Mechanical Properties Of Scaffolds Research Articles

3D Porous Edible Scaffolds from Rye Secalin for Cell-Based Pork Fat Tissue Culturing.

Cellular agriculture holds hope for a sustainable alternative to conventional meat, yet multiple technical challenges remain. These include the large-scale production of edible scaffolds and culturing methods for fat tissues, which are key to meat texture, flavor, and nutritional values. Herein. we disclose our method in the facile fabrication of sponge-like plant protein scaffolds by applying commercial sugar cubes as highly permeable templates. The prepared secalin scaffolds feature a high porosity of 85-90%, fully interconnected pores, and high water stability. The mechanical properties of scaffolds could be tuned by varying sugar-to-protein weight ratio and post-water annealing treatment. Moreover, murine preadipocytes (3T3-L1) and porcine adipose-derived stem cells (ADSCs) readily infiltrate, adhere, proliferate, and differentiate on the secalin scaffolds to develop a fat tissue morphology. A cultured fat tissue was produced by culturing porcine ADSCs for 12 days, which remarkably resembles conventional porcine subcutaneous adipose tissue in appearance, texture, flavor, and fatty acid profiles. The research demonstrates the great potential of sponge-like secalin scaffolds for cultured fat tissue production.

Engineering biomimetic scaffolds for bone regeneration: Chitosan/alginate/polyvinyl alcohol-based double-network hydrogels with carbon nanomaterials

In this study, new types of hybrid double-network (DN) hydrogels composed of polyvinyl alcohol (PVA), chitosan (CH), and sodium alginate (SA) are introduced, with the hypothesis that this combination and incorporating multi-walled carbon nanotubes (MWCNTs) and graphene nanoplatelets (GNPs) will enhance osteogenetic differentiation and the structural and mechanical properties of scaffolds for bone tissue engineering applications. Initially, the impact of varying mass ratios of the PVA/CH/SA mixture on mechanical properties, swelling ratio, and degradability was examined. Based on this investigation, a mass ratio of 4:6:6 was determined to be optimal. At this ratio, the hydrogel demonstrated a Young's modulus of 47.5 ± 5 kPa, a swelling ratio of 680 ± 6 % after 3 h, and a degradation rate of 46.5 ± 5 % after 40 days. In the next phase, following the determination of the optimal mass ratio, CNTs and GNPs were incorporated into the 4:6:6 composite resulting in a significant enhancement in the electrical conductivity and stiffness of the scaffolds. The introduction of CNTs led to a notable increase of 36 % in the viability of MG63 osteoblast cells. Additionally, the inhibition zone test revealed that GNPs and CNTs increased the diameter of the inhibition zone by 49.6 % and 52.6 %, respectively.

DESIGN AND STRUCTURAL ANALYSIS OF 3D-PRINTED POROUS POLYLACTIC ACID/HYDROXYAPATITE SCAFFOLDS

Different designs of three-dimensional (3D) structures have gained increasingly significant in bone tissue engineering. For scaffolds, having appropriate porosity and adequate mechanical properties is crucial. The porosity and mechanical properties of scaffolds are higly influenced by their 3D modeled design. By evaluating the mechanical properties of scaffolds with various designs, it can be confirmed that they could serve as an important platform for the regeneration of damaged bone tissue. In this study, a diverse range of unit cells and lattice structures featuring different pore structures of polylactic acid (PLA)/hydroxyapatite (HA) based scaffolds were modeled and designed. Structural analyses of the designed models were conducted in a simulation environment and their mechanical properties were compared with similar studies. The results suggest that PLA/HA-based scaffolds with different designs hold high potential for applications in bone tissue engineering.

Biodegradable electrospun poly(L-lactide-co-ε-caprolactone)/polyethylene glycol/bioactive glass composite scaffold for bone tissue engineering.

