scholarly journals Robocasting of Bioactive SiO2-P2O5-CaO-MgO-Na2O-K2O Glass Scaffolds

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Francesco Baino ◽  
Jacopo Barberi ◽  
Elisa Fiume ◽  
Gissur Orlygsson ◽  
Jonathan Massera ◽  
...  
Keyword(s):  
Body Fluid ◽  
Bone Repair ◽  
Glass Powder ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
In Vitro Tests ◽  
Text File ◽  
Microstructural Parameters ◽  
Scaffold Architecture

Bioactive silicate glass scaffolds were fabricated by a robocasting process in which all the movements of the printing head were programmed by compiling a script (text file). A printable ink made of glass powder and Pluronic F-127, acting as a binder, was extruded to obtain macroporous scaffolds with a grid-like three-dimensional structure. The scaffold architecture was investigated by scanning electron microscopy and microtomographic analysis, which allowed quantifying the microstructural parameters (pore size 150–180 μm and strut diameter 300 μm). In vitro tests in simulated body fluid (SBF) confirmed the apatite-forming ability (i.e., bioactivity) of the scaffolds. The compressive strength (around 10 MPa for as-produced scaffolds) progressively decreased during immersion in SBF (3.3 MPa after 4 weeks) but remains acceptable for bone repair applications. Taken together, these results (adequate porosity and mechanical strength as well as bioactivity) support the potential suitability of the prepared scaffolds for bone substitution.

scholarly journals Robocasting of SiO2-Based Bioactive Glass Scaffolds with Porosity Gradient for Bone Regeneration and Potential Load-Bearing Applications

Materials ◽  
2019 ◽  
Vol 12 (17) ◽  
pp. 2691 ◽  
Author(s):  
Barberi ◽  
Baino ◽  
Fiume ◽  
Orlygsson ◽  
Nommeots-Nomm ◽  
...  
Keyword(s):  
Glass Powder ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Great Promise ◽  
In Vitro Tests ◽  
Bioactive Scaffolds ◽  
Microstructural Parameters ◽  
Human Cancellous Bone ◽  
Bone Substitute Materials

: Additive manufacturing of bioactive glasses has recently attracted high interest in the field of regenerative medicine as a versatile class of fabrication methods to process bone substitute materials. In this study, melt-derived glass particles from the SiO2-P2O5-CaO-MgO-Na2O-K2O system were used to fabricate bioactive scaffolds with graded porosity by robocasting. A printable ink made of glass powder and Pluronic F-127 (binder) was extruded into a grid-like three-dimensional structure with bimodal porosity, i.e., the inner part of the scaffold had macropores with smaller size compared to the periphery. The crystallization behavior of the glass powder was studied by hot-stage microscopy, differential thermal analysis, and X-ray diffraction; the scaffolds were sintered at a temperature below the onset of crystallization so that amorphous structures could be obtained. Scaffold architecture was investigated by scanning electron microscopy and microtomographic analysis that allowed quantifying the microstructural parameters. In vitro tests in Kokubo’s simulated body fluid (SBF) confirmed the apatite-forming ability (i.e., bioactivity) of the scaffolds. The compressive strength was found to slightly decrease during immersion in SBF up to 4 weeks but still remained comparable to that of human cancellous bone. The pH and concentration of released ions in SBF were also measured at each time point. Taken together, these results (favorable porosity, mechanical strength, and in vitro bioactivity) show great promise for the potential application of these robocast scaffolds in bone defect repair.

scholarly journals A Nanofiber Mat With Dual Bioactive Components and a Biomimetic Matrix Structure for Improving Osteogenesis Effect

2021 ◽  
Vol 9 ◽  
Author(s):  
Yadi Han ◽  
Xiaofeng Shen ◽  
Sihao Chen ◽  
Xiuhui Wang ◽  
Juan Du ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Lactic Acid ◽  
Bone Repair ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Bioactive Components ◽  
Tissue Engineering Scaffolds ◽  
Cell Growth And Migration ◽  
Nanofiber Mats

The challenge of effectively regenerating bone tissue through tissue engineering technology is that most tissue engineering scaffolds cannot imitate the three-dimensional structure and function of the natural extracellular matrix. Herein, we have prepared the poly(L-lactic acid)–based dual bioactive component reinforced nanofiber mats which were named as poly(L-lactic acid)/bovine serum albumin/nanohydroxyapatite (PLLA/BSA/nHAp) with dual bioactive components by combining homogeneous blending and electrospinning technology. The results showed that these nanofiber mats had sufficient mechanical properties and a porous structure suitable for cell growth and migration. Furthermore, the results of cell experiments in vitro showed that PLLA/BSA/nHAp composite nanofiber mat could preferably stimulate the proliferation of mouse osteoblastic cells (MC3T3 cells) compared with pure PLLA nanofiber mats. Based on these results, the scaffolds developed in this study are considered to have a great potential to be adhibited as bone repair materials.

