis.

Healing potential of curcumin nanomicelles in cutaneous burn wounds: an in vitro and in vivo study

ABSTRACT Purpose/Aim of the study Curcumin is the active substance of turmeric and has been shown to enhance the healing potential of burn wounds. However, its high hydrophobicity and rapid degradability are great challenges for its clinical applications. The development of new curcumin formulations may provide a potential solution to these issues. Methods and Results In this study, we investigated the use of curcumin nanomicelles for wound dressing and evaluated their effects on fibroblast migration and proliferation in vitro. We found that the application of curcumin nanomicelles to the wounds significantly improved wound contraction and increased the expression of transforming growth factor-1 and basic fibroblast growth factor at day 14 of the healing process. Furthermore, curcumin nanomicelles reduced the expression of interleukin-1 at days 7 and 14 post-wounding. Histopathological analysis revealed that the curcumin nanomicelles-treated burn wounds exhibited more organized granulation tissue, improved angiogenesis, and enhanced re-epithelialization. Additionally, the curcumin treatment led to increased hydroxyproline content and enhanced TGF-β1 expression level in the wounds. The in vitro studies also demonstrated that the curcumin nanomicelles induced proliferation and migration of fibroblasts. Conclusion Overall, our findings suggest that curcumin nanomicelles can be a promising candidate for the treatment of burn wounds.

Assessment of Hydroxyproline Content in Rabbit Achilles Tendon Treated with Platelet Rich Fibrin (PRF).

Tendon is similar to rope and consists of strong, flexible, dense connective tissue. Tendon disorder healing is challenging as it is an avascular tissue. The repaired tissue appears scar-like, and its biomechanical properties never ultimately return to their pre-injury state. This study aimed to evaluate the effects of platelet-rich fibrin (PRF) on the hydroxyproline content of the Achilles tendon after injury. For this purpose, 24 adult rabbits weighing 1.5-2 kg were used in this study. The animals were divided into three groups of eight, including the advanced-PRF (A-PRF) group in which the tendon defect was treated with xenogeneic A-PRF, the leukocyte-PRF (L-PRF) group in which xenogeneic L-PRF was used for tendon defect treatment, and a control group which was treated with normal saline. Hydroxyproline concentration was measured 1 and 2 months after the operation. Clinically, lameness was improved in the A-PRF group, compared to the L-PRF and control groups at the end of the third week after the surgery. Hydroxyproline level was significantly increased in the A-PRF group (50.33±1.44), compared to the L-PRF (44.70±1.12) and control (35.97±1.05) groups 2 months after the surgery (P<0.05). Moreover, the L-PRF group showed an increase in hydroxyproline content, compared to the control at the same period. The results of the current study demonstrated that A-PRF could enhance the hydroxyproline content of rabbit Achilles tendon after injury. Xenogenic PRF can be used as an alternative biomaterial to accelerate and regenerate tendon tissue.

Wound Healing Potential of Couroupita guianensis Aubl. Fruit Pulp Investigated on Excision Wound Model.

Wound care management aims at stimulating and improving healing process without scar formation. Although various plants have been reported to possess wound healing properties in tribal and folklore medicines, there is a lack of scientific data to validate the claim. In this aspect, it becomes inevitable to prove the efficacy of naturally derived products at pharmacological levels. Couroupita guianensis as a whole plant has been reported to exhibit wound healing activity. The leaves and fruit of this plant have been utilized in folkloric medicine to cure skin diseases and infections for many years. However, to the best of our knowledge, no scientific studies have been conducted to verify the wound healing properties of C. guianensis fruit pulp. Therefore, the present study seeks to investigate the wound healing potential of C. guianensis fruit pulp using an excision wound model in Wistar albino male rats. This study indicated that the ointment prepared from crude ethanolic extract of C. guianensis fruit pulp facilitated wound contraction that were evidenced by a greater reduction in the wound area and epithelialization period and increased hydroxyproline content. The experimental groups treated with low and mid dose of C. guianensis ethanol extract (CGEE) ointments had shown a wound closure of 80.27% and 89.11% respectively within 15days, which is comparable to the standard betadine ointment which showed 91.44% healing in the treated groups. Further, the extract influenced the expression of genes VEGF and TGF-β on post wounding days that clearly explained the strong correlation between these genes and wound healing in the experimental rats. The animals treated with 10% CGEE ointment showed a significant upregulation of both VEGF and TGF-β as compared with other test and standard groups. These findings provide credence to the conventional application of this plant in the healing of wounds and other dermatological conditions, and may represent a therapeutic strategy for the treatment of wounds.