The field of tissue engineering has witnessed significant advancements in recent years, driven by the pursuit of innovative solutions to address the challenges of bone regeneration. In this study, we developed an electrospun composite scaffold for bone tissue engineering. The composite scaffold is made of a blend of poly(L-lactide-co-ε-caprolactone) (PLCL) and polyethylene glycol (PEG), with the incorporation of calcined and lyophilized silicate-chlorinated bioactive glass (BG) particles. Our investigation involved a comprehensive characterization of the scaffold's physical, chemical, and mechanical properties, alongside an evaluation of its biological efficacy employing alveolar bone-derived mesenchymal stem cells. The incorporation of PEG and BG resulted in elevated swelling ratios, consequently enhancing hydrophilicity. Thermal gravimetric analysis confirmed the efficient incorporation of BG, with the scaffolds demonstrating thermal stability up to 250°C. Mechanical testing revealed enhanced tensile strength and Young's modulus in the presence of BG; however, the elongation at break decreased. Cell viability assays demonstrated improved cytocompatibility, especially in the PLCL/PEG+BG group. Alizarin red staining indicated enhanced osteoinductive potential, and fluorescence analysis confirmed increased cell adhesion in the PLCL/PEG+BG group. Our findings suggest that the PLCL/PEG/BG composite scaffold holds promise as an advanced biomaterial for bone tissue engineering.

Interfacial blending in co-axially electrospun polymer core-shell fibers and their interaction with cells via focal adhesion point analysis

Electrospun polymer scaffolds have gained prominence in biomedical applications, including tissue engineering, drug delivery, and wound dressings, due to their customizable properties. As the interplay between cells and materials assumes fundamental significance in biomaterials research, understanding the relationship between fiber properties and cell behaviour is imperative. Nevertheless, altering fiber properties introduces complexity by intertwining mechanical and surface chemistry effects, challenging the differentiation of their individual impacts on cell behaviour. Core-shell fibers present an appealing solution, enabling the control of mechanical properties of scaffolds, flexibility in material and drug selection, efficient encapsulation, strong protection of bioactive drugs against harsh environments, and controlled, prolonged drug release. This study addresses a key challenge in core-shell fiber design related to the blending effect between core and shell polymers. Two types of fibers, PMMA and core-shell PC-PMMA, were electrospun, and thorough analyses confirmed the desired core-shell structure in PC-PMMA fibers. Surface chemistry analysis revealed PC diffusion to the PMMA shell of the core-shell fiber during electrospinning, subsequently prompting an investigation of the fiber’s surface potential. Conducting cellular studies on osteoblasts by super-resolution confocal microscopy provided insights into the direct influence of interfacial polymer blending and, consequently, altered fiber surface and mechanical properties on cell focal adhesion points, bridging the gap between material attributes and cell responses in core-shell fibers.

Tuning the properties of all natural polymeric scaffolds for tendon repair with cellulose microfibers

Rotator cuff injuries affect a large percentage of the population worldwide. Surgical repair has rates of failure of up to 70%, and existing materials used in the reinforcement of injuries often lack appropriate mechanical properties and biodegradability. There is a clinical need for tendon biomaterials that enable cell attachment and proliferation, while supporting mechanically the injury site. In this work, we develop a novel all-natural polymeric scaffold for applications in the supraspinatus tendon of the shoulder. The unidirectional freezing technique is applied to a chitosan-gelatine matrix containing varying cellulose concentrations, followed by crosslinking with genipin. The viability of the technique using a custom 3D printed mould is evaluated. The scaffolds' morphology and mechanical properties are extensively characterized: the ultimate tensile strength of the material with 20% cellulose was found to increase 6-fold compared to scaffolds without cellulose; significant effects on the microstructure of the scaffold were evidenced via scanning electron microscopy. Furthermore, the biocompatibility of the materials was characterized with both porcine tendon derived cells and normal human dermal fibroblasts. All scaffolds are highly biocompatible, the incorporation of cellulose results in higher cell metabolic activity values and density of cells on the surface of the material. Scaffolds containing 20% cellulose fibers were found to possess the optimal biomechanical properties for applications in rotator cuff tendon repair.

