Scaffold Architecture Research Articles

Anisotropic mechanical and mass-transport performance of Ti6Al4V plate-lattice scaffolds prepared by laser powder bed fusion

In bone scaffolds, the mechanical performance provides the load-bearing capability, and the mass-transport performance presented as permeability dominates the nutrients/oxygen transportation efficiency. Body-centered-cubic and face-centered-cubic plate lattice scaffolds with mechanical and mass-transport performance close to human bones are proposed in the present study. The regular periodic architecture and plane-stress state of the plate lattice scaffolds not only provide them with advanced mechanical properties but avoid stress concentration that ubiquitously exists in traditional truss lattice scaffolds. By investigating the anisotropic mechanical and mass-transport performance of plate lattice scaffolds, a valid regulation strategy is put forward to modulate their performance without changing the volume fraction and architecture, providing an alternative scheme for biomedical scaffold design. Both computational and experimental results demonstrate that body-centered-cubic and face-centered-cubic plate lattice scaffolds possess appropriate mechanical and mass-transport performance close to human bones. In addition, tuning ranges of the mechanical and mass-transport performance of plate lattice scaffolds for different orientations are up to 40% and 45%, respectively. These findings could provide valuable references for the extensive applications of plate lattice scaffolds in bone tissue engineering. Statement of significanceIn bone tissue engineering, scaffolds with low density, high strength, and proper permeability are of constant request. The present study proposes body-centered-cubic and face-centered-cubic plate lattice scaffolds with mechanical and mass-transport performance close to human bones. The deformation mechanisms and mass-transport characteristics of plate lattice scaffolds for different orientations are revealed. In addition, a valid regulation strategy is put forward to modulate the mechanical and mass-transport performance of plate lattice scaffolds without changing their volume fraction and architecture, providing an alternative scheme for biomedical scaffold design. We believe that these findings could provide significant guidance for the simultaneous improvements of advanced scaffold designing.

AI-based structure prediction empowers integrative structural analysis of human nuclear pores.

INTRODUCTION The eukaryotic nucleus pro-tects the genome and is enclosed by the two membranes of the nuclear envelope. Nuclear pore complexes (NPCs) perforate the nuclear envelope to facilitate nucleocytoplasmic transport. With a molecular weight of ∼120 MDa, the human NPC is one of the larg-est protein complexes. Its ~1000 proteins are taken in multiple copies from a set of about 30 distinct nucleoporins (NUPs). They can be roughly categorized into two classes. Scaf-fold NUPs contain folded domains and form a cylindrical scaffold architecture around a central channel. Intrinsically disordered NUPs line the scaffold and extend into the central channel, where they interact with cargo complexes. The NPC architecture is highly dynamic. It responds to changes in nuclear envelope tension with conforma-tional breathing that manifests in dilation and constriction movements. Elucidating the scaffold architecture, ultimately at atomic resolution, will be important for gaining a more precise understanding of NPC function and dynamics but imposes a substantial chal-lenge for structural biologists. RATIONALE Considerable progress has been made toward this goal by a joint effort in the field. A synergistic combination of complementary approaches has turned out to be critical. In situ structural biology techniques were used to reveal the overall layout of the NPC scaffold that defines the spatial reference for molecular modeling. High-resolution structures of many NUPs were determined in vitro. Proteomic analysis and extensive biochemical work unraveled the interaction network of NUPs. Integra-tive modeling has been used to combine the different types of data, resulting in a rough outline of the NPC scaffold. Previous struc-tural models of the human NPC, however, were patchy and limited in accuracy owing to several challenges: (i) Many of the high-resolution structures of individual NUPs have been solved from distantly related species and, consequently, do not comprehensively cover their human counterparts. (ii) The scaf-fold is interconnected by a set of intrinsically disordered linker NUPs that are not straight-forwardly accessible to common structural biology techniques. (iii) The NPC scaffold intimately embraces the fused inner and outer nuclear membranes in a distinctive topol-ogy and cannot be studied in isolation. (iv) The conformational dynamics of scaffold NUPs limits the resolution achievable in structure determination. RESULTS In this study, we used artificial intelligence (AI)-based prediction to generate an exten-sive repertoire of structural models of human NUPs and their subcomplexes. The resulting models cover various domains and interfaces that so far remained structurally uncharac-terized. Benchmarking against previous and unpublished x-ray and cryo-electron micros-copy structures revealed unprecedented accu-racy. We obtained well-resolved cryo-electron tomographic maps of both the constricted and dilated conformational states of the hu-man NPC. Using integrative modeling, we fit-ted the structural models of individual NUPs into the cryo-electron microscopy maps. We explicitly included several linker NUPs and traced their trajectory through the NPC scaf-fold. We elucidated in great detail how mem-brane-associated and transmembrane NUPs are distributed across the fusion topology of both nuclear membranes. The resulting architectural model increases the structural coverage of the human NPC scaffold by about twofold. We extensively validated our model against both earlier and new experimental data. The completeness of our model has enabled microsecond-long coarse-grained molecular dynamics simulations of the NPC scaffold within an explicit membrane en-vironment and solvent. These simulations reveal that the NPC scaffold prevents the constriction of the otherwise stable double-membrane fusion pore to small diameters in the absence of membrane tension. CONCLUSION Our 70-MDa atomically re-solved model covers >90% of the human NPC scaffold. It captures conforma-tional changes that occur during dilation and constriction. It also reveals the precise anchoring sites for intrinsically disordered NUPs, the identification of which is a prerequisite for a complete and dy-namic model of the NPC. Our study exempli-fies how AI-based structure prediction may accelerate the elucidation of subcellular ar-chitecture at atomic resolution. [Figure: see text].

