Loose Pre-Cross-Linking Mediating Cellulose Self-Assembly for 3D Printing Strong and Tough Biomimetic Scaffolds.

The lack of an effective printable ink preparation method and the usual mechanically weak performance obstruct the functional 3D printing hydrogel exploitation and application. Herein, we propose a gentle pre-cross-linking strategy to enable a loosely cross-linked cellulose network for simultaneously achieving favorable printability and a strong hydrogel network via mediating the cellulose self-assembly. A small amount of epichlorohydrin is applied to (i) slightly pre-cross-link the cellulose chains for forming the percolating network to regulate the rheological properties and (ii) form the loosely cross-linked points to mediate the cellulose chains' self-assembly for achieving superior mechanical properties. The fabrication of the complex 3D structures verifies the design flexibility. The printed cellulose hydrogels exhibit a biomimetic nanofibrous topology, remarkable tensile and compressive strength (5.22 and 11.80 MPa), as well as toughness (1.81 and 2.16 MJ/m3). As a demonstration, a bilayer scaffold (mimicking the osteochondral structure) consisting of a top pristine cellulose and a bottom cellulose/bioactive glass hydrogel is printed and exhibits superior osteochondral defect repair performance, showing a potential in tissue engineering. We anticipate that our loose pre-cross-linking 3D printing ink preparation concept can inspire the development of other polymeric inks and strong 3D printing functional hydrogels, eventually spreading the applications in diverse fields.

3D printing of Cu-doped bioactive glass composite scaffolds promotes bone regeneration through activating the HIF-1α and TNF-α pathway of hUVECs.

The increasing insight into the molecular and cellular processes within the angiogenic cascade assists in enhancing the survival and integration of engineered bone constructs. Copper-doped bioactive glass (Cu-BG) is now a potential structural component of the novel scaffolds and implants used in orthopedic and dental repairs. However, it is difficult for BG, especially micro-nano particles, to be printed into scaffolds and still retain its biological activity and ability to biodegrade. Additionally, the mechanisms of the copper-stimulating autocrine and paracrine effects of human umbilical vein endothelial cells (hUVECs) during repair and regeneration of bone are not yet clear. Therefore, in this study, we created monodispersed micro-nano spherical Cu-BG particles with varying copper content through a sol-gel process. Through in vitro tests, we found that Cu-BG enhanced angiogenesis by activating the pro-inflammatory environment and the HIF-1α pathway of hUVECs. Furthermore, 2Cu-BG diluted extracts directly promoted the osteogenic differentiation of mouse bone mesenchymal stem cells (BMSCs) in vitro. Then, a new 3D-printed tyramine-modified gelatin/silk fibroin/copper-doped bioactive glass (Gel/SF/Cu-BG) scaffold for rat bone defects was constructed, and the mechanism of the profound angiogenesis effect regulated by copper was explored in vivo. Finally, we found that hydrogel containing 1 wt% 2Cu-BG effectively regulated the spatiotemporal coupling of vascularization and osteogenesis. Therefore, Cu-BG-containing scaffolds have great potential for a wide range of bone defect repairs.

3D printed silk-gelatin hydrogel scaffold with different porous structure and cell seeding strategy for cartilage regeneration.

Hydrogel scaffolds are attractive for tissue defect repair and reorganization because of their human tissue-like characteristics. However, most hydrogels offer limited cell growth and tissue formation ability due to their submicron- or nano-sized gel networks, which restrict the supply of oxygen, nutrients and inhibit the proliferation and differentiation of encapsulated cells. In recent years, 3D printed hydrogels have shown great potential to overcome this problem by introducing macro-pores within scaffolds. In this study, we fabricated a macroporous hydrogel scaffold through horseradish peroxidase (HRP)-mediated crosslinking of silk fibroin (SF) and tyramine-substituted gelatin (GT) by extrusion-based low-temperature 3D printing. Through physicochemical characterization, we found that this hydrogel has excellent structural stability, suitable mechanical properties, and an adjustable degradation rate, thus satisfying the requirements for cartilage reconstruction. Cell suspension and aggregate seeding methods were developed to assess the inoculation efficiency of the hydrogel. Moreover, the chondrogenic differentiation of stem cells was explored. Stem cells in the hydrogel differentiated into hyaline cartilage when the cell aggregate seeding method was used and into fibrocartilage when the cell suspension was used. Finally, the effect of the hydrogel and stem cells were investigated in a rabbit cartilage defect model. After implantation for 12 and 16 weeks, histological evaluation of the sections was performed. We found that the enzymatic cross-linked and methanol treatment SF5GT15 hydrogel combined with cell aggregates promoted articular cartilage regeneration. In summary, this 3D printed macroporous SF-GT hydrogel combined with stem cell aggregates possesses excellent potential for application in cartilage tissue repair and regeneration.

A medical adhesive used in a wet environment by blending tannic acid and silk fibroin.

A multifunctional and effective medical adhesive with a combination of high toughness and superior adhesion is highly desired in biomedical fields. However, clinical application of medical adhesives is still limited due to their weak adhesion to wet tissue. In this study, a novel medical adhesive called TASK composed of tannic acid (TA) and silk fibroin (SF) based on polyphenol-gel systems was developed. TASK powder was prepared by a simple physical mixture of pyrogallol-rich tannic acid and silk fibroin in aqueous solution and further freeze drying, which was stable and convenient for sterilization before clinical application. The TASK composite gel was formed by just adding water to the TASK powder. TASK showed improved wet-adhesive properties and stability; its adhesion strength after 5 h in water reached 180.9 ± 27.4 kPa. ATR-FTIR results indicated that the plentiful phenolic hydroxyl groups in TA allowed TASK to maintain adhesion to tissue in a wet environment. Furthermore, no chemical modification or covalent cross-linking was required for this wet-adhesive TASK which may facilitate its clinical application.

Sequence analysis of F gene of SF02 isolate of goose paramyxovirus and its identification by multiplex RT-PCR.

The complete F gene of SF02 of goose paramyxovirus (GPV) has been cloned and analyzed. The sequence analysis demonstrated that the F gene of SF02 contains 1 662 nt and encodes 553 amino acids, and its cleavage activation site of F gene has the same deduced amino acid sequence, (112)R-R-Q-K-R-F(117), as the velogenic (highly pathogenic) strain of newcastle disease virus. The latter correlated with the virulence of the isolate in biological assays. The F gene of SF02 isolate with the domestic standard velogenic strain NDV, F48E9, shared 86.5% homology in nucleotide and 90.8% homology in amino acid sequences. The SF02 isolate is closer to some NDV strains prevalent in Taiwan and West-European countries in recent years. Based on the F gene sequence a multiplex RT-PCR method has been developed. It could be used for the discrimination of GPV from NDV.