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A laser-cutting-based manufacturing process for the generation of three-dimensional scaffolds for tissue engineering using Polycaprolactone/Hydroxyapatite composite polymer
摘要： A manufacturing process for sheet-based stacked scaffolds (SSCs) based on laser-cutting (LC) was developed. The sheets consist of Polycaprolactone/Hydroxyapatite (PCL/HA) composite material. Single sheets were cut from a PCL/HA foil and stacked to scaffolds with interconnecting pores of defined sizes. HA quantities up to 50% were processable with high reproducibility, while the accuracy was dependent on the applied laser power. The smallest achievable pore sizes were about 40 μm, while the smallest stable solid structures were about 125 μm. The human mesenchymal stem cell line SCP-1 was cultured on the manufactured PCL/HA scaffolds. The cells developed a natural morphology and were able to differentiate to functional osteoblasts. The generation of PCL/HA SSCs via LC offers new possibilities for tissue engineering (TE) approaches. It is reliable and fast, with high resolution. The SSC approach allows for facile cell seeding and analysis of cell fate within the three-dimensional cell culture, thus allowing for the generation of functional tissue constructs.
关键词： laser-cutting,Tissue engineering,scaffold
更新于2025-11-21 11:24:58
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Visualizing Cell-Laden Fibrin-Based Hydrogels using Cryogenic Scanning Electron Microscopy and Confocal Microscopy
摘要： The present investigation explores the microscopic aspects of cell-laden hydrogels at high resolutions, using three-dimensional cell cultures in semi-synthetic constructs that are of very-high water content (>98% water). The study aims to provide an imaging strategy for these constructs, while minimizing artifacts. Constructs of PEG-fibrinogen (PEG-Fb) and fibrin hydrogels containing embedded mesenchymal cells (human dermal fibroblasts) were first imaged by confocal microscopy. Next, high resolution scanning electron microscopy (HR-SEM) was used to provide images of the cells within the hydrogels, at submicron resolutions. Because it was not possible to obtain artifact-free images of the hydrogels using room-temperature HR-SEM, a cryogenic HR-SEM (cryo-HR-SEM) imaging methodology was employed to visualize the sample while preserving the natural hydrated state of the hydrogel. The ultrastructural details of the constructs were observed at subcellular resolutions, revealing numerous cellular components, the biomaterial in its native configuration, and the uninterrupted cell membrane as it relates with the biomaterial in the hydrated state of the construct. Constructs containing microscopic albumin microbubbles were also imaged using these methodologies to reveal fine details of the interaction between the cells, the microbubbles and the hydrogel. Taken together with the confocal microscopy, this imaging strategy provides a more complete picture of the hydrated state of the hydrogel network with cells inside. As such, this methodology addresses some of the challenges of obtaining this information in amorphous hydrogel systems containing a very-high water content (>98%) with embedded cells. Such insight may lead to better hydrogel-based strategies for tissue engineering and regeneration.
关键词： Fibrin,Electron Microscopy,Hydrogel,Tissue Engineering,Scaffold,Confocal Microscopy
更新于2025-11-21 11:08:12
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Liquid Crystal-Induced Myoblast Alignment
摘要： The ability to control cell alignment represents a fundamental requirement toward the production of tissue in vitro but also to create biohybrid materials presenting the functional properties of human organs. However, cell cultures on standard commercial supports do not provide a selective control on the cell organization morphology, and different techniques, such as the use of patterned or stimulated substrates, are developed to induce cellular alignment. In this work, a new approach toward in vitro muscular tissue morphogenesis is presented exploiting liquid crystalline networks. By using smooth polymeric films with planar homogeneous alignment, a certain degree of cellular order is observed in myoblast cultures with direction of higher cell alignment corresponding to the nematic director. The molecular organization inside the polymer determines such effects since no cell organization is observed using homeotropic or isotropic samples. These findings represent the first example of cellular alignment induced by the interaction with a nematic polymeric scaffold, setting the stage for new applications of liquid crystal polymers as active matter to control tissue growth.
