Fabrication and Characterization of Colorectal Cancer Organoids from SW1222 Cell Line in Ultrashort Self-Assembling Peptide Matrix.

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.

Fusing Peptide Epitopes for Advanced Multiplex Serological Testing for SARS-CoV-2 Antibody Detection.

The tragic COVID-19 pandemic, which has seen a total of 655 million cases worldwide and a death toll of over 6.6 million seems finally tailing off. Even so, new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continue to arise, the severity of which cannot be predicted in advance. This is concerning for the maintenance and stability of public health, since immune evasion and increased transmissibility may arise. Therefore, it is crucial to continue monitoring antibody responses to SARS-CoV-2 in the general population. As a complement to polymerase chain reaction tests, multiplex immunoassays are elegant tools that use individual protein or peptide antigens simultaneously to provide a high level of sensitivity and specificity. To further improve these aspects of SARS-CoV-2 antibody detection, as well as accuracy, we have developed an advanced serological peptide-based multiplex assay using antigen-fused peptide epitopes derived from both the spike and the nucleocapsid proteins. The significance of the epitopes selected for antibody detection has been verified by in silico molecular docking simulations between the peptide epitopes and reported SARS-CoV-2 antibodies. Peptides can be more easily and quickly modified and synthesized than full length proteins and can, therefore, be used in a more cost-effective manner. Three different fusion-epitope peptides (FEPs) were synthesized and tested by enzyme-linked immunosorbent assay (ELISA). A total of 145 blood serum samples were used, compromising 110 COVID-19 serum samples from COVID-19 patients and 35 negative control serum samples taken from COVID-19-free individuals before the outbreak. Interestingly, our data demonstrate that the sensitivity, specificity, and accuracy of the results for the FEP antigens are higher than for single peptide epitopes or mixtures of single peptide epitopes. Our FEP concept can be applied to different multiplex immunoassays testing not only for SARS-CoV-2 but also for various other pathogens. A significantly improved peptide-based serological assay may support the development of commercial point-of-care tests, such as lateral-flow-assays.

Fabrication of lumen-forming colorectal cancer organoids using a newly designed laminin-derived bioink.

160Three-dimensional (3D) bioprinting systems, which are the prominent tools for biofabrication, should evolve around the cutting-edge technologies of tissue engineering. This is the case with organoid technology, which requires a plethora of new materials to evolve, including extracellular matrices with specific mechanical and biochemical properties. For a bioprinting system to facilitate organoid growth, it must be able to recreate an organ-like environment within the 3D construct. In this study, a well-established, self-assembling peptide system was employed to generate a laminin-like bioink to provide signals of cell adhesion and lumen formation in cancer stem cells. One bioink formulation led to the formation of lumen with outperforming characteristics, which showed good stability of the printed construct.

Ecologically Friendly Biofunctional Ink for Reconstruction of Rigid Living Systems Under Wet Conditions.

The development of three-dimensional (3D)-printable inks is essential for several applications, from industrial manufacturing to novel applications for biomedical engineering. Remarkably, biomaterials for tissue engineering applications can be expanded to other new horizons; for instance, restoration of rigid living systems as coral reefs is an emergent need derived from recent issues from climate change. The coral reefs have been endangered, which can be observed in the increasing bleaching around the world. Very few studies report eco-friendly inks for matter since most conventional approaches require synthetic polymer, which at some point could be a pollutant depending on the material. Therefore, there is an unmet need for cost-effective formulations from eco-friendly materials for 3D manufacturing to develop carbonate-based inks for coral reef restoration. Our value proposition derives from technologies developed for regenerative medicine, commonly applied for human tissues like bone and cartilage. In our case, we created a novel biomaterial formulation from biopolymers such as gelatin methacrylate, poly (ethylene glycol diacrylate), alginate, and gelatin as scaffold and binder for the calcium carbonate and hydroxyapatite bioceramics needed to mimic the structure of rigid structures. This project presents evidence from 2D/3D manufacturing, chemical, mechanical, and biological characterization, which supports the hypothesis of its utility to aid in the fight to counteract the coral bleaching that affects all the marine ecosystem, primarily when this is supported by solid research in biomaterials science used for living systems, it can extend tissue engineering into new approaches in different domains such as environmental or marine sciences.