It has been well recognized that the development and use of artificial materials with high osteogenic ability is one of the most promising means to replace bone grafting that has exhibited various negative effects. The biomimetic features and unique physiochemical properties of nanomaterials play important roles in stimulating cellular functions and guiding tissue regeneration. But efficacy degree of some nanomaterials to promote specific tissue formation is still not clear. We hereby comparatively studied the osteogenic ability of our treated multi-walled carbon nanotubes (MCNTs) and the main inorganic mineral component of natural bone, nano-hydroxyapatite (nHA) in the same system, and tried to tell the related mechanism. In vitro culture of human adipose-derived mesenchymal stem cells (HASCs) on the MCNTs and nHA demonstrated that although there was no significant difference in the cell adhesion amount between on the MCNTs and nHA, the cell attachment strength and proliferation on the MCNTs were better. Most importantly, the MCNTs could induce osteogenic differentiation of the HASCs better than the nHA, the possible mechanism of which was found to be that the MCNTs could activate Notch involved signaling pathways by concentrating more proteins, including specific bone-inducing ones. Moreover, the MCNTs could induce ectopic bone formation in vivo while the nHA could not, which might be because MCNTs could stimulate inducible cells in tissues to form inductive bone better than nHA by concentrating more proteins including specific bone-inducing ones secreted from M2 macrophages. Therefore, MCNTs might be more effective materials for accelerating bone formation even than nHA.

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Hydrogels are increasingly used in medicine due to their potential to be delivered into the body in a minimally invasive manner and to be gelated at the site of introduction subsequently. The aim of this study was to develop a novel injectable and in situ-forming gel composite (GC) comprised of calcium alginate hydrogel and nano-hydroxyapatite/collagen (nHAC), assess its rheological, mechanical and in vitro degradable properties, and discuss the gelation mechanism. Injectable property test showed that the injectability of GC was tunable. Rheological results indicated that three phases of pre-gel, sol-gel phase transformation and post-gel could be found in the process of gelation. The compressive elastic modulus (E) and shear modulus (G) are in the range of 17.0-56.0 kPa and 24.7-55.0 kPa, respectively. During the in vitro degradation, the wet weight increased in the first week, then declined in the following 3 weeks, but the dry weight lost continuously during whole study. Meanwhile, the surface changed greatly after 2 weeks, but samples did not break down up to 28 days. These data indicate that GC exhibits controllable initial setting time and final setting time, tunable injectability, which provides a possible injectable material for bone repair and bone tissue engineering.

The mucus layer of the nasopharynx and bronchial epithelium has a barrier function against inhaled pathogens such as the coronavirus SARS-CoV-2. We recently found that inorganic polyphosphate (polyP), a physiological, metabolic energy (ATP)-providing polymer released from blood platelets, blocks the binding of the receptor binding domain (RBD) to the cellular ACE2 receptor in vitro. PolyP is a marine natural product and is abundantly present in marine bacteria. Now, we have approached the in vivo situation by studying the effect of polyP on the human alveolar basal epithelial A549 cells in a mucus-like mucin environment. These cells express mucins as well as the ectoenzymes alkaline phosphatase (ALP) and adenylate kinase (ADK), which are involved in the extracellular production of ATP from polyP. Mucin, integrated into a collagen-based hydrogel, stimulated cell growth and attachment. The addition of polyP to the hydrogel significantly increased cell attachment and also the expression of the membrane-tethered mucin MUC1 and the secreted mucin MUC5AC. The increased synthesis of MUC1 was also confirmed by immunostaining. This morphogenetic effect of polyP was associated with a rise in extracellular ATP level. We conclude that the nontoxic and non-immunogenic polymer polyP could possibly also exert a protective effect against SARS-CoV-2-cell attachment; first, by stimulating the innate antiviral response by strengthening the mucin barrier with its antimicrobial proteins, and second, by inhibiting virus attachment to the cells, as deduced from the reduction in the strength of binding between the viral RBD and the cellular ACE2 receptor.

Background: Three-dimensional (3D) cultivation with biomaterials was proposed to facilitate stem cell epithelial differentiation for wound healing. However, whether human adipose-derived stem cells (hASCs) on collagen sponge scaffold (CSS) better differentiate to keratinocytes remains unclear. Methods: 3D cultivation with CSS on hASC epidermal differentiation co-cultured with HaCaT cells at air-liquid interface (ALI) was compared with two-dimensional (2D) form and cultivation without "co-culture" or "ALI." Cellular morphology, cell adhesion, and growth condition were evaluated, followed by the protein and gene expression of keratin 14 (K14, keratinocyte specific marker). Results: Typical cobblestone morphology of keratinocytes was remarkably observed in co-cultured hASCs at ALI, but those seeded on the CSS exhibited more keratinocyte-like cells under an invert microscope and scanning electron microscope. Desired cell adhesion and proliferation were confirmed in 3D differentiation groups by rhodamine-labeled phalloidin staining, consistent with H&E staining. Compared with those cultured in 2D culture system or without "ALI," immunofluorescence staining and gene expression analysis revealed hASCs co-cultured over CSS expressed K14 at higher levels at day 15. Conclusions: CSS is positive to promote epithelial differentiation of hASCs, which will foster a deeper understanding of artificial dermis in skin wound healing and regeneration.

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