scholarly journals Role of Toll-like receptors in diabetic renal lesions in a miniature pig model

2015 ◽  
Vol 1 (5) ◽  
pp. e1400183 ◽  
Author(s):  
Yuanyuan Feng ◽  
Shulin Yang ◽  
Yuxiang Ma ◽  
Xue-Yuan Bai ◽  
Xiangmei Chen
Keyword(s):  
Immune System ◽  
Signaling Pathway ◽  
Inflammatory Cells ◽  
Large Animal Model ◽  
Large Animal ◽  
Toll Like Receptors ◽  
Metabolic Inflammation ◽  
Renal Lesions ◽  
Miniature Pigs ◽  
Downstream Signaling

The mechanisms of diabetic renal injury remain unclear. Recent studies have shown that immunological and inflammatory elements play important roles in the initiation and development of diabetic nephropathy (DN). Toll-like receptors (TLRs) comprise a superfamily of innate immune system receptors. The roles and mechanisms of TLRs in the pathogenesis of diabetic renal lesions are mostly unknown. Compared with rodents, miniature pigs are more similar to humans with respect to metabolism, kidney structure, and immune system, and therefore represent an ideal large-animal model for DN mechanistic studies. A diabetes model was established by feeding miniature pigs with high-sugar and high-fat diets. Functional and pathological markers, expression and activation of endogenous TLR ligands [HSP70 (heat shock protein 70) and HMGB1], TLR1 to TLR11 and their downstream signaling pathway molecules (MyD88, IRAK-1, and IRF-3), nuclear factor κB (NF-κB) signaling pathway molecules (IKKβ, IκBα, and NF-κBp65), inflammatory cytokines [IL-6 (interleukin-6), MIP-2, MCP-1, CCL5, and VCAM-1 (vascular cell adhesion molecule–1)], and infiltration of inflammatory cells were systematically evaluated. The expression of HSP70 was significantly increased in diabetic pig kidneys. The expression of MyD88-dependent TLR2, TLR4, TLR5, TLR7, TLR8, and TLR11 and their downstream signaling molecules MyD88 and phospho–IRAK-1 (activated IRAK-1), as well as that of MyD88-independent TLR3 and TLR4 and their downstream signaling molecule phospho–IRF-3 (activated IRF-3), was significantly up-regulated. The expression and activation of NF-κB pathway molecules phospho-IKKβ, phospho-IκBα, NF-κBp65, and phospho-NF-κBp65 were significantly increased. Levels of IL-6, MIP-2, MCP-1, CCL5, VCAM-1, and macrophage marker CD68 were significantly increased in diabetic pig kidneys. These results suggested that the metabolic inflammation activated by TLRs might play an important role in diabetic renal injuries.

scholarly journals Dynamic mRNA Expression Analysis of the Secondary Palatal Morphogenesis in Miniature Pigs

2019 ◽  
Vol 20 (17) ◽  
pp. 4284
Author(s):  
Jia Liu ◽  
Jing Chen ◽  
Dong Yuan ◽  
Lindong Sun ◽  
Zhipeng Fan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Cleft Lip ◽  
Integration Site ◽  
Expression Profiles ◽  
Mitogen Activated Protein Kinase ◽  
Large Animal Model ◽  
Large Animal ◽  
Palate Development ◽  
Miniature Pigs

Normal mammalian palatogenesis is a complex process that requires the occurrence of a tightly regulated series of specific and sequentially regulated cellular events. Cleft lip/palate (CLP), the most frequent craniofacial malformation birth defects, may occur if any of these events undergo abnormal interference. Such defects not only affect the patients, but also pose a financial risk for the families. In our recent study, the miniature pig was shown to be a valuable alternative large animal model for exploring human palate development by histology. However, few reports exist in the literature to document gene expression and function during swine palatogenesis. To better understand the genetic regulation of palate development, an mRNA expression profiling analysis was performed on miniature pigs, Sus scrofa. Five key developmental stages of miniature pigs from embryonic days (E) 30–50 were selected for transcriptome sequencing. Gene expression profiles in different palate development stages of miniature pigs were identified. Nine hundred twenty significant differentially expressed genes were identified, and the functional characteristics of these genes were determined by gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Some of these genes were associated with HH (hedgehog), WNT (wingless-type mouse mammary tumor virus integration site family), and MAPK (mitogen-activated protein kinase) signaling, etc., which were shown in the literature to affect palate development, while some genes, such as HIP (hedgehog interacting protein), WNT16, MAPK10, and LAMC2 (laminin subunit gamma 2), were additions to the current understanding of palate development. The present study provided a comprehensive analysis for understanding the dynamic gene regulation during palate development and provided potential ideas and resources to further study normal palate development and the etiology of cleft palate.