Evaluation of the Antioxidant and Wound Healing Properties of 80% Methanol Extract and Solvent Fractions of the Leaves of Vernonia auriculifera Hiern. (Asteraceae).

The leaves of Vernonia auriculifera (Asteraceae) have traditionally been used to treat wounds in several regions of Ethiopia. The purpose of this study was to assess the wound healing properties of the leaf extract and solvent fractions of V. auriculifera in mice. The leaf extract and solvent fractions of V. auriculifera have also been evaluated for their anti-oxidant properties because of their impact on the wound healing process. Air-dried leaves were extracted using 80% methanol. They were also successively fractionated with n-hexane, ethyl acetate, and methanol. The residue was then macerated in water for 72 hr. Simple ointment bases were formulated according to British Pharmacopoeia. Thereafter, two types of ointment formulations, 2.5% w/w and 5% w/w, were formulated. Wound healing and acute dermal toxicity studies were performed on mice. To assess free radical scavenging activity, a 2,2-diphenyl-2-picrylhydrazyl free radical (DPPH) assay was performed. In both models, wounds treated with 2.5% and 5% (w/w) of the ME, the aqueous fraction (AQF), methanol fraction (MEF), and ethyl acetate fraction (EAF) ointments demonstrated significant wound healing activity, as shown by enhanced wound contraction, a shortened epithelialization time, increased hydroxyproline content, and enhanced tissue breaking strength. The extract and solvent fractions displayed free radical scavenging activity with IC50 values of 1.2 mg/mL, 1.46 mg/mL, 1.5 mg/mL, and 2.83 mg/mL for ME, AQF, MEF, and EAF, respectively, as compared to 1.42 mg/mL for ascorbic acid. The result of this study indicates that 80% of methanol extract and solvent fractions are endowed with wound healing activity. Additionally, this study has also revealed that ME, AQF, MEF, and EAF have the capacity to scavenge free radicals. The study indicated that the wound healing effect could be attributed to the anti-inflammatory and antioxidant activities.

Q-Switched 1,064-nm Neodymium-Doped yttrium aluminum garnet laser irradiation induces collagen remodeling in SKH-1 Hairl ess mice by activating ERK1/2 and p38MAPK signaling pathway

To select the optimal condition of 1064 nm Q-switched Nd:YAG laser irradiation and study the molecular mechanism of laser irradiation for skin rejuvenation. Sixty male, 6-week-old SKH-1 hairless mice were equally divided into 6 groups: 3d, 7d, 21d, 28d, 42d and 56d of 1064 nm laser irradiation at both 0 J/cm2 (left region) and 1.5 J/cm2 (right region) fluence. The basic skin performances were analysed via quantifying hydroxyproline content and skin hydration in the dermis and epidermis. The Collagen I, MMP2 and TIMP1, as well as ERK 1/2, p38MAPK, JNK and ERK5 signaling pathways levels were detected by qRT-PCR. Laser irradiation led to a marked increase in hydroxyproline content and skin hydration in the dermis and epidermis compared to the non-irradiated area. After laser irradiation, collagen I was markedly up-regulated, MMP2 was significantly decreased and TIMP1 was unchanged. Moreover, qRT-PCR results revealed ERK 1/2 and p38MAPK signaling pathways were activated after laser irradiation. we are firstly proves that 1064 nm laser irradiation can inhibit collagen degradation by stimulating the activation of ERK1/2 and p38MAPK signaling pathways. Additionally, the condition of laser irradiation lasting 21d at fluence of 1.5 J/cm2 displays the best effect of skin rejuvenation in our study.

Investigating wound healing potential of Typha angustata L. inflorescence in albino Wistar rats

BackgroundTypha angustata L. is a common aquatic weed. Although the folkloric use of Typha in wound healing has been reported, scientific evidences are still lacking. PurposeThe present study aimed to explore the wound healing potentials of T. angustata L. MethodThe ointment was prepared from the aqueous extract of Typha inflorescence and applied on the induced wounds (viz., Excision, Incision and Burn) in albino Wistar rats. Continuous monitoring of percentage of wound contraction, epithelization duration, content of hydroxyproline and protein (from the excised wound tissues collected on 14th and 28th day of treatment) were used as parameters to know wound healing effects. In addition, catalase activity (antioxidant marker) and histopathological studies were also carried out. ResultsTopical application of the test ointment showed significant (p<0.05) wound contraction, fast epithelization, increased hydroxyproline content and increased catalase activity in comparison to the reference drug treated groups [(treated with (10%w/w) povidone iodine and (10%w/w) burnol ointment)] as well as control groups (treated with ointment base) in all wound types. Similarly, the histopathological study also revealed better reparative changes in terms of neo-vascularization and collagen tissue deposition in treated animals as compared to reference drug treated and control group animals. ConclusionThe results of the present study suggest the potentials of Typha plant in formulating wound healing agents.

