Design of advanced siRNA therapeutics for the treatment of COVID-19.

COVID-19 is a newly emerged viral disease that is currently affecting the whole globe. A variety of therapeutic approaches are underway to block the SARS-CoV-2 virus. Among these methods, siRNAs could be a safe and specific option, as they have been tested against other viruses. siRNAs are a class of inhibitor RNAs that act promisingly as mRNA expression blockers and they can be designed to interfere with viral mRNA to block virus replication. In order to do this, we designed and evaluated the efficacy of six highly specific siRNAs, which target essential viral mRNAs with no predicted human genome off-targets. We observed a significant reduction in the copy number viral mRNAs after treatment with the siRNAs, and are expected to inhibit virus replication. We propose siRNAs as a potential co-therapy for acute SARS-CoV-2 infection.

Design of hydrogel-based scaffolds for the treatment of spinal cord injuries.

Spinal cord injury (SCI) is a traumatic lesion that diminishes sensory and/or motor neuronal functionality, directly affecting the quality of the patient's life. Due to the central nervous system's (CNS) inhibitory microenvironment that presents challenges in neuron repair and regeneration, tissue engineering strategies have received significant attention to improve the quality of a patient's life. In this regard, hydrogels are attractive SC scaffolds as they can provide not only an adjustable physiologically native-like microenvironment but also an appropriate matrix for cell delivery, drug delivery, and other bioactive molecule delivery at the lesion site. This systematic review characterizes the widely used biomaterials including natural polymers; protein- and polysaccharide-based synthetic polymers; methacrylate- and polyethylene glycol-based, and self-assembling (SA) peptides. In addition, synthesis routes of hydrogels are investigated. This review is complemented by the discussion of the various techniques utilized for hydrogel scaffold designs with their in vitro and in vivo outcomes and clinical trials. The existing challenges and opportunities for SC hydrogel scaffolds are mentioned towards the end of this review.

Fabrication, characterization and cytocompatibility assessment of gelatin nanofibers coated with a polymer thin film by initiated chemical vapor deposition.

The presence of various functional groups in the structure of gelatin nanofibers (GNFs) has made it a suitable candidate for biomedical applications, yet its fast dissolution in aqueous media has been a real challenge for years. In the present work, we propose an efficient procedure to improve the durability of the GNFs. The electrospun GNFs were coated with poly(ethylene glycol dimethacrylate) (pEGDMA) using initiated chemical vapor deposition (iCVD) as a completely dry polymerization method. Morphological and chemical analysis revealed that an ultrathin layer formed around nanofibers (iCVD-GNFs) which has covalently bonded to gelatin chains. Against the instant dissolution of GNFs, the in vitro biodegradability test showed the iCVD-GNFs, to a large extent, preserve their morphology after 14 days of immersion and did not lose its integrity even after 31 days. In vitro cell culture studies, also, revealed cytocompatibility of the iCVD-GNFs for human fibroblast cells (hFC), as well as higher cell proliferation on the iCVD-GNFs compared to control made from tissue culture plate (TCP). Furthermore, contact angle measurements indicated that the hydrophilic GNFs became hydrophobic after the iCVD, yet FE-SEM images of cell-seeded iCVD-GNFs showed satisfactory cell adhesion. Taken together, the proposed method paves a promising way for the production of water-resistant GNFs utilized in biomedical applications; for instance, tissue engineering scaffolds and wound dressings.

Cytotoxicity Evaluation of The Bioresorbable and Titanium Plates/Screws Used in Maxillofacial Surgery on Gingival Fibroblasts and Human Mesenchymal Bone Marrow Stem Cells.

OBJECTIVE: Bioresorbable and titanium plates/screws are considered as a standard treatment for fixation of the bone segments of craniofacial area and paying attention to their biocompatibility is an important issue along with other aspects of application. The purpose of the study was to evaluate the cell viability of two types of plate and screw used in maxillofacial surgeries in contact with gingival fibroblasts and bone marrow stem cells. MATERIALS AND METHODS: In this experimental study after extraction and cultivation of cells from healthy human gingival tissue and alveolar bone of jaw, cytotoxicity of device was evaluated. In direct contact method, samples had near vicinity contact with the both cell lines and in indirect contact method, by-products released, like ions, from samples after 8 weeks were used to assess cytotoxicity. Then cytotoxicity was evaluated on the 2nd, 4th and 6th day with MTS tests and microscopy. The data were analyzed by one-way ANOVA and independent t tests. RESULTS: There was a statistically significant difference between the German plate and screw and all the samples studied on day 6 (P<0.05). Furthermore, a statistically significant difference was observed between both metal samples and both bio-absorbable samples on day 6 and both cell lines (P<0.05). Comparisons between the two groups with each other for both cell lines on the 6th day were statistically significant (P<0.05). CONCLUSION: Our results suggest that that cytotoxicity of biomaterial, from different brands, were not similar and some of the biomaterial showed lower degree of toxicity compared to others and specialist using these products showed be aware of this differences. Our investigation indicates more biocompatibility of bioresorbable plates and screws compared to titanium. In addition our results suggest that biomaterials were not completely neutral.

3D Titania Nanofiber-Like Webs Induced by Plasma Ionization: A New Direction for Bioreactivity and Osteoinductivity Enhancement of Biomaterials.

