scholarly journals Metformin Hydrochloride-Plga Nanoparticles in Diabetic Rats in A Periodontal Disease Experimental Model

2018 ◽  
Author(s):  
Aline de Sousa Barbosa Freitas Pereira ◽  
Gerly Anne de Castro Brito ◽  
Maria Laura de Souza Lima ◽  
Arnóbio Antônio da Silva Júnior ◽  
Emanuell dos Santos Silva ◽  
...  
Keyword(s):  
Bone Loss ◽  
Periodontal Disease ◽  
Positive Control ◽  
Plga Nanoparticles ◽  
Metformin Hydrochloride ◽  
Micro Computed Tomography ◽  
Rt Pcr ◽  
Linear Measurements ◽  
Mean Diameter

The aim of this study was synthesize and evaluate the effects of Poly (D, L-Lactide-co-glycolide) (PLGA) Nanoparticles (NPs) of metformin (PLGA+ Met) on inflammation, and bone loss in a ligature-induced periodontitis rat model. The prepared NPs were characterized by mean diameter, size particle, polydispensity index and encapsulation efficiency by Atomic force microscopy (AFM). Male albino Wistar rats were randomly divided into four groups of 20 rats in each group, and given the following treatments for 10 days to evaluate in vivo activity: (1) Sham: no ligature + water; (2) Positive control: ligature + water (with Periodontal disease and Diabetes); (3) ligature + PLGA+ 10 mg/kg Met (With Periodontal disease and Diabetes); and (4) ligature + PLGA+ 100 mg/kg Met (with Periodontal disease and Diabetes). Water or PLGA + Met was administered orally by gavage.  Maxillae were fixed and scanned using Micro-computed Tomography (μCT) to quantify linear of bone loss. Histopathological characteristics were assessed through immunohistochemical staining for Osteocalcin, Cathepsyn K, RANKL/RANK/OPG pathway. IL-1β and TNF-α from gingival tissues were analysed by Elisa immunoassay. Quantitative RT-PCR reaction was used to evaluate gene expression of AMPK, NF-κB p-65, Hmgb1 and TAK-1 from gingival tissues. Statistical analysis was performed using one-way ANOVA at 5% significance. The mean diameter of MET-loaded PLGA nanoparticles was in a range of 457.1 ± 48.9 nm with a polydispersity index of 0.285, zeta potential: 8.16 ± 1.1 mV and entrapment efficiency (EE) was 70%. The results suggest that the addition of MET in the core slightly affected the particle sizes. Treatment with PLGA+ 10 mg/kg Met showed low inflammatory cells, decreased bone loss and integrity cement and levels of IL-1β, and TNF-α (p < 0.05) were significantly reduced. Additionally, weak staining was shown by RANKL, Cathepsyn K, OPG, and osteocalcin. Radiographically, linear measurements showed a statistically significant reduction in bone loss after treatment with PLGA+ 10 mg/kg Met compared to the positive control (p < 0.05). RT-PCR showed increased AMPK expression (p < 0.05) and decreased expression of NF-κB P65, HMGB1 and TAK-1 after PLGA+ 10 mg/kg Met (p < 0.05). The PLGA nanoparticle + 10 mg/kg Met decreased glucose levels and also decreased the inflammatory response, and bone loss in ligature-induced periodontitis in rats.

scholarly journals Metformin Hydrochloride-Loaded PLGA Nanoparticle in Periodontal Disease Experimental Model Using Diabetic Rats

2018 ◽  
Vol 19 (11) ◽  
pp. 3488 ◽  
Author(s):  
Aline Pereira ◽  
Gerly Brito ◽  
Maria Lima ◽  
Arnóbio Silva Júnior ◽  
Emanuell Silva ◽  
...  
Keyword(s):  
Bone Loss ◽  
Cathepsin K ◽  
Immunohistochemical Analysis ◽  
Entrapment Efficiency ◽  
Inflammatory Cells ◽  
Plga Nanoparticles ◽  
Diabetic Rats ◽  
Metformin Hydrochloride ◽  
Tnf Α ◽  
Mean Diameter

