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Effect of ANGPTL7 on Proliferation and Differentiation of MC3T3-E1 Cells.

BACKGROUND Angiopoietin-like proteins (ANGPTL) are a family of secretory glycoproteins that are involved in many pathophysiological processes. ANGPTL7 is a newly-discovered member of the ANGPTL family and plays a role in corneal morphogenesis, angiogenesis, glaucoma, and cancer. To date, whether ANGPTL7 is involved in osteoporosis is unknown. Therefore, to discover the effects of ANGPTL7 on osteoporosis, we explored the expression of ANGPTL7 in preosteoblasts and assessed the mechanism underlying its effects on proliferation and differentiation abilities of preosteoblasts. MATERIAL AND METHODS Mouse MC3T3-E1 cells were cultured in osteogenic medium for osteogenic differentiation. The expression levels of ANGPTL7 were detected by RT-qPCR and Western blot assays. Moreover, the overexpressed plasmid of ANGPTL7 pMSCV-ANGPTL7 was transfected into MC3T3-E1 cells. CCK-8 was used to evaluate cell proliferation. ALP activity detection and alizarin red staining were performed to measure the effect of ANGPTL7 on osteogenic differentiation. The expression levels bone morphogenetic proteins (BMPs) and osteogenic markers ALP, runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and collagen I (Col I) were analyzed by Western blot. RESULTS When MC3T3-E1 cells were exposed to osteogenic medium, there was a significant increase in ANGPTL7, and overexpression of ANGPTL7 markedly promoted cell proliferation, ALP activity, and mineralization. Moreover, ANGPTL7 upregulated the levels of BMPs, especially BMP2/7, and the osteogenic markers ALP, Runx2, OCN, and Col I. CONCLUSIONS The results suggest that by regulating the expression of BMPs, ANGPTL7 directly promotes proliferation, differentiation, and mineralization of osteoblasts.

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Imidacloprid-induced liver fibrosis in quails via activation of the TGF-β1/Smad pathway.

Imidacloprid (IMI) is one of the most frequently used neonicotinoid insecticide, and its potential toxicity and environmental hazards have gradually attracted people's attention. Liver fibrosis caused by long-term inflammation or oxidative stress can lead to cirrhosis and liver failure, even death. However, the mechanism of liver fibrosis induced by neonicotinoid insecticide remains unclear. This study investigates whether IMI could induce liver fibrosis in quails and a potential mechanism. Our study used a quail 90-day IMI-induced liver fibrosis model. The results showed that IMI induced histopathological lesions, oxidative stress, inflammation, fibrosis, and changes in nuclear factor-kappa B (NF-κB), nuclear factor-E2-related factor-2 (Nrf2), and transforming growth factor (TGF-β1) levels. Furthermore, IMI enhanced the expression of liver fibrosis marker proteins, including collagen I, α-smooth muscle actin (α-SMA), and fibronectin 1 (FN-1), by activating the TGF-β1/Smad signaling pathway. In conclusion, our study demonstrated that IMI exposure induces liver fibrosis via activation of the TGF-β1/Smad signaling pathway in quails.

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Extraction and characterization of collagen from sheep slaughter by-products.

There is a growing search for alternative raw materials to obtain collagen and hydrolysates and processes that do not threaten the environment or human health. Thus, sheep slaughter residue, which doesn't yet have an adequate and sustainable destination, is an excellent source of study. The objective of this study was to investigate the technological properties of collagen extracted from sheep slaughter by-products. It was possible to produce and characterize collagens extracted from sheep slaughter by-products. The yield of collagen was 18.0% and 12.5% for lamb and sheep by-products, respectively, on a dry basis. Lamb and sheep collagens showed similar FTIR and digestibility spectra and increased solubility at acidic pH-value. Higher foaming capacity was found for lamb collagen, while the sheep collagen presented higher viscosity. The emulsifying power of the collagens was 59.1 and 69.6 m/g for lamb and sheep by-products, respectively. The collagens presented bands corresponding to α, α, and β chains, characteristic of collagen type I and a molecular weight (SDS-PAGE) between 100 and 5 kDa. The collagens of this study showed potential for application in food products, both for the technological improvement and nutrient enrichment, adding value and giving a sustainable destination to sheep slaughter by-products.

