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Cardiovascular disease (CVD) remains to be the leading cause of death globally today and therefore the need for the development of novel therapies has become increasingly important in the cardiovascular field. The mechanism(s) behind the pathophysiology of CVD have been laboriously investigated in both stem cell and bioengineering laboratories. Scientific breakthroughs have paved the way to better mimic cell types of interest in recent years, with the ability to generate any cell type from reprogrammed human pluripotent stem cells. Mimicking the native extracellular matrix using both organic and inorganic biomaterials has allowed full organs to be recapitulated in vitro. In this paper, we will review techniques from both stem cell biology and bioengineering which have been fruitfully combined and have fueled advances in the cardiovascular disease field. We will provide a brief introduction to CVD, reviewing some of the recent studies as related to the role of endothelial cells and endothelial cell dysfunction. Recent advances and the techniques widely used in both bioengineering and stem cell biology will be discussed, providing a broad overview of the collaboration between these two fields and their overall impact on tissue engineering in the cardiovascular devices and implications for treatment of cardiovascular disease., (© 2024 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals LLC.)

Bifunctional tissue engineering constructs promoting osteogenesis and angiogenesis are essential for bone regeneration. Metal ion-incorporated scaffolds and fibrin encapsulation attract much attention due to low cost, nontoxicity, and tunable control over ion and growth factor release. Herein, we investigated the effect of Cu.nHA/Cs/Gel scaffold and fibrin encapsulation on osteogenic and angiogenic differentiation of Wharton's jelly mesenchymal stem cells (WJMSCs) in vitro and in vivo. Cu-laden scaffolds were synthesized using salt leaching/freeze drying and were characterized using standard techniques. WJMSCs were isolated from the human umbilical cord and characterized. WJMSCs with or without encapsulating in fibrin were seeded onto scaffolds, followed by differentiating into the osteogenic lineage for 7 and 21 days. Osteogenic and angiogenic differentiation were evaluated using real-time polymerase chain reaction, western blot, and Alizarin red staining. Then, scaffolds were implanted into critical-sized calvarial bone defects in rats and histological assessments were performed using hematoxylin/eosin, Masson's trichrome, and CD31 immunohistochemical staining at 4 and 12 weeks. The scaffolds had good physicochemical and biological characteristics suitable for cell attachment and growth. Cu and fibrin increased the expression of ALP, RUNX2, OCN, COLI, VEGF, and HIF1α in differentiated WJMSCs. Implanted scaffolds were also biocompatible and were integrated well with the host tissue. Increased collagen condensation, mineralization, and blood vessel formation were observed in Cu-laden scaffolds. The fibrin-encapsulated groups showed the highest collagen and cell densities, immune cell infiltration, and bone trabeculae. CD31-positive cell population increased with fibrin encapsulation and seeding onto Cu-laden scaffolds. Adding Cu to scaffolds and encapsulating cells in fibrin are promising methods that guide osteogenesis and angiogenesis cellular signaling, leading to better bone regeneration., (© 2023 Wiley Periodicals LLC.)

Among the promising methods for repairing or replacing tissue defects in the human body and the hottest research topics in medical science today are regenerative medicine and tissue engineering. On the other hand, nanotechnology has been expanded into different areas of regenerative medicine and tissue engineering due to its essential benefits in improving performance in various fields. Nanotechnology, a helpful strategy in tissue engineering, offers new solutions to unsolved problems. Especially considering the excellent physicochemical properties of nanoscale structures, their application in regenerative medicine has been gradually developed, and a lot of research has been conducted in this field. In this regard, various nanoscale structures, including nanofibers, nanosheets, nanofilms, nano-clays, hollow spheres, and different nanoparticles, have been developed to advance nanotechnology strategies with tissue repair goals. Here, we comprehensively review the application of the mentioned nanostructures in constructing nanocomposite scaffolds for regenerative medicine and tissue engineering. This article is categorized under: Nanotechnology Approaches to Biology > Nanoscale Systems in Biology Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement Diagnostic Tools > Biosensing., (© 2023 Wiley Periodicals LLC.)

Graphene-based materials (GMs) have great application prospects in bone tissue engineering due to their osteoinductive ability and antimicrobial activity. GMs induce osteogenic differentiation through several mechanisms and pathways in bone tissue engineering. First of all, the surface and high hardness of the porous folds of graphene or graphene oxide (GO) can generate mechanical stimulation to initiate a cascade of reactions that promote osteogenic differentiation without any chemical inducers. In addition, change of the extracellular matrix (ECM), regulation of macrophage polarization, the oncostatin M (OSM) signaling pathway, the MAPK signaling pathway, the BMP signaling pathway, the Wnt/β-catenin signaling pathway, and other pathways are involved in GMs' regulation of osteogenesis. In bone tissue engineering, GMs prevent the formation of microbial biofilms mainly through preventing microbial adhesion and killing them. The former is mainly achieved by reducing surface free energy (SFE) and increasing hydrophobicity. The latter mainly includes oxidative stress and photothermal/photodynamic effects. Graphene and its derivatives (GDs) are mainly combined with bioactive ceramic materials, metal materials and macromolecular polymers to play an antimicrobial effect in bone tissue engineering. Concentration, number of layers, and type of GDs often affect the antimicrobial activity of GMs. In this paper, we reviewed relevant osteoinductive and antimicrobial mechanisms of GMs and their applications in bone tissue engineering., (© 2021 John Wiley & Sons Ltd.)

Since the discovery that applying electrical stimulation can promote cell growth, proliferation, and tissue regeneration, research on bio-piezoelectric materials is being actively conducted. In this study, a composite material was prepared by mixing polyvinylidene fluoride (PVDF), a conventional piezoelectric polymer, and silk fibroin (SF), a natural piezoelectric material that recently attracting attention. These two polymers were fabricated into a composite fiber mat using electrospinning technology. To find optimal conditions, SF was added in various ratios to prepare electrospun PVDF/SF mats. The characteristics of these PVDF/SF composite mats were then analyzed through various evaluations and in vitro studies. It was confirmed that PVDF and SF were successfully mixed through scanning electron microscope images and structural analysis such as x-ray diffractometer and Fourier transform infrared. The results revealed that adding an appropriate amount of SF could improve the tensile strength, enhance cell proliferation rate, and generate a voltage similar to that of a conventional PVDF-only electrospinning mat. Such fabricated electrospun PVDF/SF composite mats are expected to be useful in the bio-piezoelectric field because they can maintain piezoelectricity while compensating for the shortcomings, such as low physical properties, of a PVDF electrospun mat., (© 2021 John Wiley & Sons Ltd.)

