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Islet transplantation is currently considered the most promising method for treating insulin-dependent diabetes. The two most-studied artificial islets are alginate-encapsulated β cells or β cell spheroids. As three-dimensional (3D) models, both artificial islets have better insulin secretory functions and transplantation efficiencies than cells in two-dimensional (2D) monolayer culture. However, the effects of these two methods have not been compared yet. Therefore, in this study, cells from the mouse islet β cell line Min6 were constructed as scaffold-free spheroids or alginate-encapsulated dispersed cells.MIN6 cell spheroids were prepared by using Agarose-base microwell arrays. The insulin secretion level was determined by mouse insulin ELISA kit, and the gene and protein expression status of the MIN6 were performed by Quantitative polymerase chain reaction and immunoblot, respectively.Both 3D cultures effectively promoted the proliferation and glucose-stimulated insulin release (GSIS) of MIN6 cells compared to 2D adherent cells. Furthermore, 1% alginate-encapsulated MIN6 cells demonstrated more significant effects than the spheroids. In general, three pancreatic genes were expressed at higher levels in response to the 3D culture than to the 2D culture, and pancreatic/duodenal homeobox-1 (PDX1) expression was higher in the cells encapsulated in 1% alginate than that in the spheroids. A western blot analysis showed that 1% alginate-encapsulated MIN6 cells activated the phosphoinositide 3-kinase (PI3K)/serine/threonine protein kinase (AKT)/forkhead transcription factor FKHR (FoxO1) pathway more than the spheroids, 0.5% alginate-, or 2% alginate-encapsulated cells did. The 3D MIN6 culture, therefore, showed improved effects compared to the 2D culture, and the 1% alginate-encapsulated MIN6 cells exhibited better effects than the spheroids. The upregulation of PDX1 expression through the activation of the PI3K/AKT/FoxO1 pathway may mediate the improved cell proliferation and GSIS in 1% alginate-encapsulated MIN6 cells.This study may contribute to the construction of in vitro culture systems for pancreatic islets to meet clinical requirements.

The early stages of progressive degeneration of cartilage in articular joints are a hallmark of osteoarthritis. Healthy cartilage is lubricated by brush-like cartilage-binding nanofibres with a hyaluronan backbone and two key side chains (lubricin and lipid). Here, we show that hyaluronan backbones grafted with lubricin-like sulfonate-rich polymers or with lipid-like phosphocholine-rich polymers together enhance cartilage regeneration in a rat model of early osteoarthritis. These biomimetic brush-like nanofibres show a high affinity for cartilage proteins, form a lubrication layer on the cartilage surface and efficiently lubricate damaged human cartilage, lowering its friction coefficient to the low levels typical of native cartilage. Intra-articular injection of the two types of nanofibre into rats with surgically induced osteoarthritic joints led to cartilage regeneration and to the abrogation of osteoarthritis within 8 weeks. Biocompatible injectable lubricants that facilitate cartilage regeneration may offer a translational strategy for the treatment of early osteoarthritis. Brush-like biopolymers mimicking the lubrication properties of natural cartilage-binding complexes in articular joints enhance cartilage regeneration in a rat model of early osteoarthritis.

Coaxial bioprinting of hydrogel tubes has tremendous potential in the fabrication of highly complex large-scale vascularized structures, however, constructs with bioinks of simultaneous weak printability and perfusable networks have not been reported. Here, we report a coaxial printing method in which double-channel filaments are three-dimensional (3D) extrusion-bioprinted using a customized dual-core coaxial nozzle. The filament in one channel can perform core/shell role and the other channel can play a role in perfusion. These parallel channels within filaments are separated by an interval wall of alginate, whose thickness (∼50 μm) is beneficial to supplement nutrients via perfusion. Different cell-laden hydrogels of weak mechanics were used to test the adaptability and perfusability of our method, and the results showed that dynamic perfusion maintained higher viability and functions than static culture. By combining with a bioprinter, 8-layer perfusable double-channel constructs were fabricated, and the cell viabilities gradually decreased with the reduction in nutrients and oxygen in the downstream medium. Furthermore, the double-channel filaments were tested as a platform to mimic dynamic functions between cells through sequential perfusion by using Mouse insulinoma 6 (Min6) and Hepatocellular carcinoma (HepG2) as the model cells. These results demonstrated the insulin secreted by Min6 upstream simulated and increased the uptake of glucose by the downstream HepG2 cells. In conclusion, our study provided evidence for the probability of all-in-one fabrication of 3D double-channel perfusable constructs with high simplicity, expansibility, and versability. Our strategy has significant potential for building large-scale tissue constructs for applications in tissue engineering, possibly even in drug screening and regenerative medicine.

