scholarly journals Osteoblast-Like Cell Behavior on Porous Scaffolds Based on Poly(styrene) Fibers

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Andrada Serafim ◽  
Romain Mallet ◽  
Florence Pascaretti-Grizon ◽  
Izabela-Cristina Stancu ◽  
Daniel Chappard
Keyword(s):  
Interference Microscopy ◽  
Porous Scaffolds ◽  
Allogenic Bone ◽  
Force Microscopy ◽  
Atomic Force ◽  
Dry Spinning ◽  
Bone Trabeculae ◽  
Large Bone

Scaffolds of nonresorbable biomaterials can represent an interesting alternative for replacing large bone defects in some particular clinical cases with massive bone loss. Poly(styrene) microfibers were prepared by a dry spinning method. They were partially melted to provide 3D porous scaffolds. The quality of the material was assessed by Raman spectroscopy. Surface roughness was determined by atomic force microscopy and vertical interference microscopy. Saos-2 osteoblast-like cells were seeded on the surface of the fibers and left to proliferate. Cell morphology, evaluated by scanning electron microscopy, revealed that they can spread and elongate on the rough microfiber surface. Porous 3D scaffolds made of nonresorbable poly(styrene) fibers are cytocompatible biomaterials mimicking allogenic bone trabeculae and allowing the growth and development of osteoblast-like cellsin vitro.

scholarly journals Development and AFM study of porous scaffolds for wound healing applications

2004 ◽  
Vol 18 (4) ◽  
pp. 587-596 ◽  
Author(s):  
T. A. Doneva ◽  
H. B. Yin ◽  
P. Stephens ◽  
W. R. Bowen ◽  
D. W. Thomas
Keyword(s):  
Atomic Force Microscopy ◽  
Wound Healing ◽  
Human Fibroblasts ◽  
Three Dimensional ◽  
Positive Influence ◽  
Porous Scaffolds ◽  
Force Microscopy ◽  
Cellular Assays ◽  
Atomic Force

An engineering approach to the development of biomaterials for promotion of wound healing emphasises the importance of a well‒controlled architecture and concentrates on optimisation of morphology and surface chemistry to stimulate guidance of the cells within the wound environment. A series of three‒dimensional porous scaffolds with 80–90% bulk porosity and fully interconnected macropores were prepared from two biodegradable materials – cellulose acetate (CA) and poly (lactic‒co‒glycolic acid) (PLGA) through the phase inversion mechanism of formation. Surface morphology of obtained scaffolds was determined using atomic force microscopy (AFM) in conjunction with optical microscopy. Scanning Electron Microscopy (SEM) was applied to characterise scaffolds bulk morphology. Biocompatibility and biofunctionality of the prepared materials were assessed through a systematic study of cell/material interactions using atomic force microscopy (AFM) methodologies together within vitrocellular assays. Preliminary data with human fibroblasts demonstrated a positive influence of both scaffolds on cellular attachment and growth. The adhesion of cells on both biomaterials were quantified by AFM force measurements in conjunction with a cell probe technique since, for the first time, a fibroblast probe has been successfully developed and optimal conditions of immobilisation of the cells on the AFM cantilever have been experimentally determined.

The application of bulk fill composites in restorations of coronal part of posterior teeth

2015 ◽  
Vol 9 (2) ◽  
pp. 0-0
Author(s):  
Иванов ◽  
S. Ivanov ◽  
Шумилович ◽  
B. Shumilovich ◽  
Воробьева ◽  
...  
Keyword(s):  
Marginal Adaptation ◽  
Hybrid Layer ◽  
Posterior Teeth ◽  
Force Microscopy ◽  
Atomic Force ◽  
Dental Hard Tissues ◽  
Different Shapes ◽  
Flowable Composites

Localization of caries on posterior teeth is due to anatomical features. Fissures having different shapes and deepening are the most vulnerable point. The main causes of violations of fit composite is depressurized hybrid layer, breach conditioning technology, complex terrain the prepared cavity, manual distribution of adhesive in violation of instructions with adhesive systems, the use of eugenol-containing materials, guaiacol, aluminum chloride, ferric sulfate, aluminum sulfate, uncontrolled by power-curing unit, and condensing the composite uneven due to insufficient ductility. Not seen in the course of treatment errors, clearly visible in a few months and give rise to recurrent caries. It was investigated the quality of marginal adaptation of composites such as Bulk Fill in the restoration of posterior teeth. In vitro staining was used border composite-dental hard tissues, scanning electron microscopy and atomic force microscopy. Sealing Flowable composites SDR does not provide the necessary quality of marginal adaptation of composite (presence boundary gap seals depressurization, the presence of air pores, cohesive fracture of the composite, etc.). These facts are missing in the application of the composite with varying viscosity Sonic Fill, which provides the necessary quality and durability of the restora-tion.

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.