Evaluation of Functionally Graded Architecture using Fused Filament Fabrication Technique for Bone Scaffold Applications

Bone scaffold architecture affects the performance of bone scaffold significantly. Recently, functionally graded architecture (FGA) has received wide attention in bone scaffold application due to its architecture that mimics the cancellous bone architecture. Hence, this study aims to evaluate the mechanical properties of PLA specimens that adopt FGA fabricated using additive manufacturing via fused filament fabrication (FFF). The mechanical properties of FGA PLA specimen (1 GPa -2 Gpa) was tailored to the cancellous bone (modulus: 0.01 GPa – 2 GPa). The result shows that radially dense- in rod (RDR) has the highest Young’s Modulus, and yield strength. However, the relationship between cross section area of specimen needs to be considered in comparing the performance of FGA PLA specimen.

A Comprehensive Review of Biopolymer Fabrication in Additive Manufacturing Processing for 3D-Tissue-Engineering Scaffolds.

The selection of a scaffold-fabrication method becomes challenging due to the variety in manufacturing methods, biomaterials and technical requirements. The design and development of tissue engineering scaffolds depend upon the porosity, which provides interconnected pores, suitable mechanical strength, and the internal scaffold architecture. The technology of the additive manufacturing (AM) method via photo-polymerization 3D printing is reported to have the capability to fabricate high resolution and finely controlled dimensions of a scaffold. This technology is also easy to operate, low cost and enables fast printing, compared to traditional methods and other additive manufacturing techniques. This article aims to review the potential of the photo-polymerization 3D-printing technique in the fabrication of tissue engineering scaffolds. This review paper also highlights the comprehensive comparative study between photo-polymerization 3D printing with other scaffold fabrication techniques. Various parameter settings that influence mechanical properties, biocompatibility and porosity behavior are also discussed in detail.

Future prospects in 3-dimensional (3D) technology and Mohs micrographic surgery

The advancement of three-dimensional (3D) bioprinting has allowed for changes within the field of dermatology. Skin bioprinting is a transformative technology used to fabricate biomimetic scaffold architectures which mimic human skin. This paper briefly discusses three major types of biological printing: inkjet-based bioprinting, laser-assisted bioprinting and pressure-assisted bioprinting in regard to skin bioprinting for Mohs application. This manuscript reviews the basic principles of 3D printing (inkjet-based bioprinting, laser-assisted bioprinting and pressure-assisted bioprinting), current 3D skin bioprinting applications and methods, and perspectives on using 3D printing for Mohs surgery.

