Osteoblast Adhesion on Tissue Engineering Scaffolds Made by Bio-Manufacturing Techniques

Author(s):  
T. Dutta Roy ◽  
J. J. Stone ◽  
W. Sun ◽  
E. H. Cho ◽  
S. J. Lockett ◽  
...  

Scientific exploration into understanding and developing relationships between three-dimensional (3D) scaffolds prepared by rapid prototyping (RP) and cellular response has focused primarily on end results targeting osteoblast proliferation and differentiation. Here at the National Institute of Standards and Technology (NIST), we take a systems approach to developing relationships between material properties and quantitative biological responses. This study in particular focuses on the screening of parameters controlled by RP techniques and their ability to trigger signalling events leading to cell adhesion. This pioneering research in our group also characterizes the in vitro cell-material interactions of 2D films and 3D scaffolds. From there, one can postulate on contributory factors leading to cell migration, proliferation, and differentiation. In summary, we believe that the quantitative information from this fundamental investigation will enhance our knowledge of the interactions between cells and 3D material interfaces with respect to formation of focal adhesions. This work consists of two sections — the application of imaging techniques for 3D characterization of properties and culturing of osteoblasts for size and shape determination. This includes quantifying the number of focal adhesion sites. We are using 3D RP polycaprolactone (PCL) scaffolds as this surrogate model in which to compare 2D to 3D material performance and cell interactions. Using RP bio-manufacturing techniques to fabricate tissue engineering scaffolds allows for control of pore size, strut size, and layer thickness, therefore providing adjustable parameters to study which can potentially influence, or even dynamically modulate, cellular adhesion. Imaging results after culturing for 24 h showed differences in cell morphology and spreading relative to the different structures. The focal adhesion response also varied, indicating an apparent loss of organization in 3D scaffolds compared to 2D surfaces. See Results and Discussion for details.

Applied Nano ◽  
2021 ◽  
Vol 2 (2) ◽  
pp. 118-127
Author(s):  
Luca Zoia ◽  
Anna Binda ◽  
Laura Cipolla ◽  
Ilaria Rivolta ◽  
Barbara La Ferla

Binary nano-biocomposite 3D scaffolds of cellulose nanocrystals (CNCs)—gelatine were fabricated without using chemical crosslinking additives. Controlled oxidative treatment allowed introducing carboxyl or carbonyl functionalities on the surface of CNCs responsible for the crosslinking of gelatine polymers. The obtained composites were characterized for their physical-chemical properties. Their biocompatibility towards different cell cultures was evaluated through MTT and LDH assays, cellular adhesion and proliferation experiments. Gelatine composites reinforced with carbonyl-modified CNCs showed the most performing swelling/degradation profile and the most promising adhesion and proliferation properties towards cell lines, suggesting their potential application in the field of tissue engineering.


1993 ◽  
Vol 13 (2) ◽  
pp. 785-791
Author(s):  
M D Schaller ◽  
C A Borgman ◽  
J T Parsons

Integrins play a central role in cellular adhesion and anchorage of the cytoskeleton and participate in the generation of intracellular signals, including tyrosine phosphorylation. We have recently isolated a cDNA encoding a unique, focal adhesion-associated protein tyrosine kinase (FAK) that is a component of an integrin-mediated signal transduction pathway. Here we report the isolation of cDNAs encoding the C-terminal, noncatalytic domain of the FAK kinase, termed FRNK (FAK-related nonkinase). Both the FAK- and FRNK-encoded polypeptides, pp125FAK and p41/p43FRNK, are expressed in normal chicken embryo cells. pp125FAK and p41/p43FRNK were localized to focal adhesions, suggesting that pp125FAK is directed to the focal adhesions by sequences within its C-terminal domain. We also show that the fibronectin-dependent increase in tyrosine phosphorylation of pp125FAK is accompanied by a concomitant posttranslational modification of p41FRNK.


Pharmaceutics ◽  
2018 ◽  
Vol 10 (4) ◽  
pp. 272 ◽  
Author(s):  
Patricia Diaz-Rodriguez ◽  
Mirian Sánchez ◽  
Mariana Landin

The mimesis of biological systems has been demonstrated to be an adequate approach to obtain tissue engineering scaffolds able to promote cell attachment, proliferation, and differentiation abilities similar to those of autologous tissues. Bioceramics are commonly used for this purpose due to their similarities to the mineral component of hard tissues as bone. Furthermore, biomimetic scaffolds are frequently loaded with diverse therapeutic molecules to enhance their biological performance, leading to final products with advanced functionalities. In this review, we aim to describe the already developed bioceramic-based biomimetic systems for drug loading and local controlled release. We will discuss the mechanisms used for the inclusion of therapeutic molecules on the designed systems, paying special attention to the identification of critical parameters that modulate drug loading and release kinetics on these scaffolds.