Evidence shows that metformin is an antidiabetic drug, which can exert favorable anti-inflammatory effects and decreased bone loss. The development of nanoparticles for metformin might be useful for increased therapeutic efficacy. The aim of this study was to evaluate the effect of metformin hydrochloride-loaded Poly (d,l-Lactide-co-glycolide) (PLGA)/(MET-loaded PLGA) on a ligature-induced periodontitis model in diabetic rats. MET-loaded PLGA were characterized by mean diameter, particle size, polydispensity index, and entrapment efficiency. Maxillae were scanned using Microcomputed Tomography (µCT) and histopathological and immunohistochemical analysis. IL-1β and TNF-α levels were analyzed by ELISA immunoassay. Quantitative RT-PCR was used (AMPK, NF-κB p65, HMGB1, and TAK-1). The mean diameter of MET-loaded PLGA nanoparticles was in a range of 457.1 ± 48.9 nm (p < 0.05) with a polydispersity index of 0.285 (p < 0.05), Z potential of 8.16 ± 1.1 mV (p < 0.01), and entrapment efficiency (EE) of 66.7 ± 3.73. Treatment with MET-loaded PLGA 10 mg/kg showed low inflammatory cells, weak staining by RANKL, cathepsin K, OPG, and osteocalcin, and levels of IL-1β and TNF-α (p < 0.05), increased AMPK expression gene (p < 0.05) and decreased NF-κB p65, HMGB1, and TAK-1 (p < 0.05). It is concluded that MET-loaded PLGA decreased inflammation and bone loss in periodontitis in diabetic rats.

scholarly journals In vitro-in vivo availability of metformin hydrochloride-PLGA nanoparticles in diabetic rats in a periodontal disease experimental model

2021 ◽  
Vol 59 (1) ◽  
pp. 1576-1584
Author(s):  
Aline de Sousa Barbosa Freitas Pereira ◽  
Maria Laura de Souza Lima ◽  
Arnobio Antonio da Silva-Junior ◽  
Emanuell dos Santos Silva ◽  
Raimundo Fernandes de Araújo Júnior ◽  
...  
Keyword(s):  
Periodontal Disease ◽  
Experimental Model ◽  
Plga Nanoparticles ◽  
Diabetic Rats ◽  
Metformin Hydrochloride

scholarly journals Does systemic oral administration of curcumin effectively reduce alveolar bone loss associated with periodontal disease? A systematic review and meta-analysis of preclinical in vivo studies

2020 ◽  
Vol 75 ◽  
pp. 104226
Author(s):  
Juliana Simeão Borges ◽  
Luiz Renato Paranhos ◽  
Gabriela Leite de Souza ◽  
Felipe de Souza Matos ◽  
Ítalo de Macedo Bernardino ◽  
...  
Keyword(s):  
Systematic Review ◽  
Bone Loss ◽  
Periodontal Disease ◽  
Oral Administration ◽  
Alveolar Bone ◽  
Meta Analysis ◽  
Alveolar Bone Loss ◽  
In Vivo Studies

scholarly journals Effects of Colocasia antiquorum var. Esculenta Extract In Vitro and In Vivo against Periodontal Disease

Medicina ◽  
2021 ◽  
Vol 57 (10) ◽  
pp. 1054
Author(s):  
Seong-Hee Moon ◽  
Seong-Jin Shin ◽  
Hyun-Jin Tae ◽  
Seung-Han Oh ◽  
Ji-Myung Bae
Keyword(s):  
Bone Loss ◽  
Periodontal Disease ◽  
Pathogenic Bacteria ◽  
Alveolar Bone ◽  
Negative Control ◽  
Alveolar Bone Loss ◽  
Oral Microbiome ◽  
Control Group

Background and Objectives: Periodontal disease is a chronic inflammatory disease in which gradual destruction of tissues around teeth is caused by plaque formed by pathogenic bacteria. The purpose of this study was to evaluate the potential of 75% ethanol extract of Colocasia antiquorum var. esculenta (CA) as a prophylactic and improvement agent for periodontal disease in vitro and in vivo. Materials and Methods: The antimicrobial efficacy of CA against Porphyromonas gingivalis (P. gingivalis, ATCC 33277) was evaluated using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) test, and cytotoxicity was confirmed by CCK-8 assay. For the in vivo study, P. gingivalis was applied by oral gavage to BALB/c mice. Forty-two days after the first inoculation of P. gingivalis, intraoral swabs were taken for microbiome analysis, and the mice were sacrificed to evaluate the alveolar bone loss. Results: The MIC of CA against P. gingivalis was 31.3 μg/mL, the MBC was 62.5 μg/mL, with no cytotoxicity. The diversity of the oral microbiome decreased in the positive control group, while those of the VA (varnish) and VCA (varnish added with CA) groups increased as much as in the negative control group, although the alveolar bone loss was not induced in the mouse model. Conclusions: CA showed antibacterial effects in vitro, and the VA and VCA groups exhibited increased diversity in the oral microbiome, suggesting that CA has potential for improving periodontal disease.