The temporal bone microdissection of miniature pigs as a useful large animal model for otologic research

2013 ◽  
Vol 134 (1) ◽  
pp. 26-33 ◽  
Author(s):  
H. J. Yi ◽  
Wei Guo ◽  
N. Wu ◽  
J. N. Li ◽  
H. Z. Liu ◽  
...  
Keyword(s):  
Animal Model ◽  
Temporal Bone ◽  
Large Animal Model ◽  
Large Animal ◽  
Miniature Pigs

scholarly journals The efficacy and safety of pinocembrin in a sheep model of bleomycin-induced pulmonary fibrosis

PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0260719
Author(s):  
Habtamu B. Derseh ◽  
Jason Q. D. Goodger ◽  
Jean-Pierre Y. Scheerlinck ◽  
Chrishan S. Samuel ◽  
Ian E. Woodrow ◽  
...  
Keyword(s):  
Animal Model ◽  
Pulmonary Fibrosis ◽  
Inflammatory Cells ◽  
Lung Parenchyma ◽  
Lung Compliance ◽  
Large Animal Model ◽  
Large Animal ◽  
Lung Pathology ◽  
Sheep Model ◽  
Anti Inflammatory

The primary flavonoid, pinocembrin, is thought to have a variety of medical uses which relate to its reported anti-oxidant, anti-inflammatory, anti-microbial and anti-cancer properties. Some studies have reported that this flavonoid has anti-fibrotic activities. In this study, we investigated whether pinocembrin would impede fibrosis, dampen inflammation and improve lung function in a large animal model of pulmonary fibrosis. Fibrosis was induced in two localized lung segments in each of the 10 sheep participating in the study. This was achieved via two infusions of bleomycin delivered bronchoscopically at a two-week interval. Another lung segment in the same sheep was left untreated, and was used as a healthy control. The animals were kept for a little over 5 weeks after the final infusion of bleomycin. Pinocembrin, isolated from Eucalyptus leaves, was administered to one of the two bleomycin damaged lung segments at a dose of 7 mg. This dose was given once-weekly over 4-weeks, starting one week after the final bleomycin infusion. Lung compliance (as a measure of stiffness) was significantly improved after four weekly administrations of pinocembrin to bleomycin-damaged lung segments. There were significantly lower numbers of neutrophils and inflammatory cells in the bronchoalveolar lavage of bleomycin-infused lung segments that were treated with pinocembrin. Compared to bleomycin damaged lung segments without drug treatment, pinocembrin administration was associated with significantly lower numbers of immuno-positive CD8+ and CD4+ T cells in the lung parenchyma. Histopathology scoring data showed that pinocembrin treatment was associated with significant improvement in inflammation and overall pathology scores. Hydroxy proline analysis showed that the administration of pinocembrin did not reduce the increased collagen content that was induced by bleomycin in this model. Analyses of Masson’s Trichrome stained sections showed that pinocembrin treatment significantly reduced the connective tissue content in lung segments exposed to bleomycin when compared to bleomycin-infused lungs that did not receive pinocembrin. The striking anti-inflammatory and modest anti-fibrotic remodelling effects of pinocembrin administration were likely linked to the compound’s ability to improve lung pathology and functional compliance in this animal model of pulmonary fibrosis.

