Patterned Cellulose Nanocrystal Aerogel Films with Tunable Dimensions and Morphologies as Ultra-Porous Scaffolds for Cell Culture

2019 ◽  
Vol 2 (7) ◽  
pp. 4169-4179 ◽  
Author(s):  
Tyler Or ◽  
Sokunthearath Saem ◽  
Aurore Esteve ◽  
Daniel A. Osorio ◽  
Kevin J. De France ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cellulose Nanocrystal ◽  
Porous Scaffolds ◽  
Aerogel Films

Osteoblast Behavior In Vitro in Porous Calcium Phosphate Composite Scaffolds, Surface Activated with a Cell Adhesive Plasma Polymer Layer

2012 ◽  
Vol 706-709 ◽  
pp. 566-571 ◽  
Author(s):  
Barbara Nebe ◽  
Matthias Cornelsen ◽  
Antje Quade ◽  
Volker Weissmann ◽  
Friederike Kunz ◽  
...  
Keyword(s):  
Cell Culture ◽  
Calcium Phosphate ◽  
Trauma Surgery ◽  
Plasma Polymer ◽  
Porous Scaffolds ◽  
Temperature Plasma ◽  
Composite Surface ◽  
Initial Adhesion ◽  
Four Levels

Synthetic materials such as bone substitutes are permanently under development for applications in orthopedic and trauma surgery. Our porous scaffolds were produced from ß-tricalcium phosphate (TCP) using the three dimensional (3D)-printing technology. After sintering the porosity and the pore size of the 3D printed scaffolds reached nearly 50 % and 500 µm, respectively. TCP scaffolds were additionally stabilized by infiltration with polylactic acid (PLA). Because PLA usually impeded cell adhesion we activated the composite surface with plasma polymerized allylamine in a low temperature plasma process. For cell investigations inside the scaffold we used a module system, where two porous discs can be horizontally fixed within a clamping ring. Thereby a 3D cell culture module with four levels and a maximal height of 10 mm was generated. Human MG-63 osteoblasts (ATCC) were seeded apically and placed in serum-containing DMEM. After 14 days of a static cell culture the cell ingrowth and mobility was analyzed by scanning electron microscopy. Osteoblast's initial adhesion and short time occupation of the surface is significantly improved on plasma polymer activated TCP surfaces, which could be a precondition for an enhanced colonization inside a calcium phosphate scaffold. Interestingly, the plasma functionalization of the pure TCP scaffold was possible and successful concerning cell acceptance.

scholarly journals Porous 3D Scaffolds Enhance MSC Vitality and Reduce Osteoclast Activity

Molecules ◽  
2021 ◽  
Vol 26 (20) ◽  
pp. 6258
Author(s):  
Miriam Spreda ◽  
Nicole Hauptmann ◽  
Veronika Lehner ◽  
Christoph Biehl ◽  
Klaus Liefeith ◽  
...  
Keyword(s):  
Cell Culture ◽  
Metabolic Activity ◽  
Stromal Cells ◽  
3D Structure ◽  
Porous Scaffolds ◽  
Cytotoxic Effects ◽  
The Public ◽  
Osteoclast Activity ◽  
Biomimetic Scaffolds ◽  
Promising Solution

In the context of an aging population, unhealthy Western lifestyle, and the lack of an optimal surgical treatment, deep osteochondral defects pose a great challenge for the public health system. Biodegradable, biomimetic scaffolds seem to be a promising solution. In this study we investigated the biocompatibility of porous poly-((D,L)-lactide-ε-caprolactone)dimethacrylate (LCM) scaffolds in contrast to compact LCM scaffolds and blank cell culture plastic. Thus, morphology, cytotoxicity and metabolic activity of human mesenchymal stromal cells (MSC) seeded directly on the materials were analyzed after three and six days of culturing. Further, osteoclastogenesis and osteoclastic activity were assessed using reverse-transcriptase real-time PCR of osteoclast-specific genes, EIA and morphologic aspects after four, eight, and twelve days. LCM scaffolds did not display cytotoxic effects on MSC. After three days, metabolic activity of MSC was enhanced on 3D porous scaffolds (PS) compared to 2D compact scaffolds (CS). Osteoclast activity seemed to be reduced at PS compared to cell culture plastic at all time points, while no differences in osteoclastogenesis were detectable between the materials. These results indicate a good cytocompatibility of LCM scaffolds. Interestingly, porous 3D structure induced higher metabolic activity of MSC as well as reduced osteoclast activity.