Multi-objective Shape Optimization of Bone Scaffolds: Enhancement of Mechanical Properties and Permeability

Porous scaffolds have recently attracted attention in bone tissue engineering. The implanted scaffolds are supposed to satisfy the mechanical and biological requirements. In this study, two porous structures named MFCC-1 (modified face centered cubic-1) and MFCC-2 (modified face centered cubic-2) are introduced. The proposed porous architectures are evaluated, optimized, and tested to enhance mechanical and biological properties. The geometric parameters of the scaffolds with porosities ranging from 70% to 90% are optimized to find a compromise between the effective Young's modulus and permeability, as well as satisfying the pore size and specific surface area requirements. To optimize the effective Young's modulus and permeability, we integrated a mathematical formulation, finite element analysis, and computational fluid dynamics simulations. For validation, the optimized scaffolds were 3D-printed, tested, and compared with two different orthogonal cylindrical struts (OCS) scaffold architectures. The MFCC designs are preferred to the generic OCS scaffolds from various perspectives: a) the MFCC architecture allows scaffold designs with porosities up to 96%; b) the very porous architecture of MFCC scaffolds allows achieving high permeabilities, which could potentially improve the cell diffusion; c) despite having a higher porosity compared to the OCS scaffolds, MFCC scaffolds improve mechanical performance regarding Young's modulus, stress concentration, and apparent yield strength; d) the proposed structures with different porosities are able to cover all the range of permeability for the human trabecular bones. The optimized MFCC designs have simple architectures and can be easily fabricated and used to improve the quality of load-bearing orthopedic scaffolds. Statement of significancePorous scaffolds are increasingly being studied to repair large bone defects. A scaffold is supposed to withstand mechanical loads and provide an appropriate environment for bone cell growth after implantation. These mechanical and biological requirements are usually contradicting; improving the mechanical performance would require a reduction in porosity and a lower porosity is likely to reduce the biological performance of the scaffold. Various studies have shown that the mechanical and biological performance of bone scaffolds can be improved by internal architecture modification. In this study, we propose two scaffold architectures named MFCC-1 and MFCC-2 and provide an optimization framework to simultaneously optimize their stiffness and permeability to improve their mechanical and biological performances.

Synthetic vascular graft with spatially distinct architecture for rapid biomimetic cell organisation in a perfusion bioreactor

Access to lab-grown fully functional blood vessels would provide an invaluable resource to vascular medicine. The complex architecture and cellular makeup of native vessels, however, makes this extremely challenging to reproduce in vitro. Bioreactor systems have helped advanced research in this area by replicating many of the physiological conditions necessary for full-scale tissue growth outside of the body. A key element underpinning these technologies are 3D vascular graft templates which serve as temporary scaffolds to direct cell growth into similar cellular architectures observed in native vessels. Grafts further engineered with appropriate physical cues to accommodate the multiple cell types that reside within native vessels may help improve the production efficiency and physiological accuracy of bioreactor-grown vessel substitutes. Here, we engineered two distinct scaffold architectures into an electrospun vascular graft aiming to encourage the spatial organisation of human vascular endothelial cells (hCAECs) in a continuous luminal monolayer, co-cultured with human fibroblasts (hFBs) populating the graft wall. Using an electrospun composite of polycaprolactone and gelatin, we evaluated physical parameters including fibre diameter, fibre alignment, and porosity, that best mimicked the spatial composition and growth of hCAECs and hFBs in native vessels. Upon identifying the optimal scaffold architectures for each cell type, we constructed a custom-designed mandrel that combined these distinct architectures into a single vascular graft during a single electrospinning processing run. When connected to a perfusion bioreactor system, the dual architecture graft spatially oriented hCAECs and hFBs into the graft wall and lumen, respectively, directly from circulation. This biomimetic cell organisation was consistent with positive graft remodelling with significant collagen deposition in the graft wall. These findings demonstrate the influence of architectural cues to direct cell growth within vascular graft templates and the future potential of these approaches to more accurately and efficiency produce blood vessel substitutes in bioreactor systems.

Clinical translation of a patient-specific scaffold-guided bone regeneration concept in four cases with large long bone defects.