The use of 3D-printed hydroxyapatite (HA) scaffolds for stimulating bone healing has been increasing over the years. Although all the promising effects of these scaffolds, there are still few studies and limited understanding of their interaction with bone tissue and their effects on the process of fracture healing. In this context, this study aimed to perform a systematic literature review examining the effects of different 3D-printed HA scaffolds in bone healing. The search was made according to the preferred reporting items for systematic reviews and meta-analysis (PRISMA) orientations and Medical Subject Headings (MeSH) descriptors "3D printing," "bone," "HA," "repair," and "in vivo." Thirty-six articles were retrieved from PubMed and Scopus databases. After eligibility analyses, 20 papers were included (covering the period of 2016 and 2021). Results demonstrated that all the studies included in this review showed positive outcomes, indicating the efficacy of scaffolds treated groups in the in vivo experiments for promoting bone healing in different animal models. In conclusion, 3D-printed HA scaffolds are excellent candidates as bone grafts due to their bioactivity and good bone interaction., (© 2022 Wiley Periodicals LLC.)

Fabrication of cardiac tissue from human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) has received great interest, but a major challenge facing researchers is the alignment of cardiomyocytes in the same direction to optimize force generation. We have developed a novel method of fabricating a cardiac tissue-like construct with aligned cells based on the unidirectional stretching of an hiPS-CM sheet. A square cell sheet was harvested from a temperature-responsive culture dish and placed on a silicone surface, and an extending force was imposed on the silicone to stretch the cell sheet along one direction. To enable evaluation of cardiomyocyte morphology in vitro, a cell sheet was constructed by coculture of hiPS-CMs and human adipose-derived stem cells. In separate experiments, a stretched double-layered cell sheet constructed from hiPS-CMs alone was transplanted onto the muscle of an athymic rat, and its features were compared with those of a nonstretched (control) cell sheet. Immediately after stretching, the stretched cell sheet was significantly longer than the control cell sheet. Immunohistological analysis revealed that the cardiomyocytes showed unidirectional alignment in the stretched cell sheet but random directionality in the control cell sheet. Two weeks after transplantation, immunohistology demonstrated that the stretched cell sheet had retained the unidirectionality of its myocardial fibers and had an orientation intensity that was higher than that of the control cell sheet after transplantation or the stretched cell sheet before transplantation. Our technique provides a simple method of aligning an hiPS-CM-derived cardiac tissue-like construct without the use of a scaffold., (© 2020 John Wiley & Sons, Ltd.)

In the present work, a novel strategy was explored to fabricate nanofiber scaffolds consisting of cellulose assimilated with titanium dioxide (TiO 2 ) and silver (Ag) nanoparticles (NPs). The concentration of the TiO 2 NPs in the composite was adjusted to 1.0, 1.5, and 2.0 wt % with respect to polymer concentration used for the electrospinning of colloidal solutions. The fabricated composite scaffolds were dispensed to alkaline deacetylation using 0.05 M NaOH to remove the acetyl groups in order to generate pure cellulose nanofibers containing TiO 2 NPs. Moreover, to augment our nanofiber scaffolds with antibacterial activity, the in situ deposition approach of using Ag NPs was utilized with varied molar concentrations of 0.14, 0.42, and 0.71 M. The physicochemical properties of the nanofibers were identified by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared (FTIR) and contact angle meter studies. This demonstrated the presence of both TiO 2 and Ag NPs and complete deacetylation of nanofibers. The antibacterial efficiency of the nanofibers was scrutinized against Escherichia coli and Staphylococcus aureus, revealing proper in situ deposition of Ag NPs and confirming the nanofibers are antibacterial in nature. The biocompatibility of the scaffolds was accustomed using chicken embryo fibroblasts, which confirmed their potential role to be used as wound-healing materials. Furthermore, the fabricated scaffolds were subjected to analysis in simulated body fluid at 37°C to induce mineralization for future osseous tissue integration. These results indicate that fabricated composite nanofiber scaffolds with multifunctional characteristics will have a highest potential as a future candidate for promoting new tissues artificially., (© 2020 Wiley Periodicals, Inc.)

Osteoarthritis (OA) is a prevalent joint disorder that is characterized by the degeneration of articular cartilage in synovial joints. Most of the current treatment options for this disorder tend to focus on symptom management rather than addressing the underlying progression of the disease. Cartilage tissue engineering has emerged as a promising approach to address the limitations of current OA treatments, aiming to regenerate cartilage and restore the natural function of affected joints. Like any other tissue engineering field, cartilage tissue engineering uses different fabrication techniques and biomaterials to develop the constructs. Numerous studies over the last few years have demonstrated the preclinical efficacy of tissue-engineered constructs in promoting cartilage regeneration and highlight the potential of tissue-engineered constructs as a viable therapeutic approach for OA. This paper aims to provide a focused review of advancements in tissue-engineered constructs over the past decade. Specifically, we highlight the constructs based on natural, synthetic, and composite biomaterials and the varying conventional and advanced fabrication techniques. We also highlight the challenges in state-of-the-art cartilage tissue engineering that must be overcome in the upcoming years to fully replicate the complex anatomy of the native cartilage. We believe that continued collaborative research efforts among researchers from various facets of engineering and clinicians are required to advance the field of cartilage tissue engineering and become a viable OA therapy., (© 2025 The Author(s). Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals LLC.)

Biomaterials derived from extracellular matrices (ECMs) were extensively used for skin tissue engineering and wound healing. ECM is a complex network of biomolecules (e.g., proteins), which provide organizational support to cells for growth. Thus, ECM could be an ideal biomaterial for fabricating the scaffold. However, oxidative stress and biofilm formation at the wound site remains a major challenge that could be neutralized using herbal ingredients (e.g., curcumin). In this study, ECM was extracted from the biowaste of the goat abattoir by using decellularization. The goat small intestine submucosa (G-SIS) is decellularized to obtain the decellularized G-SIS (DG-SIS) and curcumin (in different concentrations) was incorporated in the DG-SIS to fabricate curcumin-embedded DG-SIS scaffolds. Changes brought by increasing the concentrations of the curcumin in DG-SIS were observed in various properties, including free radical scavenging and antibacterial properties. Results depicted that the scaffolds are porous, biodegradable, biocompatible, antibacterial, and hydrophilic and showed sustained release of curcumin. Besides, it showed free radicals scavenging property. The porosity and hydrophilicity of the scaffolds were decreased with an increase in the curcumin content. However, biodegradability, free radical scavenging, biocompatibility, and antibacterial properties of the scaffolds increased with an increase in the curcumin content. The DG-SIS scaffold containing 1 wt % of curcumin may be a potential biomaterial for wound-healing and skin tissue engineering., (© 2021 Wiley Periodicals LLC.)

There is a growing demand for polymer fiber scaffolds for biomedical applications and tissue engineering. Biodegradable polymers such as polycaprolactone have attracted particular attention due to their applicability to tissue engineering and optical neural interfacing. Here we report on a scalable and inexpensive fiber fabrication technique, which enables the drawing of PCL fibers in a single process without the use of auxiliary cladding. We demonstrate the possibility of drawing PCL fibers of different geometries and cross-sections, including solid-core, hollow-core, and grooved fibers. The solid-core fibers of different geometries are shown to support cell growth, through successful MCF-7 breast cancer cell attachment and proliferation. We also show that the hollow-core fibers exhibit a relatively stable optical propagation loss after submersion into a biological fluid for up to 21 days with potential to be used as waveguides in optical neural interfacing. The capacity to tailor the surface morphology of biodegradable PCL fibers and their non-cytotoxicity make the proposed approach an attractive platform for biomedical applications and tissue engineering., (© 2020 Wiley Periodicals LLC.)

