scholarly journals Influence of Nano, Micro and Macro Topography of Dental Implant Surfaces on Human Gingival Fibroblasts

2021 ◽  
Vol 22 (18) ◽  
pp. 9871
Author(s):  
Morena Petrini ◽  
Tania Vanessa Pierfelice ◽  
Emira D’Amico ◽  
Natalia Di Pietro ◽  
Assunta Pandolfi ◽  
...  
Keyword(s):  
Soft Tissues ◽  
Titanium Implant ◽  
Sem Analysis ◽  
Human Gingival Fibroblasts ◽  
Dependent Manner ◽  
Gingival Fibroblasts ◽  
Force Microscopy ◽  
Atomic Force

Current research on dental implants has mainly focused on the influence of surface roughness on the rate of osseointegration, while studies on the development of surfaces to also improve the interaction of peri-implant soft tissues are lacking. To this end, the first purpose of this study was to evaluate the response of human gingival fibroblasts (hGDFs) to titanium implant discs (Implacil De Bortoli, Brazil) having different micro and nano-topography: machined (Ti-M) versus sandblasted/double-etched (Ti-S). The secondary aim was to investigate the effect of the macrogeometry of the discs on cells: linear-like (Ti-L) versus wave-like (Ti-W) surfaces. The atomic force microscopy (AFM) and scanning electron microscopy (SEM) analysis showed that the Ti-S surfaces were characterized by a significantly higher micro and nano roughness and showed the 3D macrotopography of Ti-L and Ti-W surfaces. For in vitro analyses, the hGDFs were seeded into titanium discs and analyzed at 1, 3, and 5 days for adhesion and morphology (SEM) viability and proliferation (Cck-8 and MTT assays). The results showed that all tested surfaces were not cytotoxic for the hGDFs, rather the nano-micro and macro topography favored their proliferation in a time-dependent manner. Especially, at 3 and 5 days, the number of cells on Ti-L was higher than on other surfaces, including Ti-W surfaces. In conclusion, although further studies are needed, our in vitro data proved that the use of implant discs with Ti-S surfaces promotes the adhesion and proliferation of gingival fibroblasts, suggesting their use for in vivo applications.

scholarly journals Reelin transiently promotes N-cadherin–dependent neuronal adhesion during mouse cortical development

2017 ◽  
Vol 114 (8) ◽  
pp. 2048-2053 ◽  
Author(s):  
Yuki Matsunaga ◽  
Mariko Noda ◽  
Hideki Murakawa ◽  
Kanehiro Hayashi ◽  
Arata Nagasaka ◽  
...  
Keyword(s):  
Neural Progenitor Cells ◽  
Adhesive Force ◽  
Dependent Manner ◽  
Radial Glial Cells ◽  
Force Microscopy ◽  
Atomic Force ◽  
Rotation Culture ◽  
Neuronal Adhesion

Reelin is an essential glycoprotein for the establishment of the highly organized six-layered structure of neurons of the mammalian neocortex. Although the role of Reelin in the control of neuronal migration has been extensively studied at the molecular level, the mechanisms underlying Reelin-dependent neuronal layer organization are not yet fully understood. In this study, we directly showed that Reelin promotes adhesion among dissociated neocortical neurons in culture. The Reelin-mediated neuronal aggregation occurs in an N-cadherin–dependent manner, both in vivo and in vitro. Unexpectedly, however, in a rotation culture of dissociated neocortical cells that gradually reaggregated over time, we found that it was the neural progenitor cells [radial glial cells (RGCs)], rather than the neurons, that tended to form clusters in the presence of Reelin. Mathematical modeling suggested that this clustering of RGCs could be recapitulated if the Reelin-dependent promotion of neuronal adhesion were to occur only transiently. Thus, we directly measured the adhesive force between neurons and N-cadherin by atomic force microscopy, and found that Reelin indeed enhanced the adhesiveness of neurons to N-cadherin; this enhanced adhesiveness began to be observed at 30 min after Reelin stimulation, but declined by 3 h. These results suggest that Reelin transiently (and not persistently) promotes N-cadherin–mediated neuronal aggregation. When N-cadherin and stabilized β-catenin were overexpressed in the migrating neurons, the transfected neurons were abnormally distributed in the superficial region of the neocortex, suggesting that appropriate regulation of N-cadherin–mediated adhesion is important for correct positioning of the neurons during neocortical development.