Abstract 18: Bempedoic Acid Lowers Low Density Lipoprotein-Cholesterol and Attenuates Aortic Atherosclerosis in LDL Receptor-Deficient ( LDLR +/- and LDLR -/- ) Yucatan Miniature Pigs

2016 ◽  
Vol 36 (suppl_1) ◽  
Author(s):  
Amy C Burke ◽  
Dawn E Telford ◽  
Brian G Sutherland ◽  
Jane Y Edwards ◽  
Cynthia G Sawyez ◽  
...  
Keyword(s):  
Animal Model ◽  
Ldl Receptor ◽  
Large Animal Model ◽  
Large Animal ◽  
Aortic Atherosclerosis ◽  
Lesion Area ◽  
Aortic Lesion ◽  
Miniature Pigs ◽  
Long Term Treatment ◽  
Abdominal Lesions

Bempedoic acid (ETC-1002) is an oral investigational drug that targets hepatic adenosine triphosphate-citrate lyase to reduce cholesterol biosynthesis. In Phase 2 studies (up to 12 weeks in duration), bempedoic acid lowers elevated LDL-cholesterol (C) in patients with hypercholesterolemia. The objective of the present study was to test the ability of bempedoic acid to decrease plasma-C and LDL-C, and attenuate the development of atherosclerosis in a large animal model of familial hypercholesterolemia. Gene targeting has been used to generate Yucatan miniature pigs heterozygous ( LDLR +/- ) or homozygous ( LDLR -/- ) for LDL-receptor (R) deficiency (ExeGen). LDLR +/- pigs (n=12) and LDLR -/- pigs (n=12) were fed a high fat, cholesterol-containing diet (34% kcal fat; 0.2% cholesterol) and orally administered placebo or bempedoic acid at 120 mg/d and at 240 mg/d for 160 days. Bempedoic acid was well tolerated; weight gain, caloric intake, liver enzymes and clinical chemistry analyses were unaffected by treatment. In LDLR +/- pigs at 160 days, compared to placebo, bempedoic acid significantly decreased plasma-C and LDL-C up to 43% ( P <0.01) and 63% ( P <0.002), respectively. Compared to LDLR-/- placebo pigs, in which plasma-C and LDL-C were 5-fold higher than in LDLR +/- placebo pigs, bempedoic acid significantly decreased plasma-C and LDL-C up to 26% ( P <0.04) and 29% ( P <0.03), respectively. Plasma levels of triglycerides and HDL-C, fasting glucose and insulin, and liver lipids were not affected by treatment in either genotype. In the aorta of LDLR +/- pigs, bempedoic acid treatment robustly attenuated total Sudan IV-stained lesion area (-58%, P <0.02) and area of raised lesions in the abdominal aorta (-58%, P <0.03). In LDLR -/- pigs, in which total aortic lesion area (6-fold) and area of raised abdominal lesions (12-fold) were substantially larger compared to LDLR +/- pigs, bempedoic acid significantly decreased total aortic lesion area (-47%, P <0.01) and area of raised abdominal lesions (-50%, P <0.03). The present experiments demonstrate that in a large animal model of LDLR-deficiency and atherosclerosis, long term treatment with bempedoic acid reduces LDL-C and attenuates the progression of aortic atherosclerosis in both LDLR +/- and LDLR -/- miniature pigs.

scholarly journals Immunological and functional features of decellularized xenogeneic heart valves after transplantation into GGTA1-KO pigs

2021 ◽  
Vol 8 (4) ◽  
Author(s):  
Robert Ramm ◽  
Tobias Goecke ◽  
Peter Köhler ◽  
Igor Tudorache ◽  
Serghei Cebotari ◽  
...  
Keyword(s):  
Heart Valves ◽  
Immune Reaction ◽  
Inflammatory Cells ◽  
Giant Cells ◽  
Large Animal Model ◽  
Large Animal ◽  
Wild Type ◽  
Foreign Body Giant Cells ◽  
Antibody Titers ◽  
Functional Features