Elucidating nanoscale mechanical properties of diabetic human adipose tissue using atomic force microscopy

Obesity-related type 2 diabetes (DM) is a major public health concern. Adipose tissue metabolic dysfunction, including fibrosis, plays a central role in DM pathogenesis. Obesity is associated with changes in adipose tissue extracellular matrix (ECM), but the impact of these changes on adipose tissue mechanics and their role in metabolic disease is poorly defined. This study utilized atomic force microscopy (AFM) to quantify difference in elasticity between human DM and non-diabetic (NDM) visceral adipose tissue. The mean elastic modulus of DM adipose tissue was twice that of NDM adipose tissue (11.50 kPa vs. 4.48 kPa) to a 95% confidence level, with significant variability in elasticity of DM compared to NDM adipose tissue. Histologic and chemical measures of fibrosis revealed increased hydroxyproline content in DM adipose tissue, but no difference in Sirius Red staining between DM and NDM tissues. These findings support the hypothesis that fibrosis, evidenced by increased elastic modulus, is enhanced in DM adipose tissue, and suggest that measures of tissue mechanics may better resolve disease-specific differences in adipose tissue fibrosis compared with histologic measures. These data demonstrate the power of AFM nanoindentation to probe tissue mechanics, and delineate the impact of metabolic disease on the mechanical properties of adipose tissue.

Cardiac contractility modulation attenuates structural and electrical remodeling in a chronic heart failure rabbit model.

BackgroundCardiac contractility modulation (CCM) is non-excitatory electrical stimulation for improving cardiac function. This study aimed to evaluate the effects of CCM on structural and electrical remodeling in a rabbit model of chronic heart failure (CHF).MethodsThirty rabbits were randomly divided into the sham, CHF, and CCM groups. The CHF model was induced 12 weeks after trans-aortic constriction by pressure unloading and CCM was delivered to the myocardium for 4 weeks. Corrected QT intervals, the ventricular effective refractory period, and inducibility of ventricular tachycardia were measured by an electrophysiological examination. Connective tissue growth factor, galectin-3, Kv4.3, KCNQ1, KCNH2, and connexin 43 protein levels were measured by western blotting.ResultsThe CHF group had a significantly prolonged corrected QT interval and ventricular effective refractory period, and increased inducibility of ventricular tachycardia. Prominent myocardial fibrosis and increased hydroxyproline content were observed in the CHF group, but these were suppressed in the CCM group. Kv4.3, KCNQ1, KCNH2, and connexin 43 protein levels were significantly lower in the CHF group, but treatment with CCM partially restored their levels.ConclusionsCCM attenuates myocardial structural and electrical remodeling during CHF. These findings provide evidence for clinical use of CCM in treating CHF.

Effect of Extract of Allium stipitatum on Excisional Wound Healing in Rats

Objective- The aim of the present study was to assess the wound-healing activity of extract of Allium stipitatum. Design- Experimental study.Animals- Thirty-six male Wistar diabetic rats.Procedures- The rats weighing approximately 160-180 g and seven weeks of age were randomized into three groups of 12 rats each: Control surgery group (Control) including the creation of wounds and no treatment, base formulation groups positive (POS) with the creation of wounds and application of base formulation ointment, treatment group 1 (T1) with 2 g of powder extract of the plant material in the ointment. A wound was induced by an excisional based wound model in male rats. The mature green leaves of Allium stipitatum were collected and authenticated. Extractions of dried leaves were carried out. For wound-healing activity, the extracts were applied topically in the form of ointment and compared to control groups. The healing of the wound was assessed based on the wound area, histomorphometry, and hydroxyproline estimation studies. Results- Reduction in the wound area and hydroxyproline contents indicated that there was a significant difference (p = 0.001) between group T1 and other groups. Quantitative histological studies and mean rank of the qualitative studies demonstrated that there was a significant difference between group T1 and other groups (p = 0.001).Conclusions and Clinical Relevance- The extract of Allium stipitatum leaves enhanced wound-healing activity significantly in both the wound models studied. Enhanced wound contraction, decreased epithelialization time, increased hydroxyproline content, improved histological characteristics studies suggested Allium stipitatum leaves extract might have therapeutic benefits in wound healing.