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

2019 ◽  
Author(s):  
Priya Prakash ◽  
Travis Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Amyloid Beta ◽  
Western Blots ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
Set Up ◽  
Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

scholarly journals The Promotion of Antibacterial Effects of Ti6Al4V Alloy Modified with TiO2Nanotubes Using a Superoxidized Solution

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Ernesto Beltrán-Partida ◽  
Benjamin Valdez-Salas ◽  
Alan Escamilla ◽  
Aldo Moreno-Ulloa ◽  
Larysa Burtseva ◽  
...  
Keyword(s):  
Bacterial Adhesion ◽  
Sem Analysis ◽  
Ti6al4v Alloy ◽  
Antibacterial Efficacy ◽  
Antibacterial Effects ◽  
X Ray ◽  
Force Microscopy ◽  
Atomic Force ◽  
Bacterial Morphology

The purpose of the present study was to synthetize 80 nm diameter TiO2 nanotubes (NTs) on Ti6Al4V alloy using a commercially superoxidized water (SOW) enriched with fluoride to reduce anodization time and promote the antibacterial efficacy againstStaphylococcus aureus(S. aureus). The alloy discs were anodized for 5 min and as a result, NTs of approximately 80 nm diameters were obtained with similar morphology as reported in previous studies using longer anodization times (1-2 h). Filed emission-scanning electron microscopy (FE-SEM) and energy dispersive X-ray spectroscopy (EDX) were used to characterize the materials surfaces. The NTs showed significantly decreasedS. aureusviability after 1, 3, and 5 days of culture in comparison to nonanodized alloy. Likewise, SEM analysis also suggested lower bacterial adhesion on the NTs surface. No differences in bacterial morphology and topography were observed on both materials, as analyzed by SEM and atomic force microscopy (AFM). In conclusion, 80 nm diameter NTs were grown on Ti6Al4V alloy in 5 min by using a SOW solution enriched with fluoride, which resulted in a material with promoted antibacterial efficacy againstS. aureusfor up to 5 days of in vitro culture when compared to nonanodized alloy.

The Influence of the Operating Parameters of Titanium Electropolishing to Obtain Nanostructured Titanium Surfaces

2012 ◽  
Vol 727-728 ◽  
pp. 1638-1642
Author(s):  
Leonardo Marasca Antonini ◽  
Rafael Gomes Mielczarski ◽  
Caroline Pigatto ◽  
Iduvirges Lourdes Müller ◽  
Célia de Fraga Malfatti
Keyword(s):  
Operating Parameters ◽  
Nanostructured Titanium ◽  
Nanostructured Surfaces ◽  
Force Microscopy ◽  
Angle Measurements ◽  
Atomic Force ◽  
Different Temperatures ◽  
Titanium Surfaces

Titanium and Ti alloys have been widely used as biomaterial due to their mechanical properties and high in vitro and in vivo cytocompatibility. Studies have showed that the acceleration of the osseointegration process is associated to the modification of the surface morphology. The aim of this work is to study the influence of the operating parameters of titanium electropolishing to obtain nanostructured titanium surfaces. The titanium electropolishing was carried out with different temperatures (7°C, 18°C and 25°C), current density of 0.19 A/cm2 and electropolishing time of 8 minutes. After the electropolishing process the titanium samples were characterized by Atomic Force Microscopy, profilometry (mechanical profilometer) and contact angle measurements. Preliminary results showed that the Ti nanostructured surfaces formation, strongly depends on the control of operating parameters.

scholarly journals Ultrafiltration Segregates Tissue Regenerative Stimuli Harboured Within and Independent of Extracellular Vesicles

2020 ◽  
Author(s):  
TT Cooper ◽  
SE Sherman ◽  
T Dayarathna ◽  
GI Bell ◽  
Jun Ma ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Limb Ischemia ◽  
Murine Models ◽  
Label Free ◽  
Tubule Formation ◽  
Simple Method ◽  
Force Microscopy ◽  
Atomic Force