Biomechanically tunable scaffolds for bone tissue regeneration and testbeds of cancer bone metastasis

Current challenges in bone regeneration and design of humanoid 3D bone metastases cancer testbeds can be overcome by designing bone-mimetic tissue engineering scaffolds with tunable mechanical and biological properties. This study employs three unnatural amino acids to modify montmorillonite clay (MMT), producing polymer clay nanocomposites (PCNs) to construct 3D bone-mimetic scaffolds. The modification of MMT clay with 5-aminovaleric acid, (±)-2-aminopimelic acid, and 4-(4-Aminophenyl) butyric acid is analyzed through x-ray diffraction (XRD) and fourier-transform infrared spectroscopy (FTIR), revealing changes in the interlayer spacing and functional groups. Molecular modeling elucidates the interaction energies in the amino acid intercalated nanoclays that influence the mechanical properties of the scaffolds. 4-(4-Aminophenyl) butyric acid-modified scaffolds exhibited the highest compressive strength, highlighting the critical influence of molecular interactions on the mechanical properties of the PCN. Despite being a small fraction of the scaffold, the presence and type of amino acids significantly affect the scaffold mechanical properties. Cell viability assays affirm biocompatibility of scaffolds. The amino acids within the nanoclays also impact mineralization on scaffolds seeded with human mesenchymal stem cells (hMSCs). Amino acid type also influences the amount of mineralization in the scaffolds. A sequential culture involving hMSCs and breast cancer cells (MCF-7) on the scaffold mimics breast cancer metastasis to bone. Enhanced mineralization by bone cells in the presence of MCF-7 cells is observed, emphasizing their osteoblastic characteristics. Hence, 3D scaffolds for bone regeneration and bone metastasis cancer testbeds can exhibit tunable mechanical properties and biological responses though a simulation guided choice of unnatural amino acids.

Solvent-cast 3D printing with molecular weight polymer blends to decouple effects of scaffold architecture and mechanical properties on mesenchymal stromal cell fate.

The biochemical and physical properties of a scaffold can be tailored to elicit specific cellular responses. However, it is challenging to decouple their individual effects on cell-material interactions. Here, we solvent-cast 3D printed different ratios of high and low molecular weight (MW) poly(caprolactone) (PCL) to fabricate scaffolds with significantly different stiffnesses without affecting other properties. Ink viscosity was used to match processing conditions between inks and generate scaffolds with the same surface chemistry, crystallinity, filament diameter, and architecture. Increasing the ratio of low MW PCL resulted in a significant decrease in modulus. Scaffold modulus did not affect human mesenchymal stromal cell (hMSC) differentiation under osteogenic conditions. However, hMSC response was significantly affected by scaffold stiffness in chondrogenic media. Low stiffness promoted more stable chondrogenesis whereas high stiffness drove hMSC progression toward hypertrophy. These data illustrate how this versatile platform can be used to independently modify biochemical and physical cues in a single scaffold to synergistically enhance desired cellular response.

Immediate implantation of ultrafine fiber slow-release system based on cell electrospinning to induce osteogenesis of mesenchymal stem cells.

This study presents the development and evaluation of a poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P34HB) ultrafine fiber slow-release system for in vivo osteogenic induction of human umbilical cord mesenchymal stem cells (HUCMSCs). Utilizing dual-nozzle and cell electrospinning techniques, the system encapsulates L-ascorbic acid-2-phosphate magnesium (ASP), β-glycerophosphate sodium and dexamethasone (DEX) within the fibers, ensuring sustained osteogenic differentiation. The scaffold's morphology, characterization, hydrophilicity, mechanical properties and cellular behavior were examined. Immediate subcutaneous implantation in rabbits was conducted to observe its ectopic osteogenic induction effect. Successfully fabricated P34HB ultrafine fiber slow-release system. Characterization confirmed the uniform distribution of HUCMSCs and inducing components within the scaffold, with no chemical reactions affecting the active components. In vitro tests showcased a prolonged release of DEX and ASP, while biocompatibility assays highlighted the scaffold's suitability for cellular growth. Alizarin Red, type I collagen, and osteopontin (OPN) staining verified the scaffold's potent osteogenic induction effect on HUCMSCs. Notably, immediate implantation into New Zealand White rabbits led to significant new bone formation within 8 weeks. These findings underscore the system's potential for immediate in vivo implantation without prior in vitro induction, marking a promising advancement in bone tissue engineering.

