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

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.

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

10.20944/preprints201807.0367.v1 ◽

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.

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Histopathological Verification of Osteoimmunological Mediators in Peri-Implantitis and Correlation to Bone Loss and Implant Functional Period

Journal of Oral Implantology ◽

10.1563/aaid-joi-d-13-00355 ◽

2016 ◽

Vol 42 (1) ◽

pp. 61-68 ◽

Cited By ~ 8

Author(s):

Anna Konermann ◽

Werner Götz ◽

Michael Le ◽

Cornelius Dirk ◽

Stefan Lossdörfer ◽

...

Keyword(s):

Bone Resorption ◽

Bone Loss ◽

Retention Time ◽

Correlation Coefficients ◽

Inflammatory Cells ◽

Smoking History ◽

Tnf Α ◽

Implant Retention ◽

Higher Rank ◽

Inflammatory Infiltrates

Peri-implantitis (PI) is characterized by inflammation and bone resorption eventually leading to implant failure, but the characteristic pathologic determinants are undefined to date. This study aims to elucidate the parameters involved in PI pathogenesis, including intraoral implant retention time, extent of bone loss, smoking history, and identification of osteoimmunological markers for inflammation and bone loss. Peri-implant tissues (n = 21) displaying clinically diagnosed PI from patients with vertical bone loss ranging from 0–12 mm and implant function period between 1 and 60 months were evaluated by histochemistry and immunohistochemistry for TRAP, CD3, RANK, RANKL, OPG, and TNF-α. Statistical analyses were performed with the Welch test and correlation coefficients were calculated. Most bone resorption occurred during the first 12 months of implant function and correlated with the extent of inflammation, although histological signs of inflammation strongly varied between samples from minimal appearance of inflammatory cells to extended infiltrates. Implant function period and smoking history did not significantly affect the degree of inflammation. Higher RANK levels emerged in the first 12 months of implant function compared to longer retention times and were negatively correlated to the occurrence of RANKL. Additionally, histological signs of inflammation were about two-fold higher in specimens with bone resorption up from 5 mm compared to under 5 mm. CD3+ cells were more prevalent in extensive inflammatory infiltrates and samples derived from smokers. Our analyses proved that PI-induced bone loss is differentially influenced by the parameters evaluated in this study, but a distinct interconnection between disease severity and implant retention time can be established.

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Evolution of Periodontal Disease: Immune Response and RANK/RANKL/OPG System

Brazilian Dental Journal ◽

10.1590/0103-6440201701407 ◽

2017 ◽

Vol 28 (6) ◽

pp. 679-687 ◽

Cited By ~ 5

Author(s):

Fabrício Gibertoni ◽

Meire Ellen Ligia Sommer ◽

Marcelo Augusto Marretto Esquisatto ◽

Maria Esméria Corezola do Amaral ◽

Camila Andrea de Oliveira ◽

...

Keyword(s):

Nitric Oxide ◽

Immune Response ◽

Bone Loss ◽

Periodontal Disease ◽

Immunohistochemical Analysis ◽

Inflammatory Cells ◽

Gingival Tissue ◽

Cervical Region ◽

Nitric Oxide Concentration ◽

Gene Transcripts

Abstract The aim of this study was to evaluate markers of bone loss and immune response present in evolution of periodontal disease. One hundred and two Wistar rats were divided into three animals groups: PD0, without ligation and PD15 days and PD60 days, submitted to ligation placement with a sterile 3-0 silk cord in the cervical region of the upper first molar on both sides. Samples were obtained from the gingival tissue for histomorphometric analysis, immunohistochemical analysis of RANK, RANKL, OPG, characterization of the inflammatory infiltrate, quantification of nitric oxide, MCP-1, RANTES, IP10 chemokines, and expression of the TGF-b1, VEG, and bFGF. The number of inflammatory cells in gingival tissue was higher in PD60 samples. The collagen content and the area occupied by birefringent collagen fibers were lower for PD60. Differential leukocyte counting showed that there was a significantly higher polymorphonuclear influx in group PD15, while PD60 showed a greater number of lymphocytes. PD60 showed higher RANTES, IP-10, MCP-1 gene transcripts, as well as a higher nitric oxide concentration. Clinical evaluation revealed that the PD60 group presented an increase in furcal area. In conclusion, in this animal model the increase of RANK/RANKL and HGF markers is related to a specific immune response, and probably contributed to the evolution of periodontal disease. Investigating the effect of these biomarkers can help in targeted therapy for bone resorption, since blocking these can inhibit bone loss.

