A Transparent Polyimide Film as a Biological Cell Culture Sheet with Microstructures

Maenosono, Hirotaka; Saito, Hirofumi; Nishioka, Yasushiro

doi:10.4236/jbnb.2014.51003

Cited by 7 publications

(3 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The mHCN2-transfected MSCs preserved their viability, generated pacemaker current, grew on the porous Kapton® scaffolds with 1–3 μm diameter pore arrays, and established intercellular communication between opposite sides of the scaffold without transmigration through the pores. The polyimide films demonstrated suitable mechanical and thermal properties, and good biocompatibility, and have already been successfully applied to a vast range of biomedical investigations [ 13 – 17 ]. In parallel to the formation of functionally active HCN channels, the mHCN2-transfected MSCs expressed Cx43 necessary for communication through Cx43-based GJ channels and F-actin containing TNTs necessary for the biological pacemaker functioning.…”

Section: Discussionmentioning

confidence: 99%

Relevance of HCN2-expressing human mesenchymal stem cells for the generation of biological pacemakers

et al. 2016

View full text Add to dashboard Cite

BackgroundThe transfection of human mesenchymal stem cells (hMSCs) with the hyperpolarization-activated cyclic nucleotide-gated ion channel 2 (HCN2) gene has been demonstrated to provide biological pacing in dogs with complete heart block. The mechanism appears to be the generation of the ion current (If) by the HCN2-expressing hMSCs. However, it is not clear how the transfection process and/or the HCN2 gene affect the growth functions of the hMSCs. Therefore, we investigated survival, proliferation, cell cycle, and growth on a Kapton® scaffold of HCN2-expressing hMSCs.MethodshMSCs were isolated from the bone marrow of healthy volunteers applying a selective cell adhesion procedure and were identified by their expression of specific surface markers. Cells from passages 2–3 were transfected by electroporation using commercial transfection kits and a pIRES2-EGFP vector carrying the pacemaker gene, mouse HCN2 (mHCN2). Transfection efficiency was confirmed by enhanced green fluorescent protein (EGFP) fluorescence, quantitative real-time polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA). After hMSCs were transfected, their viability, proliferation, If generation, apoptosis, cell cycle, and expression of transcription factors were measured and compared with non-transfected cells and cells transfected with pIRES2-EGFP vector alone.ResultsIntracellular mHCN2 expression after transfection increased from 22.14 to 62.66 ng/mg protein (p < 0.05). Transfection efficiency was 45 ± 5 %. The viability of mHCN2-transfected cells was 82 ± 5 %; they grew stably for more than 3 weeks and induced If current. mHCN2-transfected cells had low mitotic activity (10.4 ± 1.24 % in G2/M and 83.6 ± 2.5 % in G1 phases) as compared with non-transfected cells (52–53 % in G2/M and 31–35 % in G1 phases). Transfected cells showed increased activation of nine cell cycle-regulating transcription factors: the most prominent upregulation was of AMP-dependent transcription factor ATF3 (7.11-fold, p = 0.00056) which regulates the G1 phase. mHCN2-expressing hMSCs were attached and made anchorage-dependent connection with other cells without transmigration through a 12.7-μm thick Kapton® HN film with micromachined 1–3 μm diameter pores.ConclusionsmHCN2-expressing hMSCs preserved the major cell functions required for the generation of biological pacemakers: high viability, functional activity, but low proliferation rate through the arrest of cell cycle in the G1 phase. mHCN2-expressing hMSCs attached and grew on a Kapton® scaffold without transmigration, confirming the relevance of these cells for the generation of biological pacemakers.

show abstract

Section: Discussionmentioning

confidence: 99%

Relevance of HCN2-expressing human mesenchymal stem cells for the generation of biological pacemakers

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Various commercial and noncommercial PI materials were proven as non-toxic by a multitude of in vitro, direct, or indirect, cytotoxicity assays. Most of them specifically declares to comply with the ISO guidelines, and investigate the viability, proliferation, degeneration, and lysis of different common cell types:L929 mouse fibroblast cells (Kapton, PI2611, Durimide 7020, Durimide 7510, PI2525, 6FDA-6FAP, EPICOLN-PPD) [27,41,44,48,51,52],3T3 mouse fibroblasts cells (Probimide 7520) [49],OP9 mouse stromal cells (Neopulim) [19],rat primary Schawnn cells (Kapton, PI2611, Durimide 7020) [27],rat astroglial cells (unknown PI trade name) [70],bovine endothelial cells (unknown PI trade name) [70],BHK-21 baby hamster kidney fibroblasts (PI2525, PI2771) [28],…”

