scholarly journals Silk Fibroin Scaffolds as Biomaterials for 3D Mesenchymal Stromal Cells Cultures

2021 ◽  
Vol 11 (23) ◽  
pp. 11345
Author(s):  
Luisa Milazzo ◽  
Francesca Vulcano ◽  
Giampiero Macioce ◽  
Giovanna Marziali ◽  
Francesca Iosi ◽  
...  
Keyword(s):  
Silk Fibroin ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Volume Ratio ◽  
Controlled Environment ◽  
Porous Scaffolds ◽  
Micro Computed Tomography ◽  
Stromal Stem Cells ◽  
Natural Characteristics

Silk fibroin (SF), a protein-based fiber extracted from Bombyx mori cocoons, has recently emerged with great potential for the biomedical field to be used as a biomaterial processable in a variety of formats and applications, due to its natural characteristics. The aims of the present study were to characterize the structural properties of the SF scaffolds, in the format of porous sponges, and to investigate their feasibility to support the adhesion of mesenchymal stromal/stem cells isolated from human Wharton’s jelly of the umbilical cord (WJ-MSC). Adhesion is a prerequisite for using the SF scaffold as biomaterial for supporting three-dimensional (3D) WJ-MSC cultures for several applications. The integration among micro-computed tomography, confocal analysis, and field emission scanning electron microscopy allowed carrying out a deep investigation based on quantitative morphological parameters and qualitative observations at high resolution. High levels of porosity, interconnection, and contact surface–volume ratio confirmed the appropriateness of the designed SF porous scaffolds as supports for cell cultures. WJ-MSC was demonstrated to be capable of adhering to and colonizing the SF scaffold applicable as a 3D cell culture system, of conducting in vitro experiments in a more controlled environment, and possibly of being used in tissue engineering, regenerative medicine, and applications in oncology.

Flow Analysis of a Porous Polymer-Based Three-Dimensional Cell Culture Device for Drug Screening

2018 ◽  
Author(s):  
Liang Ma ◽  
Lei Gao ◽  
Yichen Luo ◽  
Huayong Yang ◽  
Bin Zhang ◽  
...  
Keyword(s):  
Cell Culture ◽  
Shear Stress ◽  
Three Dimensional ◽  
Flow Simulation ◽  
3D Cell Culture ◽  
Porous Polymer ◽  
Poly Lactic Acid ◽  
Porous Scaffolds ◽  
3D Scaffold

A porous polymer-based three-dimensional (3D) cell culture device has been developed as an in vitro tissue model system for the cytotoxicity of anticancer drug test. The device had two chambers connected in tandem, each loaded with a 3D scaffold made of highly biocompatible poly (lactic acid) (PLA). Hepatoma cells (HepG2) and glioblastoma multiforme (GBM) cancer cells were cultured in the two separate porous scaffolds. A peristaltic pump was adopted to realize a perfusion cell culture. In this study, we focus on cell viability inside the 3D porous scaffolds under flow-induced shear stress effects. A flow simulation was conducted to predict the shear stress based on a realistic representation of the porous structure. The simulation results were correlated to the cell variability measurements at different flow rates. It is shown that the modeling approach presented in this paper can be useful for shear stress predication inside porous scaffolds and the computational fluid dynamics model can be an effective way to optimize the operation parameters of perfused 3D cell culture devices.

Bi-layered electrospun nanofibrous polyurethane-gelatin scaffold with targeted heparin release profiles for tissue engineering applications

2017 ◽  
Vol 37 (9) ◽  
pp. 933-941 ◽  
Author(s):  
Mohammad Mahdi Safikhani ◽  
Ali Zamanian ◽  
Farnaz Ghorbani ◽  
Azadeh Asefnejad ◽  
Mostafa Shahrezaee
Keyword(s):  
Tissue Engineering ◽  
High Surface ◽  
High Surface Area ◽  
Three Dimensional ◽  
Coagulation Factor ◽  
Volume Ratio ◽  
Fibroblast Cell ◽  
Porous Scaffolds ◽  
Nanofibrous Scaffolds

Abstract Tissue engineering is a biotechnology that is used to develop biological substitutes to restore, maintain, or improve functions. Thus, the porous scaffolds are used to accommodate cells in tissue engineering. In this research, three dimensional (3D) bi-layered polyurethane (PU)-gelatin nanofibrous scaffolds were prepared by the electrospinning method, after which the capability of the released heparin as an anti-coagulation factor was evaluated. Electrospinning has been extensively investigated for the preparation of fibers that exhibit a high surface area to volume ratio. Results showed that scanning electron microscopy (SEM) micrographs exhibited a smooth surface as well as a highly porous and bead-free structure, in which fibers were distributed in the range of 100–600 nm. The modulus and ultimate tensile strength (UTS) decreased and increased, respectively, after crosslinking the reaction of polymers. This process also reduced swelling ratio, the hydrolytic biodegradation rate, and the release rate as a function of time. Moreover, an in vitro assay demonstrated that 3D nanofibrous scaffolds supported L929 fibroblast cell viability and that cells adhered and spread on the fibers. Based on the obtained results, the heparin-loaded electrospinning nanofibrous scaffolds have initial physicochemical and mechanical properties to protect neo-tissue formation.