The protective effect of piperine against isoproterenol-induced inflammation in experimental models of myocardial toxicity

Myocardial infarction (MI) eventually exacerbates inflammatory response due to the release of inflammatory and pro-inflammatory factors. The aim of this study is to explore the protective efficacy of piperine supplementation against the inflammatory response in isoproterenol (ISO)-induced MI. Masson Trichome staining was executed to determine myocardial tissue architecture. Immunohistochemistry was performed for IL-6, TNF-α. RT-PCR studies were performed to ascertain the gene expression of IL-6, TNF-α, iNOS, eNOS, MMP-2, MMP-9, and collagen-III. Western blotting was performed to determine expression of HIF-1α, VEGF, Nrf-2, NF-ƙB, Cox-2, p-38, phospho-p38, ERK-1/2, phospho-ERK-1/2, and collagen-I. HIF-1α, VEGF, and iNOS expression were significantly upregulated with concomitant decline in eNOS expression in the heart myocardial tissue of rats received ISO alone whereas piperine pretreatment prevented these changes in ISO administered rats. Current results revealed ROS-mediated activation of MAPKs, namely, p-p38, p-ERK1/2 in the heart tissue of ISO administered group. Piperine pretreatment significantly prevented these changes in ISO treated group. NF-κB is involved in the modulation of gene expressions responsible for tissue repair. ISO-induced NF-κB-p65 expression was significantly reduced in the group pretreated with piperine and mitigated extent of myocardial inflammation. A significant increase in cardiac fibrosis upon ISO treatment was reported due to the increased hydroxyproline content, MMP-2 & 9 and upregulation of collagen-I protein compared to control group. All these cardiac hypertrophy markers were decreased in ‘piperine pretreated ISO administered group’ compared to group received ISO injection. Current findings concluded that piperine as a nutritional intervention could prevent inflammation of myocardium in ISO-induced MI.

Assessment of Healing Potential of Formulated Herbal Cream on Burn Wound

The current protocol of the study was planned to assess the healing potential of formulated herbal cream containing different medicinal plant extracts. Formulated herbal cream was containing the methanolic extract of Chenopodium album, Coccinia indica, Momordica dioica, Precitrullus fistulosus and Trichosanthes dioica with cream base. Formulated herbal cream was assessed pharmacologically for healing potential in burn wound model. The formulation showed a significantly higher contraction rate, shortened epithelialization period, better tensile strength and increase in hydroxyproline content in the burn wound model. Thus, it is concluded that formulation has the potential wound healing activity which justifying its use as wound healer.&#x0D; Keywords: Formulation, Burn, Wound, Contraction

Repeated intravenous administration of silica nanoparticles induces pulmonary inflammation and collagen accumulation via JAK2/STAT3 and TGF-β/Smad3 pathways in vivo.

BackgroundThe health hazards of silica nanoparticle (SiNP) are raising worldwide concern as SiNPs has become the second largest manufactured nanomaterial in global markets. However, insufficient data for the adverse health effects and safety evaluation of SiNPs are remaining a big question.PurposeWe evaluated the effects and related mechanism of SiNPs on pulmonary inflammation and collagen production through repeated intravenous administration in mice in a 45-day observation period.MethodsMorphological and ultrastructural change, ultradistribution of SiNPs in lungs were observed in ICR mice through intravenous administration. Oxidative damage, pro-inflammatory cytokines, hydroxyproline content, the marker of fibroblasts and epithelial-mesenchymal transition (EMT), and JAK2/STAT3 and TGF-β1/Smad3 signaling pathways were detected to explore the lung injuries and related mechanism.ResultsThe results showed repeated intravenous exposure of SiNPs increased the weight of lung tissues and destroyed pulmonary histomorphological structure. The increased MDA content, depletion of SOD and GSH-Px in lungs were observed in SiNP-treated mice. The protein expressions of JAK2/STAT3 pathway were upregulated in lungs, and the levels of inflammatory cytokines TNF-α, IL-1β, and IL-6 in serum and lungs were also elevated in SiNPs treated group. The increased hydroxyproline content indicated collagen accumulation in lungs of SiNP-treated mice. Meanwhile, the protein expressions of the marker of myofibroblast (a-SMA), the regulators in connective tissue remodeling (CTGF), TGF-β, and p-Smad3 were all upregulated in lungs. In addition, we found intravenous administration of SiNPs-induced ultrastructural changes in type II alveolar epithelial cells but without downregulation of the protein expression of the key markers of epithelial cells (E-Cadherin).ConclusionOur results revealed that oxidative stress and inflammation contributed to the collagen accumulation through activation of JAK2/STAT3 and TGF-β/Smad3 pathways. It suggests that pulmonary aberrant inflammation and collagen accumulation induced by nanoparticles should be seriously considered for the safety application in diagnostics or therapeutics.