Combinatorial Virtual Screening Revealed a Novel Scaffold for TNKS Inhibition to Combat Colorectal Cancer

The abnormal Wnt signaling pathway leads to a high expression of β-catenin, which causes several types of cancer, particularly colorectal cancer (CRC). The inhibition of tankyrase (TNKS) activity can reduce cancer cell growth, invasion, and resistance to treatment by blocking the Wnt signaling pathway. A pharmacophore search and pharmacophore docking were performed to identify potential TNKS inhibitors in the training databases. The weighted MM/PBSA binding free energy of the docking model was calculated to rank the databases. The reranked results indicated that 26.98% of TNKS inhibitors that were present in the top 5% of compounds in the database and near an ideal value ranked 28.57%. The National Cancer Institute database was selected for formal virtual screening, and 11 potential TNKS inhibitors were identified. An enzyme-based experiment was performed to demonstrate that of the 11 potential TNKS inhibitors, NSC295092 and NSC319963 had the most potential. Finally, Wnt pathway analysis was performed through a cell-based assay, which indicated that NSC319963 is the most likely TNKS inhibitor (pIC50 = 5.59). The antiproliferation assay demonstrated that NSC319963 can decrease colorectal cancer cell growth; therefore, the proposed method successfully identified a novel TNKS inhibitor that can alleviate CRC.

Development of Silk Fibroin Scaffolds by Using Indirect 3D-Bioprinting Technology

Due to the excellent biocompatibility of natural polymers, a variety of natural polymers have been widely used as biomaterials for manufacturing tissue engineered scaffolds. Despite the excellent biological activity of natural polymers, there have been obstacles in using them on their own to prepare 3D scaffolds with sufficient mechanical strength. Although multiple 3D-bioprinting technologies have recently emerged as effective manufacturing tools for scaffold preparation, scaffold preparation using only natural polymers with tunable mechanical properties is still difficult. Herein, we introduce novel scaffold fabrication methods using the natural polymer silk fibroin via indirect 3D-bioprinting technology. The developed silk fibroin scaffolds showed biocompatibility and tunable mechanical strength by changing the concentration of the silk fibroin. Furthermore, controlling the flexibility of the silk fibroin scaffolds was made possible by changing the solvent for the silk fibroin solution used to fabricate the scaffold. Consequently, silk fibroin scaffolds fabricated via our method can be considered for various applications in the bioengineering of either soft or musculoskeletal tissues.

Egg White Alginate as a Novel Scaffold Biomaterial for 3D Salivary Cell Culturing

Saliva production by salivary glands play a crucial role in oral health. The loss of salivary gland function could lead to xerostomia, a condition also known as dry mouth. Significant reduction in saliva production could lead to further complications such as difficulty in speech, mastication, and increased susceptibility to dental caries and oral infections and diseases. While some palliative treatments are available for xerostomia, there are no curative treatments to date. This study explores the use of Egg White Alginate (EWA), as an alternative scaffold to Matrigel® for culturing 3D salivary gland cells. A protocol for an optimized EWA was established by comparing cell viability using 1%, 2%, and 3% alginate solution. The normal salivary simian virus 40-immortalized acinar cell (NS-SV-AC) and the submandibular gland-human-1 (SMG-hu-1) cell lines were also used to compare the spheroid formation and cell viability properties of both scaffold biomaterials; cell viability was observed over 10 days using a Live–Dead Cell Assay. Cell viability and spheroid size in 2% EWA was significantly greater than 1% and 3%. It is evident that EWA can support salivary cell survivability as well as form larger spheroids when compared to cells grown in Matrigel®. However, further investigations are necessary as it is unclear if cultured cells were proliferating or aggregating.

High-Throughput Screening Campaign Identified a Potential Small Molecule RXFP3/4 Agonist

Relaxin/insulin-like family peptide receptor 3 (RXFP3) belongs to class A G protein-coupled receptor family. RXFP3 and its endogenous ligand relaxin-3 are mainly expressed in the brain with important roles in the regulation of appetite, energy metabolism, endocrine homeostasis and emotional processing. It is therefore implicated as a potential target for treatment of various central nervous system diseases. Since selective agonists of RXFP3 are restricted to relaxin-3 and its analogs, we conducted a high-throughput screening campaign against 32,021 synthetic and natural product-derived compounds using a cyclic adenosine monophosphate (cAMP) measurement-based method. Only one compound, WNN0109-C011, was identified following primary screening, secondary screening and dose-response studies. Although displayed agonistic effect in cells overexpressing the human RXFP3, it also showed cross-reactivity with the human RXFP4. This hit compound may provide not only a chemical probe to investigate the function of RXFP3/4, but also a novel scaffold for the development of RXFP3/4 agonists.