Prevascularization is essential to ensure the viability, functionality, and successful integration of tissue-engineered three-dimensional (3D) constructs with surrounding host tissues after transplantation. Human mesenchymal stem cell (hMSC) sheet can be prevascularized by coculturing with endothelial cells (ECs), and then be further used as building blocks for engineering 3D complex tissues. In addition, predifferentiation of hMSCs into a tissue-specific lineage in vitro has been proven to promote graft engraftment and regeneration. However, it is unclear if the prevascularized hMSC sheets can still maintain their microvascular integrity as well as the immune-regulatory properties after their tissue-specific differentiation. The objective of this study was to investigate the effects of differentiation cues on the microvascular structure, angiogenic factor secretion, and immunogenic responses of prevascularized hMSC sheets. The results showed that upon coculturing with ECs, hMSC sheets successfully formed microvascular network, while maintaining hMSCs' multi-lineage differentiation capability. The next step, osteogenic and adipogenic induction, damaged the preformed microvascular structures and compromised the angiogenic factor secretion ability of hMSCs. Nonetheless, this effect was mitigated by adjusting the concentration of differentiation factors. The subcutaneous transplantation in an immunocompetent rat model demonstrated that the osteogenic differentiated prevascularized hMSC sheet preserved its microvascular structure and immunomodulatory properties comparable to the undifferentiated prevascularized hMSC sheets. This study suggested that a balanced and optimal differentiation condition can effectively promote the tissue-specific predifferentiation of prevascularized hMSC sheet while maintaining its immunomodulatory and tissue integration properties., (© 2021 John Wiley & Sons Ltd.)

Tissue engineering is a promising approach for generating or repairing living tissues. The development of innovative biomaterials for tissue engineering has the potential to address the unmet clinical needs in certain applications. However, before these biomaterials can be used in clinical settings, they must undergo preclinical testing to ensure safety and performance. The chicken chorioallantoic membrane (CAM) assay is a preferred screening tool for studying biocompatibility, angiogenesis, and inflammation induced by biomaterials owing to ethical and economic considerations. This CAM-based platform increased the throughput of biomaterial testing for tissue engineering before in vivo testing. In this paper, we discuss the advantages of the CAM model. We also provided a step-by-step guide for implementing the CAM model in a research laboratory, along with tips and tricks for successfully running CAM assays. Finally, we present examples of biomaterials screened using CAM assays. CAM assay is a powerful in vivo model for assessing the angiogenic potential of tissue-engineered scaffolds. This guide provides a framework for conducting the assay, but specific experimental conditions should be optimized based on the scaffold material and the research question., (© 2024 The Author(s). Journal of Biomedical Materials Research Part B: Applied Biomaterials published by Wiley Periodicals LLC.)

Nanofibers are a type of nanostructures, which due to their unique properties can be used in wound dressing, and electrospinning is a good way to produce them. The benefits of wound dressings made of electrospun nanofibers include a large surface area to volume ratio, high absorption of secretions from the wound, and high air permeability, mimicking the morphology of extracellular matrix (ECM) of the damaged tissue and the possibility of the gradual release of the drug agents loaded on nanofibers. Because of the adaptability of plants to the body, low side effects and the prevalence of antibiotic resistance, interest in using plants is increasing. Combining nanofibers with plants is a way to integrate the physical properties of the structure of nanofibers and the chemical and antibacterial properties of the plants. In recent years, many plants in the forms of extracts, essential oils, and pure active ingredients have been used in the electrospininng and production of nanofiber-containing plants; some of the plants may be a good choice for wound dressings made of electrospun nanofibers., (© 2020 John Wiley & Sons, Ltd.)

The aim of this study was to develop a highly porous calcium-containing chitosan scaffold suitable for dentin regeneration. A calcium hydroxide (Ca[OH] 2 ) suspension was used to modulate the degree of porosity and chemical composition of chitosan scaffolds. The chitosan solution concentration and freezing protocol were adjusted to optimize the porous architecture using the phase-separation technique. Scanning electron microscopy/energy-dispersive spectroscopy demonstrated the fabrication of a highly porous calcium-linked chitosan scaffold (CH-Ca), with a well-organized and interconnected porous network. Scaffolds were cross-linked on glutaraldehyde (GA) vapor. Following a 28-day incubation in water, cross-linked CH scaffold had no changes on humid mass, and CH-Ca featured a controlled degradability profile since the significant humid mass loss was observed only after 21 (26.0%) and 28 days (42.2%). Fourier-transform infrared spectroscopy indicated the establishment of Schiff base on cross-linked scaffolds, along with calcium complexation for CH-Ca. Cross-linked CH-Ca scaffold featured a sustained Ca 2+ release up to 21 days in a humid environment. This porous and stable architecture allowed for human dental pulp cells (HDPCs) to spread throughout the scaffold, with cells exhibiting a widely stretched cytoplasm; whereas, the cells seeded onto CH scaffold were organized in clusters. HDPCs seeded onto CH-Ca featured significantly higher ALP activity, and gene expressions for ALP, Col1, DMP-1, and DSPP in comparison to CH, leading to a significant 3.5 times increase in calcium-rich matrix deposition. In sum, our findings suggest that CH-Ca scaffolds are attractive candidates for creating a highly porous and bioactive substrate for dentin tissue engineering., (© 2020 Wiley Periodicals, Inc.)

Polymers and scaffolds are the most significant tools in regenerative medicine. Urogenital disorders are an important group of diseases that greatly affect the patient's life expectancy and quality. Reconstruction of urogenital defects is one of the current challenges in regenerative medicine. Regenerative medicine, as well as tissue engineering, may offer suitable approaches, while the tools needed are appropriate materials and cells. Autologous urothelial cells obtained from biopsy, bone marrow-derived stem cells, adipose stem cells and urine-derived stem cells that expressed mesenchymal cell markers are the cells that mainly used. In addition, two main types of biomaterials mainly exist; synthetic polymers and composite scaffolds that are biodegradable polymers with controllable properties and naturally derived biomaterials such as extracellular matrix components and acellular tissue matrices. In this review, we present and evaluate the most appropriate and suitable scaffolds (naturally derived and synthetic polymers) and cells applied in urogenital reconstruction., (© 2020 The Authors. Journal of Biomedical Materials Research Part A published by Wiley Periodicals, Inc.)

Electrospinning technology has been widely used in the past few decades to prepare nanofibrous scaffolds that mimic extracellular matrices. However, traditional two-dimensional (2D) electrospun nanofibrous mats still have some inherent disadvantages for bone tissue engineering, such as limited cell infiltration and lack of three-dimensional (3D) structure. The development of 3D electrospun scaffolds with larger pore sizes and porosity provides new perspectives for electrospinning-based tissue engineering scaffolds. However, there are still some challenges and areas for improvement. In this review, the applications of 3D electrospun nanofibrous scaffolds in the field of bone tissue engineering from its fabrication methods to bio-functionalization are summarized, with the aim of providing new insights into the design of electrospinning-based bone tissue engineering scaffolds., (© 2019 Wiley Periodicals, Inc.)