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Physical Exercise Improves Glycemic and Inflammatory Profile and Attenuates Progression of Periodontitis in Diabetic Rats (HFD/STZ)

Nutrients ◽

10.3390/nu10111702 ◽

2018 ◽

Vol 10 (11) ◽

pp. 1702 ◽

Cited By ~ 5

Author(s):

Eric Andrade ◽

Viviam Silva ◽

Natália Moura ◽

Renata Foureaux ◽

Débora Orlando ◽

...

Keyword(s):

Bone Loss ◽

Physical Exercise ◽

Alveolar Bone ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Diabetic Rats ◽

Alveolar Bone Loss ◽

Low Density ◽

Tnf Α ◽

Inflammatory Profile

The authors aimed to evaluate the effects of physical exercise on the metabolism and progression of periodontal disease (PD), induced by ligature in diabetic rats induced by high fat diet and streptozotocin (HFD/STZ). Diabetes Mellitus (DM) was induced by four weeks of a hyperlipidic diet associated with a single low-dose of streptozotocin (35 mg/kg/animal). The exercise groups swam for 60 min/day for eight weeks (five times/week). In the last two weeks of exercise, a ligature was placed around the right and left mandibular first molars. The authors determined alveolar bone loss by morphometry. Blood biochemical profile and serum levels of IL-10 and TNF-α were evaluated by colorimetric and enzyme-linked immunosorbent assays (ELISA), respectively. The diabetic animals subjected to exercise showed decreased alveolar bone loss, lower glycemia, triacylglycerols and glycosylated hemoglobin levels than the controls. Total cholesterol and its fractions (High density lipoprotein—HDL-c, Low density lipoprotein—LDL-c and Very low density lipoprotein—VLDL-c) remained similar among the groups. Animals with PD showed higher levels of TNF-α and lower levels of IL-10, when compared to animals without PD. In diabetic animals with PD, physical exercise decreased TNF-α levels and increased IL-10 levels as well as the IL10/TNF-α ratio. In conclusion, eight weeks of physical exercise improved glycemic control and systemic inflammatory profile, and attenuated alveolar bone loss in rats with DM and PD.

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

Pharmaceutical Biology ◽

10.1080/13880209.2021.2002369 ◽

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

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Bisleuconothine A protects periodontal tissue via the regulation of RANKL expression and infiltration of inflammatory cells

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i3.6 ◽

2020 ◽

Vol 19 (3) ◽

pp. 497-503

Author(s):

Fang Wang ◽

Ping Sun ◽

Qiang Sun

Keyword(s):

