A Three-Dimensional in vitro Bone Model – Triple Culture of Primary Human Osteoblasts, Osteoclasts and Osteocytes

2021 ◽  
Author(s):  
Anne Bernhardt ◽  
Jasmin Skottke ◽  
Max von Witzleben ◽  
Michael Gelinsky

Materials ◽  
2019 ◽  
Vol 12 (17) ◽  
pp. 2739 ◽  
Author(s):  
Korbinian Benz ◽  
Andreas Schöbel ◽  
Marisa Dietz ◽  
Peter Maurer ◽  
Jochen Jackowski

The aim of this in vitro pilot study was to analyse the adhesion behaviour of human osteoblasts and fibroblasts on polyether ether ketone (PEEK) when compared with titanium surfaces in an inflammatory environment under lipopolysaccharide (LPS) incubation. Scanning electron microscopy (SEM) images of primary human osteoblasts/fibroblasts on titanium/PEEK samples were created. The gene expression of the LPS-binding protein (LBP) and the LPS receptor (toll-like receptor 4; TLR4) was measured by real-time polymerase chain reaction (PCR). Immunocytochemistry was used to obtain evidence for the distribution of LBP/TLR4 at the protein level of the extra-cellular-matrix-binding protein vinculin and the actin cytoskeleton. SEM images revealed that the osteoblasts and fibroblasts on the PEEK surfaces had adhesion characteristics comparable to those of titanium. The osteoblasts contracted under LPS incubation and a significantly increased LBP gene expression were detected. This was discernible at the protein level on all the materials. Whereas no increase of TLR4 was detected with regard to mRNA concentrations, a considerable increase in the antibody reaction was detected on all the materials. As is the case with titanium, the colonisation of human osteoblasts and fibroblasts on PEEK samples is possible under pro-inflammatory environmental conditions and the cellular inflammation behaviour towards PEEK is lower than that of titanium.



2017 ◽  
Vol 58 (5-6) ◽  
pp. 216-226
Author(s):  
Johannes Schauwecker ◽  
Mark Bock ◽  
Florian Pohlig ◽  
Heinz Mühlhofer ◽  
Jutta Tübel ◽  
...  

Background/Purpose: Polymethylmethacrylate (PMMA) and calcium phosphate (Ca-P) cements are widely used for arthroplasty surgery and augmentation of bone defects. However, aseptic implant loosening in absence of wear-induced osteolysis indicates an unfavourable interaction between the cement surface and human osteoblasts. Our underlying hypothesis is that cement surfaces directly modify cell viability, proliferation rate, and cell differentiation. Methods: To test this hypothesis, we examined primary human osteoblasts harvested from six individuals. These cells were pooled and subsequently seeded directly on cement pellets prepared from Palacos® R, Palacos® R+G, and Norian® Drillable cements. After incubation for 24 and 72 h, cell viability, proliferation rate, apoptosis rate, and cell differentiation were analysed. Results: Upon cultivation of human osteoblasts on cement surfaces, we observed a significantly reduced cell viability and DNA content compared to the control. Analysis of the apoptosis rate revealed an increase for cells on Palacos R and Norian Drillable, but a significant decrease on Palacos R+G compared to the control. Regarding osteogenic differentiation, significantly lower values of alkaline phosphatase enzyme activity were identified for all cement surfaces after 24 and 72 h compared to cultivation on tissue culture plastic, serving as control. Conclusions: In summary, these data suggest a limited biocompatibility of both PMMA and Ca-P cements, necessitating further research to reduce unfavourable cell-cement interactions and consequently extend implant survival.



2016 ◽  
Vol 45 (10) ◽  
pp. 803-809 ◽  
Author(s):  
Paul Günther Baptist Heymann ◽  
Thomas Ziebart ◽  
Peer Wolfgang Kämmerer ◽  
Robert Mandic ◽  
Akram Saydali ◽  
...  


Bone ◽  
2002 ◽  
Vol 30 (5) ◽  
pp. 718-725 ◽  
Author(s):  
D Ferrera ◽  
S Poggi ◽  
C Biassoni ◽  
G.R Dickson ◽  
S Astigiano ◽  
...  