2011 ◽  
Vol 409 ◽  
pp. 105-110 ◽  
Author(s):  
Francesca Boccafoschi ◽  
Marco Rasponi ◽  
Cecilia Mosca ◽  
Erica Bocchi ◽  
Simone Vesentini

It is well-known that cellular behavior can be guided by chemical signals and physical interactions at the cell-substrate interface. The patterns that cells encounter in their natural environment include nanometer-to-micrometer-sized topographies comprising extracellular matrix, proteins, and adjacent cells. Whether cells transduce substrate rigidity at the microscopic scale (for example, sensing the rigidity between adhesion sites) or the nanoscopic scale remains an open question. Here we report that micromolded elastomeric micropost arrays can decouple substrate rigidity from adhesive and surface properties. Arrays of poly (dimethylsiloxane) (PDMS) microposts from microfabricated silicon masters have been fabricated. To control substrate rigidity they present the same post heights but different surface area and spacing between posts. The main advantage of micropost arrays over other surface modification solutions (i.e. hydrogels) is that measured subcellular traction forces could be attributed directly to focal adhesions. This would allow to map traction forces to individual focal adhesions and spatially quantify subcellular distributions of focal-adhesion area, traction force and focal-adhesion stress. Moreover, different adhesion intracellular pathways could be used by the cells to differentiate toward a proliferative or a contractile cellular phenotype, for instance. This particular application is advantageous for vascular tissue engineering applications, where mimicking as close as possible the vessels dynamics should be a step forward in this research field.


RSC Advances ◽  
2017 ◽  
Vol 7 (29) ◽  
pp. 18046-18053 ◽  
Author(s):  
Chong Shen ◽  
Yuyan Li ◽  
Huadi Wang ◽  
Qin Meng

Hydrogels as “soft-and-wet” materials have been widely used as tissue engineering scaffolds due to their similarity to natural extracellular matrix.


2013 ◽  
Vol 750 ◽  
pp. 125-129 ◽  
Author(s):  
Yih Lin Cheng ◽  
Chin Jen Hsueh ◽  
Su Hai Hsiang

PCL is one of the popular biomaterials used in tissue engineering scaffolds, but it is seldom shaped by photo-polymerization. Layered manufacturing techniques, also known as Rapid Prototyping (RP) processes, provide a great opportunity to fabricate 3D scaffolds without problems such as limited control of pore-size and restricted geometric shapes in traditional methods. In our previous researches, the Biomedical Dynamic Masking Rapid Prototyping System was developed to photo-cure biodegradable materials through visible light. In this research, the Dynamic Masking RP System was modified to photo-polymerize cross-linkable PCL to form tissue engineering scaffolds. The cross-linkable PCL was synthesized by reacting PCL and acryloyl chloride, and dissolved in acetone mixing with photo-initiator. The tensile test and degradation test were performed on the cured PCL samples. Fabrication of single-layer pattern was first tested to understand the system’s capability and showed the errors were within 7%. Two types of scaffold design concepts were adopted—one took square, hexagon, or triangle as a basic element to create 2D grid patterns, and the interconnected pore were produced by offsetting the 2D pattern in alternating layers; the other took a double-sided trapezoid as a unit and arrayed it into tube shape with interconnected pore network. Various PCL porous tube scaffolds have been successfully fabricated in this study. In the future, they can be utilized to cell growth or mass cell duplication applications.


2020 ◽  
Vol 20 (9) ◽  
pp. 5349-5355
Author(s):  
Dong-Hwan Yang ◽  
Gwang-Min Heo ◽  
Hong-Ju Park ◽  
Hee-Kyun Oh ◽  
Min-Suk Kook

Physio-chemical surface properties to biomaterial has been attention in tissue engineering due to their properties on cell adhesion, proliferation, and differentiation. The object of this study is to evaluate the preosteoblast biological response on physio-chemical surface-layered 3D PCL scaffold and 3D PCL/β-TCP scaffold. 3D scaffolds were fabricated by FDM 3D printing. Physio-chemical surface of 3D scaffolds were prepared by oxygen plasma and amine plasma-polymerization, respectively. The results of this study demonstrated that amine plasma-treated 3D scaffold on adhesion, proliferation, and osteogenic differentiation of the MC3T3-E1 was significantly increased compared to the other scaffolds.


1993 ◽  
Vol 13 (2) ◽  
pp. 785-791 ◽  
Author(s):  
M D Schaller ◽  
C A Borgman ◽  
J T Parsons

Integrins play a central role in cellular adhesion and anchorage of the cytoskeleton and participate in the generation of intracellular signals, including tyrosine phosphorylation. We have recently isolated a cDNA encoding a unique, focal adhesion-associated protein tyrosine kinase (FAK) that is a component of an integrin-mediated signal transduction pathway. Here we report the isolation of cDNAs encoding the C-terminal, noncatalytic domain of the FAK kinase, termed FRNK (FAK-related nonkinase). Both the FAK- and FRNK-encoded polypeptides, pp125FAK and p41/p43FRNK, are expressed in normal chicken embryo cells. pp125FAK and p41/p43FRNK were localized to focal adhesions, suggesting that pp125FAK is directed to the focal adhesions by sequences within its C-terminal domain. We also show that the fibronectin-dependent increase in tyrosine phosphorylation of pp125FAK is accompanied by a concomitant posttranslational modification of p41FRNK.


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