Novel scaffold based graphene oxide doped electrospun iota carrageenan/polyvinyl alcohol for wound healing and pathogen reduction: in-vitro and in-vivo study

Wound healing is a complicated multicellular process that involves several kinds of cells including macrophages, fibroblasts, endothelial cells, keratinocytes and platelets that are leading to their differentiation towards an anti-inflammatory response for producing several chemokines, cytokine and growth factors. In this study, electrospun nanofiber scaffold named (MNS) is composed of polyvinyl alcohol (PVA)/iota carrageenan (IC) and doped with partially reduced graphene oxide (prGO) that is successfully synthesized for wound healing and skin repair. The fabricated MNS was tested in case of infection and un-infection with E. coli and Staphylococcus and in both of the presence and in the absence of yeast as a natural nutritional supplement. Numerous biochemical parameters including total protein, albumin, urea and LDH, and hematological parameters were evaluated. Results revealed that the MNS was proved to be effective on most of the measured parameters and had exhibited efficient antibacterial inhibition activity. Whereas it can be used as an effective antimicrobial agent in wound healing, however, histopathological findings confirmed that the MNS caused re-epithelialization and the presence of yeast induced hair follicles growth and subsequently it may be used to hide formed head wound scar.

Follow up coronary vessel evaluation of magnesium bioresorbable stents with coronary computed tomography angiography

Background Cardiac computed tomography angiography (CCTA) has already shown its ability to evaluate late results of polymer-based bioresorbable scaffolds (BVS) in different clinical scenarios. Recently, magnesium-based BVS (Mg-BVS) has emerged as an interesting alternative to these previous platforms due to its lower risk of thrombosis. Nonetheless, there is no systematic data about characterization of magnesium BVS with CCTA. Purpose To study the feasibility of Mg-BVS assessment with CCTA and to evaluate in-scaffold wall tissue characterization during the follow-up. Methods In this multicentre work, CCTA was performed in patients with a previously implanted Mg-BVS as a supplementary follow-up assessment. All studies were done after the theoretical resorption time of the scaffolds. A central core laboratory with an independent level 3 expert in CCTA blinded to the clinical and angiographic results analysed the studies. For this purpose, a dedicated software for coronary analysis was used to quantify coronary stenosis and evaluate coronary wall (Figure). Results Eight patients (55±6 years-old; 87.5% male) with a previously implanted Mg-BVS from 3 different centres in Spain and Italy were included. The presentation was equally distributed (2, 25%) among stable and unstable angina, NSTEMI and STEMI. Target vessels included 5 left anterior descending (62.5%), 2 left circumflex (25%) and 1 right coronary arteries (12.5%). CTCA was performed 13 [11.3–20] months after BVS implantation. In spite of the blinding, all scaffolds were accurately located through identification of proximal and distal radiopaque markers. Concordance of CCTA Mg-BVS sizing was good for diameter (ICC 0.66; p=0.09) and excellent for length (ICC 0.98; p<0,001) of the stents. Patency of all scaffolds was confirmed without significant diameter (0.14 [0–0.27]%) or area (0.39 [0.19–0.57]%) stenoses compared with proximal reference segments. Moreover, within the stent boundaries the maximum luminal diameter and area narrowing were 0.22 [0,12–0.3]% and 0.39 [0.23–0.5]% respectively, in keeping with mild in-scaffold late loss and/or underlying plaque growth. Regarding coronary wall tissue characterization of segments with BVS, there was a plaque burden of 0.37 [0.31–0.48]% and plaque volume of 87.6 [50.2–189.3] mm3. The most common component of the plaque was fibrous (69.5 [33.9–133.7]%), suggesting that Mg-BVS allows for stabilization of unstable culprit lesions (6/8 cases). Compared to the proximal and distal reference segments, there was no differences in plaque volume or composition, suggesting a good coronary vessel healing. Conclusions This short series shows for the first time the ability of CCTA to correctly locate and evaluate patency of Mg-BVS. Moreover, the lack of metal struts allows a detailed coronary plaque evaluation at treated segments. These preliminary results suggest plaque stabilization and a good coronary vessel healing with this novel scaffold. FUNDunding Acknowledgement Type of funding sources: None. Mg-BVS in LCx with mixed plaque