scholarly journals Mitogen-Activated Protein Kinase 2 Signaling Shapes Macrophage Plasticity in Aggregatibacter actinomycetemcomitans-Induced Bone Loss

2016 ◽  
Vol 85 (1) ◽  
Author(s):  
Bethany A. Herbert ◽  
Heidi M. Steinkamp ◽  
Matthias Gaestel ◽  
Keith L. Kirkwood
Keyword(s):  
Protein Kinase ◽  
Bone Loss ◽  
Periodontal Disease ◽  
Alveolar Bone ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Osteoclast Formation ◽  
Mitogen Activated Protein Kinase ◽  
Micro Computed Tomography ◽  
Content Type ◽  
Mitogen Activated Protein

ABSTRACT Aggregatibacter actinomycetemcomitans is associated with aggressive periodontal disease, which is characterized by inflammation-driven alveolar bone loss. A. actinomycetemcomitans activates the p38 mitogen-activated protein kinase (MAPK) and MAPK-activated protein kinase 2 (MK2) stress pathways in macrophages that are involved in host responses. During the inflammatory process in periodontal disease, chemokines are upregulated to promote recruitment of inflammatory cells. The objective of this study was to determine the role of MK2 signaling in chemokine regulation during A. actinomycetemcomitans pathogenesis. Utilizing a murine calvarial model, Mk2 +/+ and Mk2 −/− mice were treated with live A. actinomycetemcomitans bacteria at the midsagittal suture. MK2 positively regulated the following macrophage RNA: Emr1 (F4/80), Itgam (CD11b), Csf1r (M-CSF Receptor), Itgal (CD11a), Tnf, and Nos2. Additionally, RNA analysis revealed that MK2 signaling regulated chemokines CCL3 and CCL4 in murine calvarial tissue. Utilizing the chimeric murine air pouch model, MK2 signaling differentially regulated CCL3 and CCL4 in the hematopoietic and nonhematopoietic compartments. Bone resorption pits in calvaria, observed by micro-computed tomography, and osteoclast formation were decreased in Mk2 −/− mice compared to Mk2 +/+ mice after A. actinomycetemcomitans treatment. In conclusion, these data suggest that MK2 in macrophages contributes to regulation of chemokine signaling during A. actinomycetemcomitans-induced inflammation and bone loss.

scholarly journals Designation of a Novel DKK1 Multiepitope DNA Vaccine and Inhibition of Bone Loss in Collagen-Induced Arthritic Mice

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Xiaoqing Zhang ◽  
Sibo Liu ◽  
Shentao Li ◽  
Yuxuan Du ◽  
Yunpeng Dou ◽  
...  
Keyword(s):  
Bone Loss ◽  
Dna Vaccine ◽  
Bone Erosion ◽  
Critical Role ◽  
Serum Levels ◽  
Positive Control ◽  
Protective Effects ◽  
High Level

Dickkopf-1 (DKK1), a secretory inhibitor of canonical Wnt signaling, plays a critical role in certain bone loss diseases. Studies have shown that serum levels of DKK1 are significantly higher in rheumatoid arthritis (RA) patients and are correlated with the severity of the disease, which indicates the possibility that bone erosion in RA may be inhibited by neutralizing the biological activity of DKK1. In this study, we selected a panel of twelve peptides using the software DNASTAR 7.1 and screened high affinity and immunogenicity epitopesin vitroandin vivoassays. Furthermore, we optimized four B cell epitopes to design a novel DKK1 multiepitope DNA vaccine and evaluated its bone protective effects in collagen-induced arthritis (CIA), a mouse model of RA. High level expression of the designed vaccine was measured in supernatant of COS7 cells. In addition, intramuscular immunization of BALB/c mice with this vaccine was also highly expressed and sufficient to induce the production of long-term IgG, which neutralized natural DKK1in vivo. Importantly, this vaccine significantly attenuated bone erosion in CIA mice compared with positive control mice. These results provide evidence for the development of a DNA vaccine targeted against DKK1 to attenuate bone erosion.

scholarly journals Induction of M2 Macrophages Prevents Bone Loss in Murine Periodontitis Models