Evaluation and Biocompatibility of a New Type of Scaffold for Tissue Growth Based on Calcium Phosphate Cement

2008 ◽  
Vol 396-398 ◽  
pp. 667-670 ◽  
Author(s):  
Jéferson Luis de Moraes Machado ◽  
Luis Alberto Santos
Keyword(s):  
Body Fluid ◽  
Three Dimensional ◽  
Sodium Sulphate ◽  
Tissue Growth ◽  
Cellular Growth ◽  
Biological Molecules ◽  
In Vitro Tests ◽  
Porous Scaffolds ◽  
New Type

Growth of cells in three-dimensional porous scaffolds has gaining importance in the field of tissue engineering. The scaffolds guide cellular growth, synthesize extra-cellular matrix and other biological molecules, and make the formation of tissues and functional organs easier. The aim of this work was to use α-TCP (α-tricalcium phosphate) cement in order to obtain such scaffolds, with the aid of paraffin spheres as pores generators. These spheres were produced by suspension in an aqueous solution of polyvinylalcohol (PVA) and sodium sulphate (Na2SO4). The porosity of the scaffolds produced with paraffin spheres was analyzed and compared to the literature, and the study of scaffolds permeability using Forchenheimer equation allowed the analysis of pores interconnectivity. In vitro tests showed the behavior of scaffolds in solutions of SBF (Simulated Body Fluid), and viability and cell proliferation were also evaluated. Use of paraffin spheres showed to be appropriate to the formation of pores suitable size to allow cellular growth.

scholarly journals Nanofiber-Mâché Hollow Ball Mimicking the Three-Dimensional Structure of a Cyst

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2273
Author(s):  
Wan-Ying Huang ◽  
Norichika Hashimoto ◽  
Ryuhei Kitai ◽  
Shin-ichiro Suye ◽  
Satoshi Fujita
Keyword(s):  
Three Dimensional ◽  
Hollow Structure ◽  
Dimensional Structure ◽  
The Novel ◽  
Fibrous Scaffold ◽  
Hydrogel Beads ◽  
Inner Surface ◽  
Intracranial Epidermoid ◽  
Hollow Ball

The occasional malignant transformation of intracranial epidermoid cysts into squamous cell carcinomas remains poorly understood; the development of an in vitro cyst model is urgently needed. For this purpose, we designed a hollow nanofiber sphere, the “nanofiber-mâché ball.” This hollow structure was fabricated by electrospinning nanofiber onto alginate hydrogel beads followed by dissolving the beads. A ball with approximately 230 mm3 inner volume provided a fibrous geometry mimicking the topography of the extracellular matrix. Two ducts located on opposite sides provided a route to exchange nutrients and waste. This resulted in a concentration gradient that induced oriented migration, in which seeded cells adhered randomly to the inner surface, formed a highly oriented structure, and then secreted a dense web of collagen fibrils. Circumferentially aligned fibers on the internal interface between the duct and hollow ball inhibited cells from migrating out of the interior, similar to a fish bottle trap. This structure helped to form an adepithelial layer on the inner surface. The novel nanofiber-mâché technique, using a millimeter-sized hollow fibrous scaffold, is excellently suited to investigating cyst physiology.

scholarly journals Protein Folding: Search for Basic Physical Models

2003 ◽  
Vol 3 ◽  
pp. 623-635 ◽  
Author(s):  
Ivan Y. Torshin ◽  
Robert W. Harrison
Keyword(s):  
Protein Folding ◽  
Tertiary Structure ◽  
Three Dimensional ◽  
Physical Models ◽  
Dimensional Structure ◽  
Computational Techniques ◽  
Linear Sequence ◽  
Physical Forces

How a unique three-dimensional structure is rapidly formed from the linear sequence of a polypeptide is one of the important questions in contemporary science. Apart from biological context ofin vivoprotein folding (which has been studied only for a few proteins), the roles of the fundamental physical forces in thein vitrofolding remain largely unstudied. Despite a degree of success in using descriptions based on statistical and/or thermodynamic approaches, few of the current models explicitly include more basic physical forces (such as electrostatics and Van Der Waals forces). Moreover, the present-day models rarely take into account that the protein folding is, essentially, a rapid process that produces a highly specific architecture. This review considers several physical models that may provide more direct links between sequence and tertiary structure in terms of the physical forces. In particular, elaboration of such simple models is likely to produce extremely effective computational techniques with value for modern genomics.

scholarly journals Functional Analysis of the Mycobacterium tuberculosis FAD-Dependent Thymidylate Synthase, ThyX, Reveals New Amino Acid Residues Contributing to an Extended ThyX Motif

2008 ◽  
Vol 190 (6) ◽  
pp. 2056-2064 ◽  
Author(s):  
Jonathan E. Ulmer ◽  
Yap Boum ◽  
Christopher D. Thouvenel ◽  
Hannu Myllykallio ◽  
Carol Hopkins Sibley
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Thymidylate Synthase ◽  
Three Dimensional ◽  
Pyrimidine Ring ◽  
Dimensional Structure ◽  
Directed Mutagenesis ◽  
Catalytic Residue ◽  
Amino Acid Residues ◽  
Insight Into