Unlocking the print of poly(L-lactic acid) by melt electrowriting for medical application

Melt electrowriting (MEW) is an advanced 3D printing technology to process high-resolution scaffolds. Although MEW has printed many biodegradable polymers, most of them have low printability, including poly(L-lactic acid) (PLLA) discussed in this study. While the print of PLLA scaffolds by MEW is elementarily realized, the scaffolds' limited structural change and mechanical properties influence their application. This study aims at improving the printability and the mechanical properties of the PLLA. To this end, the printability of the PLLA regarding jet lag angle and critical translation speed is investigated; the print limitation of PLLA is discovered, and the uniaxial and cyclic tensile tests for the scaffolds in linear and nonlinear structures are carried out. The results suggest that drying the PLLA before printing can significantly restrain polymer degradation to extend the printing period to approximately one day; planar PLLA scaffolds are able to reach 25 layers, 100 µm interfiber distance, and 8 × 8 cm2 in dimension, and tubular PLLA scaffolds with 6 layers are successfully printed out; the mechanical properties of the PLLA scaffold reach the level of electrospun nonwoven fabrics. Thus, this study is a milestone work for the MEW technology to extend the printable materials.

3D Graphene/silk fibroin scaffolds enhance dental pulp stem cell osteo/odontogenic differentiation

ObjectivesThe current in vitro study aims to evaluate silk fibroin with and without the addition of graphene as a potential scaffold material for regenerative endodontics. Material and MethodsSilk fibroin (SF), Silk fibroin/graphene oxide (SF/GO) and silk fibroin coated with reduced graphene oxide (SF/rGO) scaffolds were prepared (n = 30). The microarchitectures and mechanical properties of scaffolds were evaluated using field emission scanning electron microscopy (FESEM), pore size and water uptake, attenuated total reflectance fourier transformed infrared spectroscopy (ATR-FTIR), Raman spectroscopy and mechanical compression tests. Next, the study analyzed the influence of these scaffolds on human dental pulp stem cell (hDPSC) viability, apoptosis or necrosis, cell adhesion, odontogenic differentiation marker expression and mineralized matrix deposition. The data were analyzed with ANOVA complemented with the Tukey post-hoc test (p < 0.005). ResultsSEM analysis revealed abundant pores with a size greater than 50 nm on the surface of tested scaffolds, primarily between 50 nm and 600 µm. The average value of water uptake obtained in pure fibroin scaffolds was statistically higher than that of those containing GO or rGO (p < 0.05). ATR-FTIR evidenced that the secondary structures did not present differences between pure fibroin and fibroin coated with graphene oxide, with a similar infrared spectrum in all tested scaffolds. Raman spectroscopy showed a greater number of defects in the links in SF/rGO scaffolds due to the reduction of graphene. In addition, adequate mechanical properties were exhibited by the tested scaffolds. Regarding biological properties, hDPSCs attached to scaffolds were capable of proliferating at a rate similar to the control, without affecting their viability over time. A significant upregulation of ALP, ON and DSPP markers was observed with SF/rGO and SF/GO groups. Finally, SF/GO and SF/rGO promoted a significantly higher mineralization than the control at 21 days. SignificanceData obtained suggested that SF/GO and SF/rGO scaffolds promote hDPSC differentiation at a genetic level, increasing the expression of key osteo/odontogenic markers, and supports the mineralization of the extracellular matrix. However, results from this study are to be interpreted with caution, requiring further in vivo studies to confirm the potential of these scaffolds.