关键词： liquid crystalline alignments,liquid crystalline network,cell alignment,biomaterials,muscular tissue engineering
更新于2025-11-21 11:01:37
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Photoconductive micro/nano-scale interfaces of a semiconducting polymer for wireless stimulation of neuron-like cells
摘要： We report multiscale structured fibers and patterned films based on a semiconducting polymer, poly(3-hexylthiophene) (P3HT), as photoconductive biointerfaces to promote neuronal stimulation upon light irradiation. The micro/nano scale structures of P3HT used for neuronal interfacing and stimulation include nanofibers with an average diameter of 100 nm, microfibers with an average diameter of about 1 μm, and the lithographically patterned stripes width of 3, 25 and 50 μm, respectively. The photoconductive effect of P3HT upon light irradiation provide electrical stimulation for neuronal differentiation and directed growth. Our results demonstrate that neurons on P3HT nanofibers showed a significantly higher total number of branches while neurons grown on P3HT microfibers had longer and thinner neurites. Such a combination strategy of topographical and photoconductive stimulation can be applied to further enhance neuronal differentiation and directed growth. These photoconductive polymeric micro/nano structures demonstrated their great potential for neural engineering and development of novel neural regenerative devices.
关键词： semiconductors,neurons,polymers,nanostructures,tissue engineering
更新于2025-09-23 15:23:52
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Monitoring neovascularization and integration of decellularized human scaffolds using photoacoustic imaging
摘要： Tissue engineering is a branch of regenerative medicine that aims to manipulate cells and scaffolds to create bioartificial tissues and organs for patients. A major challenge lies in monitoring the blood supply to the new tissue following transplantation: the integration and neovascularization of scaffolds in vivo is critical to their functionality. Photoacoustic imaging (PAI) is a laser-generated ultrasound-based technique that is particularly well suited to visualising the microvasculature due to the high optical absorption of haemoglobin. Here, we describe an early proof-of-concept study in which PAI in widefield tomography mode is used to image biological, decellularized human tracheal scaffolds. We found that photoacoustic imaging allowed the longitudinal tracking of scaffold integration into subcutaneous murine tissue with high spatial resolution at depth over an extended period of time. The results of the study were consistent with post-imaging histological analyses, demonstrating that photoacoustic imaging can be used to non-invasively monitor the extent of vascularization in biological tissue-engineered scaffolds. We anticipate that this technique could find application in tissue-engineering studies aimed at improving the speed and extent of scaffold neovascularization. With technological development, it could also be used to inform the clinical timing of surgical procedures following heterotopic transplantation to establish vasculature.
关键词： vascularization,angiogenesis,photoacoustic imaging,transplantation,tissue engineering,trachea
更新于2025-09-23 15:22:29
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Polybenzyl Glutamate Biocompatible Scaffold Promotes the Efficiency of Retinal Differentiation toward Retinal Ganglion Cell Lineage from Human-Induced Pluripotent Stem Cells
摘要： Optic neuropathy is one of the leading causes of irreversible blindness caused by retinal ganglion cell (RGC) degeneration. The development of induced pluripotent stem cell (iPSC)-based therapy opens a therapeutic window for RGC degeneration, and tissue engineering may further promote the efficiency of differentiation process of iPSCs. The present study was designed to evaluate the effects of a novel biomimetic polybenzyl glutamate (PBG) scaffold on culturing iPSC-derived RGC progenitors. The iPSC-derived neural spheres cultured on PBG scaffold increased the differentiated retinal neurons and promoted the neurite outgrowth in the RGC progenitor layer. Additionally, iPSCs cultured on PBG scaffold formed the organoid-like structures compared to that of iPSCs cultured on cover glass within the same culture period. With RNA-seq, we found that cells of the PBG group were differentiated toward retinal lineage and may be related to the glutamate signaling pathway. Further ontological analysis and the gene network analysis showed that the differentially expressed genes between cells of the PBG group and the control group were mainly associated with neuronal differentiation, neuronal maturation, and more specifically, retinal differentiation and maturation. The novel electrospinning PBG scaffold is beneficial for culturing iPSC-derived RGC progenitors as well as retinal organoids. Cells cultured on PBG scaffold differentiate effectively and shorten the process of RGC differentiation compared to that of cells cultured on coverslip. The new culture system may be helpful in future disease modeling, pharmacological screening, autologous transplantation, as well as narrowing the gap to clinical application.
