scholarly journals Design and Functional Testing of a Multichamber Perfusion Platform for Three-Dimensional Scaffolds

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Marco Piola ◽  
Monica Soncini ◽  
Marco Cantini ◽  
Nasser Sadr ◽  
Giulio Ferrario ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Perfusion Culture ◽  
Homogeneous Distribution ◽  
Porous Scaffolds ◽  
Cell Seeding ◽  
Flow Perfusion ◽  
Culture Viability ◽  
Compact Size ◽  
Dynamic Cell

Perfusion culture systems are widely used in tissue engineering applications for enhancing cell culture viability in the core of three-dimensional scaffolds. In this work, we present a multichamber confined-flow perfusion system, designed to provide a straightforward platform for three-dimensional dynamic cell cultures. The device comprises 6 culture chambers allowing independent and simultaneous experiments in controlled conditions. Each chamber consists of three parts: a housing, a deformable scaffold-holder cartridge, and a 7 mL reservoir, which couples water-tightly with the housing compressing the cartridge. Short-term dynamic cell seeding experiments were carried out with MC3T3-E1 cells seeded into polycaprolactone porous scaffolds. Preliminary results revealed that the application of flow perfusion through the scaffold favored the penetration of the cells to its interior, producing a more homogeneous distribution of cells with respect to dropwise or injection seeding methods. The culture chamber layout was conceived with the aim of simplifying the user operations under laminar flow hood and minimizing the risks for contamination during handling and operation. Furthermore, a compact size, a small number of components, and the use of bayonet couplings ensured a simple, fast, and sterility-promoting assembling. Finally, preliminaryin vitrotests proved the efficacy of the system in enhancing cell seeding efficiency, opening the way for further studies addressing long-term scaffold colonization.

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 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.

Development of HA-PLGA Scaffold Encapsulating Intact BMP-2 Using Solid Freeform Fabrication Technology

2011 ◽  
Author(s):  
Jin-Hyung Shim ◽  
Jong Young Kim ◽  
Kyung Shin Kang ◽  
Jung Kyu Park ◽  
Sei Kwang Hahn ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Bone Regeneration ◽  
Porous Scaffold ◽  
Solid Freeform Fabrication ◽  
Three Dimensional ◽  
Porous Scaffolds ◽  
Prolonged Release ◽  
Cellular Activity ◽  
Freeform Fabrication

Tissue engineering is an interdisciplinary field that focuses on restoring and repairing tissues or organs. Cells, scaffolds, and biomolecules are recognized as three main components of tissue engineering. Solid freeform fabrication (SFF) technology is required to fabricate three-dimensional (3D) porous scaffolds to provide a 3D environment for cellular activity. SFF technology is especially advantageous for achieving a fully interconnected, porous scaffold. Bone morphogenic protein-2 (BMP-2), an important biomolecule, is widely used in bone tissue engineering to enhance bone regeneration activity. However, methods for the direct incorporation of intact BMP-2 within 3D scaffolds are rare. In this work, 3D porous scaffolds with poly(lactic-co-glycolic acid) chemically grafted hyaluronic acid (HA-PLGA), in which intact BMP-2 was directly encapsulated, were successfully fabricated using SFF technology. BMP-2 was previously protected by poly(ethylene glycol) (PEG), and the BMP-2/PEG complex was incorporated in HA-PLGA using an organic solvent. The HAPLGA/PEG/BMP-2 mixture was dissolved in chloroform and deposited via a multi-head deposition system (MHDS), one type of SFF technology, to fabricate a scaffold for tissue engineering. An additional air blower system and suction were installed in the MHDS for the solvent-based fabrication method. An in vitro evaluation of BMP-2 release was conducted, and prolonged release of intact BMP-2, for up to 28 days, was confirmed. After confirmation of advanced proliferation of pre osteoblasts, a superior differentiation effect of the HA-PLGA/PEG/BMP-2 scaffold was validated by measuring high expression levels of bone-specific markers, such as alkaline phosphatase (ALP) and osteocalcin (OC). We show that our solvent-based fabrication is a non-toxic method for restoring cellular activity. Moreover, the HAPLGA/PEG/BMP-2 scaffold was effective for bone regeneration.

scholarly journals In vivo–mimicking microfluidic perfusion culture of pancreatic islet spheroids