In this study, polylactic acid (PLA)-based composite scaffolds with calcium carbonate (CaCO 3 ) and beta-tricalcium phosphate (β-TCP) were obtained by 3D printing. These structures were evaluated as potential 3D structures for bone tissue regeneration. Morphological, mechanical, and biological tests were carried out in order to compare the effect of each additive (added in a concentration of 5% w/w) and the combination of both (2.5% w/w of each one), on the PLA matrix. The scaffolds manufactured had a mean pore size between 400-425 μm and a porosity value in the range of 50-60%. According to the results, both additives promoted an increase of the porosity, hydrophilicity, and surface roughness of the scaffolds, leading to a significant improvement of the metabolic activity of human osteoblastic osteosarcoma cells. The best results in terms of cell attachment after 7 days were obtained for the samples containing CaCO 3 and β-TCP particles due to the synergistic effect of both additives, which results in an increase in osteoconductivity and in a microporosity that favours cell adhesion. These scaffolds (PLA:CaCO 3 :β-TCP 95:2.5:2.5) have suitable properties to be further evaluated for bone tissue engineering applications., (© 2019 John Wiley & Sons, Ltd.)

A thin plastic-like film separated from the epidermis of Cereus hildmannianus has excellent tensile strength, resistance to water and high antimicrobial activity and supports the growth of mouse fibroblast cells. Cactuses are one of the most under explored plant species with high potential for food, materials, pharmaceutical and other applications. Although studies have shown the ability of cactuses to be used for food, as a source for fibers, as reinforcement for composites and other applications, the role of individual layers and their properties has been studied to a limited extent. In this paper, a thin translucent layer was separated from the epidermis of C. hildmannianus and studied for its composition, structure and properties. The layer is composed of about 73% cellulose and 2% lignin and morphologically, shows surface with uneven and serrated edges. Films with length of up to 36 cm, strength of 6.8 MPa and elongation of 2.5% could be peeled from the cactus suggesting their suitability for food packaging and other applications. X-ray diffraction patterns and FTIR spectrums indicated that the films are similar to that of cellulose and major thermal degradation occurred above 280°C. Compared to standards, the cactus films showed about 41% and 44% inhibition against gram positive and gram negative bacteria and 67% inhibition of the common fungal strain (A. niger). Films showed high stability in water and to common chemicals. When used as substrates for mouse fibroblast cell growth, no cytotoxicity was observed and the cactus peel supported the attachment and proliferation of cells demonstrating potential to be used as a biomaterial for tissue engineering applications., (© 2023 Wiley Periodicals LLC.)

This review paper is motivated by a Back-to-Basics presentation given by the author at the 2022 Orthopaedic Research Society meeting in Tampa, Florida. I was tasked with providing a brief history of research leading up to the introduction of functional tissue engineering (FTE) for tendon and ligament repair. Beginning in the 1970s, this timeline focused on two common orthopedic soft tissue problems, anterior cruciate ligament ruptures in the knee and supraspinatus tendon injuries in the shoulder. Historic changes in the field over the next 5 decades revealed a transformation from a focus more on mechanics (called "bioMECHANICS") on a larger (tissue) scale to a more recent focus on biology (called "mechanoBIOLOGY") on a smaller (cellular and molecular) scale. Early studies by surgeons and engineers revealed the importance of testing conditions for ligaments and tendons (e.g., high strain rates while avoiding subject disuse and immobility) and the need to measure in vivo forces in these tissues. But any true tissue engineering and regeneration in these early decades was limited more to the use of auto-, allo- and xenografts than actual generation of stimulated cell-scaffold constructs in culture. It was only after the discovery of tissue engineering in 1988 and the recognition of frequent rotator cuff injuries in the early 1990s, that biologists joined surgeons and engineers to discover mechanical and biological testing criteria for FTE. This review emphasizes the need for broader and more inclusive collaborations by surgeons, biologists and engineers in the short term with involvement of those in biomaterials, manufacturing, and regulation of new products in the longer term., (© 2022 John Wiley & Sons Ltd.)

Over the last two decades, electrospun scaffolds have proved to be advantageous in the field of nerve tissue regeneration by connecting the cavity among the proximal and distal nerve stumps growth cones and leading to functional recovery after injury. Multifunctional nanofibrous structure of these scaffolds provides enormous potential by combining the advantages of nano-scale topography, and biological science. In these structures, selecting the appropriate materials, designing an optimized structure, modifying the surface to enhance biological functions and neurotrophic factors loading, and native cell-like stem cells should be considered as the essential factors. In this systematic review paper, the fabrication methods for the preparation of aligned nanofibrous scaffolds in yarn or conduit architecture are reviewed. Subsequently, the utilized polymeric materials, including natural, synthetic and blend are presented. Finally, their surface modification techniques, as well as, the recent advances and outcomes of the scaffolds, both in vitro and in vivo, are reviewed and discussed., (© 2020 Wiley Periodicals LLC.)

Articular cartilage (AC) has gradient features in both mechanics and histology as well as a poor regeneration ability. The repair of AC poses difficulties in both research and the clinic. In this paper, a gradient scaffold based on poly(lactic-co-glycolic acid) (PLGA)-extracellular matrix was proposed. Cartilage scaffolds with a three-layer gradient structure were fabricated by PLGA through three-dimensional printing, and the microstructure orientation and pore fabrication were made by decellularized extracellular matrix injection and directional freezing. The manufactured scaffold has a mechanical strength close to that of real cartilage. A quantitative optimization of the Young's modulus and shear modulus was achieved by material mechanics formulas, which achieved a more accurate mechanical bionic and a more stable interface performance because of the one-time molding process. At the same time, the scaffolds have a bionic and gradient microstructure orientation and pore size, and the stratification ratio can be quantitatively optimized by design of the freeze box and temperature simulation. In general, this paper provides a method to optimize AC scaffolds by both mechanics and histology as well as a bionic multimaterial scaffold. This paper is of significance for cell culture and clinical transplantation experiments. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1664-1676, 2018., (© 2018 Wiley Periodicals, Inc.)

Tissue engineering of corporal tissue is a new development in otherwise untreatable erectile dysfunction and in urethral reconstructions to treat hypospadias or severe urethral stricture disease. Multiple complications can arise with the current treatments, whereas engineered tissue, if well vascularized and existing of autologous cells, may lead to better results. The aim of this review was to provide an overview of literature on cell-seeded-based tissue engineering of corporal penile tissue. A literature search was performed following the PRISMA guidelines. Papers describing cell-seeded tissue engineering of corporal tissue were included. Studies using different techniques, such as intracavernous injection were excluded. Fifteen articles were included in the review. Twelve of these studies described engineering of the corpus cavernosum in animal models. Two articles were found on engineering of animal corpus spongiosum and one article on engineering of the human glans. Both synthetic scaffolds and biological scaffolds were used. The advantage of a biological, acellular scaffold was that the native, complex architecture of corporal tissue was maintained. Most studies used endothelial and smooth muscle cells from corporal origin, but stem cells were also investigated. Furthermore, dynamic culturing achieved an improved cell content and functionality. This review has summarized the developments in tissue engineering of corpus cavernosum and spongiosum tissue. Functional tissue has been developed in animal studies with the use of seeded cells on scaffolds. This knowledge will form a basis for the development of tissue engineering of corporal tissue for clinical applications., (Copyright © 2017 John Wiley & Sons, Ltd.)

