Quaternized Chitosan Derivatives as Viable Antiviral Agents: Structure–Activity Correlations and Mechanisms of Action

The authors would like to thank Ahmed and Prof. Mauri Kostiainen for help in cryo-TEM sample preparation and image acquisition and Dr. Leena Pitkänen for help toward GPC analysis. The authors would like to thank the Academy of Finland (project number 133355163 SA/COVID-19) for providing funding for this project. I.L. would like to thank Svenska Kulturfonden for the PhD scholarship. This work made use of the BIOECONOMY and OTANANO infrastructure at the Aalto University.Cationic polysaccharides have demonstrated significant antimicrobial properties and have great potential in medical applications, where the antiviral activity is of great interest. As of today, alcohols and oxidizing agents are commonly used as antiviral disinfectants. However, these compounds are not environmentally safe, have short activity periods, and may cause health issues. Therefore, this study aimed to develop metal-free and environmentally friendly quaternary chitosans (QCs) with excellent long-lasting virucidal activity. To evaluate this, both single and double QCs were obtained using AETMAC ([2-(acryloyloxy)ethyl]-trimethylammonium chloride) and GTMAC (glycidyl trimethylammonium chloride) quaternary precursors. Further, this study investigated the influence of the quaternary functional group, charge density, and molecular weight (Mw) on the antiviral properties of QCs. It is proposed that the higher charge density, along with the length of alkyl linkers, and hydrophobic interactions affected the antiviral activity of QCs. The findings demonstrated that heterogeneously functionalized chitosan exhibited excellent antiviral activity against both the enveloped virus φ6 and the nonenveloped viruses φX174 and MS2. These quaternized chitosan derivatives have promising potential as viable antiviral agents, as hand/surface sanitizers, or in other biomedical applications.Peer reviewe

Polymeric Drug Delivery Systems by Additive Manufacturing

Additive manufacturing (AM) is gaining interests in drug delivery applications, offering innovative opportunities for the design and development of systems with complex geometry and programmed controlled release profile. In addition, polymer-based drug delivery systems can improve drug safety, efficacy, patient compliance, and are the key materials in AM. Therefore, combining AM and polymers can be beneficial to overcome the existing limitations in the development of controlled release drug delivery systems. Considering these advantages, here we are focusing on the recent developments in the field of polymeric drug delivery systems prepared by AM. This review provides a comprehensive overview on a holistic polymer–AM perspective for drug delivery systems with discussion on the materials, properties, design and fabrication techniques and the mechanisms used to achieve a controlled release system. The current challenges and future perspectives for personalized medicine and clinical use of these systems are alsobriefly discussed.Peer reviewe

A minimally-invasive cryogel based approach for the development of human ectopic liver in a mouse model

Contains fulltext : 206887.pdf (publisher's version ) (Closed access)11 p

Direct ink writing of biocompatible chitosan/non-isocyanate polyurethane/cellulose nanofiber hydrogels for wound-healing applications

The demand for new biocompatible and 3D printable materials for biomedical applications is on the rise. Ideally, such materials should exhibit either biodegradability or recyclability, possess antibacterial properties, and demonstrate remarkable biocompatibility with no cytotoxic effects. In this research, we synthesized biocompatible and 3D printable hydrogels tailored for biomedical applications, such as wound healing films, by combining antibacterial double-quaternized chitosan (DQC) with cystamine-based non-isocyanate polyurethane (NIPU-Cys) - a material renowned for enhancing both the flexibility and mechanical properties of the hydrogels. To improve the rheological behavior, swelling attributes, and printability, cellulose nanofibrils were introduced into the matrix. We investigated the impact of DQC on degradability, swelling capacity, rheological behavior, printability, and cell biocompatibility. The slightly cytotoxic nature associated with quaternary chitosan was evaluated, and the optimal concentration of DQC in the hydrogel was determined to ensure biocompatibility. The resulting hydrogels were found to be suitable materials for 3D printing via a direct ink writing technique (DIW), producing porous, biocompatible hydrogels endowed with valuable attributes suitable for various wound-healing applications.Peer reviewe

Endogenous Platelet-Rich Plasma Supplements/Augments Growth Factors Delivered via Porous Collagen-Nanohydroxyapatite Bone Substitute for Enhanced Bone Formation

Polymer (acrylate) and ceramic bone cements are extensively used as bone void fillers and for implant fixation in orthopedics. These materials have micro- to nonporous architectures. Postimplantation, they may cause hypoxic and exothermic injuries to already compromised damage site. These materials also have limited interaction with surrounding tissue. In this work we have developed composite collagen-nanohydroxyapatite (CS) bone filler, mimicking porous architecture of trabecular bone. It was functionalized with clinically available bone active agents like bone morphogenetic protein-2 (rhBMP-2) and zoledronic acid (ZA). We investigated synergistic effects of the bone active molecules and endogenous platelet rich plasma (PRP), a source of growth factors on mineralization. Porous CS and collagen/gelatin/chiotosan polymer scaffold (SC) (without nanohydroxyapatite) were synthesized using cryogelation. PRP (10 μL) (∼5 × 10⁶ cells), rhBMP-2 (5 μg) and ZA (10 μg) were used to functionalize scaffolds. Bone formation was evaluated at ectopic sites in abdominal pouch and 4.0 mm critical defect in tibia metaphysis of rats. Tissue mineralization was evaluated by micro-CT and histological analysis 12 weeks postimplantation. In vitro cell based studies revealed, PRP functionalization enhances osteoblast proliferation and activity on scaffolds. In vivo BMP+ZA+PRP functionalized scaffolds had higher amount (28 mm³) of mineralized tissue formation as compared to empty defect (20 mm³), suggesting that PRP can augment the osteoinductive properties of functionalized scaffolds both in vitro and in vivo. Enhanced cell infiltration and mineralization can be achieved via CS in comparison to SC, implying their use as porous bone void fillers and substitutes for autografts

