Scaffolds For Tissue Engineering Research Articles

Nanostructured Biomaterials in 3D Tumor Tissue Engineering Scaffolds: Regenerative Medicine and Immunotherapies.

The evaluation of nanostructured biomaterials and medicines is associated with 2D cultures that provide insight into biological mechanisms at the molecular level, while critical aspects of the tumor microenvironment (TME) are provided by the study of animal xenograft models. More realistic models that can histologically reproduce human tumors are provided by tissue engineering methods of co-culturing cells of varied phenotypes to provide 3D tumor spheroids that recapitulate the dynamic TME in 3D matrices. The novel approaches of creating 3D tumor models are combined with tumor tissue engineering (TTE) scaffolds including hydrogels, bioprinted materials, decellularized tissues, fibrous and nanostructured matrices. This review focuses on the use of nanostructured materials in cancer therapy and regeneration, and the development of realistic models for studying TME molecular and immune characteristics. Tissue regeneration is an important aspect of TTE scaffolds used for restoring the normal function of the tissues, while providing cancer treatment. Thus, this article reports recent advancements in the development of 3D TTE models for antitumor drug screening, studying tumor metastasis, and tissue regeneration. Also, this review identifies the significant opportunities of using 3D TTE scaffolds in the evaluation of the immunological mechanisms and processes involved in the application of immunotherapies.

Thickening tissue by thinning electrospun scaffolds for skeletal muscle tissue engineering

AbstractElectrospun scaffolds with aligned fiber orientation are widely used in tissue engineering, such as muscle, heart, nerve, tendon, and cartilage, due to their ability to guide cell morphology and induce cellular functions. However, the dense fibrous structure of the scaffolds poses a critical obstacle to engineering highly cellular and thick 3D tissues, as it prevents cell infiltration. While many techniques have been developed to increase the pore size of electrospun scaffolds and improve cell infiltration/migration, it often leads to a decrease in direct cell‐cell contact, compromising cell differentiation and tissue maturation. This study presents an alternative approach by reducing the thickness of scaffolds to the cellular scale and stacking or rolling the cell‐scaffold complex into 3D constructs. We devise a series of novel tools to fabricate, characterize, and manipulate ultra‐thin electrospun scaffolds, which demonstrate high reproducibility, resolution, and cellularity. Our study provides a solution to the cell infiltration issue in muscle tissue engineering and is highly versatile, and can be applied to various fields that require structures with high‐resolution gradients in a layered pattern or complex spatial distribution in a rolled pattern.

Tissue Engineering Scaffolds for Dentin-Pulp Complex Regeneration

Tissue Engineering Scaffolds for Dentin-Pulp Complex Regeneration

A Biomimetic Fibrous Composite Scaffold with Nanotopography-Regulated Mineralization for Bone Defect Repair.

The effective regeneration of large bone defects via bone tissue engineering is challenging due to the difficulty in creating an osteogenic microenvironment. Inspired by the fibrillar architecture of the natural extracellular matrix, we developed a nanoscale bioengineering strategy to produce bone fibril-like composite scaffolds with enhanced osteogenic capability. To activate the surface for biofunctionalization, self-adaptive ridge-like nanolamellae were constructed on poly(ε-caprolactone) (PCL) electrospinning scaffolds via surface-directed epitaxial crystallization. This unique nanotopography with a markedly increased specific surface area offered abundant nucleation sites for Ca2+ recruitment, leading to a 5-fold greater deposition weight of hydroxyapatite than that of the pristine PCL scaffold under stimulated physiological conditions. Bone marrow mesenchymal stem cells (BMSCs) cultured on bone fibril-like scaffolds exhibited enhanced adhesion, proliferation, and osteogenic differentiation in vitro. In a rat calvarial defect model, the bone fibril-like scaffold significantly accelerated bone regeneration, as evidenced by micro-CT, histological histological and immunofluorescence staining. This work provides the way for recapitulating the osteogenic microenvironment in tissue-engineered scaffolds for bone repair.