The importance of decellularized medical products has significantly increased during the last years. In this paper, we evaluated the effects of selected physical and procedural decellularization (DC) factors with the aim to systematically assess their influence on DC results. 72 porcine aortic walls (AW) were divided into three groups and exposed to a DC solution for 4 h and 8 h, either continuously or in repeated cycles. The AW were rocked (90bpm), whirled (10 l/min), sonicated (120W, 45 kHz) or exposed to a combination of these treatments, followed by 10 washing cycles. Defining successful DC as removal of nuclei while keeping an intact extracellular matrix (ECM), we equalized the efficiency to the penetration depth (PD), obtained by DAPI fluorescence and H&E staining. Additionally, we performed scanning electron microscopy (SEM), Pentachrome and Picrosirius-Red staining. Results showed that significantly higher DC depths are achieved on outer compared to inner surfaces (61 ± 7%; p < 0.001). Furthermore, the PD showed a high time dependency for all samples. Compared to continuous rocking, we achieved a significant increase in the DC efficiency through cyclic treatments ( ∼ 43%), whirling ( ∼ 19%) and sonication ( ∼ 49%). The combined treatment supported these results. In all procedures, a skeletonized but intact Collagen fibrous network was obtained as confirmed by SEM analysis. In conclusion, we systematically identified essential factors to significantly enhance DC procedures. We highly recommend considering these factors in future DC protocols. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 153-162, 2018., (© 2016 Wiley Periodicals, Inc.)

A principal purpose of tissue engineering is the augmentation, repair or replacement of diseased or injured human tissue. This study was undertaken to determine whether human biopsies as a cell source could be utilized for successful engineering of human phalanges consisting of both bone and cartilage. This paper reports the use of cadaveric human chondrocytes and periosteum as a model for the development of phalanx constructs. Two factors, osteogenic protein-1 [OP-1/bone morphogenetic protein-7 (BMP7)], alone or combined with insulin-like growth factor (IGF-1), were examined for their potential enhancement of chondrocytes and their secreted extracellular matrices. Design of the study included culture of chondrocytes and periosteum on biodegradable polyglycolic acid (PGA) and poly-l-lactic acid (PLLA)-poly-ε-caprolactone (PCL) scaffolds and subsequent implantation in athymic nu/nu (nude) mice for 5, 20, 40 and 60 weeks. Engineered constructs retrieved from mice were characterized with regard to genotype and phenotype as a function of developmental (implantation) time. Assessments included gross observation, X-ray radiography or microcomputed tomography, histology and gene expression. The resulting data showed that human cell-scaffold constructs could be successfully developed over 60 weeks, despite variability in donor age. Cartilage formation of the distal phalanx models enhanced with both OP-1 and IGF-1 yielded more cells and extracellular matrix (collagen and proteoglycans) than control chondrocytes without added factors. Summary data demonstrated that human distal phalanx models utilizing cadaveric chondrocytes and periosteum were successfully fabricated and OP-1 and OP-1/IGF-1 accelerated construct development and mineralization. The results suggest that similar engineering and transplantation of human autologous tissues in patients are clinically feasible. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

This paper reports on the development of montmorillonite (MMT)-reinforced hydrogels, based on a peptidomimetic polyamidoamine carrying guanidine pendants (AGMA1), as substrates for the osteo-induction of osteoblast precursor cells. AGMA1 hydrogels of various degrees of crosslinking responded favourably to MMT reinforcement, giving rise to composite hydrogels with shear storage modulus G', when fully swollen in water, up to 200 kPa, i.e. 20 times higher than the virgin hydrogels and of the same order or higher than other hydrogel-based composites proposed for orthopaedic applications. This significant improvement was ascribed to the effective interpenetration between the polymer matrix and the inorganic filler. AGMA1-MMT hydrogels, when evaluated as scaffolds for the osteogenic differentiation of mouse calvaria-derived pre-osteoblastic MC3T3-E1 cells, proved able to support cell adhesion and proliferation and clearly induced differentiation towards the osteoblastic phenotype, as indicated by different markers. In addition, AGMA1-MMT hydrogels proved completely degradable in aqueous media at pH 7.4 and did not provide any evidence of cytotoxicity. The experimental evidence suggests that AGMA1-MMT composites definitely warrant potential as scaffolds for osteoblast culture and bone grafts. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

The development of new human skeletal muscle tissue is an alternative approach to the replacement of tissue after severe damage, for example in the case of traumatic injury, where surgical reconstruction is often needed following major loss of natural tissue. Treatment to date has involved the transfer of muscle tissue from other sites, resulting in a functional loss and volume deficiency of donor sites. Approaches that seek to eliminate these problems include the relatively new solution of skeletal muscle engineering. Here there are two main components to consider: (a) the cells with their regenerative potential; and (b) the polymeric structure onto which cells are seeded and where they must perform their activities. In this paper we describe well-defined two- and three-dimensional polymeric structures able to drive the myoblast process of adhesion, proliferation and differentiation. We examine a series of polymers and protein adhesions with which to functionalize the structures, and cell-seeding methods, with a view to defining the optimal protocol for engineering skeletal muscle tissue. All polymer samples were tested for their mechanical and biological properties, to support the validity of our results in the real context of muscle tissue engineering. Copyright © 2014 John Wiley & Sons, Ltd., (Copyright © 2014 John Wiley & Sons, Ltd.)

It is now widely acknowledged that implants that have been designed with an effort towards reconstructing the transition between tissues might improve their functionality and integration in vivo. This paper contributes to the development of improved treatment for articular cartilage repair by exploring the potential of the combination of electrospinning technology and cell sheet engineering to create cartilage tissue. Poly(lactic-co-glycolic acid) (PLGA) was used to create the electrospun membranes. The focus being on the cartilage-bone transition, collagen type I and hydroxyapatite (HA) were also added to the scaffolds to increase the histological biocompatibility. Human mesenchymal stem cells (hMSCs) were cultured in thermoresponsive dishes to allow non-enzymatic removal of an intact cell layer after reaching confluence. The tissue constructs were created by layering electrospun membranes with sheets of hMSCs and were cultured under chondrogenic conditions for up to 21 days. High viability was found to be maintained in the multilayered construct. Under chondrogenic conditions, reverse-transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry have shown high expression levels of collagen type X, a form of collagen typically found in the calcified zone of articular cartilage, suggesting an induction of chondrocyte hypertrophy in the PLGA-based scaffolds. To conclude, this paper suggests that layering electrospun scaffolds and cell sheets is an efficient approach for the engineering of tissue transitions, and in particular the cartilage-bone transition. The use of PLGA-based scaffold might be particularly useful for the bone-cartilage reconstruction, since the differentiated tissue constructs seem to show characteristics of calcified cartilage., (Copyright © 2013 John Wiley & Sons, Ltd.)

