Improved cell seeding efficiency and cell distribution in porous hydroxyapatite scaffolds by semi-dynamic method

2021 ◽  
Author(s):  
Feng Shi ◽  
Ke Duan ◽  
Zaijun Yang ◽  
Yumei Liu ◽  
Jie Weng
Keyword(s):  
Dynamic Method ◽  
Cell Distribution ◽  
Cell Seeding ◽  
Porous Hydroxyapatite ◽  
Seeding Efficiency

scholarly journals An optimized 3D-printed perfusion bioreactor for homogeneous cell seeding in bone substitute scaffolds for future chairside applications

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nadja Engel ◽  
Carsten Fechner ◽  
Annika Voges ◽  
Robert Ott ◽  
Jan Stenzel ◽  
...  
Keyword(s):  
In Silico ◽  
Bone Substitute ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Cell Distribution ◽  
Cell Seeding ◽  
Outlet Port ◽  
Short Period ◽  
Homogeneous Cell ◽  
Perfusion Mode

AbstractA clinical implementation of cell-based bone regeneration in combination with scaffold materials requires the development of efficient, controlled and reproducible seeding procedures and a tailor-made bioreactor design. A perfusion system for efficient, homogeneous, and rapid seeding with human adipogenic stem cells in bone substitute scaffolds was designed. Variants concerning medium inlet and outlet port geometry, i.e. cylindrical or conical diffuser, cell concentration, perfusion mode and perfusion rates were simulated in silico. Cell distribution during perfusion was monitored by dynamic [18F]FDG micro-PET/CT and validated by laser scanning microscopy with three-dimensional image reconstruction. By iterative feedback of the in silico and in vitro experiments, the homogeneity of cell distribution throughout the scaffold was optimized with adjustment of flow rates, cell density and perfusion properties. Finally, a bioreactor with a conical diffusor geometry was developed, that allows a homogeneous cell seeding (hoover coefficient: 0.24) in less than 60 min with an oscillating perfusion mode. During this short period of time, the cells initially adhere within the entire scaffold and stay viable. After two weeks, the formation of several cell layers was observed, which was associated with an osteogenic differentiation process. This newly designed bioreactor may be considered as a prototype for chairside application.

scholarly journals Electrodeposition of Hydroxyapatite on a Metallic 3D-Woven Bioscaffold

Coatings ◽  
2020 ◽  
Vol 10 (8) ◽  
pp. 715 ◽  
Author(s):  
Ju Xue ◽  
Ashley Farris ◽  
Yunfei Wang ◽  
Weiyan Yeh ◽  
Cristina Romany ◽  
...  
Keyword(s):  
Reference Electrode ◽  
Deposition Potential ◽  
Cellular Adhesion ◽  
Cell Seeding ◽  
X Ray Diffraction ◽  
X Ray ◽  
Woven Bone ◽  
Electrochemically Deposited ◽  
Seeding Efficiency ◽  
Stromal Stem Cells

In this study, we demonstrate that a uniform coating of hydroxyapatite (HAp, Ca10(PO4)6(OH)2) can be electrochemically deposited onto metallic 3D-woven bone scaffolds to enhance their bioactivity. The HAp coatings were deposited onto metallic scaffolds using an electrolyte containing Ca(NO3)2·4H2O, NH4H2PO4, and NaNO3. The deposition potential was varied to maximize the uniformity and adhesion of the coating. Using X-ray diffraction (XRD), Raman spectroscopy, and energy-dispersive spectroscopy (EDS), we found crystallized HAp on the 3D-woven lattice under all deposition potentials, while the −1.5 V mercury sulfate reference electrode potential provided the best local uniformity with a satisfactory deposition rate. The coatings generated under this optimized condition were approximately 5 µm thick and uniform throughout the internal and external sections of the woven lattice. We seeded and cultured both coated and uncoated scaffolds with human adipose-derived stromal/stem cells (ASCs) for 12 h and 4 days. We observed that the HAp coating increased the initial cell seeding efficiency by approximately 20%. Furthermore, after 4 days of culture, ASCs cultured on HAp-coated stainless-steel scaffolds increased by 32% compared to only 17% on the uncoated scaffold. Together, these results suggest that the HAp coating improves cellular adhesion.