The remarkable lubrication properties of normal articular cartilage play an essential role in daily life, providing almost frictionless movements of joints. Alterations of cartilage surface or degradation of biomacromolecules within synovial fluid increase the wear and tear of the cartilage and hence determining the onset of the most common joint disease, osteoarthritis (OA). The irreversible and progressive degradation of articular cartilage is the hallmark of OA. Considering the absence of effective options to treat OA, the mechanosensitivity of chondrocytes has captured attention. As the only embedded cells in cartilage, the metabolism of chondrocytes is essential in maintaining homeostasis of cartilage, which triggers motivations to understand what is behind the low friction of cartilage and develop biolubrication-based strategies to postpone or even possibly heal OA. This review firstly focuses on the mechanism of cartilage lubrication, particularly on boundary lubrication. Then the mechanotransduction (especially shear stress) of chondrocytes is discussed. The following summarizes the recent development of cartilage-inspired biolubricants to highlight the correlation between cartilage lubrication and OA. One might expect that the restoration of cartilage lubrication at the early stage of OA could potentially promote the regeneration of cartilage and reverse its pathology to cure OA.

The tribological properties of two novel biomimetic multihierarchical polymers, synthesized by covalently linking single bottlebrush polymers onto a hyaluronic acid (HA) backbone, were investigated in the boundary lubrication regime using the surface forces apparatus. The polymers were immobilized on flat substrates, and their lubrication properties and wear resistance were investigated in aqueous media in the absence of a polymer reservoir (i.e., no free polymer chains in the surrounding medium) in order to better reveal the underlying mechanism of surface-attached biomimetic polymers. The effects of composition, structure, and, more particularly, surface attachment (physisorbed vs chemisorbed) on the tribological properties were investigated and compared with other biomimicking systems reported in the literature. The covalently surface attached bottlebrushes allowed wear resistance between sliding surfaces to be significantly improved, compared to physisorbed bottlebrushes, with a constant coefficient of friction (10

The early stages of progressive degeneration of cartilage in articular joints are a hallmark of osteoarthritis. Healthy cartilage is lubricated by brush-like cartilage-binding nanofibres with a hyaluronan backbone and two key side chains (lubricin and lipid). Here, we show that hyaluronan backbones grafted with lubricin-like sulfonate-rich polymers or with lipid-like phosphocholine-rich polymers together enhance cartilage regeneration in a rat model of early osteoarthritis. These biomimetic brush-like nanofibres show a high affinity for cartilage proteins, form a lubrication layer on the cartilage surface and efficiently lubricate damaged human cartilage, lowering its friction coefficient to the low levels typical of native cartilage. Intra-articular injection of the two types of nanofibre into rats with surgically induced osteoarthritic joints led to cartilage regeneration and to the abrogation of osteoarthritis within 8 weeks. Biocompatible injectable lubricants that facilitate cartilage regeneration may offer a translational strategy for the treatment of early osteoarthritis.

In this study, a series of β-cyclodextrins (β-CD) polyrotaxane multi-aldehydes with different molecular weight were prepared and then were used as crosslinker for collagen. The properties of crosslinked collagen were characterized by Fourier transform infrared spectrophotometry (FTIR), collagenase degradation assay, mechanical testing, scanning electron microscope (SEM) and in vitro cell studies. Compared to crosslinked by conditional crosslinkers, such as glutaraldehyde (GA) and N-ethyl-N-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), the collagen crosslinked by the novel biocrosslinker has been improved effectively in biodegradation rate, compress modulus and show lower cytotoxicity. Besides, the modified collagen was suitable for the cell adhesion and proliferation due to the appropriate swelling property, porosity and pore size. Considering the excellent properties, β-cyclodextrins (β-CD) polyrotaxane multi-aldehydes is a novel crosslinker for collagen and the obtained collagen gel shows great potential to use as tissue engineering scaffold.