scholarly journals Mammary gland 3D cell culture systems in farm animals

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Laurence Finot ◽  
Eric Chanat ◽  
Frederic Dessauge
Keyword(s):  
Cell Culture ◽  
Mammary Gland ◽  
Bacterial Infections ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Farm Animals ◽  
Culture Systems ◽  
Cell Culture Systems

AbstractIn vivo study of tissue or organ biology in mammals is very complex and progress is slowed by poor accessibility of samples and ethical concerns. Fortunately, however, advances in stem cell identification and culture have made it possible to derive in vitro 3D “tissues” called organoids, these three-dimensional structures partly or fully mimicking the in vivo functioning of organs. The mammary gland produces milk, the source of nutrition for newborn mammals. Milk is synthesized and secreted by the differentiated polarized mammary epithelial cells of the gland. Reconstructing in vitro a mammary-like structure mimicking the functional tissue represents a major challenge in mammary gland biology, especially for farm animals for which specific agronomic questions arise. This would greatly facilitate the study of mammary gland development, milk secretion processes and pathological effects of viral or bacterial infections at the cellular level, all with the objective of improving milk production at the animal level. With this aim, various 3D cell culture models have been developed such as mammospheres and, more recently, efforts to develop organoids in vitro have been considerable. Researchers are now starting to draw inspiration from other fields, such as bioengineering, to generate organoids that would be more physiologically relevant. In this chapter, we will discuss 3D cell culture systems as organoids and their relevance for agronomic research.

A rhabdomyosarcoma hydrogel model to unveil cell-extracellular matrix interactions

2021 ◽  
Author(s):  
Mattia Saggioro ◽  
Stefania D'Agostino ◽  
Anna Gallo ◽  
Sara Crotti ◽  
Sara D'Aronco ◽  
...  
Keyword(s):  
Cell Culture ◽  
Extracellular Matrix ◽  
Three Dimensional ◽  
3D Culture ◽  
3D Cell Culture ◽  
Culture Systems ◽  
Matrix Interactions ◽  
In Vitro Systems ◽  
Extracellular Matrix Interactions

Three-dimensional (3D) culture systems are progressively getting attention given their potential in overcoming limitations of the classical 2D in vitro systems. Among different supports for 3D cell culture, hydrogels (HGs)...

scholarly journals Porous Geometry Guided Micro-mechanical Environment Within Scaffolds for Cell Mechanobiology Study in Bone Tissue Engineering

2021 ◽  
Vol 9 ◽  
Author(s):  
Feihu Zhao ◽  
Yi Xiong ◽  
Keita Ito ◽  
Bert van Rietbergen ◽  
Sandra Hofmann
Keyword(s):  
Porous Scaffold ◽  
Mechanical Strain ◽  
Three Dimensional ◽  
Pore Geometry ◽  
Cell Physiology ◽  
Porous Scaffolds ◽  
Mechanical Environment ◽  
Functional Features ◽  
Future Work

Mechanobiology research is for understanding the role of mechanics in cell physiology and pathology. It will have implications for studying bone physiology and pathology and to guide the strategy for regenerating both the structural and functional features of bone. Mechanobiological studies in vitro apply a dynamic micro-mechanical environment to cells via bioreactors. Porous scaffolds are commonly used for housing the cells in a three-dimensional (3D) culturing environment. Such scaffolds usually have different pore geometries (e.g. with different pore shapes, pore dimensions and porosities). These pore geometries can affect the internal micro-mechanical environment that the cells experience when loaded in the bioreactor. Therefore, to adjust the applied micro-mechanical environment on cells, researchers can tune either the applied load and/or the design of the scaffold pore geometries. This review will provide information on how the micro-mechanical environment (e.g. fluid-induced wall shear stress and mechanical strain) is affected by various scaffold pore geometries within different bioreactors. It shall allow researchers to estimate/quantify the micro-mechanical environment according to the already known pore geometry information, or to find a suitable pore geometry according to the desirable micro-mechanical environment to be applied. Finally, as future work, artificial intelligent – assisted techniques, which can achieve an automatic design of solid porous scaffold geometry for tuning/optimising the micro-mechanical environment are suggested.

scholarly journals COMPARATIVE ANALYSIS OF THREE-DIMENSIONAL NANOSTRUCTURE OF POROUS BIOCOMPATIBLE SCAFFOLDS MADE OF RECOMBINANT SPIDROIN AND SILK FIBROIN FOR REGENERATIVE MEDICINE