Study on the effect of dynamic chemical polishing treatment on porous titanium alloy scaffolds fabricated using laser powder bed fusion

This study addresses the issue of a significant amount of unmelted powder adhering to the surface of Laser Powder Bed Fusion (LPBF) porous titanium alloy scaffolds, and the current inability of existing technologies to effectively handle such surface defects. Porous titanium alloy scaffolds with a triple periodical minimal surface (TPMS) Gyroid structure were prepared using powder bed fusion technology for dynamic chemical polishing. The spatial and surface morphology, as well as the mechanical properties of the scaffolds, were evaluated before and after treatment. Concurrently, fluid finite element simulation was employed to analyze the flow state of the solution inside the porous scaffold during dynamic chemical polishing. Cell culture samples were prepared and subjected to dynamic chemical polishing to study the specific effects of scaffold surface morphology on cell activity. The results indicate that the surface treatment effect, internal and external homogeneity, and dynamic and static mechanical properties of the porous scaffolds with a Gyroid structure are significantly improved after dynamic chemical polishing. Additionally, it was found that dynamic chemical polishing could effectively reduce the micrometer roughness and increase the nanometer roughness of the sample surface, thereby enhancing the adhesion, proliferation, and differentiation activities of Human Bone Mesenchymal Stem cells.

Tailoring the properties of composite scaffolds with a 3D-Printed lattice core and a bioactive hydrogel shell for tissue engineering

The optimal performance of scaffolds for tissue engineering relies on a proper combination of their constituent biomaterials and on the design of their structure. In this work, composite scaffolds with a core-shell architecture are realized by grafting a gelatin-chitosan hydrogel onto a 3D-printed polylactic acid (PLA) core, aiming in particular at bone regeneration. This hydrogel was recently found to sustain osteogenic differentiation of mesenchymal stromal cells, leading to new bone tissue formation. Here, the integration with rigid PLA lattice structures provides improved mechanical support and finer control of strength and stiffness. The core is prepared by fused deposition modeling with the specific aim to study several lattice structures and thereby better tune the scaffold mechanical properties. In fact, the core architecture dictates the scaffold strength and stiffness, which are seen to match those of different types of bone tissue. For all lattice types, the hydrogel is found to penetrate throughout the entire core and to present highly interconnected pores for cell colonization. By varying the void volume fraction in the core it is possible to significantly change the bioactive shell content, as well as the mechanical properties, over a wide range of values. Looking for design guidelines, relationships between stiffness/strength and density are here outlined for scaffolds featuring different lattice parameters. Moreover, by acting on the core strut arrangement, scaffolds are reinforced along specific directions, as evaluated under compressive and bending loading conditions.

Effects of light exposure intensity and time on printing quality and compressive strength of β-TCP scaffolds fabricated with digital light processing

Digital light processing (DLP), derived from stereolithography (SL), is a lithography-based additive manufacturing (AM) process that offers the ability to tailor scaffold shape, size, material, and biometrics of fabricated scaffolds to patient requirements. The additive manufacturing of tricalcium phosphate (TCP) bioceramic scaffolds via DLP is explored in the proceeding research paper. Particularly, the role of exposure intensity and exposure time on resultant scaffold quality and mechanical properties is studied. Increased exposure intensity results in an increased deviation from the designed lattice structure and thus an increase in compressive strength. Scaffolds produced with the highest exposure intensity, resulted in a 27.6 % error increase in mean beam diameter, resulting from an increased scattering effect, penetration depth, and over-curing of peripheral slurry. TCP scaffolds with an exposure time of 3.5 s and exposure intensity of 40 mW/cm2 possessed the highest print accuracy, resulting in the lowest average compressive strength – 82 MPa for the scaffolds of cube structure and 48 MPa for the ones of I-WP structure. Statement of significanceDigital light processing (DLP) is an effective additive manufacturing (AM) method applicable for bioceramics. This study focuses on the influence of light exposure intensity on scaffold quality and compressive strength. The increase of exposure intensity can improve the compressive strength with the sacrifice of structure accuracy. The conclusions of this study can provide insights into the relationship of DLP printing parameters and scaffolds quality.

Oxygenated Scaffolds for Pancreatic Endocrine Differentiation from Induced Pluripotent Stem Cells.