Metabolite Transport Inside Channeled Porous Scaffolds for Haematopoietic Stem Cell Culture: A Computational Study

2008 ◽  
Author(s):  
Marco Cantini ◽  
Gianfranco B. Fiore ◽  
Alberto Redaelli ◽  
Monica Soncini
Keyword(s):  
Fluid Dynamics ◽  
Cell Culture ◽  
Computational Study ◽  
Three Dimensional ◽  
Polymeric Materials ◽  
Haematopoietic Stem Cell ◽  
Porous Scaffolds ◽  
A Cell ◽  
Definition Of

Porous polymeric materials play a key role in regenerative medicine, serving as three-dimensional scaffolds for cell culture. Hence, the definition of their micro-architecture should be regarded as a pivotal design issue, that has to be wittingly addressed while engineering a cell culture system. Computational fluid dynamics techniques (CFD) appear to be very valuable in this respect, since they have been appreciably applied in recent literature as a means to analyze fluid dynamics and mass transport inside scaffold or bioreactor models [1]; moreover, leading researchers in tissue engineering have acknowledged the role of numerical methodology in the issue of defining optimal flow conditions for three-dimensional dynamic culture systems.

scholarly journals Biocomposite scaffold preparation from hydroxyapatite extracted from waste bovine bone

2019 ◽  
Vol 9 (1) ◽  
pp. 37-47
Author(s):  
Quoc-Phong Ho ◽  
The-Duong Tao ◽  
Lien-Huong Huynh ◽  
Meng-Jiy Wang
Keyword(s):  
Cell Culture ◽  
Tensile Strength ◽  
Phase Separation ◽  
Pore Size ◽  
Surface Area ◽  
Cell Density ◽  
Crystal Size ◽  
Porous Scaffolds ◽  
Bovine Bone ◽  
Naoh Solution

AbstractThis study was conducted to fabricate scaffold from polylactic acid (PLA) and hydroxyapatite (HA) extracted from waste bovine bone for enhancing both mechanical and biocompatible properties. After pretreatment in dilute NaOH solution, the bone was calcined at 900°C for 6 h, ball milled and converted to HA. Factors that affect the formation of HA were investigated. Experimental results showed that HA particles with crystal size < 100 nm and 99% crystallinity could be obtained at 90°C, pH 11 and 35 mM H3PO4 solution followed by 4 h calcination at 900°C. By using non-solvent induced phase separation method, PLA scaffolds with pore size and surface area of 22.6 μm and 25.7 m2/g, respectively, containing different hydroxyapatite were successfully prepared. Tensile strength of scaffolds increased due to effective support by HA grafted collagen. PLA scaffolds containing HA were more degradable than PLA scaffolds and PLA scaffolds containing HA grafted collagen. Cell culture results showed that cell density increased significantly on porous scaffolds than that on the dense scaffolds. Moreover, cell density also increased significantly on the scaffold containing HA grafted collagen than that on the scaffold with pure HA.

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.

A T.E.M. study of mouse mammary epithelial cell secretory differentiation cultured on collagen and Matrigel

1991 ◽  
Vol 49 ◽  
pp. 188-189
Author(s):  
W.N. Bentham ◽  
V. Rocha
Keyword(s):  
Cell Culture ◽  
Mammary Epithelial ◽  
Secretory Process ◽  
Cell Culture System ◽  
Protein Maturation ◽  
Cell Culture Techniques ◽  
Culture Techniques ◽  
Mouse Mammary Epithelial Cell ◽  
Secretory Differentiation

It has been an interest of our lab to develop a mammary epethelial cell culture system that faithfully duplicates the in vivo condition of the lactating gland. Since the introduction of collagen as a matrix on which cells are cultivated other E.C.M. type matrices have been made available and are used in many cell culture techniques. We have previously demonstrated that cells cultured on collagen and Matrigel do not differentiate as they do in vivo. It seems that these cultures often produce cells that show a disruption in the secretory process. The appearance of large ribosomal studded vesicles, that specifically label with antibody to casein, suggest an interruption of both protein maturation and secretion at the E.R. to golgi transition. In this report we have examined cultures on collagen and Matrigel at relative high and low seeding densities and compared them to cells from the in vivo condition.