Targeting Breast Cancer Cells with G4 PAMAM Dendrimers and Valproic Acid Derivative Complexes

2020 ◽  
Vol 20 (15) ◽  
pp. 1857-1872
Author(s):  
Alberto M. Muñoz ◽  
Manuel J. Fragoso-Vázquez ◽  
Berenice P. Martel ◽  
Alma Chávez-Blanco ◽  
Alfonso Dueñas-González ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Cell Lines ◽  
Water Solubility ◽  
Md Simulations ◽  
Pamam Dendrimer ◽  
Pamam Dendrimers ◽  
Force Microscopy ◽  
Micromolar Concentration ◽  
Atomic Force

Background: Our research group has developed some Valproic Acid (VPA) derivatives employed as anti-proliferative compounds targeting the HDAC8 enzyme. However, some of these compounds are poorly soluble in water. Objective: Employed the four generations of Polyamidoamine (G4 PAMAM) dendrimers as drug carriers of these compounds to increase their water solubility for further in vitro evaluation. Methods: VPA derivatives were subjected to Docking and Molecular Dynamics (MD) simulations to evaluate their affinity on G4 PAMAM. Then, HPLC-UV/VIS, 1H NMR, MALDI-TOF and atomic force microscopy were employed to establish the formation of the drug-G4 PAMAM complexes. Results: The docking results showed that the amide groups of VPA derivatives make polar interactions with G4 PAMAM, whereas MD simulations corroborated the stability of the complexes. HPLC UV/VIS experiments showed an increase in the drug water solubility which was found to be directly proportional to the amount of G4 PAMAM. 1H NMR showed a disappearance of the proton amine group signals, correlating with docking results. MALDI-TOF and atomic force microscopy suggested the drug-G4 PAMAM dendrimer complexes formation. Discussion: In vitro studies showed that G4 PAMAM has toxicity in the micromolar concentration in MDAMB- 231, MCF7, and 3T3-L1 cell lines. VPA CF-G4 PAMAM dendrimer complex showed anti-proliferative properties in the micromolar concentration in MCF-7 and 3T3-L1, and in the milimolar concentration in MDAMB- 231, whereas VPA MF-G4 PAMAM dendrimer complex didn’t show effects on the three cell lines employed. Conclusion: These results demonstrate that G4 PAMAM dendrimers are capableof transporting poorly watersoluble aryl-VPA derivate compounds to increase its cytotoxic activity against neoplastic cell lines.

Structural and Optical Properties of InAsSbBi Grown by Molecular Beam Epitaxy on Offcut GaSb Substrates

Photonics ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 215
Author(s):  
Rajeev R. Kosireddy ◽  
Stephen T. Schaefer ◽  
Marko S. Milosavljevic ◽  
Shane R. Johnson
Keyword(s):  
Molecular Beam Epitaxy ◽  
Molecular Beam ◽  
Optical Quality ◽  
X Ray Diffraction ◽  
Interface Quality ◽  
Force Microscopy ◽  
Atomic Force ◽  
Transmission Electron ◽  
Lateral Variations

Three InAsSbBi samples are grown by molecular beam epitaxy at 400 °C on GaSb substrates with three different offcuts: (100) on-axis, (100) offcut 1° toward [011], and (100) offcut 4° toward [011]. The samples are investigated using X-ray diffraction, Nomarski optical microscopy, atomic force microscopy, transmission electron microscopy, and photoluminescence spectroscopy. The InAsSbBi layers are 210 nm thick, coherently strained, and show no observable defects. The substrate offcut is not observed to influence the structural and interface quality of the samples. Each sample exhibits small lateral variations in the Bi mole fraction, with the largest variation observed in the on-axis growth. Bismuth rich surface droplet features are observed on all samples. The surface droplets are isotropic on the on-axis sample and elongated along the [011¯] step edges on the 1° and 4° offcut samples. No significant change in optical quality with offcut angle is observed.

Mechanical Interlocking on Leadframe Surface for Bondability of Au Wedge Bond

2016 ◽  
Vol 857 ◽  
pp. 79-82
Author(s):  
Roslina Ismail ◽  
Fuaida Harun ◽  
Azman Jalar ◽  
Shahrum Abdullah
Keyword(s):  
Optical Microscope ◽  
Bonding Process ◽  
Mechanical Interlocking ◽  
Average Roughness ◽  
Force Microscopy ◽  
Atomic Force ◽  
Surface Morphologies ◽  
Scanning Electron ◽  
Effect Of Surface

This work is a contribution towards the understanding of wire bond integrity and reliability in relation to their microstructural and mechanical properties in semiconductor packaging.The effect of surface roughness and hardness of leadframe on the bondability of Au wedge bond still requires detail analysis. Two type of leadframes namely leadframe A and leadframe B were chosen and scanning electron microscope (SEM) and optical microscope were used to inspect the surface morphology of leadframes and the quality of created Au wedge bond after wire bonding process. It was found that there were significant differences in the surface morphologies between these two leadframes. The atomic force microscopy (AFM) which was utilized to measure the average roughness, Ra of lead finger confirms that leadframe A has the highest Ra with value of 166.46 nm compared to that of leadframe B with value of 85.89 nm. While hardness value of different lead finger from the selected leadframe A and B obtained using Vicker microhardness tester are 180.9 VH and 154.2VH respectively.