Bone Loss ◽

Alveolar Bone ◽

Inflammatory Cells ◽

Alveolar Bone Loss ◽

Gingival Tissue ◽

Enzyme Linked Immunosorbent Assay ◽

Periodontal Tissue ◽

Rankl Expression ◽

Periodontal Tissues ◽

Tnf Α

Purpose: To investigate the protective effect of bisleuconothine A on periodontal tissue in rats and the mechanism involved. Methods: Adult male Sprague Dawley rats (n = 32) weighing 180 - 200 g (mean weight, 190 ± 10 g) were randomly assigned to four groups of eight rats each: control group, periodontitis group, bisleuconothine A (50 mg/kg) group and bisleuconothine A (100 mg/kg) group. Rats in the treatment groups received bisleuconothine intraperitoneally for two weeks. Periodontitis was induced in the rats using standard procedures. Serum and tissue samples were used for biochemical analysis. Alveolar bone loss was measured in rat maxillae, while the activity of bone alkaline phosphatase (BALP) was determined in serum. Tumor necrosis factor α (TNF-α) interleukin-1β and interleukin-6 (IL-1β and IL-6) were determined in gingival tissue using enzyme-linked immunosorbent assay (ELISA) kit. Gene and protein expressions of receptor activator of nuclear factor kappa-Β ligand (RANKL), osteoprotegerin (OPG), and matrix metallopeptidase-9 (MMP-9) were measured in gingival tissue using real-time quantitative polymerase chain reaction (qRT-PCR) and Western blotting, respectively. Results: Bisleuconothine A treatment significantly and dose-dependently reduced alveolar bone loss, as well as serum levels of TNF-α, IL-1β and IL-6, but increased BALP activity in periodontitis rats (p < 0.05). It also significantly and dose-dependently reduced mRNA expressions of RANKL and MMP-9, but significantly increased OPG mRNA expression (p < 0.05). Similarly, treatment with bisleuconothine A significantly and dose-dependently down-regulated RANKL, p-NF-kB, p-IkBα and iNOS proteins in gingival tissue of periodontitis rats (p < 0.05). The results of histopathological examination indicated that bisleuconothine A treatment significantly reversed histological changes in periodontal tissues of periodontitis rats. It also significantly reduced the degree of polymorphonuclear (PMN) cell infiltration in periodontal tissue. Conclusion: The results obtained show that bisleuconothine A protects periodontal tissue via the regulation of RANKL expression and infiltration of inflammatory cells. Keywords: Bisleuconothine A, Expression, Inflammation, Periodontitis, RANKL

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A PPARψ agonist improved diabetic state via decreasing TNF-α without any change including obesity in genetically obese diabetic rats

Gastroenterology ◽

10.1016/s0016-5085(01)83353-9 ◽

2001 ◽

Vol 120 (5) ◽

pp. A674-A674 ◽

Cited By ~ 1

Author(s):

A FUNAKOSHI ◽

M ICHIKAWA ◽

Y SATO ◽

S KANAI ◽

M OHTA ◽

...

Keyword(s):

Diabetic Rats ◽

Diabetic State ◽

Tnf Α ◽

Obese Diabetic Rats

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Evaluation of Pulp Repair after BiodentineTM Full Pulpotomy in a Rat Molar Model of Pulpitis

Biomedicines ◽

10.3390/biomedicines9070784 ◽

2021 ◽

Vol 9 (7) ◽

pp. 784

Author(s):

Sandra Minic ◽

Marion Florimond ◽

Jérémy Sadoine ◽

Anne Valot-Salengro ◽

Catherine Chaussain ◽

...

Keyword(s):

Immunohistochemical Analysis ◽

Tricalcium Silicate ◽

Male Rats ◽

Enzyme Linked Immunosorbent Assay ◽

E Coli ◽

Inflammatory Conditions ◽

Tnf Α ◽

Pulp Inflammation ◽

Characteristics Analysis ◽

Pulp Repair

Dental pulp is a dynamic tissue able to heal after injury under moderate inflammatory conditions. Our study aimed to evaluate pulp repair under inflammatory conditions in rats. For this purpose, we developed a rat model of controlled pulpitis followed by pulpotomy with a tricalcium silicate-based cement. Fifty-four cavities were prepared on the occlusal face of the maxillary upper first molar of 27 eight-week-old male rats. E. coli lipopolysaccharides at 10 mg/mL or phosphate-buffered saline PBS was injected after pulp injury. Non-inflamed molars were used as controls. Levels of inflammation-related molecules were measured 6 and 24 h after induction by enzyme-linked immunosorbent assay of coronal pulp samples. Pulp capping and coronal obturation after pulpotomy were performed with tricalcium silicate-based cement. Four and fifteen days after pulpotomy, histological and immunohistochemical analysis was performed to assess pulp inflammation and repair processes. Our results showed significantly higher levels of innate inflammatory proteins (IL-1β, IL-6, TNF-α and CXCL-1) compared with those in controls. Moderate residual inflammation near the capping material was demonstrated by histology and immunohistochemistry, with the presence of few CD68-positive cells. We showed that, in this model of controlled pulpitis, pulpotomy with BiodentineTM allowed the synthesis at the injury site of a mineralized bridge formed from mineralized tissue secreted by cells displaying odontoblastic characteristics. Analysis of these data suggests overall that, with the limitations inherent to findings in animal models, pulpotomy with a silicate-based cement is a good treatment for controlling inflammation and enhancing repair in cases of controlled pulpitis.

