scholarly journals MicroCT analysis of connectivity in porous structures: optimizing data acquisition and analytical methods in the context of tissue engineering

2020 ◽  
Vol 17 (165) ◽  
pp. 20190833
Author(s):  
Malavika Nair ◽  
Jennifer H. Shepherd ◽  
Serena M. Best ◽  
Ruth E. Cameron
Keyword(s):  
Tissue Engineering ◽  
Analytical Methods ◽  
Large Scale ◽  
Structural Parameters ◽  
Three Dimensional ◽  
Data Capture ◽  
Pixel Size ◽  
Scale Invariant ◽  
Structural Anisotropy

Micro-computed X-ray tomography (MicroCT) is one of the most powerful techniques available for the three-dimensional characterization of complex multi-phase or porous microarchitectures. The imaging and analysis of porous networks are of particular interest in tissue engineering due to the ability to predict various large-scale cellular phenomena through the micro-scale characterization of the structure. However, optimizing the parameters for MicroCT data capture and analyses requires a careful balance of feature resolution and computational constraints while ensuring that a structurally representative section is imaged and analysed. In this work, artificial datasets were used to evaluate the validity of current analytical methods by considering the effect of noise and pixel size arising from the data capture, and intrinsic structural anisotropy and heterogeneity. A novel ‘segmented percolation method’ was developed to exclude the effect of anomalous, non-representative features within the datasets, allowing for scale-invariant structural parameters to be obtained consistently and without manual intervention for the first time. Finally, an in-depth assessment of the imaging and analytical procedures are presented by considering percolation events such as micro-particle filtration and cell sieving within the context of tissue engineering. Along with the novel guidelines established for general pixel size selection for MicroCT, we also report our determination of 3 μm as the definitive pixel size for use in analysing connectivity for tissue engineering applications.

scholarly journals Characterization of Polycaprolactone Nanohydroxyapatite Composites with Tunable Degradability Suitable for Indirect Printing

Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 295
Author(s):  
Stephanie E. Doyle ◽  
Lauren Henry ◽  
Ellen McGennisken ◽  
Carmine Onofrillo ◽  
Claudia Di Bella ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Large Scale ◽  
Three Dimensional ◽  
Cell Number ◽  
Compressive Modulus ◽  
Degradation Rates ◽  
Scaffold Materials ◽  
Natural Tissue ◽  
Complete Regeneration

Degradable bone implants are designed to foster the complete regeneration of natural tissue after large-scale loss trauma. Polycaprolactone (PCL) and hydroxyapatite (HA) composites are promising scaffold materials with superior mechanical and osteoinductive properties compared to the single materials. However, producing three-dimensional (3D) structures with high HA content as well as tuneable degradability remains a challenge. To address this issue and create homogeneously distributed PCL-nanoHA (nHA) scaffolds with tuneable degradation rates through both PCL molecular weight and nHA concentration, we conducted a detailed characterisation and comparison of a range of PCL-nHA composites across three molecular weight PCLs (14, 45, and 80 kDa) and with nHA content up to 30% w/w. In general, the addition of nHA results in an increase of viscosity for the PCL-nHA composites but has little effect on their compressive modulus. Importantly, we observe that the addition of nHA increases the rate of degradation compared to PCL alone. We show that the 45 and 80 kDa PCL-nHA groups can be fabricated via indirect 3D printing and have homogenously distributed nHA even after fabrication. Finally, the cytocompatibility of the composite materials is evaluated for the 45 and 80 kDa groups, with the results showing no significant change in cell number compared to the control. In conclusion, our analyses unveil several features that are crucial for processing the composite material into a tissue engineered implant.