2015 ◽  
Vol 17 (2) ◽  
pp. 37-44 ◽  
Author(s):  
O. I. Agapova ◽  
A. E. Efimov ◽  
M. M. Moisenovich ◽  
V. G. Bogush ◽  
I. I. Agapov
Keyword(s):  
Volume Fraction ◽  
Three Dimensional ◽  
Scanning Probe ◽  
Porous Scaffolds ◽  
Volume Porosity ◽  
Integral Density ◽  
Regenerative Activity ◽  
Calculated Volume ◽  
Recombinant Spidroin

Aim.To perform a comparison of three-dimensional nanostructure of porous biocompatible scaffolds made of fibroinBombix moriand recombinant spidroin rS1/9.Materials and methods.Three-dimensional porous scaffolds were produced by salt leaching technique. The comparison of biological characteristics of the scaffolds shows that adhesion and proliferation of mouse fibroblastsin vitroon these two types of scaffolds do not differ significantly. Comparative experimentsin vivoshow that regeneration of bone tissue of rats is faster with implantation of recombinant spidroin scaffolds. Three-dimensional nanostructure of scaffolds and interconnectivity of nanopores were studied with scanning probe nanotomography (SPNT) to explain higher regenerative activity of spidroin-based scaffolds.Results.Significant differences were detected in the integral density and volume of pores: the integral density of nanopores detected on 2D AFM images is 46 μm–2    and calculated volume porosity is 24% in rS1/9-based scaffolds; in fibroin-based three-dimensional structures density of nanopores and calculated volume porosity were 2.4 μm–2  and 0.5%, respectively. Three-dimensional reconstruction system of nanopores and clusters of interconnected nanopores in rS1/9-based scaffolds showed that volume fraction of pores interconnected in percolation clusters is 35.3% of the total pore volume or 8.4% of the total scaffold volume.Conclusion.Scanning probe nanotomography method allows obtaining unique information about topology of micro – and nanopore systems of artificial biostructures. High regenerative activity of rS1/9-based scaffolds can be explained by higher nanoporosity of the scaffolds.

scholarly journals iTRAQ-Based Quantitative Proteomic Comparison of 2D and 3D Adipocyte Cell Models Co-cultured with Macrophages Using Online 2D-nanoLC-ESI-MS/MS

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Sun Young Lee ◽  
Sung Bum Park ◽  
Young Eun Kim ◽  
Hee Min Yoo ◽  
Jongki Hong ◽  
...  
Keyword(s):  
Metabolic Disorders ◽  
Cultured Cells ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Proteomic Profiling ◽  
Esi Ms ◽  
Mitochondrial Fatty Acid ◽  
2D And 3D

AbstractThe demand for novel three-dimensional (3D) cell culture models of adipose tissue has been increasing, and proteomic investigations are important for determining the underlying causes of obesity, type II diabetes, and metabolic disorders. In this study, we performed global quantitative proteomic profiling of three 3D-cultured 3T3-L1 cells (preadipocytes, adipocytes and co-cultured adipocytes with macrophages) and their 2D-cultured counterparts using 2D-nanoLC-ESI-MS/MS with iTRAQ labelling. A total of 2,885 shared proteins from six types of adipose cells were identified and quantified in four replicates. Among them, 48 proteins involved in carbohydrate metabolism (e.g., PDHα, MDH1/2, FH) and the mitochondrial fatty acid beta oxidation pathway (e.g., VLCAD, ACADM, ECHDC1, ALDH6A1) were relatively up-regulated in the 3D co-culture model compared to those in 2D and 3D mono-cultured cells. Conversely, 12 proteins implicated in cellular component organisation (e.g., ANXA1, ANXA2) and the cell cycle (e.g., MCM family proteins) were down-regulated. These quantitative assessments showed that the 3D co-culture system of adipocytes and macrophages led to the development of insulin resistance, thereby providing a promising in vitro obesity model that is more equivalent to the in vivo conditions with respect to the mechanisms underpinning metabolic syndromes and the effect of new medical treatments for metabolic disorders.