AbstractThe release of extracellular vesicles (EVs) from human multipotent stromal cells (MSC) has been proposed as a mechanism by which MSC mediate regenerative functions in vivo. Our recent work has characterized MSC derived from human pancreatic tissues (Panc-MSC) that generated a tissue regenerative secretome. Despite these advancements, it remains unknown whether regenerative stimuli are released independent or within extracellular vesicles. Herein, this study demonstrates ultrafiltration is a simple method to enrich for EVs which can be injected in murine models of tissue regeneration. The enrichment of EVs from Panc-MSC conditioned media (CM) was validated using nanoscale flow cytometry and atomic force microscopy; in addition to the exclusive detection of classical EV-markers CD9, CD81, CD63 using label-free mass spectrometry. Additionally, we identified several pro-regenerative stimuli, such as WNT5A or ANGPT1, exclusive to EV-enriched CM. Endothelial cell tubule formation was enhanced in response to both Panc-MSC CM fractions in vitro yet only intramuscular injection of EV-enriched CM demonstrated vascular regenerative functions in NOD/SCID mice with unilateral hind-limb ischemia (*<p<0.05). Furthermore, both EV-depleted and EV-enriched CM reduced hyperglycemia following intrapancreatic injection in hyperglycemic mice (**p<0.01). Collectively, understanding the functional synergy between compartments of the secretome is required to advance cell-free biotherapeutics into applications of regenerative medicine.

scholarly journals Nanosized Particles Assembled by a Recombinant Virus Protein Are Able to Encapsulate Negatively Charged Molecules and Structured RNA

Polymers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 858
Author(s):  
Hemalatha Mani ◽  
Yi-Cheng Chen ◽  
Yen-Kai Chen ◽  
Wei-Lin Liu ◽  
Shih-Yen Lo ◽  
...  
Keyword(s):  
Self Assembly ◽  
Rna Binding ◽  
Core Protein ◽  
Average Diameter ◽  
Virus Protein ◽  
Force Microscopy ◽  
Atomic Force ◽  
Hcv Core Protein

RNA-based molecules have recently become hot candidates to be developed into therapeutic agents. However, successful applications of RNA-based therapeutics might require suitable carriers to protect the RNA from enzymatic degradation by ubiquitous RNases in vivo. Because of their better biocompatibility and biodegradability, protein-based nanoparticles are considered to be alternatives to their synthetic polymer-based counterparts for drug delivery. Hepatitis C virus (HCV) core protein has been suggested to be able to self-assemble into nucleocapsid-like particles in vitro. In this study, the genomic RNA-binding domain of HCV core protein consisting of 116 amino acids (p116) was overexpressed with E. coli for investigation. The recombinant p116 was able to assemble into particles with an average diameter of approximately 27 nm, as visualized by electron microscopy and atomic force microscopy. Measurements with fluorescence spectroscopy, flow cytometry, and fluorescence quenching indicated that the p116-assembled nanoparticles were able to encapsulate small anionic molecules and structured RNA. This study demonstrates methods that exploit the self-assembly nature of a virus-derived protein for nanoparticle production. This study also suggests that the virus-derived protein-assembled particles could possibly be developed into potential carriers for anionic molecular drugs and structured RNA-based therapeutics.

scholarly journals Antibacterial Properties of Nano-Ag Coating on Healing Abutment: An In Vitro and Clinical Study

Antibiotics ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 347 ◽  
Author(s):  
Tetsurou Odatsu ◽  
Shinichiro Kuroshima ◽  
Mika Sato ◽  
Kazuma Takase ◽  
Alireza Valanezhad ◽  
...  
Keyword(s):  
Clinical Trial ◽  
Dental Plaque ◽  
Antibacterial Effect ◽  
Antibacterial Properties ◽  
Pure Titanium ◽  
Human Gingival Fibroblasts ◽  
Microwave Assisted Synthesis ◽  
Gingival Fibroblasts