The discovery of the interesting intrinsic properties of graphene, a two-dimensional nanomaterial, has boosted further research and development for various types of applications from electronics to biomedicine. During the last decade, graphene and several graphene-derived materials, such as graphene oxide, carbon nanotubes, activated charcoal composite, fluorinated graphenes and three-dimensional graphene foams, have been extensively explored as components of biosensors or theranostics, or to remotely control cell-substrate interfaces, because of their remarkable electro-conductivity. To date, despite the intensive progress in human stem cell research, only a few attempts to use carbon nanotechnology in the stem cell field have been reported. Interestingly, most of the recent in vitro studies indicate that graphene-based nanomaterials (i.e. mainly graphene, graphene oxide and carbon nanotubes) promote stem cell adhesion, growth, expansion and differentiation. Although cell viability in vitro is not affected, their potential nanocytoxicity (i.e. nanocompatibility and consequences of uncontrolled nanobiodegradability) in a clinical setting using humans remains unknown. Therefore, rigorous internationally standardized clinical studies in humans that would aim to assess their nanotoxicology are requested. In this paper we report and discuss the recent and pertinent findings about graphene and derivatives as valuable nanomaterials for stem cell research (i.e. culture, maintenance and differentiation) and tissue engineering, as well as for regenerative, translational and personalized medicine (e.g. bone reconstruction, neural regeneration). Also, from scarce nanotoxicological data, we also highlight the importance of functionalizing graphene-based nanomaterials to minimize the cytotoxic effects, as well as other critical safety parameters that remain important to take into consideration when developing nanobionanomaterials., (Copyright © 2014 John Wiley & Sons, Ltd.)

Vascular tissue engineering is a relevant research field aimed at elaborating and proposing innovative solutions to overcome the drawbacks related to the use of conventional blood vessel substitutes, especially referring to small-diameter grafts. For this aim, electrospinning can be regarded as a valuable technique to produce novel scaffolds with several functional characteristics that can be usefully tailored for the application discussed here. The reproduction of the natural extracellular matrix obtained by processing bioresorbable polymers, either functionalized or not, is driving the biomedical research towards technical solutions that can lead to an actual therapeutic improvement. In this context, this paper reviews those studies focused on the selection of suitable biomaterials for vascular applications, their microstructure, the cell response to polymeric fibres and the strategies considered so far to modify and therefore enhance the performance of final electrospun scaffolds., (Copyright © 2013 John Wiley & Sons, Ltd.)

The repair of osteochondral defects requires a tissue engineering approach that aims at mimicking the physiological properties and structure of two different tissues (cartilage and bone) using specifically designed scaffold-cell constructs. Biphasic and triphasic approaches utilize two or three different architectures, materials, or composites to produce a multilayered construct. This article gives an overview of some of the current strategies in multiphasic/gradient-based scaffold architectures and compositions for tissue engineering of osteochondral defects. In addition, the application of finite element analysis (FEA) in scaffold design and simulation of in vitro and in vivo cell growth outcomes has been briefly covered. FEA-based approaches can potentially be coupled with computer-assisted fabrication systems for controlled deposition and additive manufacturing of the simulated patterns. Finally, a summary of the existing challenges associated with the repair of osteochondral defects as well as some recommendations for future directions have been brought up in the concluding section of this article., (© 2014 Wiley Periodicals, Inc.)

Low back pain is one of the major health problems in industrialized countries, as a leading source of disability in the working population. Intervertebral disc degeneration has been identified as its main cause, being a progressive process mainly characterized by alteration of extracellular matrix composition and water content. Many factors are involved in the degenerative cascade, such as anabolism/catabolism imbalance, reduction of nutrition supply and progressive cell loss. Currently available treatments are symptomatic, and surgical procedures consisting of disc removal are often necessary. Recent advances in our understanding of intervertebral disc biology led to an increased interest in the development of novel biological treatments aimed at disc regeneration. Growth factors, gene therapy, stem cell transplantation and biomaterials-based tissue engineering might support intervertebral disc regeneration by overcoming the limitation of the self-renewal mechanism. The aim of this paper is to overview the literature discussing the current status of our knowledge from the degenerative cascade of the intervertebral disc to the latest molecular, cell-based therapies and tissue-engineering strategies for disc regeneration., (Copyright © 2013 John Wiley & Sons, Ltd.)

Tissue engineering (TE) has been introduced for more than 25 years without a boom in clinical trials. More than 70 TE-related start-up companies spent more than $600 million/year, with only two FDA-approved tissue-engineered products. Given the modest performance in clinically approved organs, TE is a tenaciously promising field. The TE community is advocating the application of clinically driven methodologies in large animal models enabling clinical translation. This challenge is hindered by the scarcity of tissue biopsies and the absence of standardized evaluation tools, but can be negated through non-invasive assessment of growth and integration, with reduced sample size and low cost. Solving this issue will speed the transition to cost-efficient clinical studies. In this paper we: (a) introduce magnetic resonance elastography to the tissue-engineering and regenerative medicine (TERM) community; (b) review recent MRE applications in TERM; and (c) discuss future directions of MRE in TERM. We have used MRE to study engineered tissues both in vitro and in vivo, where the mechanical properties of mesenchymally derived constructs were progressively monitored before and after tissues were implanted in mouse models. This study represents a stepping stone toward the applications of MRE in directing clinical trials with low cost and likely expediting the translation to more relevantly large animal models and clinical trials., (Copyright © 2013 John Wiley & Sons, Ltd.)

The clinical use of biological scaffold materials has become commonplace. Such scaffolds are composed of extracellular matrix (ECM), or components of ECM, derived from allogeneic or xenogeneic tissues. Such scaffold materials vary widely in their source tissue, processing methods and sterilization methods. The success or failure of an ECM scaffold for a given application is dependent on the host response following implantation; a response that is largely mediated by the innate immune system and which is influenced by a numerous factors, including the processing methods used in the preparation of biological scaffolds. The present paper reviews various aspects of the host response to biological scaffolds and factors that affect this response. In addition, some of the logistical, regulatory and reconstructive implications associated with the use of biological scaffolds are discussed., (Copyright © 2014 John Wiley & Sons, Ltd.)