Modification of Thai Silk Fibroin Scaffolds by Gelatin Conjugation for Tissue Engineering

2008 ◽  
Vol 55-57 ◽  
pp. 685-688 ◽  
Author(s):  
J. Chamchongkaset ◽  
Sorada Kanokpanont ◽  
David L. Kaplan ◽  
Siriporn Damrongsakkul
Keyword(s):  
Tissue Engineering ◽  
Bombyx Mori ◽  
Silk Fibroin ◽  
Textile Industry ◽  
Three Dimensional ◽  
Biological Properties ◽  
Salt Leaching ◽  
Biological Compatibility ◽  
Salt Crystals

Silk has been used commercially as biomedical sutures for decades. Recently silk fibroin, especially from Bombyx mori silkworm, has been explored for many tissue engineering applications such as bone and cartilage due to its impressive biological compatibility and mechanical properties. In Thailand, Thai native silkworms have been long cultivated. Distinct characteristics of cocoon Thai silk are its yellow color and coarse filament. There is more sericin in Thai silk than in other Bombyx mori silks. These characteristics provide Thai silk a unique texture for textile industry. It is therefore the aim of this study to develop three-dimensional silk fibroin-based scaffolds from Thai yellow cocoon “Nangnoi-Srisaket” of Bombyx mori silkworms using salt-leaching method. To enhance the biological properties, type A gelatin, the denature form of collagen having good biocompactibility, was used to conjugate with silk fibroin scaffolds. The pore size of salt-leached silk fibroin scaffold structure represented the size of salt crystals used (600-710µm). After gelatin conjugation, gelatin was partly formed fibers inside the pores of silk fibroin scaffolds resulting in fiber-like structure with highly interconnection. Gelatin conjugation enhanced the compressive modulus of silk fibroin scaffolds by 93%. The results on in vitro culture using mouse osteoblast-like cells (MC3T3-E1) showed that gelatin conjugation could promote the cell proliferation in silk fibroin scaffolds. Moreover, the observed morphology of cells proliferated inside the scaffold after 14 days of culture showed the larger spreading area of cells on conjugated gelatin/silk fibroin scaffolds, compared to round-shaped cells on silk fibroin scaffolds. The results implied that Thai silk fibroin looked promising to be applied in tissue engineering and gelatin conjugation on Thai silk fibroin scaffolds could enhance the biological properties of scaffolds.

scholarly journals The Secretome Analysis of Activated Human Renal Fibroblasts Revealed Beneficial Effect of the Modulation of the Secreted Peptidyl-Prolyl Cis-Trans Isomerase A in Kidney Fibrosis

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1724
Author(s):  
Gry H. Dihazi ◽  
Marwa Eltoweissy ◽  
Olaf Jahn ◽  
Björn Tampe ◽  
Michael Zeisberg ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Renal Fibrosis ◽  
Cell Activation ◽  
Cell Transformation ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Kidney Fibrosis ◽  
Matrix Accumulation ◽  
The Impact

The secretome is an important mediator in the permanent process of reciprocity between cells and their environment. Components of secretome are involved in a large number of physiological mechanisms including differentiation, migration, and extracellular matrix modulation. Alteration in secretome composition may therefore trigger cell transformation, inflammation, and diseases. In the kidney, aberrant protein secretion plays a central role in cell activation and transition and in promoting renal fibrosis onset and progression. Using comparative proteomic analyses, we investigated in the present study the impact of cell transition on renal fibroblast cells secretome. Human renal cell lines were stimulated with profibrotic hormones and cytokines, and alterations in secretome were investigated using proteomic approaches. We identified protein signatures specific for the fibrotic phenotype and investigated the impact of modeling secretome proteins on extra cellular matrix accumulation. The secretion of peptidyl-prolyl cis-trans isomerase A (PPIA) was demonstrated to be associated with fibrosis phenotype. We showed that the in-vitro inhibition of PPIA with ciclosporin A (CsA) resulted in downregulation of PPIA and fibronectin (FN1) expression and significantly reduced their secretion. Knockdown studies of PPIA in a three-dimensional (3D) cell culture model significantly impaired the secretion and accumulation of the extracellular matrix (ECM), suggesting a positive therapeutic effect on renal fibrosis progression.

scholarly journals Three-Dimensional Cell Cultures as an In Vitro Tool for Prostate Cancer Modeling and Drug Discovery

2020 ◽  
Vol 21 (18) ◽  
pp. 6806 ◽  
Author(s):  
Fabrizio Fontana ◽  
Michela Raimondi ◽  
Monica Marzagalli ◽  
Michele Sommariva ◽  
Nicoletta Gagliano ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Culture ◽  
Drug Discovery ◽  
Cancer Cells ◽  
Cell Cultures ◽  
Antitumor Agents ◽  
Three Dimensional ◽  
3D Cell Culture ◽  
Cancer Modeling

In the last decade, three-dimensional (3D) cell culture technology has gained a lot of interest due to its ability to better recapitulate the in vivo organization and microenvironment of in vitro cultured cancer cells. In particular, 3D tumor models have demonstrated several different characteristics compared with traditional two-dimensional (2D) cultures and have provided an interesting link between the latter and animal experiments. Indeed, 3D cell cultures represent a useful platform for the identification of the biological features of cancer cells as well as for the screening of novel antitumor agents. The present review is aimed at summarizing the most common 3D cell culture methods and applications, with a focus on prostate cancer modeling and drug discovery.

Sign in / Sign up

Export Citation Format

Share Document