Metallic Vessel with Mesh Culture Surface Fabricated Using Three-dimensional Printing Engineers Tissue Culture Environment

Imashiro, Chikahiro; Morikura, Takashi; Hayama, Motoaki; Ezura, Atsushi; Komotori, Jun; Miyata, Shogo; Sakaguchi, Katsuhisa; Shimizu, Tatsuya

doi:10.1007/s12257-022-0227-1

Cited by 3 publications

(4 citation statements)

References 55 publications

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“…2 E to H, the cells were cultured without exhibiting cytotoxicity caused by the vessel. Although a different cell line was used in a previous study, Casp9, which is expressed in the early stage of apoptosis, was not upregulated owing to cell culture characteristics in pure titanium vessels [ 26 ]. Thus, it can be argued that employing a metallic culture vessel is effective for evaluating thermal tolerance.…”

Section: Discussionmentioning

confidence: 99%

Titanium Culture Vessel Presenting Temperature Gradation for the Thermotolerance Estimation of Cells

et al. 2023

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Hyperthermia can be induced to exploit the thermal intolerance of cancer cells, which is worse than that of normal cells, as a potential noninvasive cancer treatment. To develop an effective hyperthermia treatment, thermal cytotoxicity of cells should be comprehensively investigated. However, to conduct such investigations, the culture temperature must be accurately regulated. We previously reported a culture system in which the culture temperature could be accurately regulated by employing metallic culture vessels. However, appropriate temperature conditions for hyperthermia depend on the cell species. Consequently, several experiments need to be conducted, which is a bottleneck of inducing hyperthermia. Hence, we developed a cell culture system with temperature gradation on a metallic culture surface. Michigan Cancer Foundation-7 cells and normal human dermal fibroblasts were used as cancer and normal cell models, respectively. Normal cells showed stronger thermal tolerance; this was because the novel system immediately exhibited a temperature gradation. Thus, the developed culture system can be used to investigate the optimum thermal conditions for effective hyperthermia treatment. Furthermore, as the reactions of cultured cells can be effectively assessed with the present results, further research involving the thermal stimulation of cells is possible.

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Section: Discussionmentioning

confidence: 99%

Titanium Culture Vessel Presenting Temperature Gradation for the Thermotolerance Estimation of Cells

et al. 2023

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“…On the other hand, TETs are involved in an active DNA demethylation pathway by converting 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) [ 50 , 51 , 52 ] and they are important to demethylate DNA of cells responding to environmental changes, such as titanium or other metallic biomaterials used in medicine or dentistry. Additionally, this might be explored in predictable tools such as bioreactors responding to implantable biomaterials [ 53 , 54 , 55 , 56 ] as cultures of blood vessels in bioreactors have been reported [ 57 , 58 ]. Additionally, to know the effects of titanium in cells and humans within these fields is also relevant in considering novel propositions in bioengineering research and, as these enzymes are druggable, they are targets for specific design of small molecules able to modulate their activities, aiming at enhancing angiogenesis, and they might accelerate the recovery of patients by improving the osteogenesis surrounding titanium-based implants.…”

Section: Discussionmentioning

confidence: 99%

Titanium-Enriched Medium Promotes Environment-Induced Epigenetic Machinery Changes in Human Endothelial Cells

Fernandes

Silva

Wood

et al. 2023

JFB

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It is important to understand whether endothelial cells are epigenetically affected by titanium-enriched media when angiogenesis is required during bone development and it is expected to be recapitulated during osseointegration of biomaterials. To better address this issue, titanium-enriched medium was obtained from incubation of titanium discs for up to 24 h as recommended by ISO 10993-5:2016, and further used to expose human umbilical vein endothelial cells (HUVECs) for up to 72 h, when the samples were properly harvested to allow molecular analysis and epigenetics. In general, our data show an important repertoire of epigenetic players in endothelial cells responding to titanium, reinforcing protein related to the metabolism of acetyl and methyl groups, as follows: Histone deacetylases (HDACs) and NAD-dependent deacetylase sirtuin-1 (Sirt1), DNA methyltransferases (DNMTs) and ten-eleven translocation (TET) methylcytosine dioxygenases, which in conjunction culminate in driving chromatin condensation and the methylation profile of DNA strands, respectively. Taking our data into consideration, HDAC6 emerges as important player of this environment-induced epigenetic mechanism in endothelial cells, while Sirt1 is required in response to stimulation of reactive oxygen species (ROS) production, as its modulation is relevant to vasculature surrounding implanted devices. Collectively, all these findings support the hypothesis that titanium keeps the surrounding microenvironment dynamically active and so affects the performance of endothelial cells by modulating epigenetics. Specifically, this study shows the relevance of HDAC6 as a player in this process, possibly correlated with the cytoskeleton rearrangement of those cells. Furthermore, as those enzymes are druggable, it opens new perspectives to consider the use of small molecules to modulate their activities as a biotechnological tool in order to improve angiogenesis and accelerate bone growth with benefits of a fast recovery time for patients.

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“…As such, there's a pressing need to refine and advance the bioengineering workflows for both researchers and industries. In recent work, we introduced reusable culture vessels crafted from biocompatible materials like titanium and SUS316L, commonly used in the human body [5][6][7]. These materials not only show excellent cytocompatibility but also demonstrate robust heat resistance in autoclaves, a prevalent sterilization technique.…”

Section: Introductionmentioning

confidence: 99%

“…Earlier studies have tailored the wettability of metallic culture surfaces using surface morphology, maintaining stability even in an autoclave [7,[9][10][11]. Building on this, we present a novel approach: employing stripe-patterned metallic culture surfaces with differential wettabilities, derived from varied surface morphologies, to guide cell orientation.…”

Section: Introductionmentioning

confidence: 99%

Regulating Cell Orientation with a Femtosecond Laser-Induced Macro Stripe Design on Metallic Culture Surfaces

Imashiro,

Ezura,

Yamada

et al. 2023

Preprint

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Controlling cell orientation is paramount in bioengineering processes. While several surface modification techniques have emerged to guide cell alignment, they often involve complex, repetitive procedures for each experiment. A streamlined approach for cell orientation is thus necessary. In this study, we present a reusable metallic culture surface that induces an anisotropic cell orientation, attributed to its unique geometric morphology. By employing a femtosecond laser, periodic nanostructures were produced on the metallic culture surface, leading to a distinctive macro-stripe design composed of both laser-treated and mirrored areas. Myoblast cells cultured on this surface displayed a pronounced orientation. Initial random cell adherence was followed by migration towards the mirror surface, culminating in the desired orientation. This shift can be credited to the mirrored sections offering superior cell adhesion and reduced wettability compared to the laser-treated sections. This innovative culture surface holds significant potential for advancing bioengineering endeavors, especially in the realm of tissue engineering.

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Metallic Vessel with Mesh Culture Surface Fabricated Using Three-dimensional Printing Engineers Tissue Culture Environment

Cited by 3 publications

References 55 publications

Titanium Culture Vessel Presenting Temperature Gradation for the Thermotolerance Estimation of Cells

Titanium Culture Vessel Presenting Temperature Gradation for the Thermotolerance Estimation of Cells

Titanium-Enriched Medium Promotes Environment-Induced Epigenetic Machinery Changes in Human Endothelial Cells

Regulating Cell Orientation with a Femtosecond Laser-Induced Macro Stripe Design on Metallic Culture Surfaces

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