The study investigated the potential of electrospun fiber assembled hydrogel, with physical gradients of chondroitin sulfate (CS) and sol-gel-derived bioactive glass (BG), to engineer hyaline and mineralized cartilage in a single 3D system. Electrospun poly(caprolactone) (PCL) fibers incorporated with 0.1% w/w of CS (CSL) and 0.5% w/w of CS (CSH), 2.4% w/w of BG (BGL) and 12.5% w/w of BG (BGH) were fabricated. The CS showed a sustained release up to 3 days from CSL and 14 days from CSH fibers. Chondrocytes secreted hyaline like matrix with higher sulfated glycosaminoglycans (sGAG), collagen type II and aggrecan on CSL and CSH fibers. Mineralization was observed on BGL and BGH fibers when incubated in simulated body fluid for 14 days. Chondrocytes cultured on these fibers secreted a mineralized matrix that consisted of sGAG, hypertrophic proteins, collagen type X, and osteocalcin. The CS and BG incorporated PCL fiber mats were assembled in an agarose-gelatin hydrogel to generate a 3D hybrid scaffold. The signals in the fibers diffused and generated continuous opposing gradients of CS (chondrogenic signal) and BG (mineralization) in the hydrogel. The chondrocytes were encapsulated in hybrid scaffolds; live dead assay at 48 h showed viable cells. Cells maintained their phenotype and secreted specific extracellular matrix (ECM) in response to signals within the hydrogel. Continuous opposing gradients of sGAG enriched and mineralized ECM were observed surrounding each cell clusters on gradient hydrogel after 14 days of culture in response to the physical gradients of raw materials CS and BG. A construct with gradient mineralization might accelerate integration to subchondral bone during in vivo regeneration.

The advent of 3D printing technology has made remarkable progress in the field of tissue engineering. Yet, it has been challenging to reproduce the desired mechanical properties of certain tissues by 3D printing. This was majorly due to the lack of 3D printable materials possessing mechanical properties similar to the native tissue. In this study, we have synthesized four different ratios of poly(caprolactone- co -lactide (PLCL) and tested their 3D printing capabilities. The physicochemical properties of the material were characterized using Fourier-transform infrared (FTIR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography (GPC), and differential scanning calorimetry (DSC). Furthermore, the mechanical properties were assessed using the universal testing machine (UTM). The ratio with the higher lactide content was found to have better printability. Out of the different ratios assessed, a suitable ratio having the desired mechanical properties and printability was identified and 3D printed into a tracheal scaffold. Thus, PLCL can be a potential material for 3D printing of tissues like the trachea.

Stem cell‐derived islet‐like clusters (ILCs) are an alternative source of pancreatic beta cells for the treatment of diabetic mellitus. An ideal 3D culture platform for the generation of ILCs of desired cluster size is a challenge due to the clustering of islet cells in the 2D culture systems. The islet cells cultured in 2D conditions produce clusters of large size, which are less efficient in terms of insulin secretion and viability. In this study, we report that ILCs formed on a PCL‐based wet electrospun fibrous scaffold with larger pore size produced clusters of the desired size, compared to that cultured on a conventional electrospun sheet. The collagen functionalization on this wet electrospun polycaprolactone (PCL) scaffold showed enhanced insulin secretion and cell viability compared to the non‐functionalized or conventionally electrospun PCL scaffold. The collagen‐coated wet electrospun 3D scaffold produced ILCs of cluster diameter 70 ± 20 μm and the conventionally electrospun PCL sheet produced larger ILC clusters of diameter 300 ± 10 μm. Hence the results indicate the collagen‐functionalized wet electrospun scaffold system could be a potential scaffold for islet tissue engineering.

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