scholarly journals Nanostructural and mechanical changes in the sclera following proteoglycan depletion

2018 ◽  
Vol 2 (2) ◽  
pp. 14-17
Author(s):  
Zhuola Zhuola ◽  
Steve Barrett ◽  
Yalda Ashraf Kharaz ◽  
Riaz Akhtar
Keyword(s):  
Mechanical Properties ◽  
Collagen Fibril ◽  
Enzymatic Degradation ◽  
Ocular Tissues ◽  
Force Microscopy ◽  
Nanomechanical Properties ◽  
Atomic Force ◽  
Fibril Morphology

The mechanical properties of ocular tissues, such as the sclera, have a major impact on healthy eye function, and are governed by the properties and composition of the microstructural components. For example, biomechanical degradation associated with myopia occurs alongside a reduction of proteoglycans (PGs). In this study, the role of PG degradation in the nanomechanical properties of the porcine sclera is explored. In-vitro enzymatic degradation of PGs was conducted with α-amylase and chondroitinase ABC enzymes. Collagen fibril morphology and nanomechanical stiffness were measured with atomic force microscopy (AFM). The elastic modulus of the tissue was reduced in all enzyme-treated samples relative to controls. In addition, collagen fibril organization was disrupted by PG depletion. Our data demonstrate that PGs play an important role in determining not only the mechanical properties at these length scales, but also collagen fibril arrangement.

scholarly journals Visualization by atomic force microscopy of tobacco mosaic virus movement protein–RNA complexes formed in vitro

2001 ◽  
Vol 82 (6) ◽  
pp. 1503-1508 ◽  
Author(s):  
O. I. Kiselyova ◽  
I. V. Yaminsky ◽  
E. M. Karger ◽  
O. Yu. Frolova ◽  
Y. L. Dorokhov ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Movement Protein ◽  
Structural Conversion ◽  
Force Microscopy ◽  
Rna Transcripts ◽  
Atomic Force ◽  
Rna Complexes

The structure of complexes formed in vitro by tobacco mosaic virus (TMV)-coded movement protein (MP) with TMV RNA and short (890 nt) synthetic RNA transcripts was visualized by atomic force microscopy on a mica surface. MP molecules were found to be distributed along the chain of RNA and the structure of MP–RNA complexes depended on the molar MP:RNA ratios at which the complexes were formed. A rise in the molar MP:TMV RNA ratio from 20:1 to 60–100:1 resulted in an increase in the density of the MP packaging on TMV RNA and structural conversion of complexes from RNase-sensitive ‘beads-on-a-string’ into a ‘thick string’ form that was partly resistant to RNase. The ‘thick string’-type RNase-resistant complexes were also produced by short synthetic RNA transcripts at different MP:RNA ratios. The ‘thick string’ complexes are suggested to represent clusters of MP molecules cooperatively bound to discrete regions of TMV RNA and separated by protein-free RNA segments.

scholarly journals Dynamic mechanisms of CRISPR interference by Escherichia coli CRISPR-Cas3

2021 ◽  
Author(s):  
Kazuto Yoshimi ◽  
Kohei TAKESHITA ◽  
Noriyuki Kodera ◽  
Satomi Shibumura ◽  
Yuko Yamauchi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Speed ◽  
Dna Helicase ◽  
Type I ◽  
Loop Formation ◽  
Force Microscopy ◽  
Atomic Force ◽  
Target Strand ◽  
Target Dna

Type I CRISPR-Cas3 uses an RNA-guided multi Cas-protein complex, Cascade, which detects and degrades foreign nucleic acids via the helicase-nuclease Cas3 protein. Despite many studies using cryoEM and smFRET, the precise mechanism of Cas3-mediated cleavage and degradation of target DNA remains elusive. Here we reconstitute the CRISPR-Cas3 system in vitro to show how the Escherichia coli Cas3 (EcoCas3) with EcoCascade exhibits collateral non-specific ssDNA cleavage and target specific DNA degradation. Partial binding of EcoCascade to target DNA with tolerated mismatches within the spacer sequence, but not the PAM, elicits collateral ssDNA cleavage activity of recruited EcoCas3. Conversely, stable binding with complete R-loop formation drives EcoCas3 to nick the non-target strand (NTS) in the bound DNA. Helicase-dependent unwinding then combines with trans ssDNA cleavage of the target strand and repetitive cis cleavage of the NTS to degrade the target dsDNA substrate. High-speed atomic force microscopy demonstrates that EcoCas3 bound to EcoCascade repeatedly reels and releases the target DNA, followed by target fragmentation. Together, these results provide a revised model for collateral ssDNA cleavage and target dsDNA degradation by CRISPR-Cas3, furthering understanding of type I CRISPR priming and interference and informing future genome editing tools.

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