BackgroundBone defects after trauma, infection, or tumour resection present a challenge for patients and clinicians. To date, autologous bone graft (ABG) is the gold standard for bone regeneration. To address the limitations of ABG such as limited harvest volume as well as overly fast remodelling and resorption, a new treatment strategy of scaffold-guided bone regeneration (SGBR) was developed. In a well-characterized sheep model of large to extra-large tibial segmental defects, three-dimensional (3D) printed composite scaffolds have shown clinically relevant biocompatibility and osteoconductive capacity in SGBR strategies. Here, we report four challenging clinical cases with large complex posttraumatic long bone defects using patient-specific SGBR as a successful treatment.MethodsAfter giving informed consent computed tomography (CT) images were used to design patient-specific biodegradable medical-grade polycaprolactone-tricalcium phosphate (mPCL-TCP, 80:20 ​wt%) scaffolds. The CT scans were segmented using Materialise Mimics to produce a defect model and the scaffold parts were designed with Autodesk Meshmixer. Scaffold prototypes were 3D-printed to validate robust clinical handling and bone defect fit. The final scaffold design was additively manufactured under Food and Drug Administration (FDA) guidelines for patient-specific and custom-made implants by Osteopore International Pte Ltd.ResultsFour patients (age: 23–42 years) with posttraumatic lower extremity large long bone defects (case 1: 4 ​cm distal femur, case 2: 10 ​cm tibia shaft, case 3: complex malunion femur, case 4: irregularly shaped defect distal tibia) are presented. After giving informed consent, the patients were treated surgically by implanting a custom-made mPCL-TCP scaffold loaded with ABG (case 2: additional application of recombinant human bone morphogenetic protein-2) harvested with the Reamer-Irrigator-Aspirator system (RIA, Synthes®). In all cases, the scaffolds matched the actual anatomical defect well and no perioperative adverse events were observed. Cases 1, 3 and 4 showed evidence of bony ingrowth into the large honeycomb pores (pores >2 ​mm) and fully interconnected scaffold architecture with indicative osseous bridges at the bony ends on the last radiographic follow-up (8–9 months after implantation). Comprehensive bone regeneration and full weight bearing were achieved in case 2 ​at follow-up 23 months after implantation.ConclusionThis study shows the bench to bedside translation of guided bone regeneration principles into scaffold-based bone tissue engineering. The scaffold design in SGBR should have a tissue-specific morphological signature which stimulates and directs the stages from the initial host response towards the full regeneration. Thereby, the scaffolds provide a physical niche with morphology and biomaterial properties that allow cell migration, proliferation, and formation of vascularized tissue in the first one to two months, followed by functional bone formation and the capacity for physiological bone remodelling. Great design flexibility of composite scaffolds to support the one to three-year bone regeneration was observed in four patients with complex long bone defects.The translational potential of this articleThis study reports on the clinical efficacy of SGBR in the treatment of long bone defects. Moreover, it presents a comprehensive narrative of the rationale of this technology, highlighting its potential for bone regeneration treatment regimens in patients with any type of large and complex osseous defects.

Fused filament fabrication of polycaprolactone bioscaffolds: Influence of fabrication parameters and thermal environment on geometric fidelity and mechanical properties

