Oriented Collagen Scaffolds for Tissue Engineering

Oriented collagen scaffolds were developed in the form of sheet, mesh and tube by arraying flow-oriented collagen string gels and dehydrating the arrayed gels. The developed collagen scaffolds can be any practical size with any direction of orientation for tissue engineering applications. The birefringence of the collagen scaffolds was quantitatively analyzed by parallel Nicols method. Since native collagen in the human body has orientations such as bone, cartilage, tendon and cornea, and the orientation has a special role for the function of human organs, the developed various types of three-dimensional oriented collagen scaffolds are expected to be useful biomaterials for tissue engineering and regenerative medicines.     

Fabrication of Piezoelectric Electrospun Termite Nest-like 3D Scaffolds for Tissue Engineering

A high piezoelectric coefficient polymer and biomaterial for bone tissue engineering— poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP)—has been successfully fabricated into 3D scaffolds using the wet electrospinning method. Three-dimensional (3D) scaffolds have significant advantages for tissue engineering applications. Electrospinning is an advanced method and can fabricate 3D scaffolds. However, it has some limitations and is difficult to fabricate nanofibers into 3D shapes because of the low controllability of porosity and internal pore shape. The PVDF-HFP powders were dissolved in a mixture of acetone and dimethylformamide with a ratio of 1:1 at various concentrations of 10, 13, 15, 17, and 20 wt%. However, only the solutions at 15 and 17 wt% with optimized electrospinning parameters can be fabricated into biomimetic 3D shapes. The produced PVDF-HFP 3D scaffolds are in the cm size range and mimic the structure of the natural nests of termites of the genus Apicotermes. In addition, the 3D nanofiber-based structure can also generate more electrical signals than the conventional 2D ones, as the third dimension provides more compression. The cell interaction with the 3D nanofibers scaffold was investigated. The in vitro results demonstrated that the NIH 3T3 cells could attach and migrate in the 3D structures. While conventional electrospinning yields 2D (flat) structures, our bio-inspired electrospun termite nest-like 3D scaffolds are better suited for tissue engineering applications since they can potentially mimic native tissues as they have biomimetic structure, piezoelectric, and biological properties.     

Cartilage Tissue Engineering with Silk Fibroin Scaffolds Fabricated by Indirect Additive Manufacturing Technology

Advanced tissue engineering (TE) technology based on additive manufacturing (AM) can fabricate scaffolds with a three-dimensional (3D) environment suitable for cartilage regeneration. Specifically, AM technology may allow the incorporation of complex architectural features. The present study involves the fabrication of 3D TE scaffolds by an indirect AM approach using silk fibroin (SF). From scanning electron microscopic observations, the presence of micro-pores and interconnected channels within the scaffold could be verified, resulting in a TE scaffold with both micro- and macro-structural features. The intrinsic properties, such as the chemical structure and thermal characteristics of SF, were preserved after the indirect AM manufacturing process. In vitro cell culture within the SF scaffold using porcine articular chondrocytes showed a steady increase in cell numbers up to Day 14. The specific production (per cell basis) of the cartilage-specific extracellular matrix component (collagen Type II) was enhanced with culture time up to 12 weeks, indicating the re-differentiation of chondrocytes within the scaffold. Subcutaneous implantation of the scaffold-chondrocyte constructs in nude mice also confirmed the formation of ectopic cartilage by histological examination and immunostaining.     

Additive Manufactured Magnesium-Based Scaffolds for Tissue Engineering

Additive manufacturing (AM) is an important technology that led to a high evolution in the manufacture of personalized implants adapted to the anatomical requirements of patients. Due to a worldwide graft shortage, synthetic scaffolds must be developed. Regarding this aspect, biodegradable materials such as magnesium and its alloys are a possible solution because the second surgery for implant removal is eliminated. Magnesium (Mg) exhibits mechanical properties, which are similar to human bone, biodegradability in human fluids, high biocompatibility, and increased ability to stimulate new bone formation. A current research trend consists of Mg-based scaffold design and manufacture using AM technologies. This review presents the importance of biodegradable implants in treating bone defects, the most used AM methods to produce Mg scaffolds based on powder metallurgy, AM-manufactured implants properties, and in vitro and in vivo analysis. Scaffold properties such as biodegradation, densification, mechanical properties, microstructure, and biocompatibility are presented with examples extracted from the recent literature. The challenges for AM-produced Mg implants by taking into account the available literature are also discussed.     