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Hydroxylapatite-collagen hybrid scaffold induces human adipose-derived mesenchymal stem cells to osteogenic differentiation in vitro and bone regrowth in patients.

Tissue engineering-based bone graft is an emerging viable treatment modality to repair and regenerate tissues damaged as a result of diseases or injuries. The structure and composition of scaffolds should modulate the classical osteogenic pathways in human stem cells. The osteoinductivity properties of the hydroxylapatite-collagen hybrid scaffold named Coll/Pro Osteon 200 were investigated in an in vitro model of human adipose mesenchymal stem cells (hASCs), whereas the clinical evaluation was carried out in maxillofacial patients. Differentially expressed genes (DEGs) induced by the scaffold were analyzed using the Osteogenesis RT PCR Array. The osteoinductivity potential of the scaffold was also investigated by studying the alkaline phosphatase (ALP) activity, matrix mineralization, osteocalcin (OCN), and CLEC3B expression protein. Fifty patients who underwent zygomatic augmentation and bimaxillary osteotomy were evaluated clinically, radiologically, and histologically during a 3-year follow-up. Among DEGs, osteogenesis-related genes, including BMP1/2, ALP, BGLAP, SP7, RUNX2, SPP1, and EGFR, which play important roles in osteogenesis, were found to be upregulated. The genes to cartilage condensation SOX9, BMPR1B, and osteoclast cells TNFSF11 were detected upregulated at every time point of the investigation. This scaffold has a high osteoinductivity revealed by the matrix mineralization, ALP activity, OCN, and CLEC3B expression proteins. Clinical evaluation evidences that the biomaterial promotes bone regrowth. Histological results of biopsy specimens from patients showed prominent ossification. Experimental data using the Coll/Pro Osteon 200 indicate that clinical evaluation of bone regrowth in patients, after scaffold implantation, was supported by DEGs implicated in skeletal development as shown in "in vitro" experiments with hASCs.

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Decreased Uterine Vascularization and Uterine Arterial Expansive Remodeling with Reduced Matrix Metalloproteinase-2 and -9 in Hypertensive Pregnancy.

Preeclampsia is a pregnancy-related disorder characterized by hypertension-in-pregnancy (HTN-Preg), and inadequate trophoblast invasion of the uterine wall could be initiating events. We hypothesized that HTN-Preg involves decreased uterine vascularization and arterial remodeling by MMPs, and accumulation of collagen. Blood pressure (BP) and fetal parameters were assessed in normal Preg rats and Preg rats with reduced uterine perfusion pressure (RUPP), and uterine vascularity, MMP levels and collagen deposition were measured. BP was higher, and the uterus weight, litter size, and pup weight were reduced in RUPP vs Preg rats. Uterine tissue sections showed reduced number (5.75±0.95 vs 11.50±0.87) and size (0.05±0.01 vs 0.12±0.02mm) of uterine arterioles, and diminished intima, media and adventitia of the uterine arterial wall in RUPP vs Preg rats. Immunohistochemistry showed localization of endothelial cell marker CD31 and smooth muscle marker a-actin in uterine arterial intima and media, respectively, and confirmed reduction in number and size of uterine arterioles in RUPP vs Preg rats. The cytotrophoblast marker cytokeratin-7 showed less immunostaining and invasion of uterine spiral arterioles of RUPP vs Preg rats. Uterine arterioles showed less expansion in response to increases in intraluminal pressure in RUPP vs Preg rats. Western blots, gelatin zymography and immunohistochemistry showed decreases in MMP-2 and MMP-9 and increases in the MMP substrate collagen-IV in the uterus and uterine spiral arteries of RUPP vs Preg rats. The results decreased uterine vascularization and uterine arteriolar expansive remodeling with decreased MMP-2 and MMP-9 and increased collagen-IV could be underlying mechanisms of uteroplacental ischemia in HTN-Preg.

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Effects of Toll-Like Receptor 4 Inhibition on Transforming Growth Factor-β2 Signaling in the Human Trabecular Meshwork.