Abstract Decellularization of xenogeneic heart valves might lead to excellent regenerative implants, from which many patients could benefit. However, this material carries various xenogeneic epitopes and thus bears a considerable inherent immunological risk. Here, we investigated the regenerative and immunogenic potential of xenogeneic decellularized heart valve implants using pigs deficient for the galactosyltransferase gene (GGTA1-KO) as novel large animal model. Decellularized aortic and pulmonary heart valves obtained from sheep, wild-type pigs or GGTA1-KO pigs were implanted into GGTA1-KO pigs for 3, or 6 months, respectively. Explants were analyzed histologically, immunhistologically (CD3, CD21 and CD172a) and anti-αGal antibody serum titers were determined by ELISA. Xenogeneic sheep derived implants exhibited a strong immune reaction upon implantation into GGTA1-KO pigs, characterized by massive inflammatory cells infiltrates, presence of foreign body giant cells, a dramatic increase of anti-αGal antibody titers and ultimately destruction of the graft, whereas wild-type porcine grafts induced only a mild reaction in GGTA1-KO pigs. Allogeneic implants, wild-type/wild-type and GGTA1-KO/GGTA1-KO valves did not induce a measurable immune reaction. Thus, GGTA1-KO pigs developed a ‘human-like’ immune response toward decellularized xenogeneic implants showing that immunogenicity of xenogeneic implants is not sufficiently reduced by decellularization, which detracts from their regenerative potential.

scholarly journals Regeneration of Subcutaneous Cartilage in a Swine Model Using Autologous Auricular Chondrocytes and Electrospun Nanofiber Membranes Under Conditions of Varying Gelatin/PCL Ratios

2021 ◽  
Vol 9 ◽  
Author(s):  
Rui Zheng ◽  
Xiaoyun Wang ◽  
Jixin Xue ◽  
Lin Yao ◽  
Gaoyang Wu ◽  
...  
Keyword(s):  
Cartilage Regeneration ◽  
Inflammatory Cells ◽  
Large Animal Model ◽  
Swine Model ◽  
Large Animal ◽  
Nude Mouse Model ◽  
Cartilage Formation ◽  
Nanofiber Membranes ◽  
Large Animals

The scarcity of ideal biocompatible scaffolds makes the regeneration of cartilage in the subcutaneous environment of large animals difficult. We have previously reported the successful regeneration of good-quality cartilage in a nude mouse model using the electrospun gelatin/polycaprolactone (GT/PCL) nanofiber membranes. The GT/PCL ratios were varied to generate different sets of membranes to conduct the experiments. However, it is unknown whether these GT/PCL membranes can support the process of cartilage regeneration in an immunocompetent large animal model. We seeded swine auricular chondrocytes onto different GT/PCL nanofiber membranes (GT:PCL = 30:70, 50:50, and 70:30) under the sandwich cell-seeding mode. Prior to subcutaneously implanting the samples into an autologous host, they were cultured in vitro over a period of 2 weeks. The results revealed that the nanofiber membranes with different GT/PCL ratios could support the process of subcutaneous cartilage regeneration in an autologous swine model. The maximum extent of homogeneity in the cartilage tissues was achieved when the G5P5 (GT: PC = 50: 50) group was used for the regeneration of cartilage. The formed homogeneous cartilage tissues were characterized by the maximum cartilage formation ratio. The extents of the ingrowth of the fibrous tissues realized and the extents of infiltration of inflammatory cells achieved were found to be the minimum in this case. Quantitative analyses were conducted to determine the wet weight, cartilage-specific extracellular matrix content, and Young’s modulus. The results indicated that the optimal extent of cartilage formation was observed in the G5P5 group. These results indicated that the GT/PCL nanofiber membranes could serve as a potential scaffold for supporting subcutaneous cartilage regeneration under clinical settings. An optimum GT/PCL ratio can promote cartilage formation.

scholarly journals Splenic Nerve Neuromodulation Reduces Inflammation and Promotes Resolution in Chronically Implanted Pigs

2021 ◽  
Vol 12 ◽  
Author(s):  
David M. Sokal ◽  
Alex McSloy ◽  
Matteo Donegà ◽  
Joseph Kirk ◽  
Romain A. Colas ◽  
...  
Keyword(s):  
Immune System ◽  
Pulse Generator ◽  
Large Animal Model ◽  
Lipid Mediator ◽  
Autonomic Nerves ◽  
Large Animal ◽  
Necrosis Factor Alpha ◽  
Safety Testing ◽  
Inflammatory Monocytes ◽  
Stimulation Parameters