Auxetic Structures for Tissue Engineering Scaffolds and Biomedical Devices

An auxetic structure utilizing a negative Poisson’s ratio, which can expand transversally when axially expanded under tensional force, has not yet been studied in the tissue engineering and biomedical area. However, the recent advent of new technologies, such as additive manufacturing or 3D printing, has showed prospective results aimed at producing three-dimensional structures. Auxetic structures are fabricated by additive manufacturing, soft lithography, machining technology, compressed foaming, and textile fabrication using various biomaterials, including poly(ethylene glycol diacrylate), polyurethane, poly(lactic-glycolic acid), chitosan, hydroxyapatite, and using a hard material such as a silicon wafer. After fabricating the scaffold with an auxetic effect, researchers have cultured fibroblasts, osteoblasts, chondrocytes, myoblasts, and various stem cells, including mesenchymal stem cells, bone marrow stem cells, and embryonic stem cells. Additionally, they have shown new possibilities as scaffolds through tissue engineering by cell proliferation, migration, alignment, differentiation, and target tissue regeneration. In addition, auxetic structures and their unique deformation characteristics have been explored in several biomedical devices, including implants, stents, and surgical screws. Although still in the early stages, the auxetic structure, which can create mechanical properties tailored to natural tissue by changing the internal architecture of the structure, is expected to show an improved tissue reconstruction ability. In addition, continuous research at the cellular level using the auxetic micro and nano-environment could provide a breakthrough for tissue reconstruction.     

组织工程心脏瓣膜支架材料进展

当前应用于临床的瓣膜置换物(生物瓣和机械瓣)都不同程度的存在着一定缺陷。组织工程心脏瓣膜为理想生物瓣膜替代物的出现带来了希望。组织工程心脏瓣膜的构建包括:⑴支架的选材及制备;⑵种子细胞的选择、分离与培养;⑶种子细胞在支架上的种植与培养三部分。现重点综述近年来国内外在组织工程心脏瓣膜支架材料研究上的新进展新突破,展望了支架材料发展的方向和前景。     

Novel Textile Scaffolds Generated by Flock Technology for Tissue Engineering of Bone and Cartilage

Textile scaffolds can be found in a variety of application areas in regenerative medicine and tissue engineering. In the present study we used electrostatic flocking—a well-known textile technology—to produce scaffolds for tissue engineering of bone. Flock scaffolds stand out due to their unique structure: parallel arranged fibers that are aligned perpendicularly to a substrate, resulting in mechanically stable structures with a high porosity. In compression tests we demonstrated good mechanical properties of such scaffolds and in cell culture experiments we showed that flock scaffolds allow attachment and proliferation of human mesenchymal stem cells and support their osteogenic differentiation. These matrices represent promising scaffolds for tissue engineering.     

Mesenchymal Stem Cells in Combination with Scaffolds for Bone Tissue Engineering

This article reviews past and current strategies of the use of bone graft substitutes along with the future biologic alternatives that can enhance the functional capabilities of those grafts. Many of these bone graft substitute alternatives include ceramic-based, allograft-based, factor-based and polymer-based whereas others are cell-based. The ways of achieving the goal of tissue engineering using stem cells and their lineage to regenerate tissue have been detailed with regard to both the generation of sufficient vascular invasion of the tissue to improve oxygen and nutrient supply, and the development of innovative physical/chemical stimuli to induce bone formation with the proper biomaterial to carry the cells. It is imperative to integrate basic polymer science with molecular biology and stem cell biology, in the design of new materials that perform very sophisticated signaling needed for integration and function.     

Experimental and Numerical Simulations of 3D-Printed Polycaprolactone Scaffolds for Bone Tissue Engineering Applications

Ideal bone scaffolds for tissue engineering should be highly porous allowing cell attachment, spreading, and differentiation and presenting appropriate biomechanical properties. These antagonistic characteristics usually require extensive experimental work to achieve optimised balanced properties. This paper presents a simulation approach to determine the mechanical behaviour of bone scaffolds allowing the compressive modulus and the deformation mechanisms to be predicted. Polycaprolactone scaffolds with regular square pores and different porosities were considered. Scaffolds were also printed using an extrusion-based additive manufacturing and assessed under compressive loads. Similar designs were used for both simulation and fabrication steps. A good correlation between numerical and experimental results was obtained, highlighting the suitability of the simulation tool for the mechanical design of 3D-printed bone scaffolds.     