X-ray topographic determination of the granular structure in a graphite mosaic crystal: a three-dimensional reconstruction

2000 ◽  
Vol 33 (4) ◽  
pp. 1023-1030 ◽  
Author(s):  
M. Ohler ◽  
M. Sanchez del Rio ◽  
A. Tuffanelli ◽  
M. Gambaccini ◽  
A. Taibi ◽  
...  
Keyword(s):  
Structural Parameters ◽  
Pyrolytic Graphite ◽  
Three Dimensional ◽  
X Ray ◽  
Dimensional Reconstruction ◽  
Mosaic Crystals ◽  
Spatial Locations ◽  
Crystalline Domains

Section topographs recorded at different spatial locations and at different rocking angles of a highly oriented pyrolytic graphite (HOPG) crystal allow three-dimensional maps of the local angular-dependent scattering power to be obtained. This is performed with a direct reconstruction from the intensity distribution on such topographs. The maps allow the extraction of information on local structural parameters such as size, form and internal mosaic spread of crystalline domains. This data analysis leads to a new method for the characterization of mosaic crystals. Perspectives and limits of applicability of this method are discussed.

scholarly journals A Customizable, Low-Cost Perfusion System for Sustaining Tissue Constructs

2018 ◽  
Vol 23 (6) ◽  
pp. 592-598
Author(s):  
Brian J. O’Grady ◽  
Jason X. Wang ◽  
Shannon L. Faley ◽  
Daniel A. Balikov ◽  
Ethan S. Lippmann ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Cell Viability ◽  
Large Scale ◽  
Low Cost ◽  
Three Dimensional ◽  
Tissue Scaffolds ◽  
Perfusion System ◽  
Pump System ◽  
Tissue Constructs ◽  
Engineered Tissue

The fabrication of engineered vascularized tissues and organs requiring sustained, controlled perfusion has been facilitated by the development of several pump systems. Currently, researchers in the field of tissue engineering require the use of pump systems that are in general large, expensive, and generically designed. Overall, these pumps often fail to meet the unique demands of perfusing clinically useful tissue constructs. Here, we describe a pumping platform that overcomes these limitations and enables scalable perfusion of large, three-dimensional hydrogels. We demonstrate the ability to perfuse multiple separate channels inside hydrogel slabs using a preprogrammed schedule that dictates pumping speed and time. The use of this pump system to perfuse channels in large-scale engineered tissue scaffolds sustained cell viability over several weeks.

Requirements for the Manufacturing of Scaffold Biomaterial With Features at Multiple Scales

2005 ◽  
Author(s):  
I. M. Sebastine ◽  
D. J. Williams
Keyword(s):  
Tissue Engineering ◽  
Multiple Scales ◽  
Cell Attachment ◽  
Structural Parameters ◽  
Complex Function ◽  
Three Dimensional ◽  
Cell Orientation ◽  
Length Scales

Tissue engineering aims to restore the complex function of diseased tissue using cells and scaffold materials. Tissue engineering scaffolds are three-dimensional (3D) structures that assist in the tissue engineering process by providing a site for cells to attach, proliferate, differentiate and secrete an extra-cellular matrix, eventually leading cells to form a neo-tissue of predetermined, three-dimensional shape and size. For a scaffold to function effectively, it must possess the optimum structural parameters conducive to the cellular activities that lead to tissue formation; these include cell penetration and migration into the scaffold, cell attachment onto the scaffold substrate, cell spreading and proliferation and cell orientation. In vivo, cells are organized in functional tissue units that repeat on the order of 100 μm. Fine scaffold features have been shown to provide control over attachment, migration and differentiation of cells. In order to design such 3D featured constructs effectively understanding the biological response of cells across length scales from nanometer to millimeter range is crucial. Scaffold biomaterials may need to be tailored at three different length scales: nanostructure (<1μm), microstructure (<20–100μm), and macrostructure (>100μm) to produce biocompatible and biofunctional scaffolds that closely resemble the extracellular matrix (ECM) of the natural tissue environment and promote cell adhesion, attachment, spreading, orientation, rate of movement, and activation. Identification of suitable fabrication techniques for manufacturing scaffolds with the required features at multiple scales is a significant challenge. This review highlights the effect and importance of the features of scaffolds that can influence the behaviour of cells/tissue at different length scales in vitro to increase our understanding of the requirements for the manufacture of functional 3D tissue constructs.