Neuromodulation of the immune system has been proposed as a novel therapeutic strategy for the treatment of inflammatory conditions. We recently demonstrated that stimulation of near-organ autonomic nerves to the spleen can be harnessed to modulate the inflammatory response in an anesthetized pig model. The development of neuromodulation therapy for the clinic requires chronic efficacy and safety testing in a large animal model. This manuscript describes the effects of longitudinal conscious splenic nerve neuromodulation in chronically-implanted pigs. Firstly, clinically-relevant stimulation parameters were refined to efficiently activate the splenic nerve while reducing changes in cardiovascular parameters. Subsequently, pigs were implanted with a circumferential cuff electrode around the splenic neurovascular bundle connected to an implantable pulse generator, using a minimally-invasive laparoscopic procedure. Tolerability of stimulation was demonstrated in freely-behaving pigs using the refined stimulation parameters. Longitudinal stimulation significantly reduced circulating tumor necrosis factor alpha levels induced by systemic endotoxemia. This effect was accompanied by reduced peripheral monocytopenia as well as a lower systemic accumulation of CD16+CD14high pro-inflammatory monocytes. Further, lipid mediator profiling analysis demonstrated an increased concentration of specialized pro-resolving mediators in peripheral plasma of stimulated animals, with a concomitant reduction of pro-inflammatory eicosanoids including prostaglandins. Terminal electrophysiological and physiological measurements and histopathological assessment demonstrated integrity of the splenic nerves up to 70 days post implantation. These chronic translational experiments demonstrate that daily splenic nerve neuromodulation, via implanted electronics and clinically-relevant stimulation parameters, is well tolerated and is able to prime the immune system toward a less inflammatory, pro-resolving phenotype.

Beneficial Therapeutic Approach of Acellular PLGA Implants Coupled With Rehabilitation Exercise for Osteochondral Repair: A Proof of Concept Study in a Minipig Model

2020 ◽  
Vol 48 (11) ◽  
pp. 2796-2807
Author(s):  
Chih-Chan Lin ◽  
Chih-Jou Chu ◽  
Pei-Hsi Chou ◽  
Chun-Hao Liang ◽  
Peir-In Liang ◽  
...  
Keyword(s):  
Treadmill Exercise ◽  
Total Duration ◽  
Cartilage Regeneration ◽  
Inflammatory Cells ◽  
Large Animal Model ◽  
Large Animal ◽  
Trochlear Groove ◽  
Plga Scaffold ◽  
Medial Condyle ◽  
Osteochondral Repair

Background: Osteochondral (OC) repair presents a significant challenge to clinicians. However, whether the use of acellular spongy poly(lactic-co-glycolic acid) (PLGA) scaffolding plus treadmill exercise as a rehabilitation program regenerates OC defects in a large-animal model has yet to be determined. Hypothesis: PLGA scaffolding plus treadmill exercise may offer improved OC repair for both high and low weightbearing regions in a minipig model. Study Design: Controlled laboratory study. Methods: A total of 9 mature minipigs (18 knees) were randomly divided into the treadmill exercise (TRE) group or sedentary (SED) group. All pigs received critically sized OC defects in a higher weightbearing region of the medial condyle and a lower weightbearing region of the trochlear groove. In each minipig, a PLGA scaffold was placed in the defect of the right knee (PLGA subgroup), and the defect of the left knee was untreated (empty defect [ED] subgroup). The TRE group performed exercises in 3 phases: warm-up, 3 km/h for 5 minutes; main exercise, 4 km/h for 20 minutes; and cool-down, 3 km/h for 5 minutes. The total duration was about 30 minutes whenever possible. The SED group was allowed free cage activity. Results: At 6 months, the TRE-PLGA group showed the highest gross morphology scores and regenerated a smooth articular surface covered with new hyaline-like tissue, while the defects of the other groups remained and contained nontransparent tissue. Histologically, the TRE-PLGA group also revealed sound OC integration, chondrocyte-like cells embedded in lacunae, abundant glycosaminoglycans, a sound collagen structure, and modest inflammatory cells with an inflammatory response (ie, tumor necrosis factor–α, interleukin-6). In addition, in the medial condyle region, the TRE-PLGA group (31.80 ± 3.03) had the highest total histological scores (TRE-ED: 20.20 ± 5.76; SED-PLGA: 10.25 ± 6.24; SED-ED: 11.75 ± 6.50; P = .004). In the trochlear groove region, the TRE-PLGA group (30.20 ± 6.42) displayed significantly higher total histological scores (TRE-ED: 19.60 ± 7.00; SED-PLGA: 10.00 ± 5.42; SED-ED: 11.25 ± 5.25; P = .006). In contrast, the SED-PLGA and SED-ED groups revealed an irregular surface with abrasion, fibrotic tissue with an empty void and inflammatory cells, disorganized collagen fibers, and less glycosaminoglycan deposition. Micro–computed tomography analysis revealed that the TRE-PLGA group had integrated OC interfaces with continued remodeling in the subchondral bone. Furthermore, comparing the 2 defect regions, no statistically significant differences in cartilage regeneration were detected, indicating the suitability of this regenerative approach for both high and low weightbearing regions. Conclusion: Implanting an acellular PLGA scaffold plus treadmill exercise promoted articular cartilage regeneration for both high and low weightbearing regions in minipigs. Clinical Relevance: This study suggests the use of a cell-free porous PLGA scaffold and treadmill exercise rehabilitation as an alternative therapeutic strategy for OC repair in a large-animal knee joint model. This combined effect may pave the way for biomaterials and exercise regimens in the application of OC repair.