Glucosamine (GlcN) is a component of native cartilage extracellular matrix and useful in cartilage repair, but it was limited by toxicity in high concentrations. With the aim of altering bioactive properties of GlcN to reduce the toxicity and to facilitate chondrogenesis for hyaline cartilage formation, we introduced an amino-group modification with double bond into GlcN to produce N-acryloyl-glucosamine (AGA). The cell ATDC5 was chosen to evaluate its cytotoxicity and chondrogenesis capability. Cell proliferation and cytotoxicity assay showed that AGA had significantly reduced the cytotoxicity compared to GlcN, and promoted ATDC5 proliferation. Alcian blue staining and biochemical analysis indicated that AGA enhanced extracellular matrix deposition. Both the mRNA and protein levels of articular cartilage markers, like Collagen II and Aggrecan were up-regulated, as shown by quantitative real-time PCR and immunofluorescence staining. Moreover, the level of fibrocartilage marker Collagen I and hypertrophic marker Collagen Χ weren't significantly changed. Overall, these results demonstrated that the AGA achieved the functional double-bond, reduction in toxicity and enhancement in chondrogenesis could be more potential in cartilage repair.

Glucosamine (GA) is an important cartilage matrix precursor for the glycosaminoglycan biochemical synthesis, and has positive effects on cartilage regeneration, particularly in osteoarthritis therapy. However, it has not been used as a bioactive group in scaffolds for cartilage repair widely. In this study, we synthesized modified polyethylene glycol (PEG) hydrogel with glucosamine and then encapsulated human bone mesenchymal stem cells (hBMSCs) in the hydrogel to induce the differentiation of hBMSCs into chondrocytes in three-dimensional culture. The GA-modified PEG hydrogels promoted the chondrogenesis of hBMSCs, particularly in the concentration of 5mM and 10mM. The subcutaneous transplantation of 10mM GA-modified hydrogels with hBMSCs formed cartilage-like blocks in vivo for 8weeks. Importantly, with glucosamine increase, the modified hydrogels down-regulated the fibrosis and hypertrophic cartilage markers in protein level. Therefore, glucosamine modified PEG hydrogels facilitated the chondrogenesis of hBMSCs, which might represent a new method for cartilage repair using a tissue-engineering approach.

Cartilage repairing grafts have been widely studied, and osteochondral replacement hydrogels have proven to be an excellent method in research and clinical fields. However, it has been difficult to simultaneously solve three main issues in osteochondral replacement preparation: surface lubrication, overall mechanical support and good simulations of cell regeneration. A novel integrated bilayered hydrogel osteochondral replacement was constructed by blending polyvinyl alcohol (PVA) and β-tricalcium phosphate (β-TCP) in this study. Separated nano-ball milling with ultrasound dispersion prepared β-TCP demonstrated suitable properties of tiny particle size, high purity and ideal distribution, improving the mechanical properties of the novel integrated hydrogel, and providing a cartilage-like lubrication effect and high biocompatibility, including cytocompatibility and osteogenesis. The reinforcement of β-TCP and integrated molding technology enabled the hydrogel to demonstrate excellent component compatibility and good bonding between the two layers, which promoted the strengthening of the compression modulus and tensile modulus up to three times by mechanical testing. The surface lubrication properties of the novel osteochondral hydrogel were similar to the natural cartilage by friction coefficient characterization. The two layers of the novel integrated graft provided a considerable bio-function by co-culturing with chondrocytes and synovium mesenchymal stem cells: chondrocytes promoted adherence achieved by the upper density layer and better osteogenesis performance of the porous lower layer. The design of the bilayered β-TCP/PVA osteochondral hydrogel is promising for use in articular cartilage repair.