In this study, we describe the formation method of web-like three-dimensional (3-D) titania nanofibrous structures coated on transparent substrate via a high intensity laser induced reverse transfer (HILIRT) process. First, we demonstrate the mechanism of ablation and deposition of Ti on the glass substrates using multiple picosecond laser pulses at ambient air in an explicit analytical form and compare the theoretical results with the experimental results of generated nanofibers. We then examine the performance of the developed glass samples coated by titania nanofibrous structures at varied laser pulse durations by electron microscopy and characterization methods. We follow this by exploring the response of human bone-derived mesenchymal stem cells (BMSCs) with the specimens, using a wide range of in-vitro analyses including MTS assay (colorimetric method for assessing cell metabolic activity), immunocytochemistry, mineralization, ion release examination, gene expression analysis, and protein adsorption and absorption analysis. Our results from the quantitative and qualitative analyses show a significant biocompatibility improvement in the laser treated samples compared to untreated substrates. By decreasing the pulse duration, more titania nanofibers with denser structures can be generated during the HILIRT technique. The findings also suggest that the density of nanostructures and concentration of coated nanofibers play critical roles in the bioreactivity properties of the treated samples, which results in early osteogenic differentiation of BMSCs.

A ternary nanofibrous scaffold potential for central nerve system tissue engineering.

In the present research, a ternary polycaprolactone (PCL)/gelatin/fibrinogen nanofibrous scaffold for tissue engineering application was developed. Through this combination, PCL improved the scaffold mechanical properties; meanwhile, gelatin and fibrinogen provided more hydrophilicity and cell proliferation. Three types of nanofibrous scaffolds containing different fibrinogen contents were prepared and characterized. Morphological study of the nanofibers showed that the prepared nanofibers were smooth, uniform without any formation of beads with a significant reduction in nanofiber diameter after incorporation of fibrinogen. The chemical characterization of the scaffolds confirmed that no chemical reaction occurred between the scaffold components. The tensile test results of the scaffolds showed that increasing in fibrinogen content led to a decrease in mechanical properties. Furthermore, adipose-derived stem cells were employed to evaluate cell-scaffold interaction. Cell culture results indicated that higher cell proliferation occurred for the higher amount of fibrinogen. Statistical analysis was also carried out to evaluate the significant difference for the obtained results of water droplet contact angle and cell culture. Therefore, the results confirmed that PCL/gel/fibrinogen scaffold has a good potential for tissue engineering applications including central nerve system tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A:2394-2401, 2018.

Activated platelet-rich plasma improves cartilage regeneration using adipose stem cells encapsulated in a 3D alginate scaffold.

In the current study, the effect of superimposing platelet-rich plasma (PRP) on different culture mediums in a three-dimensional alginate scaffold encapsulated with adipose-derived mesenchymal stem cells for cartilage tissue repair is reported. The three-dimensional alginate scaffolds with co-administration of PRP and/or chondrogenic supplements had a significant effect on the differentiation of adipose mesenchymal stem cells into mature cartilage, as assessed by an evaluation of the expression of cartilage-related markers of Sox9, collagen II, aggrecan and collagen, and glycosaminoglycan assays. For in vivo studies, following induction of osteochondral lesion in a rabbit model, a high degree of tissue regeneration in the alginate plus cell group (treated with PRP plus chondrogenic medium) compared with other groups of cell-free alginate and untreated groups (control) were observed. After 8 weeks, in the alginate plus cell group, functional chondrocytes were observed, which produced immature matrix, and by 16 weeks, the matrix and hyaline-like cartilage became completely homogeneous and integrated with the natural surrounding cartilage in the defect site. Similar effect was also observed in the subchondral bone. The cell-free scaffolds formed fibrocartilage tissue, and the untreated group did not form a continuous cartilage over the defect by 16 weeks.

In vivo integration of poly(ε-caprolactone)/gelatin nanofibrous nerve guide seeded with teeth derived stem cells for peripheral nerve regeneration.

Artificial nanofiber nerve guides have gained huge interest in bridging nerve gaps and associated peripheral nerve regeneration due to its high surface area, flexibility and porous structure. In this study, electrospun poly (ε-caprolactone)/gelatin (PCL/Gel) nanofibrous mats were fabricated, rolled around a copper wire and fixed by medical grade adhesive to obtain a tubular shaped bio-graft, to bridge 10 mm sciatic nerve gap in in vivo rat models. Stem cells from human exfoliated deciduous tooth (SHED) were transplanted to the site of nerve injury through the nanofibrous nerve guides. In vivo experiments were performed in animal models after creating a sciatic nerve gap, such that the nerve gap was grafted using (i) nanofiber nerve guide (ii) nanofiber nerve guide seeded with SHED (iii) suturing, while an untreated nerve gap remained as the negative control. In vitro cell culture study was carried out for primary investigation of SHED-nanofiber interaction and its viability within the nerve guides after 2 and 16 weeks of implantation time. Walking track analysis, plantar test, electrophysiology and immunohistochemistry were performed to evaluate functional recovery during nerve regeneration. Vascularization was also investigated by hematoxilin/eosine (H&E) staining. Overall results showed that the SHED seeded on nanofibrous nerve guide could survive and promote axonal regeneration in rat sciatic nerves, whereby the biocompatible PCL/Gel nerve guide with cells can support axonal regeneration and could be a promising tissue engineered graft for peripheral nerve regeneration.