Polycaprolactone (PCL) is frequently used in the fabrication of porous scaffold architectures for tissue engineering applications. PCL's biocompatibility, bio-resorption profile and thermoplasticity make it an exceptionally versatile polymer for the fabrication of 3D printed lattice bioscaffold architectures. However, despite its ubiquitous use, optimal parameters for printing PCL lattice bioscaffolds have not been fully elucidated. In this study, fused filament fabrication (FFF) and a custom temperature-regulated enclosure were used to fabricate cubic PCL lattice porous solids. It was hypothesised that the cooling rate of the extruded scaffold material plays a key role in ensuring geometrical fidelity. Samples were fabricated at 3 different layer heights 0.12, 0.13 and 0.14 mm within two temperature regulated envelopes at 17.5 ± 1.5 °C for sample group ET17.5 °C and 27.5 ± 1.5 °C for sample group ET27.5 °C.Scaffold architectures fabricated within the 17.5 ± 1.5 °C envelope at the respective layer heights presented consistently less geometric deviation from the original CAD model than those scaffolds fabricated within a 27.5 ± 1.5 °C envelope. Echoed in the mechanical performance evaluation, the ET17.5 °C group performed with greater consistency and less deviation in Young's modulus and ultimate compressive strength. Samples from ET17.5 °C were also found to possess lower levels of crystallinity compared to ET27.5 °C which correlates with the extended cooling cycle within a 27.5 ± 1.5 °C envelope. Optical profilometry also revealed less undulation in the surface topography of polymer struts from ET17.5 °C when compared to ET27.5 °C. These results could assist tissue engineers in further optimising FFF of PCL bioscaffolds by reducing geometric variability between the design and fabrication of bioscaffold architectures; reducing the potential for inconsistent mechanical performance and improving yield rates as this technology transitions from prototyping to mass manufacturing environments.

Structural Aspects of Electrospun Scaffolds Intended for Prosthetics of Blood Vessels.

Electrospinning is a popular method used to fabricate small-diameter vascular grafts. However, the importance of structural characteristics of the scaffold determining interaction with endothelial cells and their precursors and blood cells is still not exhaustively clear. This review discusses current research on the significance and impact of scaffold architecture (fiber characteristics, porosity, and surface roughness of material) on interactions between cells and blood with the material. In addition, data about the effects of scaffold topography on cellular behaviour (adhesion, proliferation, and migration) are necessary to improve the rational design of electrospun vascular grafts with a long-term perspective.

Bilayered extracellular matrix derived scaffolds with anisotropic pore architecture guide tissue organization during osteochondral defect repair

While some clinical advances in cartilage repair have occurred, osteochondral (OC) defect repair remains a significant challenge, with current scaffold-based approaches failing to recapitulate the complex, hierarchical structure of native articular cartilage (AC). To address this need, we fabricated bilayered extracellular matrix (ECM)-derived scaffolds with aligned pore architectures. By modifying the freeze-drying kinetics and controlling the direction of heat transfer during freezing, it was possible to produce anisotropic scaffolds with larger pores which supported homogenous cellular infiltration and improved sulfated glycosaminoglycan deposition. Neo-tissue organization in vitro could also be controlled by altering scaffold pore architecture, with collagen fibres aligning parallel to the long-axis of the pores within scaffolds containing aligned pore networks. Furthermore, we used in vitro and in vivo assays to demonstrate that AC and bone ECM derived scaffolds could preferentially direct the differentiation of mesenchymal stromal cells (MSCs) towards either a chondrogenic or osteogenic lineage respectively, enabling the development of bilayered ECM scaffolds capable of spatially supporting unique tissue phenotypes. Finally, we implanted these scaffolds into a large animal model of OC defect repair. After 6 months in vivo, scaffold implantation was found to improve cartilage matrix deposition, with collagen fibres preferentially aligning parallel to the long axis of the scaffold pores, resulting in a repair tissue that structurally and compositionally was more hyaline-like in nature. These results demonstrate how scaffold architecture and composition can be spatially modulated to direct the regeneration of complex interfaces such as the osteochondral unit, enabling their use as cell-free, off-the-shelf implants for joint regeneration. Statement of significanceThe architecture of the extracellular matrix, while integral to tissue function, is often neglected in the design and evaluation of regenerative biomaterials. In this study we developed a bilayered scaffold for osteochondral defect repair consisting of tissue-specific extracellular matrix (ECM)-derived biomaterials to spatially direct stem/progenitor cell differentiation, with a tailored pore microarchitecture to promote the development of a repair tissue that recapitulates the hierarchical structure of native AC. The use of this bilayered scaffold resulted in improved tissue repair outcomes in a large animal model, specifically the ability to guide neo-tissue organization and therefore recapitulate key aspects of the zonal structure of native articular cartilage. These bilayer scaffolds have the potential to become a new therapeutic option for osteochondral defect repair.

Graphene Oxide-Protein-Based Scaffolds for Tissue Engineering: Recent Advances and Applications.