Transforming growth factor-β2 (TGFβ2) and Toll-like receptor 4 (TLR4) crosstalk have been implicated in extracellular matrix regulation in the trabecular meshwork (TM) and ocular hypertension in mice. We investigated TLR4 expression in normal and glaucomatous human trabecular meshwork (HTM) sections and utilized a human perfusion organ culture model to determine TGFβ2-TLR4 signaling crosstalk in glaucoma. Expression of TLR4 was determined in TM of normal and glaucomatous human eyes. Anterior segments of paired human eyes were perfused at a constant flow rate (2.5 μL/min) for 4 days to acquire stable baseline intraocular pressures (IOPs). We treated paired eyes with two different treatment paradigms: (1) TGFβ2 in one eye and vehicle control in the paired eye, (2) TGFβ2 in one eye and TGFβ2 + TLR4 inhibitor TAK-242 in the paired eye. Perfusate and TM tissue were collected and analyzed for fibronectin (FN) and collagen IV (COLIV) expression. We observed increased TLR4 expression in glaucomatous HTM sections compared to normal (age-matched) ( < 0.05). Significant elevation of IOP was detected in 47% of TGFβ2-treated anterior segments ( < 0.01) compared to control, and in TGFβ2 treated compared with co-treatment with TGFβ2 + TLR4 inhibitor ( < 0.0001). An increase in FN and COLIV expression was observed after TGFβ2 treatment, and inhibition of TLR4 signaling decreased TGFβ2-induced FN and COLIV expression in perfusate ( < 0.05). These studies identify TGFβ2-TLR4 crosstalk as a novel pathway in glaucoma. They provide a potential new target to lower IOP and explore glaucoma pathogenesis.

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Photobiostimulation activity of different low-level laser dosage on masticatory muscles and temporomandibular joint in an induced arthritis rat model.

This study aimed to investigate the anti-inflammatory effects of different dosage of low-level laser therapy (LLLT) in an experimental model of temporomandibular joint (TMJ) arthritis. One hundred male Wistar rats were used and divided into the following groups: CG, control group; AG, animals group with left TMJ arthritis induced by intra-articular injection of Complete Freund's adjuvant - CFA; LG5, LG10 and LG20 - animals with arthritis and treated with LLLT at doses 5, 10, and 20 J/cm, respectively. Morphological analysis was performed by TMJ histological sections stained with hematoxylin-eosin (HE), picrosirius (PSR), and toluidine blue (TB), as well as histomorphometric evaluation of cartilage, articular disc, and masticatory muscles. The amount of feed consumed within 3 weeks was evaluated, and biochemical analysis of TMJ tissues included measurement of sulfated glycosaminoglycans (GAGs), matrix metalloproteinases (MMPs) 2 and 9 zymography, and ELISA for cytokines IL-6, TNF-α, and IL-1β. Only the 20 J/cm dose promoted higher feed intake compared to AG. On the other hand, all LLLT doses promoted better organization of articular disc collagen fibers, greater number of proteoglycans in articular cartilage, increased area and diameter of left lateral pterygoid fibers, reduced latent and active MMP 9 and 2 activity, and lower IL-1β concentration compared to AG. Considering the study limitations, it was observed that LLLT treatments were effective in protecting and tissue cleansing joint structures, accelerating tissue repair, especially at lower doses.

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Discussion: Correction of Lower Eyelid Retraction with En Glove Placement of Porcine Dermal Collagen Matrix Implant.


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CD34 Staining in Disorders of Collagen Degeneration Other Than Morphea.


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Amphiregulin as a driver of tissue fibrosis.

Most deaths in Western countries can currently be associated with fibrotic diseases. During fibrosis excessive amounts of extra-cellular matrix components, such as collagen, are deposited in tissues, which can lead to tissue stiffness and organ failure. The underlying mechanisms leading to tissue fibrosis remain poorly understand and, consequently, our capacities to treat fibrotic diseases remain limited. Currently approved drugs only delay the progression of fibrosis and are often associated with severe side-effects, which limit their clinical application. Thus, a better understanding of the ontology of fibrotic diseases and the development of more specific and more efficient drugs for the treatment of tissue fibrosis is an urgent clinical meet.

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