Chitosan-incorporated Bioceramic-based Nanomaterials for Localized Release of Therapeutics and Bone Regeneration: An Overview of Recent Advances and Progresses

Abstract: The usage of nanoparticles in tissue engineering applications has increased significantly in the last several years. Functional tissues are developed by regulating cell proliferation, differentiation, and migration on nanostructured scaffolds containing cells. These scaffolds provide an environment that is more structurally supportive than the microarchitecture of natural bone. Given its exceptional properties, such as its osteogenic potential, biocompatibility, and biodegradability, chitosan is a good and promising biomaterial. Unfortunately, chitosan's low mechanical strength makes it unsuitable for load-bearing applications. By mixing chitosan with other biomaterials, this drawback might be mitigated. Bone tissue engineering uses both bioresorbable materials like tricalcium phosphate and bioactive materials like hydroxyapatite and bioglass. Alumina and titanium are examples of bioinert materials that are part of these bioceramics. When produced at nanoscale scales, these materials have a larger surface area and better cell adhesion. This review paper will go into great detail on the bioinert, bioresorbable, and bioactive nanoceramics-reinforced chitosan scaffolds for bone tissue engineering.

Engineering Cell Instructive Microenvironments for In Vitro Replication of Functional Barrier Organs.

Multicellular organisms exhibit synergistic effects among their components, giving rise to emergent properties crucial for their genesis and overall functionality and survival. Morphogenesis involves and relies upon intricate and biunivocal interactions among cells and their environment, that is, the extracellular matrix (ECM). Cells secrete their own ECM, which in turn, regulates their morphogenetic program by controlling time and space presentation of matricellular signals. The ECM, once considered passive, is now recognized as an informative space where both biochemical and biophysical signals are tightly orchestrated. Replicating this sophisticated and highly interconnected informative media in a synthetic scaffold for tissue engineering is unattainable with current technology and this limits the capability to engineer functional human organs in vitro and in vivo. This review explores current limitations to in vitro organ morphogenesis, emphasizing the interplay of gene regulatory networks, mechanical factors, and tissue microenvironment cues. In vitro efforts to replicate biological processes for barrier organs such as the lung and intestine, are examined. The importance of maintaining cells within their native microenvironmental context is highlighted to accurately replicate organ-specific properties. The review underscores the necessity for microphysiological systems that faithfully reproduce cell-native interactions, for advancing the understanding of developmental disorders and disease progression.

Scaffold-based microsphere in drug delivery system

Microspheres are free-flowing powders having a synthetic and natural polymer. A targeted drug delivery system can overcome some of the problems of conventional therapy and enhance the therapeutic efficacy of the drug. This is a biodegradable and non-biodegradable efficacy of a given drug. There are various approaches to delivering a therapeutic substance to the target site in a sustained controlled release fashion. One approach is a scaffold-based microsphere for drug delivery, where the target site is very specific if modified, and the proper concentration is maintained at the site of interest without causing toxic effects. Microspheres received much attention not only for control release but also in drug targeting for example in anticancer therapy. Moreover, the joining of medications (i.e., incendiary inhibitors and additionally anti-microbial) into platforms might be utilized to forestall contamination after medical procedures and other infections for the longer term. The framework additionally can be utilized to give sufficient signs to the cells, to initiate and keep them in their coveted separation organization, and to keep up their survival and development. The present survey gives an itemized record of the requirement for the advancement of frameworks alongside the materials utilized and systems embraced to fabricate platforms for tissue designing and delay. The present review gives a detailed account of the need for the development of scaffolds along with the materials used and techniques adopted to manufacture scaffolds for tissue engineering and microspheres for drug delivery systems.

Recent Applications of Chitosan and Its Derivatives in Antibacterial, Anticancer, Wound Healing, and Tissue Engineering Fields.