2018 ◽  
Vol 98 (2) ◽  
pp. 200-208 ◽  
Author(s):  
Z. Zhuang ◽  
S. Yoshizawa-Smith ◽  
A. Glowacki ◽  
K. Maltos ◽  
C. Pacheco ◽  
...  
Keyword(s):  
Bone Loss ◽  
Periodontal Disease ◽  
Mrna Expression ◽  
Alveolar Bone ◽  
Quantitative Polymerase Chain Reaction ◽  
Interleukin 1 ◽  
Alveolar Bone Loss ◽  
Micro Computed Tomography ◽  
M2 Phenotype

Periodontitis is characterized by the progressive destruction of tooth-supporting alveolar bone, which is mainly caused by chronic inflammation in response to persistent bacterial insult. It has recently become clear that the pathogenesis of periodontitis is associated with a high ratio of proinflammatory M1 (classically activated) macrophages to anti-inflammatory M2 (alternatively activated). To decrease the inflammatory activity, we locally delivered the C-C motif chemokine ligand 2 (CCL2) using controlled-release microparticles (MPs). CCL2 is known to promote chemotaxis of M0 or M2 phenotype macrophages to the inflamed site and induce M2 phenotype polarization locally. Our in vitro data showed that CCL2 increased the number of M2 phenotype macrophages, decreased TNF-α secretion, and enhanced chemotaxis of RAW264.7 cells toward CCL2 MPs. Moreover, we induced periodontal disease in 2 animal models through inoculation of Porphyromonas gingivalis and ligature around the murine molar. Micro–computed tomography analysis showed significant reduction of alveolar bone loss in the CCL2 MP treatment group when compared with a blank MP group and a no-treatment periodontitis group in both models. Immunohistologic analysis showed a significant increase in the M2 phenotype subset and a decrease in the M1 phenotype subset in the CCL2 MP group of the P. gingivalis–induced model. Also, in both models, tartrate-resistant acidic phosphatase staining showed significantly fewer numbers of osteoclasts in the CCL2 MP group in alveolar bone area. Moreover, quantitative polymerase chain reaction results showed a significant increase in IL-1RA (interleukin 1 receptor antagonist) mRNA expression and a decrease in RANKL (receptor activator of nuclear factor kappa-Β ligand) mRNA expression in the CCL2 MP group in the ligature model. In summary, manipulation of endogenous M2 phenotype macrophages with CCL2 MPs decreased the M1 phenotype:M2 phenotype ratio and prevented alveolar bone loss in mouse periodontitis models. The delivery of CCL2 MPs provides a novel approach to treat periodontal disease.

3D In Vivo Bone Dynamic Imaging of PTH’s Anabolic Action

2013 ◽  
Author(s):  
Allison R. Altman ◽  
Beom Kang Huh ◽  
Abhishek Chandra ◽  
Wei-Ju Tseng ◽  
Ling Qin ◽  
...  
Keyword(s):  
Bone Loss ◽  
Bone Formation ◽  
Human Life ◽  
Combined Therapy ◽  
Term Treatment ◽  
Dynamic Imaging ◽  
Combined Treatments ◽  
Micro Computed Tomography ◽  
Long Term Treatment

Aging shifts bone remodeling toward a negative balance between bone formation and resorption, causing bone loss and increased fracture risk. Anti-resorptive agents are commonly used to inhibit bone resorption and stabilize bone mass. While they are effective to prevent further bone loss, there is also a great need for anabolic agents which can reverse bone deterioration and regain lost skeletal integrity. Intermittent parathyroid hormone (PTH) treatment is the only FDA-approved anabolic treatment for osteoporosis, which greatly stimulates bone formation. Combined therapy of anti-resorptive drugs, such as alendronate (ALN), and PTH have been proposed and are expected to further stimulate bone formation. However, studies show conflicting results regarding the effectiveness of combined treatments: some have reported the addition of ALN to impair PTH function [1, 2], while others suggest an improvement over PTH monotherapy [3, 4]. The first objective of this study is to document the immediate changes of individual trabecular structures due to PTH and combined therapy within 12 days using in vivo micro computed tomography (μCT). As PTH is typically prescribed for 1 to 3 years to osteoporotic patients, a treatment of 12 days for rats (approximately equivalent to one year of human life) may be more clinically relevant than long-term treatment studies on rats. The secondary purpose of this study was to gain insight into the mechanism of combined versus PTH treatments through a bone dynamic imaging strategy to track events over an individual remodeling site. We hypothesized that PTH and combined treatments would immediately enhance bone formation on the trabecular surface.

