Enhancing alginate dialdehyde-gelatin (ADA-GEL) based hydrogels for biofabrication by addition of phytotherapeutics and mesoporous bioactive glass nanoparticles (MBGNs).

This study explores the 3D printing of alginate dialdehyde-gelatin (ADA-GEL) inks incorporating phytotherapeutic agents, such as ferulic acid (FA), and silicate mesoporous bioactive glass nanoparticles (MBGNs) at two different concentrations. 3D scaffolds with bioactive properties suitable for bone tissue engineering (TE) were obtained. The degradation and swelling behaviour of films and 3D printed scaffolds indicated an accelerated trend with increasing MBGN content, while FA appeared to stabilize the samples. Determination of the degree of crosslinking validated the increased stability of hydrogels due to the addition of FA and 0.1% (w/v) MBGNs. The incorporation of MBGNs not only improved the effective moduli and conferred bioactive properties through the formation of hydroxyapatite (HAp) on the surface of ADA-GEL-based samples but also enhanced VEGF-A expression of MC3T3-E1 cells. The beneficial impact of FA and low concentrations of MBGNs in ADA-GEL-based inks for 3D (bio)printing applications was corroborated through various printing experiments, resulting in higher printing resolution, as also confirmed by rheological measurements. Cytocompatibility investigations revealed enhanced MC3T3-E1 cell activity and viability. Furthermore, the presence of mineral phases, as confirmed by an in vitro biomineralization assay, and increased ALP activity after 21 days, attributed to the addition of FA and MBGNs, were demonstrated. Considering the acquired structural and biological properties, along with efficient drug delivery capability, enhanced biological activity, and improved 3D printability, the newly developed inks exhibit promising potential for biofabrication and bone TE.

3D bioprinting of multifunctional alginate dialdehyde (ADA)-gelatin (GEL) (ADA-GEL) hydrogels incorporating ferulic acid.

The present work explores the 3D extrusion printing of ferulic acid (FA)-containing alginate dialdehyde (ADA)-gelatin (GEL) scaffolds with a wide spectrum of biophysical and pharmacological properties. The tailored addition of FA (≤0.2 %) increases the crosslinking between FA and GEL in the presence of calcium chloride (CaCl2) and microbial transglutaminase, as confirmed using trinitrobenzenesulfonic acid (TNBS) assay. In agreement with an increase in crosslinking density, a higher viscosity of ADA-GEL with FA incorporation was achieved, leading to better printability. Importantly, FA release, enzymatic degradation and swelling were progressively reduced with an increase in FA loading to ADA-GEL, over 28 days. Similar positive impact on antibacterial properties with S. epidermidis strains as well as antioxidant properties were recorded. Intriguingly, FA incorporated ADA-GEL supported murine pre-osteoblast proliferation with reduced osteosarcoma cell proliferation over 7 days in culture, implicating potential anticancer property. Most importantly, FA-incorporated and cell-encapsulated ADA-GEL can be extrusion printed to shape fidelity-compliant multilayer scaffolds, which also support pre-osteoblast cells over 7 days in culture. Taken together, the present study has confirmed the significant potential of 3D bioprinting of ADA-GEL-FA ink to obtain structurally stable scaffolds with a broad spectrum of biophysical and therapeutically significant properties, for bone tissue engineering applications.

Alginate Core-Shell Capsules for 3D Cultivation of Adipose-Derived Mesenchymal Stem Cells.

Mesenchymal stem cells (MSCs) are primary candidates in tissue engineering and stem cell therapies due to their intriguing regenerative and immunomodulatory potential. Their ability to self-assemble into three-dimensional (3D) aggregates further improves some of their therapeutic properties, e.g., differentiation potential, secretion of cytokines, and homing capacity after administration. However, high hydrodynamic shear forces and the resulting mechanical stresses within commercially available dynamic cultivation systems can decrease their regenerative properties. Cells embedded within a polymer matrix, however, lack cell-to-cell interactions found in their physiological environment. Here, we present a "semi scaffold-free" approach to protect the cells from high shear forces by a physical barrier, but still allow formation of a 3D structure with in vivo-like cell-to-cell contacts. We highlight a relatively simple method to create core-shell capsules by inverse gelation. The capsules consist of an outer barrier made from sodium alginate, which allows for nutrient and waste diffusion and an inner compartment for direct cell-cell interactions. Next to capsule characterization, a harvesting procedure was established and viability and proliferation of human adipose-derived MSCs were investigated. In the future, this encapsulation and cultivation technique might be used for MSC-expansion in scalable dynamic bioreactor systems, facilitating downstream procedures, such as cell harvest and differentiation into mature tissue grafts.

Oxidized Hyaluronic Acid-Gelatin-Based Hydrogels for Tissue Engineering and Soft Tissue Mimicking.

Hydrogels are ideal materials for mimicking and engineering soft tissue. Hyaluronic acid is a linear polysaccharide native to the human extracellular matrix. In this study, we first develop and characterize two hydrogel compositions built from oxidized HA and gelatin with and without alginate-di-aldehyde (ADA) crosslinked by ionic and enzymatic agents with potential applications in soft tissue engineering and tissue mimicking structures. The stability under incubation conditions was improved by adjusting crosslinking times. Through large-strain mechanical measurements, the hydrogels' properties were compared to human brain tissue and the samples containing ADA revealed similar mechanical properties to the native tissue specimens in cyclic compression-tension. In vitro characterization demonstrated a high viability of encapsulated mouse embryonic fibroblasts and a spreading of the cells in case of ADA-free samples. Impact statement Brain mimicking materials are required in several medical and industrial fields for the development of safety gear, testing of medical imaging techniques, surgical training, tissue engineering, and modeling of the mechanical behavior of tissues. The materials must resemble the microstructure, chemistry, and mechanical properties of the native tissue extracellular matrix while being adjustable in degradation to suit the various applications. In this article, different methods are used to evaluate a novel hydrogel material and its suitability as brain mimicking matrix.

3D printing and characterization of human nasoseptal chondrocytes laden dual crosslinked oxidized alginate-gelatin hydrogels for cartilage repair approaches.

As cartilage is one of the few tissues in the human body that is not vascularized, the body has very limited capabilities to repair cartilage defects. Hence, novel condro-instructive biomaterials facilitating cartilage formation by implanted chondrocytes are required. In this work, an oxidized alginate-gelatin hydrogel system, alginate-di-aldehyde (ADA) and gelatin (GEL), was used to fabricate 3D printed grid-like structures for cartilage tissue engineering. Enzymatic and ionic crosslinking techniques using microbial transglutaminase (mTG) and divalent ions (CaCl2) were combined to ensure long-term stability of the 3D printed structures. Human nasoseptal chondrocytes were embedded in ADA-GEL prior to 3D printing. Cell viability, proliferation, and metabolic activity were analyzed after 7 and 14 days. The influence of the enzymatic crosslinking and the 3D printing process on the primary human chondrocytes were investigated. It was found that neither the 3D printing process nor the crosslinking by mTG impaired chondrocyte viability. The formation of the main cartilage-specific extracellular matrix components collagen type II and cartilage proteoglycans was shown by immunohistochemical staining. The combination of enzymatic and ionic crosslinking for the 3D printing of ADA-GEL hydrogels is therefore a promising approach for the 3D cultivation of primary human chondrocytes for cartilage tissue engineering.