Coral reef degradation is a rising problem, driven by marine heatwaves, the spread of coral diseases, and human impact by overfishing and pollution. Our capacity to restore coral reefs lags behind in ...terms of scale, effectiveness, and cost-efficiency. While common restoration efforts rely on the formation of carbonate skeletons on structural frames for supported coral growth, this technique is a rate-limiting step in the growth of scleractinian corals. Reverse engineering and additive manufacturing technologies offer an innovative shift in approach from the use of concrete blocks and metal frames to sophisticated efforts that use scanned geometries of harvested corals to fabricate artificial coral skeletons for installation in coral gardens and reefs. Herein, we present an eco-friendly and sustainable approach for coral fabrication by merging three-dimensional (3D) scanning, 3D printing, and molding techniques. Our method, 3D CoraPrint, exploits the 3D printing technology to fabricate artificial natural-based coral skeletons, expediting the growth rate of live coral fragments and quickening the reef transplantation process while minimizing nursery costs. It allows for flexibility, customization, and fast return time with an enhanced level of accuracy, thus establishing an environmentally friendly, scalable model for coral fabrication to boost restorative efforts around the globe.
Ultrashort self-assembling peptides (SAPs) can spontaneously form nanofibers that resemble the extracellular matrix. These fibers allow the formation of hydrogels that are biocompatible, ...biodegradable, and non-immunogenic. We have previously proven that SAPs, when biofunctionalized with protein-derived motifs, can mimic the extracellular matrix characteristics that support colorectal organoid formation. These biofunctional peptide hydrogels retain the original parent peptide's mechanical properties, tunability, and printability while incorporating cues that allow cell-matrix interactions to increase cell adhesion. This paper presents the protocols needed to evaluate and characterize the effects of various biofunctional peptide hydrogels on cell adhesion and lumen formation using an adenocarcinoma cancer cell line able to form colorectal cancer organoids cost-effectively. These protocols will help evaluate biofunctional peptide hydrogel effects on cell adhesion and luminal formation using immunostaining and fluorescence image analysis. The cell line used in this study has been previously utilized for generating organoids in animal-derived matrices.
This roadmap, through the contributions of ten groups worldwide, contains different techniques, methods and materials devoted to sensing in nanomedicine. Optics is used in different ways in the ...detection schemes. Raman, fluorescence and infrared spectroscopies, plasmonics, second harmonic generation and optical tweezers are all used in applications from single molecule detection (both in highly diluted and in highly concentrated solutions) to single cell manipulation. In general, each optical scheme, through device miniaturization and electromagnetic field localization, exploits an intrinsic optical enhancement mechanism in order to increase the sensitivity and selectivity of the device with respect to the complex molecular construct. The materials used for detection include nanoparticles and nanostructures fabricated with different 2D and 3D lithographic methods. It is shown that sensitivity to a single molecule is already accessible whether the system under study is a single cell or a multitude of cells in a molecular mixture. Throughout the roadmap there is an attempt to foresee and to suggest future directions in this interdisciplinary field.
Millions of people worldwide suffer from skin injuries, which create significant problems in their lives and are costly to cure. Tissue engineering is a promising approach that aims to fabricate ...functional organs using biocompatible scaffolds. We designed ultrashort tetrameric peptides with promising properties required for skin tissue engineering. Our work aimed to test the efficacy of these scaffolds for the fabrication of dermal grafts and 3D vascularized skin tissue models. We found that the direct contact of keratinocytes and fibroblasts enhanced the proliferation of the keratinocytes. Moreover, the expression levels of TGF-β1, b-FGF, IL-6, and IL-1α is correlated with the growth of the fibroblasts and keratinocytes in the co-culture. Furthermore, we successfully produced a 3D vascularized skin co-culture model using these peptide scaffolds. We believe that the described results represent an advancement in the fabrication of skin tissue equivalent, thereby providing the opportunity to rebuild missing, failing, or damaged parts.
Graphical abstract
An alarming increase in antibiotic-resistant bacterial strains is driving clinical demand for new antibacterial agents. One of the oldest antimicrobial agents is elementary silver (Ag), which has ...been used for thousands of years. Even today, elementary Ag is used for medical purposes such as treating burns, wounds, and microbial infections. In consideration of the effectiveness of elementary Ag, the present researchers generated effective antibacterial/antibiofilm agents by combining elementary Ag with biocompatible ultrashort peptide compounds. The innovative antibacterial agents comprised a hybrid peptide bound to Ag nanoparticles (IVFK/Ag NPs). These were generated by photoionizing a biocompatible ultrashort peptide, thus reducing Ag ions to form Ag NPs with a diameter of 6 nm. The IVFK/Ag NPs demonstrated promising antibacterial/antibiofilm activity against reference Gram-positive and Gram-negative bacteria compared with commercial Ag NPs. Through morphological changes in Escherichia coli and Staphylococcus aureus, we proposed that the mechanism of action for IVFK/Ag NPs derives from their ability to disrupt bacterial membranes. In terms of safety, the IVFK/Ag NPs demonstrated biocompatibility in the presence of human dermal fibroblast cells, and concentrations within the minimal inhibitory concentration had no significant effect on cell viability. These results demonstrated that hybrid peptide/Ag NPs hold promise as a biocompatible material with strong antibacterial/antibiofilm properties, allowing them to be applied across a wide range of applications in tissue engineering and regenerative medicine.