关键词： induced pluripotent stem cells,retinal ganglion cells,tissue engineering,glaucoma,optic neuropathy,polybenzyl glutamate,electrospinning scaffold
更新于2025-09-23 15:22:29
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[IEEE 2017 21st National Biomedical Engineering Meeting (BIYOMUT) - Istanbul (2017.11.24-2017.12.26)] 2017 21st National Biomedical Engineering Meeting (BIYOMUT) - Three Dimensional Bioprinting of Tissue Engineered Artificial Heart Valves by Stereolithography
摘要： Over the past several decades, there has been an ever-increasing demand for organ transplants. There are no significant increase observed in organ donations while the need for organ transplantation is keep increasing. Organ donation emerges as the most feasible solution especially in heart valve diseases. There are two types of artificial heart valves are utilized for heart valve replacement operations. These are mechanical heart valves and bioprosthetic heart valves. However, there are severe intrinsic problems exist for both type of artificial heart valves that are hard to be solved. Due to these reasons, growing or fabricating tissue engineered heart valves using biomaterial scaffolds with a person’s own cells with high biocompatability. The purpose of this study is; fabricating 3D artificial heart valves by stereolithography that has potential to be obtained for heart valve replacements. Stereolithography has proven potential for fabricating 3D models with high structural integrity and strong mechanical properties, yet it has not been evaluated as a bioprinter. In this study, this purpose is aimed to be succeeded.
关键词： 3D bioprinting,artificial heart valves,stereolithography,tissue engineering
更新于2025-09-23 15:21:21
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Femtosecond laser surface engineering of biopolymer ceramic scaffolds coated with ZnO by low temperature atomic layer deposition method
摘要： Surface femtosecond laser texturing of biopolymer and biopolymer/ceramic composites with subsequent ZnO film deposition on the samples by low temperature Atomic Layer Deposition (ALD) method was performed. In the current study, the deposition of ZnO layers was implemented at temperature 50 °C under pressure of 2 mbar. In order to investigate the effect of diverse ZnO thin films thickness, 100 or 500 repeating ALD cycles were applied. The samples were exposed to ultra-short laser pulses of 130 fs duration generated by a CPA (chirped pulse amplifier) Ti:Sapphire laser system. The artificial scaffolds were irradiated by varying the laser fluence (0.2 J/cm2, 0.41 J/cm2 and 2.07 J/cm2) and the number of applied laser pulses (N = 1, 2, 5 and 10). The morphology and chemical properties of the treated samples were evaluated by Scanning Electronic Microscopy, Energy-Dispersive X-ray Spectroscopy and X-ray photoelectron spectroscopy. By combining fs laser modification with low temperature ALD method, essentially improved bioactivity properties of hybrid organic–inorganic bone tissue scaffolds could be achieved, which is of great importance for future tissue engineering application of the samples.
关键词： Atomic layer deposition,Biopolymer/ceramic composites,Femtosecond laser modification,Tissue engineering
更新于2025-09-23 15:21:01
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Shedding light on novel approach to artificial cornea
摘要： Arti?cial human corneas that could circumvent the problem of limited donors or ethical concerns have been developed by scientists at Newcastle University. These are the ?rst human corneas to have been 3D printed. The cornea is outermost, transparent layer of the eye, which covers the iris and pupil and plays a role in focusing light on to the retina. The team demonstrated a proof of principle experiment in which they have taken human corneal stromal cells from a healthy donor cornea, mixed them with alginate and collagen and created 3D bio printer ink. Using a conventional commercially available 3D bio-printer, they were able to extrude the mixture to form increasing concentric circles allowing them to fabricate an arti?cial human cornea from actual cells within ten minutes.
关键词： artificial cornea,hydrogel,tissue engineering,bio-ink,3D printing
更新于2025-09-23 15:21:01
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Bone quality assessment of osteogenic cell cultures by Raman microscopy
摘要： The use of autologous stem/progenitor cells represents a promising approach to the repair of craniofacial bone defects. The calvarium is recognized as a viable source of stem/progenitor cells that can be transplanted in vitro to form bone. However, it is unclear if bone formed in cell culture is similar in quality to that found in native bone. In this study, the quality of bone mineral formed in osteogenic cell cultures were compared against calvarial bone from postnatal mice. Given the spectroscopic resemblance that exists between cell and collagen spectra, the feasibility of extracting information on cell activity and bone matrix quality were also examined. Stem/progenitor cells isolated from fetal mouse calvaria were cultured onto fused‐quartz slides under osteogenic differentiation conditions for 28 days. At specific time intervals, slides were removed and analyzed by Raman microscopy and mineral staining techniques. We show that bone formed in culture at Day 28 resembled calvarial bone from 1‐day‐old postnatal mice with comparable mineralization, mineral crystallinity, and collagen crosslinks ratios. In contrast, bone formed at Day 28 contained a lower degree of ordered collagen fibrils compared with 1‐day‐old postnatal bone. Taken together, bone formed in osteogenic cell culture exhibited progressive matrix maturation and mineralization but could not fully replicate the high degree of collagen fibril order found in native bone.
关键词： Raman microscopy,osteogenic differentiation,tissue engineering,stem cells,bone quality
更新于2025-09-23 15:21:01