Chitosan, a versatile biopolymer derived from chitin, has garnered significant attention in various biomedical applications due to its unique properties, such as biocompatibility, biodegradability, and mucoadhesiveness. This review provides an overview of the diverse applications of chitosan and its derivatives in the antibacterial, anticancer, wound healing, and tissue engineering fields. In antibacterial applications, chitosan exhibits potent antimicrobial properties by disrupting microbial membranes and DNA, making it a promising natural preservative and agent against bacterial infections. Its role in cancer therapy involves the development of chitosan-based nanocarriers for targeted drug delivery, enhancing therapeutic efficacy while minimising side effects. Chitosan also plays a crucial role in wound healing by promoting cell proliferation, angiogenesis, and regulating inflammatory responses. Additionally, chitosan serves as a multifunctional scaffold in tissue engineering, facilitating the regeneration of diverse tissues such as cartilage, bone, and neural tissue by promoting cell adhesion and proliferation. The extensive range of applications for chitosan in pharmaceutical and biomedical sciences is not only highlighted by the comprehensive scope of this review, but it also establishes it as a fundamental component for forthcoming research in biomedicine.

Quantitative Evaluation of the Pore and Window Sizes of Tissue Engineering Scaffolds on Scanning Electron Microscope Images Using Deep Learning.

The morphological characteristics of tissue engineering scaffolds, such as pore and window diameters, are crucial, as they directly impact cell-material interactions, attachment, spreading, infiltration of the cells, degradation rate and the mechanical properties of the scaffolds. Scanning electron microscopy (SEM) is one of the most commonly used techniques for characterizing the microarchitecture of tissue engineering scaffolds due to its advantages, such as being easily accessible and having a short examination time. However, SEM images provide qualitative data that need to be manually measured using software such as ImageJ to quantify the morphological features of the scaffolds. As it is not practical to measure each pore/window in the SEM images as it requires extensive time and effort, only the number of pores/windows is measured and assumed to represent the whole sample, which may cause user bias. Additionally, depending on the number of samples and groups, a study may require measuring thousands of samples and the human error rate may increase. To overcome such problems, in this study, a deep learning model (Pore D2) was developed to quantify the morphological features (such as the pore size and window size) of the open-porous scaffolds automatically for the first time. The developed algorithm was tested on emulsion-templated scaffolds fabricated under different fabrication conditions, such as changing mixing speed, temperature, and surfactant concentration, which resulted in scaffolds with various morphologies. Along with the developed model, blind manual measurements were taken, and the results showed that the developed tool is capable of quantifying pore and window sizes with a high accuracy. Quantifying the morphological features of scaffolds fabricated under different circumstances and controlling these features enable us to engineer tissue engineering scaffolds precisely for specific applications. Pore D2, an open-source software, is available for everyone at the following link: https://github.com/ilaydakaraca/PoreD2.

Can self-powered piezoelectric materials be used to treat disc degeneration by means of electrical stimulation?

Intervertebral disc degeneration (IDD) due to multiple causes is one of the major causes of low back pain (LBP). A variety of traditional treatments and biologic therapies are currently used to delay or even reverse IDD; however, these treatments still have some limitations. Finding safer and more effective treatments is urgent for LBP patients. With increasing reports it has been found that the intervertebral disc (IVD) can convert pressure loads from the spine into electrical stimulation in a variety of ways, and that this electrical stimulation is of great importance in modulating cell behavior, the immune microenvironment and promoting tissue repair. However, when intervertebral disc degeneration occurs, the normal structures within the IVD are destroyed. This eventually leads to a weakening or loss of self-powered. Currently various piezoelectric materials with unique crystal structures can mimic the piezoelectric effect of normal tissues. Based on this, tissue-engineered scaffolds prepared using piezoelectric materials have been widely used for regenerative repair of various types of tissues, however, there are no reports of their use for the treatment of IDD. For this reason, we propose to utilize tissue-engineered scaffolds prepared from piezoelectric biomaterials with excellent biocompatibility and self-powered properties to be implanted into degenerated IVD to help restore cell type and number, restore extracellular matrix, and modulate immune responses. It provides a feasible and novel therapeutic approach for the clinical treatment of IDD.