2015 ◽  
pp. 1 ◽  
Author(s):  
Vincenzo Luca Zizzari ◽  
Guya Diletta Marconi ◽  
Marianna De Colli ◽  
Susi Zara ◽  
Barbara Zavan ◽  
...  


2010 ◽  
Vol 638-642 ◽  
pp. 506-511 ◽  
Author(s):  
Claudia Bergemann ◽  
Ernst Dieter Klinkenberg ◽  
Frank Lüthen ◽  
Arne Weidmann ◽  
Regina Lange ◽  
...  

Porous tantalum (Ta) biomaterial is designed to function as a scaffold for osseous ingrowths and has found applications in orthopedics. Integration of this Ta foam into the neighboring bone requires that osteoprogenitor cells attach to the implant, grow into the scaffold, proliferate and differentiate to osteoblasts. The aim of the present study was to create an in vitro 3D model system to investigate the interaction of human osteoblasts with porous Ta in the depth of the corpus. To explore active migration of osteoblasts into the Ta scaffold two porous Ta discs (Zimmer, Poland) were horizontally fixed within a clamping ring. Thereby a 3D Ta module with 4 levels is generated, which is placed into a cell culture well with the appropriate medium. Osteoblast-like cells were seeded apical onto the Ta module and cultured for 7 days in humidified atmosphere. Active migration of cells into the scaffold was monitored by field emission scanning electron microscopy (FESEM) imaging of the apical, medial and basal layers. A problem in 3D cell culture is the nutrition of cells inside of the scaffold. Therefore morphological changes and differentiation of the cells in distinct layers were analyzed.



Metals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 1681
Author(s):  
Fiedler Jörg ◽  
Katmer Amet Betül ◽  
Michels Heiner ◽  
Kappelt Gerhard ◽  
Brenner Rolf Erwin

(1) In order to enable a more widespread use of uncemented titanium-based endoprostheses to replace cobalt-containing cemented endoprostheses for joint replacement, it is essential to achieve optimal osseointegrative properties and develop economic fabrication processes while retaining the highest biomedical quality of titanium materials. One approach is the usage of an optimized form of Ti6Al4V-precision casting for manufacturing. Besides the chemical and physical properties, it is necessary to investigate possible biological influences in order to test whether the new manufacturing process is equivalent to conventional methods. (2) Methods: Primary human osteoblasts were seeded on discs, which were produced by a novel Ti6Al4V centrifugal-casting process in comparison with standard machined discs of the same titanium alloy. In a second step, the surfaces were modified by calcium or phosphorus ion beam implantation. In vitro, we analyzed the effects on proliferation, differentiation, and apoptotic processes. (3) Results: SEM analysis of cells seeded on the surfaces showed no obvious differences between the reference material and the cast material with or without ion implantation. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) proliferation assay also did not reveal any significant differences. Additionally, the osteogenic differentiation process tested by quantitative polymerase chain reactions (PCR), Alizarin red S assay, and C-terminal collagen type I propeptide (CICP) Elisa was not significantly modified. No signs of induced apoptosis were observed. (4) Conclusions: In this study, we could show that the newly developed process of centrifugal casting generated a material with comparable surface features to standard machined Ti6Al4V material. In terms of biological impact on primary human osteoblasts, no significant differences were recognized. Additional Ca- or P-ion implantation did not improve or impair these characteristics in the dosages applied. These findings indicate that spin casting of Ti6Al4V may represent an interesting alternative to the production of geometrically complex orthopedic implants.



Injury ◽  
2014 ◽  
Vol 45 (8) ◽  
pp. 1156-1164 ◽  
Author(s):  
Markus Prause ◽  
Claudine Seeliger ◽  
Marina Unger ◽  
Martijn van Griensven ◽  
Alexander Tobias Haug


2018 ◽  
Vol 106 (7) ◽  
pp. 2565-2575 ◽  
Author(s):  
Anja Klein ◽  
Andreas Baranowski ◽  
Ulrike Ritz ◽  
Hermann Götz ◽  
Sascha Heinemann ◽  
...  


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