125 Presence of melatonin receptors in ovine blastocysts

2019 ◽  
Vol 31 (1) ◽  
pp. 188
Author(s):  
A. Casao ◽  
R. Pérez-Pé ◽  
J. A. Cebrián-Pérez ◽  
T. Muiño-Blanco ◽  
F. Forcada ◽  
...  
Keyword(s):  
Positive Control ◽  
Melatonin Receptors ◽  
Penicillin G ◽  
Rt Pcr ◽  
Embryo Viability ◽  
Alexa Fluor ◽  
Streptomycin Sulfate ◽  
Positive Effects

Melatonin increases in vivo embryo viability in sheep and improves the blastocyst rate on in vitro sheep embryo culture. To determine whether this melatonin effect is receptor-mediated, we evaluated the presence of melatonin receptors MT1 and MT2 on in vitro-obtained sheep embryos by means of RT-PCR and indirect immunofluorescence (IIF). For in vitro embryo production, oocytes were collected from ovaries of adult ewes and matured with TCM-199; 10% oestrus sheep serum; FSH and LH, 10μg mL−1 each; cysteamine, 100 μM; sodium pyruvate, 0.3 mM; penicillin G, 100IU mL−1; and streptomycin sulfate, 100μg mL−1 for 24h at 39°C and 5% CO2. Matured oocytes were transferred to a fertilization medium (SOF without glucose, with 2% oestrus sheep serum; heparin, 10μg mL−1; and hypotaurine, 1μg mL−1) and fertilized with swim-up selected spermatozoa at a final concentration of 106 cells mL−1. After 36h, the cleaved embryos were incubated in culture medium (SOF with amino acids, 0.4% BSA; l-glutamine, 1 mM; penicillin G, 100IU mL−1; and streptomycin sulfate, 100μg mL−1) for 8 days at 39°C with 5% CO2, 5% O2, and 90% N2. In vitro fertilization was repeated 3 times. Day 8 hatched blastocysts were selected for RT-PCR or IIF. For RT-PCR assays, RNA from 5 to 10 blastocysts was extracted and reverse transcribed with SuperScript™ III CellsDirect™ cDNA Synthesis Kit (Thermo Fisher Scientific Inc., Waltham, MA, USA). The PCR amplification was carried out in a Step One Plus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The PCR mix contained 2μL of embryo cDNA, iTaq Universal SYBR Green Supermix (Bio-Rad Laboratories Inc., Waltham, MA, USA) and 200nM of the MT1 (forward: 5′-CTCCATCCTCATCTTCACCATC-3′, reverse: 5′-GGCTCACCACAAACACATTC-3′, 113bp) or MT2 (forward: 5′-GCTGAGAGAATGGAGCGATATG-3′, reverse: 5′-GTCCACAGTGAGAAGCCATC-3′, 81bp) primers. The RT-PCR products were visualised under ultraviolet light on 2% agarose gel in a TBE buffer (Tris, 0.9 M; boric acid, 0.9 M; and EDTA, 20mM) with 0.5μL mL−1 ethidium bromide. Ovine testis was used as positive control. For IIF, 5 blastocysts were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% BSA in PBS for 2h at RT. Embryos were incubated overnight at 4°C with MTNR1A mouse polyclonal antibody (Abnova, Taipei City, Taiwan) for MT1, and rabbit melatonin receptor 1B antibody (Acris Antibodies, Atlanta, GA, USA) for MT2, both diluted 1/50 in PBS with 1% BSA. Secondary antibodies were Alexa Fluor 594 chicken anti-mouse (Invitrogen, Carlsbad, CA, USA) for MT1 and Alexa Fluor 488 chicken anti-rabbit for MT2, diluted 1/800 in PBS containing 1% BSA. Embryos were mounted in slides and visualised on a Nikon Eclipse E-400 microscope (Nikon, Tokyo, Japan). The RT-PCR revealed a single band of 113bp for MT1 and another one of 81bp for MT2 in both the embryo samples and the positive control, which confirmed the gene expression of melatonin receptors in the ovine embryo. The IIF located the MT1 receptor around the nucleus of the trophoblastic cells, whereas MT2 was over the nucleus in the same cells. These results indicate the presence of melatonin receptors in sheep blastocyst, which could mediate the positive effects of this hormone on embryo viability.

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