A 3D microenvironment is known to endorse pancreatic islet development from human induced pluripotent stem cells (iPSCs). However, oxygen supply becomes a limiting factor in a scaffold culture. In this study, oxygen-releasing biomaterials are fabricated and an oxygenated scaffold culture platform is developed to offer a better oxygen supply during 3D iPSC pancreatic differentiation. It is found that the oxygenation does not alter the scaffold's mechanical properties. The in situ oxygenation improves oxygen tension within the scaffolds. The unique 3D differentiation system enables the generation of islet organoids with enhanced expression of islet signature genes and proteins. Additionally, it is discovered that the oxygenation at the early stage of differentiation has more profound impacts on islet development from iPSCs. More C-peptide+ /MAFA+ β and glucagon+ /MAFB+ α cells formed in the iPSC-derived islet organoids generated under oxygenated conditions, suggesting enhanced maturation of the organoids. Furthermore, the oxygenated 3D cultures improve islet organoids' sensitivity to glucose for insulin secretion. It is herein demonstrated that the oxygenated scaffold culture empowers iPSC islet differentiation to generate clinically relevant tissues for diabetes research and treatment.

Optimization of 3D Synthetic Scaffolds for Neuronal Tissue Engineering Applications.

The increasing prevalence of neurodegenerative diseases has spurred researchers to develop advanced 3D models that accurately mimic neural tissues. Hydrogels stand out as ideal candidates as their properties closely resemble those of the extracellular matrix. A critical challenge in this regard is to comprehend the influence of the scaffold's mechanical properties on cell growth and differentiation, thus enabling targeted modifications. In light of this, a synthesis and comprehensive analysis of acrylamide-based hydrogels incorporating a peptide has been conducted. Adequate cell adhesion and development is achieved due to their bioactive nature and specific interactions with cellular receptors. The integration of a precisely controlled physicochemical hydrogel matrix and inclusion of the arginine-glycine-aspartic acid peptide sequence has endowed this system with an optimal structure, thus providing a unique ability to interact effectively with biomolecules. The analysis fully examined essential properties governing cell behavior, including pore size, mechanical characteristics, and swelling ability. Cell-viability experiments were performed to assess the hydrogel's biocompatibility, while the incorporation of grow factors aimed to promote the differentiation of neuroblastoma cells. The results underscore the hydrogel's ability to stimulate cell viability and differentiation in the presence of the peptide within the matrix.

Bone repair potential of collagen-poly(3-hydroxybutyrate)-carbon nanotubes scaffold loaded with mesenchymal stem cells for the reconstruction of critical-sized mandibular defects

The poor structural stability of collagen (COL) upon hydration poses a significant challenge in tissue engineering (TE). To overcome this limitation, the incorporation of hydrophobic polymers such as poly(3-hydroxybutyrate) (PHB), and nanomaterials such as carbon nanotubes (CNTs) has been explored. In this study, we investigated the physical, chemical, and biological characteristics of COL-based scaffolds modified with PHB and CNTs for bone tissue engineering (BTE) applications. The tensile strength analysis revealed a substantial improvement in the ultimate tensile strength with the addition of 10 % PHB and 4 % CNTs. Scanning electron microscopy (SEM) images depicted a denser and more compact structure resulting from the presence of PHB and CNTs, enhancing the scaffold's mechanical properties. Fourier-transform infrared spectroscopy (FTIR) confirmed the successful incorporation of PHB and CNTs into the composite scaffold, maintaining the chemical integrity of COL. Stereological studies also conducted in a rat model with induced critical-sized bone defects in the mandibular bone further emphasize the substantial increase in bone formation and reduction in defect volume achieved by the scaffold loaded with stem cells. These findings underscore the promising approach to enhance bone healing, using COL-based scaffolds loaded with stem cells, and the favorable results obtained in this study can contribute to the advancement of BTE strategies.

Multimodal Three-Dimensional Printing for Micro-Modulation of Scaffold Stiffness Through Machine Learning.