Successful Application of a Galanin-Coated Scaffold for Periodontal Regeneration

The nervous system exerts finely tuned control over all aspects of the life of an organism, including pain, sensation, growth, and development. Recent developments in tissue regeneration research have increasingly turned to small molecule peptides to tailor and augment the biological response following tissue loss or injury. In the present study, we have introduced the small molecule peptide galanin (GAL) as a novel scaffold-coating agent for the healing and regeneration of craniofacial tissues. Using immunohistochemistry, we detected GAL and GAL receptors in healthy periodontal tissues and in the proximity of blood vessels, while exposure to our periodontal disease regimen resulted in a downregulation of GAL. In a 3-dimensional bioreactor culture, GAL coating of collagen scaffolds promoted cell proliferation and matrix synthesis. Following subcutaneous implantation, GAL-coated scaffolds were associated with mineralized bone-like tissue deposits, which reacted positively for alizarin red and von Kossa, and demonstrated increased expression and protein levels of RUNX2, OCN, OSX, and iBSP. In contrast, the GAL receptor antagonist galantide blocked the effect of GAL on Runx2 expression and inhibited mineralization in our subcutaneous implantation model. Moreover, GAL coating promoted periodontal regeneration and a rescue of the periodontal defect generated in our periodontitis model mice. Together, these data demonstrate the efficacy of the neuropeptide GAL as a coating material for tissue regeneration. They are also suggestive of a novel role for neurogenic signaling pathways in craniofacial and periodontal regeneration.

Synthesis, characterization and molecular docking studies on some new N-substituted 2-phenylpyrido[2,3-d]pyrimidine derivatives

We report a novel scaffold of N-substituted 2-phenylpyrido(2,3-d)pyrimidine derivatives with potent antibacterial activity by targeting this biotin carboxylase enzyme. The series of eighteen N-substituted 2-phenylpyrido(2,3-d)pyrimidine derivatives were synthesized, characterized and further molecular docking studied to determine the mode of binding and energy changes with the crystal structure of biotin carboxylase (PDB ID: 2V58) was employed as the receptor with compounds 6a-r as ligands. The results obtained from the simulation were obtained in the form of dock score; these values represent the minimum energies. Compounds 6d, 6l, 6n, 6o, 6r and 6i showed formation of hydrogen bonds with the active site residues and van Der Walls interactions with the biotin carboxylase enzyme in their molecular docking studies. This compound can be studied further and developed into a potential antibacterial lead molecule.

Fabrication and Characterization of Silk Fibroin-Based Nanofibrous Scaffolds Supplemented with Gelatin for Corneal Tissue Engineering

Tissue engineering is a promising approach to overcome the severe worldwide shortage of healthy donor corneas. In this work, we have developed a silk-gelatin composite scaffold using electrospinning and permeation techniques to achieve the properties comparable to cornea analog. In particular, we present the fabrication and comparative evaluation of the novel gelatin sheets consisting of silk fibroin nanofibers, which are prepared using silk fibroin (SF) (in formic acid) and SF (in aqueous) electrospun scaffolds, for its suitability as corneal stromal analogs. All the fabricated samples were treated with ethanol vapor (T) to physically crosslink the silk nanofibers. Micro/nano-scale features of the fabricated scaffolds were analyzed using scanning electron microscopy micrographs. Fourier transform infrared spectroscopy revealed characteristic peaks of polymeric functional groups and modifications upon ethanol vapor treatment. Transparency of the scaffolds was determined using UV-visible spectra. Among all the fabricated samples, the gelatin-permeated SF (in formic acid; T) scaffold showed the highest level of transparency, i.e., 77.75 ± 2.3%, which is similar to that of the native cornea (∼70%–90% [variable with age group]) with healthy acute vision. Contact angle of the samples was studied to estimate the hydrophilicity of the scaffolds. All the scaffolds except non-treated SF (in aqueous; NT) were found to be significantly stable up to 14 days when incubated in phosphate buffered saline at 37°C. Treated samples showed significantly better stability, both physically and microscopically, in comparison to nontreated samples. Proliferation and viability assays of rabbit corneal fibroblast cells (SIRC) and mouse fibroblast cells (L929 RFP) when cultured on fabricated scaffolds revealed remarkable cellular compatibility with gelatin-permeated SF (in formic acid; T) scaffolds compared to SF (in aqueous; T). Unlike other reports in the existing literature, this work presents the design and development of a nanofibrous silk-gelatin composite that exhibits acceptable transparency, cellular biocompatibility, as well as improved mechanical stability comparable to that of native cornea. Therefore, we anticipate that the fabricated novel scaffold is likely to be a good candidate for corneal tissue construct. Moreover, among the fabricated scaffolds, the outcomes depict gelatin-permeated SF (in formic acid; T) composite scaffold to be a better candidate as a corneal stromal analog that carries properties of both the silk and gelatin, i.e., optimal transparency, better stability, and enhanced cytocompatibility.