Incorporation of Human Osteoblast Cells and Osteoblast-Like Cells into Porous Hydroxyapatite Scaffolds

2005 ◽  
Vol 284-286 ◽  
pp. 627-630
Author(s):  
Meera Q. Arumugam ◽  
Roger A. Brooks ◽  
Neil Rushton ◽  
William Bonfield
Keyword(s):  
Cell Attachment ◽  
Osteosarcoma Cell ◽  
Osteosarcoma Cells ◽  
Mechanical Agitation ◽  
Human Osteoblasts ◽  
Porous Hydroxyapatite ◽  
Adhesion Protein ◽  
Seeding Efficiency ◽  
Osteosarcoma Cell Line Mg63 ◽  
Initial Seeding

The object of this study was to investigate methods of seeding cells onto porous hydroxyapatite granules with the aim of optimising cell attachment. Two cells types were used; an osteosarcoma cell line, MG63, and human osteoblasts (HOBs) isolated from trabecular bone. Several conditions were investigated to determine their effect on cell attachment. These included varying the initial seeding concentration, pre-adsorption of the granules with the adhesion protein fibronectin and the use of mechanical agitation. Human osteoblasts and MG63 osteosarcoma cells attached to both dense and porous HA granules but with a low seeding efficiency while seeding was not significantly improved by pre-coating scaffolds with fibronectin or by introducing fluid flow.

Oscillating perfusion of cell suspensions through three-dimensional scaffolds enhances cell seeding efficiency and uniformity

2003 ◽  
Vol 84 (2) ◽  
pp. 205-214 ◽  
Author(s):  
D. Wendt ◽  
A. Marsano ◽  
M. Jakob ◽  
M. Heberer ◽  
I. Martin
Keyword(s):  
Three Dimensional ◽  
Cell Suspensions ◽  
Cell Seeding ◽  
Seeding Efficiency

scholarly journals The Influence of Scaffold Designs on Cell Seeding Efficiency in Establishing A Three-Dimensional Culture

2018 ◽  
Vol 1082 ◽  
pp. 012072
Author(s):  
Yanny Marliana Baba Ismail ◽  
Yvonne Reinwald ◽  
Ian Wimpenny ◽  
Oana Bretcanu ◽  
Kenneth Dalgarno ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Seeding ◽  
Three Dimensional Culture ◽  
Seeding Efficiency

Polarized protein membrane for high cell seeding efficiency

2007 ◽  
Vol 83B (2) ◽  
pp. 472-480 ◽  
Author(s):  
Björn Atthoff ◽  
Cecilia Aulin ◽  
Catharina Adelöw ◽  
Jöns Hilborn
Keyword(s):  
Cell Seeding ◽  
High Cell ◽  
Protein Membrane ◽  
Seeding Efficiency

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.

scholarly journals Novel Cell Seeding System into a Porous Scaffold Using a Modified Low-Pressure Method to Enhance Cell Seeding Efficiency and Bone Formation

2007 ◽  
Vol 16 (7) ◽  
pp. 729-739 ◽  
Author(s):  
Ichiro Torigoe ◽  
Shinichi Sotome ◽  
Akio Tsuchiya ◽  
Toshitaka Yoshii ◽  
Makoto Takahashi ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Bone Formation ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Low Pressure ◽  
Porous Scaffolds ◽  
Cell Seeding ◽  
Modified Technique ◽  
Significant Difference ◽  
Seeding Efficiency

The efficient seeding of cells into porous scaffolds is important in bone tissue engineering techniques. To enhance efficiency, we modified the previously reported cell seeding techniques using low-pressure conditions. In this study, the effects of low pressure on bone marrow-derived stromal cells (BMSCs) of rats and the usefulness of the modified technique were assessed. There was no significant difference found in the proliferative and osteogenic capabilities among various low-pressure (50–760 mmHg, 1–10 min) conditions. To analyze the efficacies of the cell seeding techniques, BMSCs suspended in the plasma of rats were seeded into porous β-tricalcium phosphate (β-TCP) blocks by the following three procedures: 1) spontaneous penetration of cell suspension under atmospheric pressure (SP); 2) spontaneous penetration and subsequent low pressure treatment (SPSL), the conventional technique; and 3) spontaneous penetration under low pressure conditions (SPUL), the modified technique. Subsequently, these BMSCs/β-TCP composites were used for the analysis of cell seeding efficiency or in vivo bone formation capability. Both the number of BMSCs seeded into β-TCP blocks and the amount of bone formation of the SPUL group were significantly higher than those of the other groups. The SPUL method with a simple technique permits high cell seeding efficiency and is useful for bone tissue engineering using BMSCs and porous scaffolds.

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