Three-dimensional (3D) hepatocyte cultures have attracted much attention to obtain high biological functions of hepatocyte for pharmaceutical drug assessment. However, maintaining the high functions for over one month is still a key challenge although many approaches have been reported. In this study, we demonstrate for the first time simple and rapid construction of 3D-hepatocyte constructs by our cell accumulation technique and their high biological functions for one month, without any medium change. The human hepatocyte carcinoma (HepG2) cells were coated with ∼ 7 nm-sized extracellular matrix (ECM) films consisting of fibronectin (FN) and gelatin (G), and then incubated in cell culture insert to construct 3D-tissue constructs for 24 h. The thickness of obtained 3D-HepG2 constructs was easily controlled by altering seeding cell number and the maximum is over 100 μm. When a large volume of culture media was employed, the 3D-constructs showed higher mRNA expression of albumin and some cytochrome P450 (CYP) enzymes as compared to general two-dimensional (2D) culture. Surprisingly, their high cell viabilities (over 80%) and high mRNA expressions were successfully maintained without medium change for at least 27 days. These results demonstrate novel easy and rapid technique to construct 3D-human liver tissue models which can maintain their high functions and viability for 1 month without medium change., (© 2014 Wiley Periodicals, Inc.)

'Additive manufacturing' (AM) refers to a class of manufacturing processes based on the building of a solid object from three-dimensional (3D) model data by joining materials, usually layer upon layer. Among the vast array of techniques developed for the production of tissue-engineering (TE) scaffolds, AM techniques are gaining great interest for their suitability in achieving complex shapes and microstructures with a high degree of automation, good accuracy and reproducibility. In addition, the possibility of rapidly producing tissue-engineered constructs meeting patient's specific requirements, in terms of tissue defect size and geometry as well as autologous biological features, makes them a powerful way of enhancing clinical routine procedures. This paper gives an extensive overview of different AM techniques classes (i.e. stereolithography, selective laser sintering, 3D printing, melt-extrusion-based techniques, solution/slurry extrusion-based techniques, and tissue and organ printing) employed for the development of tissue-engineered constructs made of different materials (i.e. polymeric, ceramic and composite, alone or in combination with bioactive agents), by highlighting their principles and technological solutions., (Copyright © 2012 John Wiley & Sons, Ltd.)

Poly(D,L-lactide-co-glycolide) (PLGA) is widely used in a variety of tissue engineering and drug delivery applications due to its biodegradability and biocompatibility. But PLGA surfaces are usually hydrophobic which limited the loading and seeding capacities for cells, especially semiadherent immune cells. In this paper we described an attempt to improve the hydrophilicity and surface architecture for accommodating dendritic cells (DCs) that are widely used as professional antigen presenting cells in immune therapy of cancer and other diseases. The 3D porous PLGA scaffold was made by solvent casting/salt leaching of PLGA blended with surface functionalized multiwalled carbon nanotubes (F-MWCNTs). The incorporation and dispersion of F-MWCNT in the scaffold structures resulted in not only improved surface hydrophilicity but also nanoscale surface structure that would provide a preferable microenvironment for DCs attachment. We think such a scaffold material may be more desirable for immune cell delivery for immunotherapy., (© 2014 Wiley Periodicals, Inc.)

Viruses are no longer recognized purely for being ubiquitous pathogens, but have served as building blocks for material chemistry and nanotechnology. Thousands of coat protein subunits of a viral particle can be modified chemically and/or genetically. We have previously shown that the three-dimensional porous hydrogels can easily be functionalized by Tobacco mosaic virus (TMV), a rod-like plant virus, using its mutant, RGD-TMV. RGD-TMV hosted bioadhesive peptide (RGD) in the hydrogel, which was shown to enhance cell attachment and promote osteogenic differentiation of cultured stem cell. To translate this technology to potential clinical applications, we sought to study the biocompatibility of the hydrogel. In this paper, the hydrogels were implanted in vivo and assessed for their immunogenicity, toxicity, and biodegradability. Immune response for TMV substantially decreased when incorporated in the hydrogel implants. The implanted TMV hydrogels exhibited no apparent toxicity and were degradable in mice. The results highlighted the feasibility of using TMV incorporated hydrogels as scaffolding materials for regenerative medicine in terms of biocompatibility and biodegradability., (© 2014 Wiley Periodicals, Inc.)

Tissue engineering makes use of 3D scaffolds to sustain three-dimensional growth of cells and guide new tissue formation. To meet the multiple requirements for regeneration of biological tissues and organs, a wide range of scaffold fabrication techniques have been developed, aiming to produce porous constructs with the desired pore size range and pore morphology. Among different scaffold fabrication techniques, thermally induced phase separation (TIPS) method has been widely used in recent years because of its potential to produce highly porous scaffolds with interconnected pore morphology. The scaffold architecture can be closely controlled by adjusting the process parameters, including polymer type and concentration, solvent composition, quenching temperature and time, coarsening process, and incorporation of inorganic particles. The objective of this review is to provide information pertaining to the effect of these parameters on the architecture and properties of the scaffolds fabricated by the TIPS technique., (© 2014 Wiley Periodicals, Inc.)

Several stem cell-based therapeutic tools are currently being investigated for the regeneration of central nervous system (CNS) injuries. This review focuses on innovative approaches for CNS tissue repair via the use of implantable cellular devices. These devices are supported by biopharmaceuticals and conventional physiotherapy for the restoration of lost neuronal circuits and CNS function. This paper further reviews new and promising tools currently in pre-clinical and clinical tests for the treatment of CNS diseases where substantial loss of cellular and extracellular components of neural tissue has occurred such as stroke, encephalopathy and traumatic neural injuries. We also discuss selected 3D bioscaffolds co-cultured with clinically applicable human mesenchymal stem cells. Recent advances in neural tissue engineering and stem cell differentiation methods have shown promise for their clinical application in treating yet incurable CNS deficits., (Copyright © 2012 John Wiley & Sons, Ltd.)

This paper describes a non-invasive approach for efficient detachment of cells adhered to a gold substrate via a specific oligopeptide. Detachment is effected by an electrical stimulus. The oligopeptide contains cysteine, which spontaneously forms a gold-thiolate bond on a gold surface. This chemical adsorption reaches > 95% equilibrium within 10 min after immersion of a gold-coated substrate in a solution containing the peptide. The peptide is reversibly desorbed from the surface within 5 min of application of a negative electrical potential. By taking advantage of this simple adsorption and desorption mechanism, cells can be grown on an oligopeptide-functionalized gold surface and can be efficiently detached as single cells or cell sheets by application of a negative electrical potential. This approach was also applied to the surface of gold-coated microrods. Capillary-like microchannels were formed in collagen gel by transferring endothelial cells to the internal surfaces of the microchannels. During subsequent perfusion culture, the enveloped endothelial cells migrated into the collagen gel and formed luminal structures, which sprouted from the microchannels. This technique has the potential to provide a fundamental tool for the engineering of thick cell sheets as well as vascularized tissues and organs., (Copyright © 2011 John Wiley & Sons, Ltd.)