The field of tissue engineering is constantly evolving as it aims to develop bioengineered and functional tissues and organs for repair or replacement. Due to their large surface area and ability to interact with proteins and peptides, graphene oxides offer valuable physiochemical and biological features for biomedical applications and have been successfully employed for optimizing scaffold architectures for a wide range of organs, from the skin to cardiac tissue. This review critically focuses on opportunities to employ protein–graphene oxide structures either as nanocomposites or as biocomplexes and highlights the effects of carbonaceous nanostructures on protein conformation and structural stability for applications in tissue engineering and regenerative medicine. Herein, recent applications and the biological activity of nanocomposite bioconjugates are analyzed with respect to cell viability and proliferation, along with the ability of these constructs to sustain the formation of new and functional tissue. Novel strategies and approaches based on stem cell therapy, as well as the involvement of the extracellular matrix in the design of smart nanoplatforms, are discussed.

Three-dimensional printing of polycaprolactone/hydroxyapatite bone tissue engineering scaffolds mechanical properties and biological behavior

Controlled pore size and desirable internal architecture of bone scaffolds play a significant role in bone regeneration efficiency. In addition to choosing appropriate materials, the manufacturing method is another significant factor in fabricating the ideal scaffold. In this study, scaffolds were designed and fabricated by the fused filament fabrication (FFF) technique. Polycaprolactone (PCL) and composites films with various percentages of hydroxyapatite (HA) (up to 20%wt) were used to fabricate filaments. The influence of (HA) addition on the mechanical properties of filaments and scaffolds was investigated. in vitro biological evaluation was examined as well as the apatite formation in simulated body fluid (SBF). The addition of HA particles increased the compressive strength and Young’s modulus of filaments and consequently the scaffolds. Compared to PCL, Young’s modulus of PCL/HA20% filament and three-dimensional (3D) printed scaffold has increased by 30% and 50%, respectively. Also, Young’s modulus for all scaffolds was in the range of 30–70 MPa, which is appropriate to use in spongy bone. Besides, the MTT assay was utilized to evaluate cell viability on the scaffolds. All the samples had qualified cytocompatibility, and it would be anticipated that addition of HA particles raise the biocompatibility in vivo. Alkaline phosphatase (ALP) evaluation shows that the addition of HA caused higher ALP activity in the PCL/HA scaffolds than PCL. Furthermore, calcium deposition in the PCL/HA specimens is higher than control. In conclusion, the addition of HA particles into the PCL matrix, as well as utilizing an inexpensive commercial FFF device, lead to the fabrication of scaffolds with proper mechanical and biological properties for bone tissue engineering applications.Graphical abstract

Modified Histopathological Protocol for Poly-ɛ-Caprolactone Scaffolds Preserving Their Trabecular, Honeycomb-like Structure.

Poly-ɛ-caprolactone (PCL) is now widely studied in relation to the engineering of bone, cartilage, tendons, and other tissues. Standard histological protocols can destroy the carefully created trabecular and honeycomb-like architecture of PCL scaffolds, and could lead to scaffold fibers swelling, resulting in the displacement or compression of tissues inside the scaffold. The aim of this study was to modify a standard histopathological protocol for PCL scaffold preparation and evaluate it on porous cylindrical PCL scaffolds in a rat model. In 16 inbred Wag rats, 2 PCL scaffolds were implanted subcutaneously to both inguinal areas. Two months after implantation, harvested scaffolds were first subjected to μCT imaging, and then to histopathological analysis with standard (left inguinal area) and modified histopathological protocols (right inguinal area). To standardize the results, soft tissue percentages (STPs) were calculated on scaffold cross-sections obtained from both histopathological protocols and compared with corresponding µCT cross-sections. The modified protocol enabled the assessment of almost 10× more soft tissues on the scaffold cross-section than the standard procedure. Moreover, STP was only 1.5% lower than in the corresponding µCT cross-sections assessed before the histopathological procedure. The presented modification of the histopathological protocol is cheap, reproducible, and allows for a comprehensive evaluation of PCL scaffolds while maintaining their trabecular, honeycomb-like structure on cross-sections.