Role of OncoTherad immunotherapy in the regulation of toll-like receptors-mediated immune system and RANK/RANKL signaling: New therapeutic perspective for non-muscle invasive bladder cancer.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. e16004-e16004
Author(s):  
Wagner José Fávaro ◽  
Sonia Regina Iantas ◽  
Juliana Mattoso Gonçalves ◽  
Queila Cristina Dias ◽  
Ianny Brum Reis ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Immune System ◽  
Signaling Pathway ◽  
Invasive Bladder Cancer ◽  
Interferon Signaling ◽  
Toll Like Receptors ◽  
Rankl Expression ◽  
Muscle Invasive Bladder Cancer ◽  
New Perspective ◽  
Muscle Invasive

e16004 Background: The new modalities for treating patients with non-muscle invasive bladder cancer (NMIBC) for whom Bacillus Calmette-Guerin (BCG) has failed or is contraindicated are recently increasing due to the development of new drugs. In this scenario, a new perspective is represented by OncoTherad immunomodulator. OncoTherad is a nanostructured inorganic phosphate complex associated to glycosidic protein, developed by University of Campinas/ Brazil, which exhibits antitumor properties. This study detailed and characterized the therapeutic effects of OncoTherad based on activation of Toll-Like Receptors (TLRs) signaling pathways and regulation of receptor activator of nuclear factor kβ (RANK)/RANK ligand (RANKL) cytokine system in an animal model of NMIBC, as well as, compared these effects with BCG treatment. Methods: Fisher 344 female rats were submitted to NMIBC induction with N-methyl-N-nitrosourea (MNU). MNU treated animals were further divided into 3 groups (10 animals per group): the NMIBC group received 0.30 mL of saline solution; the NMIBC-BCG group received of 2 mg/mL of BCG; the NMIBC-OncoTherad group received 20 mg/Kg dose of OncoTherad. All animals were treated intravesically every other week for 6 weeks. Results: Our results demonstrated that OncoTherad intravesical immunotherapy led to distinct activation of TLRs 2 and 4-mediated innate immune system, resulting in increased interferons signaling pathway, which was more effective (80.0%) in the NMIBC treatment, when compared to BCG treatment (20.0%). Interferon signaling pathway activation induced by OncoTherad led to increase of iNOS expression, resulting in apoptosis and histopathological recovery. Also, OncoTherad immunotherapy decreased RANK/RANKL expression, resulting in reduced regulatory T (Treg) cells. Conclusions: The decreased of RANK/RANKL expression by OncoTherad was fundamental to up-regulation interferon signaling pathway and in suppresion of abnormal cell proliferation. Thus, OncoTherad immunotherapy could be considered an important therapeutic strategy for NMIBC, as well as, opens a new perspective for treatment of BCG-refractory or relapsed patients.

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