Solid-phase immunoelectron microscopy for rapid diagnosis of enteroviruses

1984 ◽  
Vol 42 ◽  
pp. 226-227 ◽  
Author(s):  
K. Pegg-Feige ◽  
F. W. Doane
Keyword(s):  
Electron Microscopy ◽  
Cell Culture ◽  
Immunoelectron Microscopy ◽  
Detection Method ◽  
Solid Phase ◽  
Protein A ◽  
Plant Viruses ◽  
Rapid Diagnosis ◽  
Virus Diagnosis ◽  
Antiviral Antibody

Immunoelectron microscopy (IEM) applied to rapid virus diagnosis offers a more sensitive detection method than direct electron microscopy (DEM), and can also be used to serotype viruses. One of several IEM techniques is that introduced by Derrick in 1972, in which antiviral antibody is attached to the support film of an EM specimen grid. Originally developed for plant viruses, it has recently been applied to several animal viruses, especially rotaviruses. We have investigated the use of this solid phase IEM technique (SPIEM) in detecting and identifying enteroviruses (in the form of crude cell culture isolates), and have compared it with a modified “SPIEM-SPA” method in which grids are coated with protein A from Staphylococcus aureus prior to exposure to antiserum.

Automated cell counting of astrocytes on patterned substrates containing aliphatic and charged properties

1995 ◽  
Vol 53 ◽  
pp. 974-975
Author(s):  
W. Shain ◽  
H. Ancin ◽  
H.C. Craighead ◽  
M. Isaacson ◽  
L. Kam ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Culture Method ◽  
Cell Counting ◽  
Data Sets ◽  
Nuclear Staining ◽  
Double Positive ◽  
A Cell ◽  
Wafer Test ◽  
Cell Densities

Neural protheses have potential to restore nervous system functions lost by trauma or disease. Nanofabrication extends this approach to implants for stimulating and recording from single or small groups of neurons in the spinal cord and brain; however, tissue compatibility is a major limitation to their practical application. We are using a cell culture method for quantitatively measuring cell attachment to surfaces designed for nanofabricated neural prostheses.Silicon wafer test surfaces composed of 50-μm bars separated by aliphatic regions were fabricated using methods similar to a procedure described by Kleinfeld et al. Test surfaces contained either a single or double positive charge/residue. Cyanine dyes (diIC18(3)) stained the background and cell membranes (Fig 1); however, identification of individual cells at higher densities was difficult (Fig 2). Nuclear staining with acriflavine allowed discrimination of individual cells and permitted automated counting of nuclei using 3-D data sets from the confocal microscope (Fig 3). For cell attachment assays, LRM5 5 astroglial cells and astrocytes in primary cell culture were plated at increasing cell densities on test substrates, incubated for 24 hr, fixed, stained, mounted on coverslips, and imaged with a 10x objective.

Environmental microscopy of capillary stress-induced strain behavior in ambient-pressure aerogels

1996 ◽  
Vol 54 ◽  
pp. 850-851
Author(s):  
Sudeep M. Rao ◽  
Joshua Samuel ◽  
Sai S. Prakash ◽  
C. Jeffrey Brinker
Keyword(s):  
Silica Aerogel ◽  
Ambient Pressure ◽  
Drying Shrinkage ◽  
Pore Filling ◽  
Silica Network ◽  
Strain Behavior ◽  
Partially Saturated ◽  
Capillary Stress ◽  
Wetting Liquid ◽  
Aerogel Films

Ambient pressure silica aerogel thin films have recently been prepared by exploiting reversible drying shrinkage caused by derivatization of the internal gel surface. Aerogels have porosities of upto 99.9% and due to the small size of the pores (few nanometers), large capillary stresses are produced in gels that are partially saturated with a wetting liquid. As a result of these capillary stresses, the flexible silica network undergoes strain which has been observed using environmental microscopy. This technique allows variation of the equilibrium vapor pressure and temperature, and a simultaneous monitoring of the deformation of the unconstrained film thickness. We have observed >600% deformation during the pore-filling and pore-emptying cycles. In this presentation, we discuss the unique stress-strain behavior of these films.Ref.: Sai S. Prakash, C. Jeffrey Brinker, Alan J. Hurd & Sudeep M. Rao, "Silica aerogel films prepared at ambient pressure by using surface derivatization to induce reversible drying shrinkage", Nature. Vol. 374, 30 March, 1995, 439-443.

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