How fishes are able to detect trace molecules in large bodies of water is not understood. It is plausible that they use olfactory receptors to detect water-soluble compounds. How the zebra fish Danio ...Rerio, an organism with only 98 functional olfactory receptors, is able to selectively detect and recognize numerous compounds in water remains a puzzling phenomenon. We are interested in studying the biochemical and molecular mechanisms of olfaction in fish. Here, we report on the study of a bioengineered zebra fish olfactory receptor OR131-2, affinity-purified from a HEK293S tetracycline-inducible system. This receptor was expressed and translocated to the cell plasma membrane as revealed by confocal microscopy. Circular dichroism spectroscopy showed that the purified zebra fish receptor folded into an α-helical structure, as observed for other G-protein coupled receptors (GPCRs). Our study shows that it is possible to produce viable quantities of the zebra fish olfactory receptor. This will not only enable detailed structural and functional analyses, but also aid in the design of biosensor devices in order to detect water-soluble metabolites or its intermediates, which are associated with human health.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Amyloid‐like peptides are an ideal model for the mechanistic study of amyloidosis, which may lead to many human diseases, such as Alzheimer disease. This study reports a strong second harmonic ...generation (SHG) effect of amyloid‐like peptides, having a signal equivalent to or even higher than those of endogenous collagen fibers. Several amyloid‐like peptides (both synthetic and natural) were examined under SHG microscopy and shown they are SHG‐active. These peptides can also be observed inside cells (in vitro). This interesting property can make these amyloid‐like peptides second harmonic probes for bioimaging applications. Furthermore, SHG microscopy can provide a simple and label‐free approach to detect amyloidosis. Lattice corneal dystrophy was chosen as a model disease of amyloidosis. Morphological difference between normal and diseased human corneal biopsy samples can be easily recognized, proving that SHG can be a useful tool for disease diagnosis.
Ultrashort peptides undergo self‐assembling and form nanofibers. In this schematic diagram, each blue triangle together with an orange circle represents an ultrashort peptide. The orange circle represents a hydrophilic headgroup and the blue triangle represents a hydrophobic tail with an increasing hydrophobicity. The ultrashort peptide nanofibers are able to generate second harmonic light. When excited at 810 nm, ultrashort peptide nanofibers emitted a sharp peak at 405 nm.
Purpose: Next-generation sequencing (NGS) technology detects specific mutations that can provide treatment opportunities for colorectal cancer (CRC) patients. Patients and Methods: We analyzed the ...mutation frequencies of common actionable genes and their association with clinicopathological characteristics and oncologic outcomes using targeted NGS in 107 Saudi Arabian patients without a family history of CRC. Results: Approximately 98% of patients had genetic alterations. Frequent mutations were observed in BRCA2 (79%), CHEK1 (78%), ATM (76%), PMS2 (76%), ATR (74%), and MYCL (73%). The APC gene was not included in the panel. Statistical analysis using the Cox proportional hazards model revealed an unusual positive association between poorly differentiated tumors and survival rates (p = 0.025). Although no significant univariate associations between specific mutations or overall mutation rate and overall survival were found, our preliminary analysis of the molecular markers for CRC in a predominantly Arab population can provide insights into the molecular pathways that play a significant role in the underlying disease progression. Conclusions: These results may help optimize personalized therapy when drugs specific to a patient’s mutation profile have already been developed.
The field of three-dimensional (3D) bioprinting is rapidly emerging as an additive manufacturing method for tissue and organ fabrication. The demand for tissues and organ transplants is ever ...increasing, although donors are not as readily available. Consequently, tissue engineering is gaining much attention to alleviate this problem. The process of achieving well-structured 3D bioprinted constructs using hydrogel bioinks depends on symmetrical precision, regulated flow rates, and viability of cells. Even with the mentioned parameters optimized, the printed structures need additional refining by removing excessive liquids, as peptide hydrogel bioprints encapsulate water. However, it is challenging to eliminate the confined fluids without compromising the printing process. In this paper, we introduced a vacuum system to our 3D bioprinting robotic arm and thus optimized the printing quality for complex and refined 3D scaffolds. Moreover, the proposed vacuum system supports printing with cells. Our results show improved printing resolution which facilitates the printing of higher and more stable structures.
62Articular cartilage is a nonvascularized and poorly cellularized tissue with a low self-repair capacity. Therefore, damage to this tissue due to trauma or degenerative joint diseases such as ...osteoarthritis needs a high-end medical intervention. However, such interventions are costly, have limited healing capacity, and could impair patients' quality of life. In this regard, tissue engineering and three-dimensional (3D) bioprinting hold great potential. However, identifying suitable bioinks that are biocompatible, with the desired mechanical stiffness, and can be used under physiological conditions is still a challenge. In this study, we developed two tetrameric self-assembling ultrashort peptide bioinks that are chemically well-defined and can spontaneously form nanofibrous hydrogels under physiological conditions. The printability of the two ultrashort peptides was demonstrated; different shape constructs were printed with high shape fidelity and stability. Furthermore, the developed ultrashort peptide bioinks gave rise to constructs with different mechanical properties that could be used to guide stem cell differentiation toward specific lineages. Both ultrashort peptide bioinks demonstrated high biocompatibility and supported the chondrogenic differentiation of human mesenchymal stem cells. Additionally, the gene expression analysis of differentiated stem cells with the ultrashort peptide bioinks revealed articular cartilage extracellular matrix formation preference. Based on the different mechanical stiffness of the two ultrashort peptide bioinks, they can be used to fabricate cartilage tissue with different cartilaginous zones, including the articular and calcified cartilage zones, which are essential for engineered tissue integration.
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