The Clinical Application of Porous Tantalum and Its New Development for Bone Tissue Engineering

Porous tantalum (Ta) is a promising biomaterial and has been applied in orthopedics and dentistry for nearly two decades. The high porosity and interconnected pore structure of porous Ta promise fine bone ingrowth and new bone formation within the inner space, which further guarantee rapid osteointegration and bone–implant stability in the long term. Porous Ta has high wettability and surface energy that can facilitate adherence, proliferation and mineralization of osteoblasts. Meanwhile, the low elastic modulus and high friction coefficient of porous Ta allow it to effectively avoid the stress shield effect, minimize marginal bone loss and ensure primary stability. Accordingly, the satisfactory clinical application of porous Ta-based implants or prostheses is mainly derived from its excellent biological and mechanical properties. With the advent of additive manufacturing, personalized porous Ta-based implants or prostheses have shown their clinical value in the treatment of individual patients who need specially designed implants or prosthesis. In addition, many modification methods have been introduced to enhance the bioactivity and antibacterial property of porous Ta with promising in vitro and in vivo research results. In any case, choosing suitable patients is of great importance to guarantee surgical success after porous Ta insertion.     

Bioactive Ceramic Scaffolds for Bone Tissue Engineering by Powder Bed Selective Laser Processing: A Review

The implementation of a powder bed selective laser processing (PBSLP) technique for bioactive ceramics, including selective laser sintering and melting (SLM/SLS), a laser powder bed fusion (L-PBF) approach is far more challenging when compared to its metallic and polymeric counterparts for the fabrication of biomedical materials. Direct PBSLP can offer binder-free fabrication of bioactive scaffolds without involving postprocessing techniques. This review explicitly focuses on the PBSLP technique for bioactive ceramics and encompasses a detailed overview of the PBSLP process and the general requirements and properties of the bioactive scaffolds for bone tissue growth. The bioactive ceramics enclosing calcium phosphate (CaP) and calcium silicates (CS) and their respective composite scaffolds processed through PBSLP are also extensively discussed. This review paper also categorizes the bone regeneration strategies of the bioactive scaffolds processed through PBSLP with the various modes of functionalization through the incorporation of drugs, stem cells, and growth factors to ameliorate critical-sized bone defects based on the fracture site length for personalized medicine.     

Comparison of CAD and Voxel-Based Modelling Methodologies for the Mechanical Simulation of Extrusion-Based 3D Printed Scaffolds

Porous structures are of great importance in tissue engineering. Most scaffolds are 3D printed, but there is no single methodology to model these printed parts and to apply finite element analysis to estimate their mechanical behaviour. In this work, voxel-based and geometry-based modelling methodologies are defined and compared in terms of computational efficiency, dimensional accuracy, and mechanical behaviour prediction of printed parts. After comparing the volumes and dimensions of the models with the theoretical and experimental ones, they are more similar to the theoretical values because they do not take into account dimensional variations due to the printing temperature. This also affects the prediction of the mechanical behaviour, which is not accurate compared to reality, but it makes it possible to determine which geometry is stiffer. In terms of comparison of modelling methodologies, based on process efficiency, geometry-based modelling performs better for simple or larger parts, while voxel-based modelling is more advantageous for small and complex geometries.     

3D Printing of PLLA/Biomineral Composite Bone Tissue Engineering Scaffolds

Tissue engineering is one of the most effective ways to treat bone defects in recent years. However, current highly active bone tissue engineering (BTE) scaffolds are mainly based on the addition of active biological components (such as growth factors) to promote bone repair. High cost, easy inactivation and complex regulatory requirements greatly limit their practical applications. In addition, conventional fabrication methods make it difficult to meet the needs of personalized customization for the macroscopic and internal structure of tissue engineering scaffolds. Herein, this paper proposes to select five natural biominerals (eggshell, pearl, turtle shell, degelatinated deer antler and cuttlebone) with widely available sources, low price and potential osteo-inductive activity as functional particles. Subsequently compounding them into L-polylactic acid (PLLA) biomaterial ink to further explore 3D printing processes of the composite scaffold, and reveal their potential as biomimetic 3D scaffolds for bone tissue repair. The research results of this project provide a new idea for the construction of a 3D scaffold with growth-factor-free biomimetic structure, personalized customization ability and osteo-inductive activity.     