Hydroxyapatite/Biopolymers Composite Scaffolds for Bone Tissue Engineering

2011 ◽  
Vol 493-494 ◽  
pp. 826-831
Author(s):  
A.C.B.M. Fook ◽  
Thiago Bizerra Fideles ◽  
R.C. Barbosa ◽  
G.T.F.S. Furtado ◽  
G.Y.H. Sampaio ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Freeze Drying ◽  
Three Dimensional ◽  
Porous Scaffolds ◽  
X Ray Diffraction ◽  
X Ray ◽  
Interconnected Pores

The application of a hybrid composite consisting of biopolymer and calcium phosphate, similar morphology and properties of natural bone, may be a way to solve the problem of the fragility of ceramics without reducing its mechanical properties, retaining the properties of biocompatibility and high bioactivity. This work aims at the preparation and characterization of three-dimensional scaffolds composite HA / biopolymers (chitosan and gelatin). The freeze-drying technique was employed in this study to obtain these frameworks and partial results showed the effectiveness of this method. This involved the study of structural, chemical and morphological frameworks, in order to direct the research suggested the application. The X Ray Diffraction (XRD) and infrared spectroscopy and Fourier transform (FTIR) results confirmed the formation of hydroxyapatite (HA) phase and the presence of characteristic bands of HA and biopolymers in all compositions. The microstructure of the scaffolds study conducted by Scanning Electron Microscopy (SEM) revealed the formation of longitudinally oriented microchannels with interconnected pores. In all compositions the porous scaffolds showed varying sizes and mostly larger than 100μm, and is therefore considered materials with potential for application in bone tissue engineering.

scholarly journals Hybrid Tissue Engineering Scaffolds by Combination of Three-Dimensional Printing and Cell Photoencapsulation

2015 ◽  
Vol 6 (2) ◽  
Author(s):  
Marica Markovic ◽  
Jasper Van Hoorick ◽  
Katja Hölzl ◽  
Maximilian Tromayer ◽  
Peter Gruber ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Fused Deposition Modeling ◽  
Structural Parameters ◽  
Three Dimensional ◽  
Poly Lactic Acid ◽  
Tissue Engineering Scaffolds ◽  
Ecm Remodeling ◽  
Initial Cell ◽  
Fused Deposition ◽  
Hydrogel Matrix

Three-dimensional (3D) printing offers versatile possibilities for adapting the structural parameters of tissue engineering scaffolds. However, it is also essential to develop procedures allowing efficient cell seeding independent of scaffold geometry and pore size. The aim of this study was to establish a method for seeding the scaffolds using photopolymerizable cell-laden hydrogels. The latter facilitates convenient preparation, and handling of cell suspension, while distributing the hydrogel precursor throughout the pores, before it is cross-linked with light. In addition, encapsulation of living cells within hydrogels can produce constructs with high initial cell loading and intimate cell-matrix contact, similar to that of the natural extra-cellular matrix (ECM). Three dimensional scaffolds were produced from poly(lactic) acid (PLA) by means of fused deposition modeling. A solution of methacrylamide-modified gelatin (Gel-MOD) in cell culture medium containing photoinitiator Li-TPO-L was used as a hydrogel precursor. Being an enzymatically degradable derivative of natural collagen, gelatin-based matrices are biomimetic and potentially support the process of cell-induced remodeling. Preosteoblast cells MC3T3-E1 at a density of 10 × 106 cells per 1 mL were used for testing the seeding procedure and cell proliferation studies. Obtained results indicate that produced constructs support cell survival and proliferation over extended duration of our experiment. The established two-step approach for scaffold seeding with the cells is simple, rapid, and is shown to be highly reproducible. Furthermore, it enables precise control of the initial cell density, while yielding their uniform distribution throughout the scaffold. Such hybrid tissue engineering constructs merge the advantages of rigid 3D printed constructs with the soft hydrogel matrix, potentially mimicking the process of ECM remodeling.

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