Additional file 1: of Mesenchymal stromal cell-derived exosome-rich fractionated secretome confers a hepatoprotective effect in liver injury

presents additional information on methods to isolate and characterize MSCs; how to characterize the exosome-rich fractionated secretome using microscopy, flow cytometry, ELISA and western blot techniques; quantifying ROS activity in cells; performing qualitative fluorescence microscopy analysis for the in-vitro experiments; and calculation of the liver regeneration rat

Gelatin-Modified Bone Substitute with Bioactive Molecules Enhance Cellular Interactions and Bone Regeneration

In this work, we have synthesized injectable bone cement incorporated with gelatin to enhance cellular interaction. Human osteosarcoma Saos-2 cells derived bone morphogenetic proteins (BMP's) and a bisphosphonate (zoledronic acid (0.2 mM)) were also incorporated to cement. In vitro studies conducted using Saos-2 demonstrated enhanced cell proliferation on gelatin (0.2%w/v) cement. The differentiation of C2C12 mouse myoblast cells into bone forming cells showed 6-fold increase in ALP levels on gelatin cement. Polymerase chain reaction (PCR) for bone biomarkers showed osteoinductive potential of gelatin cement. We investigated efficacy for local delivery of these bioactive molecules in enhancing bone substitution qualities of bone cements by implanting in 3.5 mm critical size defect in tibial metaphysis of wistar rats. The rats were sacrificed after 12 weeks and 16 weeks post implantation. X-ray, micro-CT, histology, and histomorphometry analysis were performed to check bone healing. The cement materials slowly resorbed from the defect site leaving HAP creating porous matrix providing surface for bone formation. The materials showed high biocompatibility and initial bridging was observed in all the animals but maximum bone formation was observed in animals implanted with cement incorporated with zoledronic acid followed by cement with BMP's compared to other groups

Characterisation of porous knitted titanium for replacement of intervertebral disc nucleus pulposus

Effective restoration of human intervertebral disc degeneration is challenged by numerous limitations of the currently available spinal fusion and arthroplasty treatment strategies. Consequently, use of artificial biomaterial implant is gaining attention as a potential therapeutic strategy. Our study is aimed at investigating and characterizing a novel knitted titanium (Ti6Al4V) implant for the replacement of nucleus pulposus to treat early stages of chronic intervertebral disc degeneration. Specific knitted geometry of the scaffold with a porosity of 67.67 ± 0.824% was used to overcome tissue integration failures. Furthermore, to improve the wear resistance without impairing original mechanical strength, electro-polishing step was employed. Electro-polishing treatment changed a surface roughness from 15.22 ± 3.28 to 4.35 ± 0.87 μm without affecting its wettability which remained at 81.03 ± 8.5°. Subsequently, cellular responses of human mesenchymal stem cells (SCP1 cell line) and human primary chondrocytes were investigated which showed positive responses in terms of adherence and viability. Surface wettability was further enhanced to super hydrophilic nature by oxygen plasma treatment, which eventually caused substantial increase in the proliferation of SCP1 cells and primary chondrocytes. Our study implies that owing to scaffolds physicochemical and biocompatible properties, it could improve the clinical performance of nucleus pulposus replacement

Additional file 2: Figure S1. of Mesenchymal stromal cell-derived exosome-rich fractionated secretome confers a hepatoprotective effect in liver injury

showing characterization of rat bone marrow-derived mesenchymal stem cells. Phase-contrast microscopic image of cultured MSCs showing fibroblast-like spindle-shaped morphology (A), fluorescence microscopic image of MSCs stained with fluorescein diacetate (FDA) (B), digital image of crystal violet staining of colonies of MSCs formed (C), microscopic image of MSCs stained with crystal violet (D), Alizarin Red staining of undifferentiated MSCs (E) and osteogenically differentiated MSCs (F) and Oil Red O staining of undifferentiated MSCs (G) and adipogenically differentiated MSCs (H). Scale bar for all microscopic images: 100 Οm (TIF 2112 kb

Biocomposite macroporous cryogels as potential carrier scaffolds for bone active agents augmenting bone regeneration

Osteoinduction can be enhanced by combining scaffolds with bone morphogenic protein-2 (BMP-2). However, BMP's are known to also cause bone resorption. This can be controlled using bisphosphonates like zoledronic acid (ZA). In this study, we produced two different scaffolds containing silk-fibroin, chitosan, agarose and hydroxyapatite (HA) with and without bioactive glass. The aims of the study were to fabricate, physico-chemically characterize and evaluate the carrier properties of the scaffolds for recombinant human BMP-2 (rhBMP-2) and ZA. Scaffolds were characterized using various methods to confirm their composition. During cell-material interactions, both scaffolds exhibited gradual but sustained proliferation of both C2C12 and MSCs for a period of 6 weeks with augmentative effects on their phenotype indicated by elevated levels of alkaline phosphatase (ALP) cuing towards osteogenic differentiation. In-vitro effects of rhBMP-2 and ZA contained within both the scaffolds was assessed on MC3T3 preosteoblast cells and the results show a significant increase in the ALP activity of the cells seeded on scaffolds with rhBMP-2. Further, the scaffold with both HA and bioactive glass was considered for the animal study. In-vitro, this scaffold released nearly 25% rhBMP-2 in 21-days and the addition of ZA did not affect the release. In the animal study, the scaffolds were combined with rhBMP-2 and ZA, rhBMP-2 or implanted alone in an ectopic muscle pouch model. Significantly higher bone formation was observed in the scaffold loaded with both rhBMP-2 and ZA as seen from micro-computed tomography, histomorphometry and energy dispersive X-ray spectroscopy