Assessment of Electrospun Pellethane-Based Scaffolds for Vascular Tissue Engineering

We examined the physicochemical properties and the biocompatibility and hemocompatibility of electrospun 3D matrices produced using polyurethane Pellethane 2363-80A (Pel-80A) blends Pel-80A with gelatin or/and bivalirudin. Two layers of vascular grafts of 1.8 mm in diameter were manufactured and studied for hemocompatibility ex vivo and functioning in the infrarenal position of Wistar rat abdominal aorta in vivo (n = 18). Expanded polytetrafluoroethylene (ePTFE) vascular grafts of similar diameter were implanted as a control (n = 18). Scaffolds produced from Pel-80A with Gel showed high stiffness with a long proportional limit and limited influence of wetting on mechanical characteristics. The electrospun matrices with gelatin have moderate capacity to support cell adhesion and proliferation (~30–47%), whereas vascular grafts with bivalirudin in the inner layer have good hemocompatibility ex vivo. The introduction of bivalirudin into grafts inhibited platelet adhesion and does not lead to a change hemolysis and D-dimers concentration. Study in vivo indicates the advantages of Pel-80A grafts over ePTFE in terms of graft occlusion, calcification level, and blood velocity after 6 months of implantation. The thickness of neointima in Pel-80A–based grafts stabilizes after three months (41.84 ± 20.21 µm) and does not increase until six months, demonstrating potential for long-term functioning without stenosis and as a suitable candidate for subsequent preclinical studies in large animals.     

Design of Materials for Bone Tissue Scaffolds

The strong impulse recently experienced by the manufacturing technologies as well as the development of innovative biocompatible materials has allowed the fabrication of high-performing scaffolds for bone tissue engineering. The design process of materials for bone tissue scaffolds represents, nowadays, an issue of crucial importance and the object of study of many researchers throughout the world. A number of studies have been conducted, aimed at identifying the optimal material, geometry, and surface that the scaffold must possess to stimulate the formation of the largest amounts of bone in the shortest time possible. This book presents a collection of 10 research articles and 2 review papers describing numerical and experimental design techniques definitively aimed at improving the scaffold performance, shortening the healing time, and increasing the success rate of the scaffold implantation process.     

心脏瓣膜组织工程研究新进展

瓣膜置换术是目前临床上治疗心脏瓣膜病的最终手段。但临床应用的各种人工心脏瓣膜在持久性和远期疗效上,都存在着难以克服的缺陷。而随着组织工程技术的发展,组织工程瓣膜以良好的组织相容性、经久耐用且具有生物活性等多方面的优势而成为近年来心脏外科领域的研究热点之一。现综述了近年来国外组织工程心脏瓣膜在瓣膜材料、培养方式及种子细胞等方面的最新进展并展望了该领域的研究未来。     

Marine Plankton-Derived Whitlockite Powder-Based 3D-Printed Porous Scaffold for Bone Tissue Engineering

Powder-based 3D printing is an excellent technique for the fabrication of complex structural shapes. The outstanding bone remodeling capacity of calcium phosphate bioceramics is a desirable characteristic for such fabrication. Whitlockite (WH) is a calcium phosphate-based ceramic that contains Mg ions and possesses good mechanical properties, rapid resorbability, and promotes osteogenesis. The aim of this study was to fabricate 3D-printed scaffolds using marine plankton-derived WH (MP-WH) powder. The surface morphology and composition of the fabricated scaffolds were characterized by scanning electron microscopy and X-ray diffraction. The biocompatibility and osteogenic effects were evaluated using human mesenchymal stem cells. We successfully obtained a 3D porous scaffold using MP-WH. The MP-WH 3D scaffold showed improved compressive strength compared to the tricalcium phosphate (TCP) 3D scaffold. The in vitro results showed that compared with TCP 3D scaffolds, MP-WH 3D scaffolds were biocompatible and enhanced cell proliferation and adhesion. In addition, alkaline phosphatase activity and real-time polymerase chain reaction assays demonstrated that osteoblast differentiation was improved on the MP-WH scaffold. These results suggest that marine plankton-derived WH is useful for fabricating 3D-printed scaffolds for bone tissue engineering applications.     