Section: Biocompatibility Screening Of Polyimides By In Vitro Methodsmentioning

confidence: 99%

“…This section is dedicated to aromatic polyimides, which are planar imidic macromolecules containing rigid aromatic rings and various flexible, heteroaliphatic, unsymmetrical, bulky or pendant structural motifs. We chose to focus on the aromatic polymers since the majority of polyimidic materials tested for biomedical uses subscribe to this structural architecture, even if some commercial or in-house aliphatic backbones have found their way into the field [19,20]. Moreover, the review is mainly centered on polyimidic films (also encompassing foils and coatings) since, with some noticeable exceptions which will be underlined at their rightful place, this is the format chosen by most bio-related applications and devices.…”

Section: Polyimides: Synthesis General Features and Biomedical Amentioning

confidence: 99%

Biocompatibility of Polyimides: A Mini-Review

Constantin

Aflori

Damian³

et al. 2019

Materials

141

View full text Add to dashboard Cite

Polyimides (PIs) represent a benchmark for high-performance polymers on the basis of a remarkable collection of valuable traits and accessible production pathways and therefore have incited serious attention from the ever-demanding medical field. Their characteristics make them suitable for service in hostile environments and purification or sterilization by robust methods, as requested by most biomedical applications. Even if PIs are generally regarded as “biocompatible”, proper analysis and understanding of their biocompatibility and safe use in biological systems deeply needed. This mini-review is designed to encompass some of the most robust available research on the biocompatibility of various commercial or noncommercial PIs and to comprehend their potential in the biomedical area. Therefore, it considers (i) the newest concepts in the field, (ii) the chemical, (iii) physical, or (iv) manufacturing elements of PIs that could affect the subsequent biocompatibility, and, last but not least, (v) in vitro and in vivo biocompatibility assessment and (vi) reachable clinical trials involving defined polyimide structures. The main conclusion is that various PIs have the capacity to accommodate in vivo conditions in which they are able to function for a long time and can be judiciously certified as biocompatible.

show abstract

Femtosecond laser micro‐machined polyimide films for cell scaffold applications

Antanavičiūtė

Šimatonis

Ulčinas

et al. 2017

J Tissue Eng Regen Med

View full text Add to dashboard Cite

Engineering of sophisticated synthetic 3D scaffolds that allow controlling behaviour and location of the cells requires advanced micro/nano-fabrication techniques. Ultrafast laser micro-machining employing a 1030-nm wavelength Yb:KGW femtosecond laser and a micro-fabrication workstation for micro-machining of commercially available 12.7 and 25.4 μm thickness polyimide (PI) film was applied. Mechanical properties of the fabricated scaffolds, i.e. arrays of differently spaced holes, were examined via custom-built uniaxial micro-tensile testing and finite element method simulations. We demonstrate that experimental micro-tensile testing results could be numerically simulated and explained by two-material model, assuming that 2-6 μm width rings around the holes possessed up to five times higher Young's modulus and yield stress compared with the rest of the laser intacted PI film areas of 'dog-bone'-shaped specimens. That was attributed to material modification around the micro-machined holes in the vicinity of the position of the focused laser beam track during trepanning drilling. We demonstrate that virgin PI films provide a suitable environment for the mobility, proliferation and intercellular communication of human bone marrow mesenchymal stem cells, and discuss how cell behaviour varies on the micro-machined PI films with holes of different diameters (3.1, 8.4 and 16.7 μm) and hole spacing (30, 35, 40 and 45 μm). We conclude that the holes of 3.1 μm diameter were sufficient for metabolic and genetic communication through membranous tunneling tubes between cells residing on the opposite sides of PI film, but prevented the trans-migration of cells through the holes. Copyright © 2016 John Wiley & Sons, Ltd.

show abstract

A Transparent Polyimide Film as a Biological Cell Culture Sheet with Microstructures

Cited by 7 publications

References 34 publications

Relevance of HCN2-expressing human mesenchymal stem cells for the generation of biological pacemakers

Relevance of HCN2-expressing human mesenchymal stem cells for the generation of biological pacemakers

Biocompatibility of Polyimides: A Mini-Review

Femtosecond laser micro‐machined polyimide films for cell scaffold applications

Contact Info

Product

Resources

About