This paper is concerned with reliable and physically sound elasticity determination of rapid-prototyped tissue engineering scaffolds made of poly-L-lactide (PLLA), with and without small portions of tricalcium phosphate (TCP) inclusions. At the level of overall scaffolds, that is, that of several millimeters, multiple uniaxial loading-unloading (quasistatic) tests were performed, giving access to the scaffolds' Young's moduli, through stress-strain characteristics during unloading. In addition, acoustic tests with 0.05 MHz frequency delivered an independent access to elastic properties, in terms of the normal components of the scaffolds' stiffness tensors. The latter strongly correlate, in a linear fashion, with the Young's moduli from the unloading tests, revealing porosity independence of Poisson's ratio. The magnitude of the latter is in full agreement with literature data on polymers. Both of these facts underline that both ultrasound tests and quasistatic unloading tests reliably provide the elastic properties of tissue engineering scaffolds., (Copyright © 2012 Wiley Periodicals, Inc.)

Tissue engineering was proposed approximately 15 years ago as an alternative and innovative way to address tissue regeneration problems. During the development of this field, researchers have proposed a variety of ways of looking into the regeneration and engineering of tissues, using different types of materials coupled with a wide range of cells and bioactive agents. This trilogy is commonly considered the basis of a tissue-engineering strategy, meaning by this the use of a support material, cells and bioactive agents. Different researchers have been adding to these basic approaches other parameters able to improve the functionality of the tissue-engineered construct, such as specific mechanical environments and conditioned gaseous atmospheres, among others. Nowadays, tissue-engineering principles have been applied, with different degrees of success, to almost every tissue lacking efficient regeneration ability and the knowledge and intellectual property produced since then has experienced an immense growth. Materials for regenerating tissues, namely cartilage, have also been continuously increasing and most of the theoretical requirements for a tissue engineering support have been addressed by a single material or a mixture of materials. Due to their intrinsic features, polysaccharides are interesting for cartilage tissue-engineering approaches and as a result their exploitation for this purpose has been increasing. The present paper intends to provide an overview of some of the most relevant polysaccharides used in cartilage tissue-engineering research., (Copyright © 2010 John Wiley & Sons, Ltd.)

Cell culture is one of the critical bioprocessing steps required to generate sufficient human-derived cellular material for most cell-based therapeutic applications in regenerative medicine. Automated cell expansion is fundamental to the development of scaled, robust and cost effective commercial production processes for cell-based therapeutic products. This paper describes the first application of process capability analysis to establish and compare the short-term process capability of manual and automated processes for the in vitro expansion of a selected anchorage-dependent cell line. Estimates of the process capability indices (Cp, Cpk) have been used to assess the ability of both processes to consistently meet the requirements for a selected productivity output and to direct process improvement activities. Point estimates of Cp and Cpk show that the manual process has poor capability (Cp = 0.55, Cpk = 0.26) compared to the automated process (Cp = 1.32, Cpk = 0.25), resulting from excess variability. Comparison of point estimates, which shows that Cpk < Cp, indicates that the automated process mean was off-centre and that intervention is required to adjust the location of the process mean. A process improvement strategy involving an adjustment to the automated process settings has demonstrated in principle that the process mean can be shifted closer to the centre of the specification to achieve an estimated seven-fold improvement in process performance. In practice, the 90% confidence bound estimate of Cp (Cp = 0.90) indicates that that once the process is centred within the specification, a further reduction of process variation is required to attain an automated process with the desired minimum capability requirement., (2009 John Wiley & Sons, Ltd.)

The quest for novel materials as scaffolds with suitable micro-architecture for supporting tissue neogenesis in tissue engineering and regenerative medicine (TERM) is continuing. In this paper we report an Antheraea assama silk-based non-woven fibroin scaffold for applications in TERM. The novel three-dimensional scaffold is highly interconnected and porous, with a pore size of 150 microm, porosity of 90% and water uptake capacity of 85%. FTIR revealed a typical beta-sheet structure of fibroin. The scaffold has thermal and mechanical properties superior to those of Bombyx mori, as revealed by DSC, TGA and tensile tests. The scaffold exhibited satisfactory blood compatibility, as determined by thrombogenicity, haemolysis, platelet/leukocyte count, platelet adhesion and protein adsorption studies. The scaffold was found to be cytocompatible with human cell lines A549, KB, HepG2 and HeLa for a period of up to 4 weeks. SEM analysis revealed excellent attachment, spreading and migration of cells in the scaffold. MTT assay was performed to estimate the viability and growth of cells in the matrix. Quantification of collagen in cell-scaffold constructs was done by picro-Sirius red assay. Ex ovo chorioallantoic membrane assay and nitric oxide estimations in spent culture medium showed the scaffold's ability to promote angiogenesis. Finally, the biodegradability of the scaffold was determined by the weight loss observed upon treatment with trypsin over a period of 4 weeks. The results reveal that the fibroin from A. assama is a promising candidate as a biocompatible, biomimetic and biodegradable biomaterial of natural origin for applications in TERM., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

Modern tissue engineering concepts integrate cells, scaffolds, signalling molecules and growth factors. For the purposes of regenerative medicine, fetal development is of great interest because it is widely accepted that regeneration recapitulates in part developmental processes. In tissue engineering of cartilage the growth plate of the long bone represents an interesting, well-organized developmental structure with a spatial distribution of chondrocytes in different proliferation and differentiation stages, embedded in a scaffold of extracellular matrix components. The proliferation and differentiation of these chondrocytes is regulated by various hormonal and paracrine factors. Thus, members of the TGFbeta superfamily, the parathyroid hormone-related peptide-Indian hedgehog loop and a number of transcription factors, such as Sox and Runx, are involved in the regulation of chondrocyte proliferation and differentiation. Furthermore, adhesion molecules, homeobox genes, metalloproteinases and prostaglandins play a role in the complex regulation mechanisms. The present paper summarizes the morphological organization of the growth plate and provides a short but not exhaustive overview of the regulation of growth plate development, giving interesting insights for tissue engineering of cartilage.

Silks have been proposed as potential scaffold materials for tissue engineering, mainly because of their physical properties. They are stable at physiological temperatures, flexible and resist tensile and compressive forces. Bombyx mori (silkworm) cocoon silk has been used as a suture material for over a century, and has proved to be biocompatible once the immunogenic sericin coating is removed. Spider silks have a similar structure to silkworm silk but do not have a sericin coating. This paper provides a general overview on the use of silk protein in biomaterials, with a focus on skeletal tissue engineering.

Heart valve disease is a significant cause of mortality worldwide. However, to date, a nonthrombogenic, noncalcific prosthetic, which maintains normal valve mechanical properties and hemodynamic flow, and exhibits sufficient fatigue properties has not been designed. Current prosthetic designs have not been optimized and are unsuitable treatment for congenital heart defects. Research is therefore moving towards the development of a tissue engineered heart valve equivalent. Two approaches may be used in the creation of a tissue engineered heart valve, the traditional approach, which involves seeding a scaffold in vitro, in the presence of specific signals prior to implantation, and the guided tissue regeneration approach, which relies on autologous reseeding in vivo. Regardless of the approach taken, the design of a scaffold capable of supporting the growth of cells and extracellular matrix generation and capable of withstanding the unrelenting cardiovascular environment while forming a tight seal during closure, is critical to the success of the tissue engineered construct. This paper focuses on the quest to design, such a scaffold.