ABSTRACT A novel FAD-dependent thymidylate synthase, ThyX, is present in a variety of eubacteria and archaea, including the mycobacteria. A short motif found in all thyX genes, RHRX7-8S, has been identified. The three-dimensional structure of the Mycobacterium tuberculosis ThyX enzyme has been solved. Building upon this information, we used directed mutagenesis to produce 67 mutants of the M. tuberculosis thyX gene. Each enzyme was assayed to determine its ability to complement the defect in thymidine biosynthesis in a ΔthyA strain of Escherichia coli. Enzymes from selected strains were then tested in vitro for their ability to catalyze the oxidation of NADPH and the release of a proton from position 5 of the pyrimidine ring of dUMP. The results defined an extended motif of amino acids essential to enzyme activity in M. tuberculosis (Y44X24 H69X25R95HRX7 S105XRYX90R199 [with the underlined histidine acting as the catalytic residue and the underlined serine as the nucleophile]) and provided insight into the ThyX reaction mechanism. ThyX is found in a variety of bacterial pathogens but is absent in humans, which depend upon an unrelated thymidylate synthase, ThyA. Therefore, ThyX is a potential target for development of antibacterial drugs.

scholarly journals In Vitro Selection and Characterization of Hepatitis C Virus Serine Protease Variants Resistant to an Active-Site Peptide Inhibitor

2003 ◽  
Vol 77 (6) ◽  
pp. 3669-3679 ◽  
Author(s):  
Caterina Trozzi ◽  
Linda Bartholomew ◽  
Alessandra Ceccacci ◽  
Gabriella Biasiol ◽  
Laura Pacini ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Serine Protease ◽  
In Vitro Selection ◽  
Three Dimensional ◽  
Host Cells ◽  
Dimensional Structure ◽  
In Vitro Systems

ABSTRACT The hepatitis C virus (HCV) serine protease is necessary for viral replication and represents a valid target for developing new therapies for HCV infection. Potent and selective inhibitors of this enzyme have been identified and shown to inhibit HCV replication in tissue culture. The optimization of these inhibitors for clinical development would greatly benefit from in vitro systems for the identification and the study of resistant variants. We report the use HCV subgenomic replicons to isolate and characterize mutants resistant to a protease inhibitor. Taking advantage of the replicons' ability to transduce resistance to neomycin, we selected replicons with decreased sensitivity to the inhibitor by culturing the host cells in the presence of the inhibitor and neomycin. The selected replicons replicated to the same extent as those in parental cells. Sequence analysis followed by transfection of replicons containing isolated mutations revealed that resistance was mediated by amino acid substitutions in the protease. These results were confirmed by in vitro experiments with mutant enzymes and by modeling the inhibitor in the three-dimensional structure of the protease.

Synthesis of sol-gel derived glass powder and in vitro bioactivity property tested in simulated body fluid

2016 ◽  
Author(s):  
S. A. Syed Nuzul Fadzli ◽  
S. Roslinda ◽  
Firuz Zainuddin ◽  
Hamisah Ismail
Keyword(s):  
Simulated Body Fluid ◽  
Body Fluid ◽  
Glass Powder ◽  
Sol Gel ◽  
In Vitro Bioactivity

Rational humanization of the powerful antithrombotic anti-GPIbα antibody: 6B4

2006 ◽  
Vol 96 (11) ◽  
pp. 671-684 ◽  
Author(s):  
Alexandre Fontayne ◽  
Karen Vanhoorelbeke ◽  
Inge Pareyn ◽  
Isabel Van Rompaey ◽  
Muriel Meiring ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Three Dimensional ◽  
Vector System ◽  
Dimensional Structure ◽  
Fab Fragment ◽  
Variable Regions ◽  
Significant Prolongation ◽  
Von Willebrand ◽  
First Time

SummaryFab-fragments of the monoclonal antibody 6B4, raised against human glycoprotein Ibα (GPIbα), have a powerful antithrombotic effect in baboons by blocking the GPIbα binding site for von Willebrand factor (VWF), without significant prolongation of the skin bleeding time. In order to bring this antibody to the clinic,we here humanized for the first time an anti-human GPIbα by variable-domain resurfacing guided by computer modeling. First, the genes coding for the variable regions of the heavy and light chains of 6B4 were cloned and sequenced. Based on this,a three-dimensional structure of the Fv-fragment was constructed by using homology-based modeling, and with this and comparison with antibodies with known structure,”murine” putative immunogenic residues which are exposed, were changed for “human-like” residues. The humanized Fab-fragment, h6B4-Fab, was constructed in the pKaneo vector system, expressed and purified and showed in vitro an unaltered, even slightly higher binding affinity for its antigen than the murine form as determined by different ELISA set-ups and surface plasmon resonance. Finally, injection of doses of 0.1 to 1.5 mg/kg of h6B4-Fab in baboons showed that both pharmacokinetics and ex-vivo bio-activity of the molecule were to a large extent preserved.In conclusion, the method used here to humanize 6B4 by resurfacing resulted in a fully active derivative, which is now ready for further development.

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