Chitosan in association with osteogenic factors as a cell-homing platform for dentin regeneration: Analysis in a pulp-in-a-chip model

ObjectiveIn this paper we propose the association of β-glycerophosphate (βGP) and calcium-hydroxide with chitosan (CH) to formulate a porous bioactive scaffold suitable as a cell-homing platform for dentin regeneration. MethodsCalcium hydroxide and βGP solutions were incorporated into chitosan to modulate scaffold architecture and composition by a phase separation technique. Architecture, chemical composition, and degradability were evaluated, and biological characterizations were performed by the seeding of dental pulp cells (DPCs) onto scaffolds, or by cultivating them in contact with leachable components (extracts), to determine cytocompatibility and odontoblastic differentiation. Cell-free scaffolds were then positioned in intimate contact with a 3D culture of DPCs in a pulp-in-a-chip platform under simulated pulp pressure. Cell mobilization and odontoblastic marker expression were evaluated. Deposition of mineralized matrix was assessed in direct contact with dentin, in the absence of osteogenic factors. ResultsIncorporation of calcium hydroxide and βGP generated a stable porous chitosan scaffold containing Ca-P nanoglobule topography (CH-Ca-βGP), which favored cell viability, alkaline phosphatase activity, and mineralized matrix deposition by cells seeded onto the scaffold structure and at a distance. The pulp-in-a-chip assay denoted its chemotactic and bioactive potential, since dentin sialoprotein-positive DPCs from 3D culture adhered to CH-Ca-βGP more than to plain chitosan. The higher deposition of mineralized matrix onto the scaffold and surrounding dentin was also observed. SignificanceA CH-Ca-βGP scaffold creates a microenvironment capable of mobilizing DPC migration toward its structure, harnessing the odontogenic potential and culminating in the expression of a highly mineralizing phenotype, key factors for a cell-homing strategy.

Polysucrose hydrogel and nanofiber scaffolds for skin tissue regeneration: Architecture and cell response

Scaffolds capable of mediating overlapping multi-cellular activities to support the different phases of wound healing while preventing scarring are essential for tissue regeneration. The potential of polysucrose as hydrogels and electrospun mats for wound healing was evaluated in vitro by seeding fibroblasts, endothelial cells and macrophages either singly or in combination. It was found that the scaffold architecture impacted cell behaviour. Electrospun mats promoted fibroblasts flattened morphology while polysucrose methacrylate (PSucMA) hydrogels promoted fibroblast spheroids formation, accentuated in the presence of endothelial cells. Hydrogels exhibited lower inflammatory response than mats and curcumin loaded scaffolds reduced TNF-α production. In vivo biocompatibility of the hydrogels tested on Wistar rats was superior to electrospun mats. In vivo wound healing studies indicated that PSucMA hydrogels integrated the surrounding tissue with better cellular infiltration and proliferation throughout the entire wound region. PSucMA hydrogels led to scarless wound closure comparable with commercially available gels.

Towards Clinical Translation of Melt Electrowritten Scaffolds: Visualisation of USPIO Labelled Polycaprolactone Scaffolds by Magnetic Resonance Imaging

Objective: Melt electrowriting (MEW) is an advanced fibre forming technology for 3D printing of complex scaffold architectures, which have convincingly shown their potential in cardiovascular constructs.1,2 Driven by electrohydrodynamic forces, a polymer fibre jet is digitally controlled to form microfibrous scaffolds via a direct writing approach. Clinical translation of such grafts requires establishing monitoring options, such as with magnetic resonance imaging (MRI). Consequently, this work proposes the labelling of medical grade polycaprolactone (PCL) with ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles as contrast agents to render the melt electrowritten composite scaffolds detectable via MRI.

Additive manufacturing of cartilage-mimetic scaffolds as off-the-shelf implants for joint regeneration

Biomimetic scaffolds that provide a tissue-specific environment to cells are particularly promising for tissue engineering and regenerative medicine applications. The goal of this study was to integrate emerging additive manufacturing and biomaterial design strategies to produce articular cartilage (AC) mimetic scaffolds that could be used as ‘off-the-shelf’ implants for joint regeneration. To this end alginate sulfate, a sulfated glycosaminoglycan (sGAG) mimic, was used to functionalize porous alginate-based scaffolds and to support the sustained release of transforming growth factor-β3 (TGF-β3). Covalent crosslinking dramatically improved the elasticity of the alginate/alginate sulfate scaffolds, while scaffold architecture could be tailored using a directional freezing technique. Introducing such an anisotropic architecture was found to promote mesenchymal stem cell (MSC) infiltration into the scaffold and to direct the orientation of the deposited extracellular matrix, leading to the development of cartilage tissue with a biomimetic zonal architecture. In vitro experiments also demonstrated the capacity of the sulfated scaffolds to both enhance chondrogenesis of MSCs and to control the release of TGF-β3, leading to the development of a tissue rich in sGAG and type II collagen. The scaffolds were further reinforced with a 3D printed poly(lactide-co-ϵ-caprolactone) (PLCL) framework, leading to composite implants that were more elastic than those reinforced with polycaprolactone, and which better mimicked the bulk mechanical properties of native cartilage tissue. The ability of this composite scaffold to support chondrogenesis was then confirmed within a dynamic culture system. Altogether, these findings demonstrate the potential of such biomimetic scaffolds as putative ‘single-stage’ or ‘off-the-shelf’ strategies for AC regeneration.

Novel 3D Bioglass Scaffolds for Bone Tissue Regeneration.

The design of scaffolds with optimal biomechanical properties for load-bearing applications is an important topic of research. Most studies have addressed this problem by focusing on the material composition and not on the coupled effect between the material composition and the scaffold architecture. Polymer–bioglass scaffolds have been investigated due to the excellent bioactivity properties of bioglass, which release ions that activate osteogenesis. However, material preparation methods usually require the use of organic solvents that induce surface modifications on the bioglass particles, compromising the adhesion with the polymeric material thus compromising mechanical properties. In this paper, we used a simple melt blending approach to produce polycaprolactone/bioglass pellets to construct scaffolds with pore size gradient. The results show that the addition of bioglass particles improved the mechanical properties of the scaffolds and, due to the selected architecture, all scaffolds presented mechanical properties in the cortical bone region. Moreover, the addition of bioglass indicated a positive long-term effect on the biological performance of the scaffolds. The pore size gradient also induced a cell spreading gradient.

Electrospun Scaffold Micro-Architecture Induces an Activated Transcriptional Phenotype within Tendon Fibroblasts.

Biomaterial augmentation of surgically repaired rotator cuff tendon tears aims to improve the high failure rates (∼40%) of traditional repairs. Biomaterials that can alter cellular phenotypes through the provision of microscale topographical cues are now under development. We aimed to systematically evaluate the effect of topographic architecture on the cellular phenotype of fibroblasts from healthy and diseased tendons. Electrospun polydioxanone scaffolds with fiber diameters ranging from 300 to 4000 nm, in either a highly aligned or random configuration, were produced. Healthy tendon fibroblasts cultured for 7 days on scaffolds with highly aligned fibers demonstrated a distinctive elongated morphology, whilst those cultured on randomly configured fibers demonstrated a flattened and spread morphology. The effect of scaffold micro-architecture on the transcriptome of both healthy and diseased tendon fibroblasts was assessed with bulk RNA-seq. Both healthy (n = 3) and diseased tendon cells (n = 3) demonstrated a similar transcriptional response to architectural variants. Gene set enrichment analysis revealed that large diameter (≥2000 nm) aligned scaffolds induced an upregulation of genes involved in cellular replication and a downregulation of genes defining inflammatory responses and cell adhesion. Similarly, PDPN and CD248, markers of inflammatory or “activated” fibroblasts, were downregulated during culture of both healthy and diseased fibroblasts on aligned scaffolds with large (≥2000 nm) fiber diameters. In conclusion scaffold architectures resembling that of disordered type III collagen, typically present during the earlier phases of wound healing, resulted in tendon fibroblast activation. Conversely, scaffolds mimicking aligned diameter collagen I fibrils, present during tissue remodelling, did not activate tendon derived fibroblasts. This has implications for the design of scaffolds used during rotator cuff repair augmentation.