2019 ◽  
Vol 5 (11) ◽  
pp. eaax4520 ◽  
Author(s):  
Yesl Jun ◽  
JaeSeo Lee ◽  
Seongkyun Choi ◽  
Ji Hun Yang ◽  
Maike Sander ◽  
...  
Keyword(s):  
Drug Testing ◽  
Cell Damage ◽  
Three Dimensional ◽  
Perfusion Culture ◽  
Interstitial Flow ◽  
Static Culture ◽  
Culture Models ◽  
Main Component

Native pancreatic islets interact with neighboring cells by establishing three-dimensional (3D) structures, and are surrounded by perfusion at an interstitial flow level. However, flow effects are generally ignored in islet culture models, although cell perfusion is known to improve the cell microenvironment and to mimic in vivo physiology better than static culture systems. Here, we have developed functional islet spheroids using a microfluidic chip that mimics interstitial flow conditions with reduced shear cell damage. Dynamic culture, compared to static culture, enhanced islet health and maintenance of islet endothelial cells, reconstituting the main component of islet extracellular matrix within spheroids. Optimized flow condition allowed localization of secreted soluble factors near spheroids, facilitating diffusion-mediated paracrine interactions within islets, and enabled long-term maintenance of islet morphology and function for a month. The proposed model can aid islet preconditioning before transplantation and has potential applications as an in vitro model for diabetic drug testing.

Microscopic Analysis of Porous Biodegradable Scaffolds for Tissue Engineering

2000 ◽  
Vol 6 (S2) ◽  
pp. 988-989
Author(s):  
F. Buevich ◽  
S. Pulapura ◽  
J. Kohn
Keyword(s):  
Tissue Regeneration ◽  
Combinatorial Library ◽  
Three Dimensional ◽  
Microscopic Analysis ◽  
Porous Scaffolds ◽  
Biodegradable Scaffolds ◽  
Condensation Polymers ◽  
Scaffold Architecture ◽  
Useful Lifetime

Introduction: There is considerable interest in the use of three-dimensional porous scaffolds for tissue regeneration. The presence of an interconnected framework of pores with large surface area facilitates the formation of extracellular matrix and permits cellular ingrowth into implanted structures. For scaffolds to be useful for tissue regeneration, they must maintain good dimensional stability during the lifetime of the implant. While the initial scaffold architecture is often well characterized, a systematic study of the influence of incubation on the scaffold architecture is critical to ensure that the scaffolds retain their interconnected network of pores during their useful lifetime. Herein, we report on the evaluation of the architecture of polyarylate scaffolds and their stability under in vitro conditions using scanning electron microscopy (SEM).The polymers used in this study were selected from a library of degradable polyarylates. This library is the first reported combinatorial library of biodegradable condensation polymers.

scholarly journals The importance of factorial design in tissue engineering and biomaterials science: Optimisation of cell seeding efficiency on dermal scaffolds as a case study

2018 ◽  
Vol 9 ◽  
pp. 204173141878169 ◽  
Author(s):  
Alexandra Levin ◽  
Vaibhav Sharma ◽  
Lilian Hook ◽  
Elena García-Gareta
Keyword(s):  
Tissue Engineering ◽  
Experimental Design ◽  
Factorial Design ◽  
Three Dimensional ◽  
Cell Types ◽  
Cell Seeding ◽  
Factorial Experimental Design ◽  
Seeding Efficiency

This article presents a case study to show the usefulness and importance of using factorial design in tissue engineering and biomaterials science. We used a full factorial experimental design (2 × 2 × 2 × 3) to solve a routine query in every biomaterial research project: the optimisation of cell seeding efficiency for pre-clinical in vitro cell studies, the importance of which is often overlooked. In addition, tissue-engineered scaffolds can be cellularised with relevant cell type(s) to form implantable tissue constructs, where the cell seeding method must be reliable and robust. Our results show the complex relationship between cells and scaffolds and suggest that the optimum seeding conditions for each material may be different due to different material properties, and therefore, should be investigated for individual scaffolds. Our factorial experimental design can be easily translated to other cell types and three-dimensional biomaterials, where multiple interacting variables can be thoroughly investigated for better understanding of cell–biomaterial interactions.

Sign in / Sign up

Export Citation Format

Share Document