Continuous fabrication of braided composite nanofibrous surgical yarns using advanced AC electrospinning and braiding technology

Composite nanofibrous yarns (CNY), featuring a submicron electrospun sheath coated on a micro-scale core yarn have gained attention within the academic and industrial communities due to their unique structure. When braided, these CNYs hold great promise as next-generation surgical yarns. However, large-scale production of CNYs using conventional electrospinning techniques remains technologically challenging, and maintaining the integrity of the fibrous sheath during braiding presents further difficulties. Here, we propose a novel approach for continuously fabricating functional braided CNYs using collectorless alternating current (AC) electrospinning and braiding technology. Our approach utilized polycaprolactone-polylactic acid (PCL-PLA) blends and PCL-PLA with chlorhexidine (CHX) or triclosan (TRC) electrospun fibers for the sheath layer, while PLA micro-yarns acted as the core layer. Morphological analyses confirmed the successful fabrication of braided CNYs. Additionally, infrared spectroscopy validated the presence of CHX or TRC in the resulting yarns. The resulting braided CNYs exhibited excellent breaking force (29 N) and thermal stability (270 °C). Cytotoxicity and antibacterial assessments demonstrated that CHX-loaded braided CNYs could serve as biocompatible antibacterial surgical sutures. The proposed method offers a versatile approach for producing various functional braided CNYs applicable in tissue engineering scaffolds, filters, wearable electronics, and sensors.

Challenges in Application: Gelation Strategies of DAT-Based Hydrogel Scaffolds.

Decellularized adipose tissue (DAT) has great clinical applicability, owing to its abundant source material, natural extracellular matrix microenvironment, and nonimmunogenic attributes, rendering it a versatile resource in the realm of tissue engineering. However, practical implementations are confronted with multifarious limitations. Among these, the selection of an appropriate gelation strategy serves as the foundation for adapting to diverse clinical contexts. The cross-linking strategies under varying physical or chemical conditions exert profound influences on the ultimate morphology and therapeutic efficacy of DAT. This review sums up the processes of DAT decellularization and subsequent gelation, with a specific emphasis on the diverse gelation strategies employed in recent experimental applications of DAT. The review expounds upon methodologies, underlying principles, and clinical implications of different gelation strategies, aiming to offer insights and inspiration for the application of DAT in tissue engineering and advance research for tissue engineering scaffold development.

A Pump-Free Strategy for the Controllable Generation of Alginate Microgels as Cellular Microcarriers.

Microgels are advanced scaffolds for tissue engineering due to their proper biodegradability, good biocompatibility, and high specific surface area for effective oxygen and nutrient transfer. However, most of the current monodispersed microgel fabrication systems rely heavily on various precision pumps, which highly increase the cost and complexity of their downstream application. In this work, we developed a simple and facile system for the controllable generation of uniform alginate microgels by integrating a gas-shearing strategy into a glass microfluidic device. Importantly, the cell-laden microgels can be rapidly prepared in a pump-free manner under an all-aqueous environment. The three-dimensional cultured green fluorescent protein-human A549 cells in alginate microgels exhibited enhanced stemness and drug resistance compared to those under two-dimensional conditions. The pancreatic cancer organoids in alginate microgels exhibited some of the key features of pancreatic cancer. The proposed microgels showed decent monodispersity, biocompatibility, and versatility, providing great opportunities in various biomedical applications such as microcarrier fabricating, organoid engineering, and high-throughput drug screening.

CT-Visible Microspheres Enable Whole-Body In Vivo Tracking of Injectable Tissue Engineering Scaffolds.

Targeted delivery and retention are essential requirements for implantable tissue-engineered products. Non-invasive imaging methods that can confirm location, retention, and biodistribution of transplanted cells attached to implanted tissue engineering scaffolds will be invaluable for the optimization and enhancement of regenerative therapies. To address this need, an injectable tissue engineering scaffold consisting of highly porous microspheres compatible with transplantation of cells is modified to contain the computed tomography (CT) contrast agent barium sulphate (BaSO4). The trackable microspheres show high x-ray absorption, with contrast permitting whole-body tracking. The microspheres are cellularized with GFP+ Luciferase+ mesenchymal stem cells and show in vitro biocompatibility. In vivo, cellularized BaSO4-loaded microspheres are delivered into the hindlimb of mice where they remain viable for 14 days. