Design Of Tissue Engineering Scaffolds Research Articles

Bioactive polymer composite scaffolds fabricated from 3D printed negative molds enable bone formation and vascularization

Scaffolds for bone defect treatment should ideally support vascularization and promote bone formation, to facilitate the translation into biomedical device applications. This study presents a novel approach utilizing 3D-printed water-dissolvable polyvinyl alcohol (PVA) sacrificial molds to engineer polymerized High Internal Phase Emulsion (polyHIPE) scaffolds with microchannels and distinct multiscale porosity. Two sacrificial mold variants (250 µm and 500 µm) were generated using fused deposition modeling, filled with HIPE, and subsequently dissolved to create polyHIPE scaffolds containing microchannels. In vitro assessments demonstrated significant enhancement in cell infiltration, proliferation, and osteogenic differentiation, underscoring the favorable impact of microchannels on cell behavior. High loading efficiency and controlled release of the osteogenic factor BMP-2 were achieved, with microchannels facilitating release of the growth factor. Evaluation in a mouse critical-size calvarial defect model revealed enhanced vascularization and bone formation in microchanneled scaffolds containing BMP-2. This study not only introduces an accessible method for creating multiscale porosity in polyHIPE scaffolds but also emphasizes its capability to enhance cellular infiltration, controlled growth factor release, and in vivo performance. The findings suggest promising applications in bone tissue engineering and regenerative medicine, and are expected to facilitate the translation of this type of biomaterial scaffold. Statement of significanceThis study holds significance in the realm of biomaterial scaffold design for bone tissue engineering and regeneration. We demonstrate a novel method to introduce controlled multiscale porosity and microchannels into polyHIPE scaffolds, by utilizing 3D-printed water-dissolvable PVA molds. The strategy offers new possibilities for improving cellular infiltration, achieving controlled release of growth factors, and enhancing vascularization and bone formation outcomes. This microchannel approach not only marks a substantial stride in scaffold design but also demonstrates its tangible impact on enhancing osteogenic cell differentiation and fostering robust bone formation in vivo. The findings emphasize the potential of this methodology for bone regeneration applications, showcasing an interesting advancement in the quest for effective and innovative biomaterial scaffolds to regenerate bone defects.

Optimizing Filament-Based TCP Scaffold Design for Osteoconduction and Bone Augmentation: Insights from In Vivo Rabbit Models.

Additive manufacturing has emerged as a transformative tool in biomedical engineering, offering precise control over scaffold design for bone tissue engineering and regenerative medicine. While much attention has been focused on optimizing pore-based scaffold architectures, filament-based microarchitectures remain relatively understudied, despite the fact that the majority of 3D-printers generate filament-based structures. Here, we investigated the influence of filament characteristics on bone regeneration outcomes using a lithography-based additive manufacturing approach. Three distinct filament-based scaffolds (Fil050, Fil083, and Fil125) identical in macroporosity and transparency, crafted from tri-calcium phosphate (TCP) with varying filament thicknesses and distance, were evaluated in a rabbit model of bone augmentation and non-critical calvarial defect. Additionally, two scaffold types differing in filament directionality (Fil and FilG) were compared to elucidate optimal design parameters. Distance of bone ingrowth and percentage of regenerated area within scaffolds were measured by histomorphometric analysis. Our findings reveal filaments of 0.50 mm as the most effective filament-based scaffold, demonstrating superior bone ingrowth and bony regenerated area compared to larger size filament (i.e., 0.83 mm and 1.25 mm scaffolds). Optimized directionality of filaments can overcome the reduced performance of larger filaments. This study advances our understanding of microarchitecture's role in bone tissue engineering and holds significant implications for clinical practice, paving the way for the development of highly tailored, patient-specific bone substitutes with enhanced efficacy.

Design and Additive Manufacturing of Nerve Guide Conduits Using Triple Periodic Minimal Surface Structures

Scaffold design is a key study area in tissue engineering. A scaffold is a three-dimensional framework that provides temporary support for the formation of new tissue before being implanted with isolated cells. The aim of tissue engineering scaffolds is to be colonized by cells. To ensure sufficient tissue growth, scaffolds need to transmit the necessary chemical and physical signals. The design of the scaffold determines its functionality. The design and manufacturing of tissue engineering scaffolds is a highly complex procedure. Scaffolds must have the necessary qualities to create an optimal architecture for cell growth, proliferation, and differentiation in order to form tissue. However, constrained structural designs and outdated manufacturing procedures impede the enhancement of scaffold qualities. To address these restrictions, researchers are merging computer-aided scaffold design with 3D printing processes during production. This method permits the design and manufacture of scaffolds with extremely intricate microstructures. The literature shows that computer-aided design combined with 3D printing technology is often utilized to design and manufacture nerve guide conduits for nerve regeneration. In this study, three different nerve guide conduit structures were designed and produced. Two of them are based on triple periodic minimal surfaces derived from Gyroid, schwarz. Although triple periodic minimal surfaces used as the basis for scaffold designs offer promising advantages for tissue engineering applications, limited information is available regarding their manufacturability. The designs created in this study, as well as their fabrication, will add to the literature on the manufacturability of triple periodic minimum surfaces.

Silk fibroin incorporated electrospun aliphatic polyester nanofiber scaffolds with excellent dimensional stability and cytocompatibility

Poly (lactide-co-glycolide) (PLGA) has been extensively utilized to fabricate electrospun fibers for tissue engineering scaffolds due to its good biocompatibility and biodegradability. However, such scaffolds in biological media shrink and distort severely, which is undesired for practical application. In this study, a silk fibroin incorporated electrospun aliphatic polyester nanofiber scaffold (PLGA/PCL/SF) was developed via facile one-step electrospinning, with PLGA as the substrate and incorporating polycaprolactone (PCL) and silk fibroin (SF). The resulting scaffold exhibited fine fibers and water absorbency. Particularly, the addition of SF into scaffolds improved the water adsorption ability, and that of PCL effectively prevented scaffold shrinkage behaviors. In vitro degradation analyses showed that it was accelerating for the obtained scaffolds with SF added, without any undesirable acidic environment induced. Additionally, the PLGA/PCL/SF scaffold exhibited the most promising result in cell proliferation, presenting a valuable insight into the design of tissue engineering scaffolds.

Electrospun Scaffolds are Not Necessarily Always Made of Nanofibers as Demonstrated by Polymeric Heart Valves for Tissue Engineering.

In the last 30 years, there are ≈60000 publications about electrospun nanofibers, but it is still unclear whether nanoscale fibers are really necessary for electrospun tissue engineering scaffolds. The present report puts forward this argument and reveals that compared with electrospun nanofibers, microfibers with diameter of ≈3 µm (named as "oligo-micro fiber") are more appropriate for tissue engineering scaffolds owing to their better cell infiltration ability caused by larger pores with available nuclear deformation. To further increase pore sizes, electrospun poly(ε-caprolactone) (PCL) scaffolds are fabricated using latticed collectors with meshes. Fiber orientation leads to sufficient mechanical strength albeit increases porosity. The latticed scaffolds exhibit good biocompatibility and improve cell infiltration. Under aortic conditions in vitro, the performances of latticed scaffolds are satisfactory in terms of the acute systolic hemodynamic functionality, except for the higher regurgitation fraction caused by the enlarged pores. This hierarchical electrospun scaffold with sparse fibers in macropores and oligo-micro fibers in filaments provides new insights into the design of tissue engineering scaffolds, and tissue engineering may provide living heart valves with regenerative capabilities for patients with severe valve disease in the future.

Design and Fabricating of Structured Tissue Engineering Scaffolds With Different Porosity Architecture

Design and Fabricating of Structured Tissue Engineering Scaffolds With Different Porosity Architecture

Nanodiamonds for tissue engineering and regeneration

Tissue engineering mainly mimics the native cell environment for cell growth and regeneration. In regard to this strategy, different materials have been used for fabrication of cell support in an in vivo resembling ambience. An ideal tissue engineering scaffold provides biocompatible, three-dimensional (3D), porous structure with tissue-specific properties such as the electrical conductivity in neural tissue and controlled biodegradability. Therefore, the precise selection of biopolymers and nanomaterials with properties of non-interfering biological processes, like cell proliferation and differentiation plays an important role in the successful scaffold design and final outcome in tissue engineering. Nanodiamonds (NDs) are carbon-based nanomaterials and possess remarkable features including biocompatibility, high surface area, optical properties, chemical stability, excellent mechanical and physical properties, highly functional surfaces, and highly resistant to harsh environments, which make them highly validated as multi-functional materials for biomedical applications. These characteristics have recently attracted much attention in the area of regenerative medicine from the scaffold to prosthesis designing. This review will discuss the extensive applications of NDs in different scopes of regenerative medicine, including tissue engineering, however, the main focus will be on neural regeneration.

Ligand Composition and Coating Density Co-Modulate the Chondrocyte Function on Poly(glycerol-dodecanedioate).

Inducing chondrocyte redifferentiation and promoting cartilaginous matrix accumulation are key challenges in the application of biomaterials in articular cartilage repair. Poly(glycerol-dodecanedioate) (PGD) is a viable candidate for scaffold design in cartilage tissue engineering (CTE). However, the surface properties of PGD are not ideal for cell attachment and growth due to its relative hydrophobicity compared with natural extracellular matrix (ECM). In this study, PGD was coated with various masses of collagen type I or hyaluronic acid, individually or in combination, to generate a cell-material interface with biological cues. The effects of ligand composition and density on the PGD surface properties and shape, metabolic activity, cell phenotype, and ECM production of human articular chondrocytes (hACs) were evaluated. Introducing ECM ligands on PGD significantly improved its hydrophilicity and promoted the chondrocyte's anabolic activity. The morphology and anabolic activity of hACs on PGD were co-modulated by ligand composition and density, suggesting a combinatorial effect of both coating parameters on chondrocyte function during monolayer culture. Hyaluronic acid and its combination with collagen maintained a round cell shape and redifferentiated phenotype. This study demonstrated the complex mechanism of ligand-guided interactions between cell and biomaterial substrate and the potential of PGD as a scaffold material in the field of CTE.

Feasibility of caffeic acid as a crosslinking agent in modifying acellular extracellular matrices

Acellular extracellular matrices (aECM) are commonly utilized, both experimentally and clinically, in the regenerative medicine field. However, some disadvantages such as rapid degradation, poor mechanical properties, chronic inflammatory reactions and low antioxidant activity have limited their further application. In this study the feasibility of caffeic acid as a crosslinking agent in fixing small intestinal submucosa (SIS) was evaluated. The ninhydrin assay, swelling ratio and FTIR spectra indicated that caffeic acid can efficiently react with free amino groups to crosslink SIS and the highest crosslinking index reached 21.60 ± 1.37%. Moreover, the shrinkage temperature of SIS remarkably increased from 59 °C to about 80 °C and the degradation rate of CA-SIS was all lower than 6%, demonstrating their improved biostability and hydrothermal stability. Importantly, the antioxidant activity of CA-SIS ranged from 55% to 90%, statistically higher than that of native SIS (37.33 ± 2.94%). Additionally the cytotoxicity test presented that the cytotoxicity grade of CA-SIS was 1 or 0, whilst large numbers of living HUVECs were attached to the surface of the material and exhibited high cell viability. These results indicated their excellent cytocompatibility. The data of subcutaneous implant displayed that the number of inflammatory cells in 2%- and 2.5%CA-SIS groups remained at a low level (below 100 cells/field) while that of the native SIS group continued increasing, finally reaching 142.33 ± 30.92 cells/field. In conclusion, caffeic acid is a promising candidate for modifying aECM and may play a vital role in the design and fabrication of tissue engineering scaffolds.

Toward a New Generation of Bio-Scaffolds for Neural Tissue Engineering: Challenges and Perspectives.

Neural tissue engineering presents a compelling technological breakthrough in restoring brain function, holding immense promise. However, the quest to develop implantable scaffolds for neural culture that fulfill all necessary criteria poses a remarkable challenge for material science. These materials must possess a host of desirable characteristics, including support for cellular survival, proliferation, and neuronal migration and the minimization of inflammatory responses. Moreover, they should facilitate electrochemical cell communication, display mechanical properties akin to the brain, emulate the intricate architecture of the extracellular matrix, and ideally allow the controlled release of substances. This comprehensive review delves into the primary requisites, limitations, and prospective avenues for scaffold design in brain tissue engineering. By offering a panoramic overview, our work aims to serve as an essential resource, guiding the creation of materials endowed with bio-mimetic properties, ultimately revolutionizing the treatment of neurological disorders by developing brain-implantable scaffolds.

Stiffness-dependent dynamic effect of inflammation on keratocyte phenotype and differentiation

Although extensive studies have evaluated the regulation effect of microenvironment on cell phenotype and cell differentiation, further investigations in the field of the cornea are needed to gain sufficient knowledge for possible clinical translation. This study aims to evaluate the regulation effects of substrate stiffness and inflammation on keratocyte phenotype of corneal fibroblasts, as well as the differentiation from stem cells towards keratocytes. Soft and stiff substrates were prepared based on polydimethylsiloxane. HTK and stem cells were cultured on these substrates to evaluate the effects of stiffness. The possible synergistic effects between substrate stiffness and inflammatory factor IL-1β were examined by qPCR and immunofluorescence staining. In addition, macrophages were cultured on soft and stiff substrates to evaluate the effect of substrate stiffness on the synthesis of inflammatory factors. The conditioned medium of macrophages (Soft-CM and Stiff-CM) was collected to examine the effects on HTK and stem cells. It was found that inflammatory factor IL-1β promoted keratocyte phenotype and differentiation when cells were cultured on soft substrate (∼130 kPa), which were different from cells cultured on stiff substrate (∼2 × 103 kPa) and TCP (∼106 kPa). Besides, macrophages cultured on stiff substrates had significantly higher expression of IL-1β and Tnf-α as compared to the cells cultured on soft substrates. And Stiff-CM decreased the expression of keratocyte phenotype markers as compared to Soft-CM. The results of our study indicate a stiffness-dependent dynamic effect of inflammation on keratocyte phenotype and differentiation, which is of significance not only in gaining a deeper knowledge of corneal pathology and repair, but also in being instructive for scaffold design in corneal tissue engineering and ultimate regeneration.

Material Stiffness in Cooperation with Macrophage Paracrine Signals Determines the Tenogenic Differentiation of Mesenchymal Stem Cells.

Stiffness is an important physical property of biomaterials that determines stem cell fate. Guiding stem cell differentiation via stiffness modulation has been considered in tissue engineering. However, the mechanism by which material stiffness regulates stem cell differentiation into the tendon lineage remains controversial. Increasing evidence demonstrates that immune cells interact with implanted biomaterials and regulate stem cell behaviors via paracrine signaling; however, the role of this mechanism in tendon differentiation is not clear. In this study, polydimethylsiloxane (PDMS) substrates with different stiffnesses are developed, and the tenogenic differentiation of mesenchymal stem cells (MSCs) exposed to different stiffnesses and macrophage paracrine signals is investigated. The results reveal that lower stiffnesses facilitates tenogenic differentiation of MSCs, while macrophage paracrine signals at these stiffnesses suppress the differentiation. When exposed to these two stimuli, MSCs still exhibit enhanced tendon differentiation, which is further elucidated by global proteomic analysis. Following subcutaneous implantation in rats for 2 weeks, soft biomaterial induces only low inflammation and promotes tendon-like tissue formation. In conclusion, the study demonstrates that soft, rather than stiff, material has a greater potential to guide tenogenic differentiation of stem cells, which provides comprehensive evidence for optimized bioactive scaffold design in tendon tissue engineering.

3D printing of cell-delivery scaffolds for tissue regeneration.

Tissue engineering strategy that combine biomaterials with living cells has shown special advantages in tissue regeneration and promoted the development of regenerative medicine. In particular, the rising of 3D printing technology further enriched the structural design and composition of tissue engineering scaffolds, which also provided convenience for cell loading and cell delivery of living cells. In this review, two types of cell-delivery scaffolds for tissue regeneration, including 3D printed scaffolds with subsequent cell-seeding and 3D cells bioprinted scaffolds, are mainly reviewed. We devote a major part to present and discuss the recent advances of two 3D printed cell-delivery scaffolds in regeneration of various tissues, involving bone, cartilage, skin tissues etc. Although two types of 3D printed cell-delivery scaffolds have some shortcomings, they do have generally facilitated the exploration of tissue engineering scaffolds in multiple tissue regeneration. It is expected that 3D printed cell-delivery scaffolds will be further explored in function mechanism of seeding cells in vivo, precise mimicking of complex tissues and even organ reconstruction under the cooperation of multiple fields in future.

Maintenance of multipotency of bone marrow mesenchymal stem cells on poly(ε-caprolactone) nanoneedle arrays through the enhancement of cell-cell interaction.

Mesenchymal stem cells (MSCs), with high self-renewal ability and multipotency, are commonly used as the seed cells for tissue engineering. However, the reduction and loss of multipotential ability after necessary expansion in vitro set up a heavy obstacle to the clinical application of MSCs. Here in this study, we exploit the autologous crystallization ability of biocompatible poly (ε-caprolactone) (PCL) to obtain uniformly distributed nanoneedle arrays. By controlling the molecular weight of PCL, nanoneedle with a width of 2μm and height of 50nm, 80nm, and 100nm can be successfully fabricated. After surface chemical modification with polydopamine (PDA), the water contact angle of the fabricated PCL nanoneedle arrays are reduced from 84° to almost 60° with no significant change of the nanostructure. All the fabricated substrates are cultured with bone marrow MSCs (BMMSCs), and the adhesion, spreading, proliferation ability and multipotency of cells on different substrates are investigated. Compared with the BMMSCs cultured on pure PCL nanoneedle arrays, the decoration of PDA can improve the adhesion and spreading of cells and further change them from aggregated distribution to laminar distribution. Nevertheless, the laminar distribution of cultured cells leads to a weak cell-cell interaction, and hence the multipotency of BMMSCs cultured on the PCL-PDA substrates is decimated. On the contrary, the pure PCL nanoneedle arrays can be used to maintain the multipotency of BMMSCs via clustered growth, and the PCL1 nanoneedle array with a height of 50nm is more promising than the other 2 with regard to the highest proliferation rate and best multipotential differentiation ability of cultured cells. Interestingly, there is a positive correlation between the strength of cell-cell interaction and the multipotency of stem cells in vitro. In conclusion, we have successfully maintained the multipotency of BMMSCs by using the PCL nanoneedle arrays, especially the PCL1 nanoneedle array with a height of 50nm, as the substrates for in vitro extension, and further revealed the importance of cell-cell interaction on the multipotency of MSCs. The study provides a theoretical basis for the behavioral regulation of MSCs, and is instructive to the design of tissue engineering scaffolds.

DESIGNING OF DIFFERENT TYPES OF GYROID SCAFFOLD ARCHITECTURE TO ACHIEVE PATIENT-SPECIFIC OSSEOINTEGRATION FRIENDLY MECHANICAL ENVIRONMENT

Porous three-dimensional scaffolds provide a favorable environment for the regeneration of tissues and organs to recover the injured tissue in terms of structure and biological function. Proper design of scaffold architecture is a crucial factor for tissue engineering. The mechanical properties and performance of the scaffold depend on the scaffold architecture, material, and geometry. In this study, numerical analysis is performed on four types of gyroids to see the variation of effective elastic modules in three different directions. The fluid flow induces wall shear stress (WSS), considering the Newtonian fluid is also investigated. Initially, the WSS is evaluated through computational fluid dynamics simulation and compared to previous literature. A good agreement between the results of published literature and those initial works encourages dealing with several cases by varying different parameters. The effect of scaffold architecture, pore size, and flow direction on WSS is studied using 16 different gyroid models. A significant reduction of scaffold stiffness can be achieved with porosity, and WSS increases with a decrease in pore size. Inlet flow direction also has a good influence on WSS. Fluid flow perpendicular to the gyroid cross-sectional area gives maximum WSS and gradually decreases with an increase in direction angle. The present study can be used to reference a specific scaffold design in tissue engineering.

Modular-based gradient scaffold design and experimental studies for tissue engineering: enabling customized structures and mechanical properties

Modular-based gradient scaffold design and experimental studies for tissue engineering: enabling customized structures and mechanical properties

Geometry-Based Computational Fluid Dynamic Model for Predicting the Biological Behavior of Bone Tissue Engineering Scaffolds.

The use of biocompatible and biodegradable porous scaffolds produced via additive manufacturing is one of the most common approaches in tissue engineering. The geometric design of tissue engineering scaffolds (e.g., pore size, pore shape, and pore distribution) has a significant impact on their biological behavior. Fluid flow dynamics are important for understanding blood flow through a porous structure, as they determine the transport of nutrients and oxygen to cells and the flushing of toxic waste. The aim of this study is to investigate the impact of the scaffold architecture, pore size and distribution on its biological performance using Computational Fluid Dynamics (CFD). Different blood flow velocities (BFV) induce wall shear stresses (WSS) on cells. WSS values above 30 mPa are detrimental to their growth. In this study, two scaffold designs were considered: rectangular scaffolds with uniform square pores (300, 350, and 450 µm), and anatomically designed circular scaffolds with a bone-like structure and pore size gradient (476–979 µm). The anatomically designed scaffolds provided the best fluid flow conditions, suggesting a 24.21% improvement in the biological performance compared to the rectangular scaffolds. The numerical observations are aligned with those of previously reported biological studies.

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.

A focused review of short electrospun nanofiber preparation techniques for composite reinforcement

Abstract Short nanofibers have been of interest in preparing 3D porous structures, aerosol filters, and nanocomposites. These materials require nanofiber retrieval and application in short form with simultaneous control over aspect ratio. Electrospinning, conventionally, offers minimal control over short nanofiber yield as nonwoven mat is the default configuration of collected sample. High surface area to volume ratio nanofiber, however, can offer new vistas in material design if standardization of short nanofiber preparation practices, offering control over aspect ratio, can be attained. It will provide novel insights into design of tissue engineering scaffolds, filtration membranes, and nanocomposite properties. This work summarizes reported efforts to prepare short nanofiber through mechanical, chemical, material, and operational variables. It aims to provide comparative glance at attempts to control aspect ratio along with pros and cons of the adopted techniques. Lastly, discussion shares generalized conclusions and insights gathered while reviewing material and operational variables adopted for short nanofiber preparation.

Application of microcarrier technology in cartilage repairing: a review

Cartilage has poor self-recovery because of its characteristics of no blood vessels and high extracellular matrix. In clinical treatment, physical therapy or drug therapy is usually used for mild cartilage defects, and surgical treatment is needed for severe ones. In recent years, cartilage tissue engineering technology provides a new way for the treatment of cartilage defects. Compared with the traditional surgical treatment, cartilage tissue engineering technology has the advantages of small wound and good recovery. The application of microcarrier technology in the design of tissue engineering scaffolds further expands the function of scaffolds and promotes cartilage regeneration. This review summarized the main preparation methods and development of microcarrier technology in recent years. Subsequently, the properties and specific application scenarios of microcarriers with different materials and functions were introduced according to the materials and functions of microcarriers used in cartilage repair. Based on our research on osteochondral integrated layered scaffolds, we proposed an idea of optimizing the performance of layered scaffolds through microcarriers, which is expected to prepare bionic scaffolds that are more suitable for the structural characteristics of natural cartilage.