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Hyaluronic Acid-Modified and TPCA-1-Loaded Gold Nanocages Alleviate Inflammation

Pharmaceutics ◽

10.3390/pharmaceutics11030143 ◽

2019 ◽

Vol 11 (3) ◽

pp. 143 ◽

Cited By ~ 4

Author(s):

Jingnan Zhao

Keyword(s):

Hyaluronic Acid ◽

Inflammatory Cells ◽

Oxygen Species ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Nanogold Particles ◽

Gold Nanocages ◽

Factor Alpha ◽

Anti Inflammatory ◽

Necrosis Factor

Gold nanocages (AuNCs) are biocompatible and porous nanogold particles that have been widely used in biomedical fields. In this study, hyaluronic acid (HA) and peptide- modified gold nanocages (HA-AuNCs/T/P) loaded with 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1) were prepared to investigate their potential for combating inflammation. TPCA-1 was released from AuNCs, intracellularly when HA was hydrolyzed by hyaluronidase. HA-AuNCs/T/P show a much higher intracellular uptake than AuNCs/T/P, and exhibit a much higher efficacy on the suppression of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) than free TPCA-1, suggesting great improvement to the anti-inflammatory efficacy of TPCA-1 through the application of AuNCs. HA-AuNCs/T/P can also reduce the production of reactive oxygen species in inflammatory cells. This study suggests that HA-AuNCs/T/P may be potential agents for anti-inflammatory treatment, and are worthy of further investigation.

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Formulation and development of metformin-loaded microspheres using Khaya senegalensis (Meliaceae) gum as co-polymer

Future Journal of Pharmaceutical Sciences ◽

10.1186/s43094-020-00139-6 ◽

2020 ◽

Vol 6 (1) ◽

Author(s):

Chukwuebuka H. Ozoude ◽

Chukwuemeka P. Azubuike ◽

Modupe O. Ologunagba ◽

Sejoro S. Tonuewa ◽

Cecilia I. Igwilo

Keyword(s):

Controlled Release ◽

Sodium Alginate ◽

Release Kinetics ◽

Drug Carrier ◽

In Vitro Release ◽

Entrapment Efficiency ◽

Metformin Hydrochloride ◽

Scanning Calorimetry ◽

Khaya Senegalensis

Abstract Background Khaya gum is a bark exudate from Khaya senegalensis (Maliaecae) that has drug carrier potential. This study aimed to formulate and comparatively evaluate metformin-loaded microspheres using blends of khaya gum and sodium alginate. Khaya gum was extracted and subjected to preformulation studies using established protocols while three formulations (FA; FB and FC) of metformin (1% w/v)-loaded microspheres were prepared by the ionic gelation method using 5% zinc chloride solution as the cross-linker. The formulations contained 2% w/v blends of khaya gum and sodium alginate in the ratios of 2:3, 9:11, and 1:1, respectively. The microspheres were evaluated by scanning electron microscopy, Fourier transform-infrared spectroscopy, differential scanning calorimetry, entrapment efficiency, swelling index, and in vitro release studies. Results Yield of 28.48%, pH of 4.00 ± 0.05, moisture content (14.59% ± 0.50), and fair flow properties (Carr’s index 23.68 ± 1.91 and Hausner’s ratio 1.31 ± 0.03) of the khaya gum were obtained. FTIR analyses showed no significant interaction between pure metformin hydrochloride with excipients. Discrete spherical microspheres with sizes ranging from 1200 to 1420 μm were obtained. Drug entrapment efficiency of the microspheres ranged from 65.6 to 81.5%. The release of the drug from microspheres was sustained for the 9 h of the study as the cumulative release was 62% (FA), 73% (FB), and 80% (FC). The release kinetics followed Korsmeyer-Peppas model with super case-II transport mechanism. Conclusion Blends of Khaya senegalensis gum and sodium alginate are promising polymer combination for the preparation of controlled-release formulations. The blend of the khaya gum and sodium alginate produced microspheres with controlled release properties. However, the formulation containing 2:3 ratio of khaya gum and sodium alginate respectively produced microspheres with comparable controlled release profiles to the commercial brand metformin tablet.

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