Polymeric Orthosis with Electromagnetic Stimulator Controlled by Mobile Application for Bone Fracture Healing: Evaluation of Design Concepts for Medical Use

Background: The occurrence of bone fractures is increasing worldwide, mainly due to the health problems that follow the aging population. The use of additive manufacturing and electrical stimulators can be applied for bioactive achievements in bone healing. However, such technologies are difficult to be transferred to medical practice. This work aims to develop an orthosis with a combined magnetic field (CFM) electrostimulator that demonstrates concepts and design aspects that facilitate its use in a real scenario. Methods: A 3D-printed orthosis made of two meshes was manufactured using PLA for outer mechanical stabilization mesh and TPU for inner fixation mesh to avoid mobilization. A CFM stimulator of reduced dimension controlled by a mobile application was coupled onto the orthosis. The design concepts were evaluated by health professionals and their resistance to chemical agents commonly used in daily activities were tested. Their thermal, chemical and electrical properties were also characterized. Results: No degradation was observed after exposure to chemical agents. The CMF achieved proper intensity (20–40 µT). The thermal analysis indicated its appropriate use for being modelled during clinical assessment. Conclusion: An orthosis with a coupled electrostimulator that works with a combined magnetic field and is controlled by mobile application was developed, and it has advantageous characteristics when compared to traditional techniques for application in real medical environments.     

Novel Electrospun Polycaprolactone/Calcium Alginate Scaffolds for Skin Tissue Engineering

After decades of research, fully functional skin regeneration is still a challenge. Skin is a multilayered complex organ exhibiting a cascading healing process affected by various mechanisms. Specifically, nutrients, oxygen, and biochemical signals can lead to specific cell behavior, ultimately conducive to the formation of high-quality tissue. This biomolecular exchange can be tuned through scaffold engineering, one of the leading fields in skin substitutes and equivalents. The principal objective of this investigation was the design, fabrication, and evaluation of a new class of three-dimensional fibrous scaffolds consisting of poly(ε-caprolactone) (PCL)/calcium alginate (CA), with the goal to induce keratinocyte differentiation through the action of calcium leaching. Scaffolds fabricated by electrospinning using a PCL/sodium alginate solution were treated by immersion in a calcium chloride solution to replace alginate-linked sodium ions by calcium ions. This treatment not only provided ion replacement, but also induced fiber crosslinking. The scaffold morphology was examined by scanning electron microscopy and systematically assessed by measurements of the pore size and the diameter, alignment, and crosslinking of the fibers. The hydrophilicity of the scaffolds was quantified by contact angle measurements and was correlated to the augmentation of cell attachment in the presence of CA. The in vitro performance of the scaffolds was investigated by seeding and staining fibroblasts and keratinocytes and using differentiation markers to detect the evolution of basal, spinous, and granular keratinocytes. The results of this study illuminate the potential of the PCL/CA scaffolds for tissue engineering and suggest that calcium leaching out from the scaffolds might have contributed to the development of a desirable biological environment for the attachment, proliferation, and differentiation of the main skin cells (i.e., fibroblasts and keratinocytes).     

Characterization of Porous Scaffolds Fabricated by Joining Stacking Based Laser Micro-Spot Welding (JS-LMSW) for Tissue Engineering Applications

A novel manufacturing approach was used to fabricate metallic scaffolds. A calibration of the laser cutting process was performed using the kerf width compensation in the calculations of the tool trajectory. Welding defects were studied through X-ray microtomography. Penetration depth and width resulted in relative errors of 9.4%, 1.0%, respectively. Microhardness was also measured, and the microstructure was studied in the base material. The microhardness values obtained were 400 HV, 237 HV, and 215 HV for the base material, HAZ, and fusion zone, respectively. No significant difference was found between the microhardness measurement along with different height positions of the scaffold. The scaffolds’ dimensions and porosity were measured, their internal architecture was observed with micro-computed tomography. The results indicated that geometries with dimensions under 500 µm with different shapes resulted in relative errors of ~2.7%. The fabricated scaffolds presented an average compressive modulus ~13.15 GPa, which is close to cortical bone properties. The proposed methodology showed a promising future in bone tissue engineering applications.