The ability to precisely control a scaffold's microstructure and geometry with light-based three-dimensional (3D) printing has been widely demonstrated. However, the modulation of scaffold's mechanical properties through prescribed printing parameters is still underexplored. This study demonstrates a novel 3D-printing workflow to create a complex, elastomeric scaffold with precision-engineered stiffness control by utilizing machine learning. Various printing parameters, including the exposure time, light intensity, printing infill, laser pump current, and printing speed were modulated to print poly (glycerol sebacate) acrylate (PGSA) scaffolds with mechanical properties ranging from 49.3 ± 3.3 kPa to 2.8 ± 0.3 MPa. This enables flexibility in spatial stiffness modulation in addition to high-resolution scaffold fabrication. Then, a neural network-based machine learning model was developed and validated to optimize printing parameters to yield scaffolds with user-defined stiffness modulation for two different vat photopolymerization methods: a digital light processing (DLP)-based 3D printer was utilized to rapidly fabricate stiffness-modulated scaffolds with features on the hundreds of micron scale and a two-photon polymerization (2PP) 3D printer was utilized to print fine structures on the submicron scale. A novel 3D-printing workflow was designed to utilize both DLP-based and 2PP 3D printers to create multiscale scaffolds with precision-tuned stiffness control over both gross and fine geometric features. The described workflow can be used to fabricate scaffolds for a variety of tissue engineering applications, specifically for interfacial tissue engineering for which adjacent tissues possess heterogeneous mechanical properties (e.g., muscle-tendon).

3D-printed biomimetic scaffolds with precisely controlled and tunable structures guide cell migration and promote regeneration of osteochondral defect

Untreated osteochondral defects will develop into osteoarthritis, affecting patients’ quality of life. Since articular cartilage and subchondral bone exhibit distinct biological characteristics, repairing osteochondral defects remains a major challenge. Previous studies have tried to fabricate multilayer scaffolds with traditional methods or 3D printing technology. However, the efficacy is unsatisfactory because of poor control over internal structures or a lack of integrity between adjacent layers, severely compromising repair outcomes. Therefore, there is a need for a biomimetic scaffold that can simultaneously boost osteochondral defect regeneration in both structure and function. Herein, an integrated bilayer scaffold with precisely controlled structures is successfully 3D-printed in one step via digital light processing (DLP) technology. The upper layer has both ‘lotus- and radial-’ distribution pores, and the bottom layer has ‘lotus-’ pores to guide and facilitate the migration of chondrocytes and bone marrow mesenchymal stem cells, respectively, to the defect area. Tuning pore sizes could modulate the mechanical properties of scaffolds easily. Results show that 3D-printed porous structures allow significantly more cells to infiltrate into the area of ‘lotus- and radial-’ distribution pores during cell migration assay, subcutaneous implantation, and in situ transplantation, which are essential for osteochondral repair. Transplantation of this 3D-printed bilayer scaffold exhibits a promising osteochondral repair effect in rabbits. Incorporation of Kartogenin into the upper layer of scaffolds further induces better cartilage formation. Combining small molecules/drugs and precisely size-controlled and layer-specific porous structure via DLP technology, this 3D-printed bilayer scaffold is expected to be a potential strategy for osteochondral regeneration.

Smart and sustainable: Exploring the future of PHAs biopolymers for 3D printing in tissue engineering

Polyhydroxyalkanoates (PHAs) are biodegradable biopolymers (polyesters), produced by a wide range of bacterial strains. They are gaining increasing interest in different research fields, due to their sustainability and environmental-friendly properties. Additionally, PHAs are also biocompatible, which makes them interesting for tissue engineering and regenerative medicine. At the same time, they are characterized by properties ideal for 3D printing processing, such as high tensile strength, easy processability and thermoplasticity. To date, the techniques employed in PHAs printing mostly include fused deposition modeling (FDM), selective laser sintering (SLS), electrospinning (ES), and melt electrospinning (MES). In this review, we provide a comprehensive summary of the versatile and sustainably sourced bacterial PHAs, also modified by blending with natural and synthetic polymers (e.g., PLA, PGA) or combining them with inorganic fillers (e.g., nanoparticles, glass), used for 3D printing in biomedical applications. We specify focus on the printing conditions and the properties of the obtained scaffolds with a focus on the print resolution and scaffolds mechanical and biological properties. New perspectives in the emerging field of PHAs biofabrication process, characterized by sustainability and efficiency of the scaffold production, are demonstrated. The use of alternative printing techniques, i.e. melt electrowriting (MEW), and producing smart and functional materials degrading on demand under in vitro and in vivo conditions is proposed.