Peri-implantitis is an inflammatory disease with a relevant focus on the long-term success of dental implants and implant-supported prostheses. The present study focuses on the antibacterial effect of the silver nanoparticle and investigated the suppression of dental plaque adhesion on implant abutment and/or superstructure by micro-wave assistant nanosilver coating in vivo and in vitro. Nanosilver coating on pure titanium was prepared by microwave-assisted synthesis, and characterized by scanning electron microscopy and energy-dispersive X-ray spectroscopy. In vitro studies were conducted to analyze biocompatibility using MTS assay and fluorescence microscopy with human gingival fibroblasts to evaluate antibacterial activity. During the in vivo study, nanosilver coating was applied to the healing abutments, and the prevention of plaque accumulation on nanosilver coating was confirmed by a split-mouth randomized clinical trial. The aggregation of nano-sized particles was found on the titanium surface with an antibacterial effect. The coating had no cytotoxic effect on human gingival fibroblasts. The result of the clinical trial showed that the coating suppressed the dental plaque adhesion on the healing abutments. Nanosilver coating is a promising material with antibacterial properties and can be used for implant abutments and prostheses for preventing peri-implantitis.

scholarly journals Computational model for nanocarrier binding to endothelium validated using in vivo, in vitro, and atomic force microscopy experiments

2010 ◽  
Vol 107 (38) ◽  
pp. 16530-16535 ◽  
Author(s):  
J. Liu ◽  
G. E. R. Weller ◽  
B. Zern ◽  
P. S. Ayyaswamy ◽  
D. M. Eckmann ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Computational Model ◽  
Force Microscopy ◽  
Atomic Force

scholarly journals Analysis of long dsRNA produced in vitro and in vivo using atomic force microscopy in conjunction with ion-pair reverse-phase HPLC

The Analyst ◽  
2019 ◽  
Vol 144 (16) ◽  
pp. 4985-4994
Author(s):  
Alison O. Nwokeoji ◽  
Sandip Kumar ◽  
Peter M. Kilby ◽  
David E. Portwood ◽  
Jamie K. Hobbs ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
High Performance ◽  
Ion Pair ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Force Microscopy ◽  
Atomic Force ◽  
Insight Into

Atomic force microscopy (AFM) in conjunction with ion-pair reverse-phase high performance liquid chromatography (IP-RP-HPLC) provides novel insight into dsRNA for RNAi applications.

scholarly journals Exploring the action of RGDV-gemcitabine on tumor metastasis, tumor growth and possible action pathway

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Xiaoyi Zhang ◽  
Jinhuan Zhang ◽  
Wenchao Liu ◽  
Yaonan Wang ◽  
Jianhui Wu ◽  
...  
Keyword(s):  
Tumor Metastasis ◽  
A549 Cells ◽  
Force Microscopy ◽  
Atomic Force ◽  
Tnf Α ◽  
First Time ◽  
Inhibit Tumor Metastasis ◽  
Necrosis Factor

Abstract The coupling of Arg-Gly-Asp-Val (RGDV) and gemcitabine led to a hypothesis that the conjugate (RGDV-gemcitabine) could inhibit tumor metastasis. To confirm this hypothesis the activities of RGDV-gemcitabine inhibiting tumor metastasis in vitro and in vivo were presented for the first time. AFM (atomic force microscopy) imaged that RGDV-gemcitabine was able to adhere onto the surface of serum-starved A549 cells, to block the extending of the pseudopodia. Thereby RGDV-gemcitabine was able to inhibit the invasion, migration and adhesion of serum-starved A549 cells in vitro. On C57BL/6 mouse model RGDV-gemcitabine dose dependently inhibited the metastasis of planted tumor towards the lung and the minimal dose was 0.084 µmol/kg/3 days. The decrease of serum TNF-α (tumor necrosis factor), IL-8 (interleukin-8), MMP-2 (matrix metalloprotein-2) and MMP-9 (matrix metalloprotein-9) of the treated C57BL/6 mice was correlated with the action pathway of RGDV-gemcitabine inhibiting the metastasis of the planted tumor towards lung.

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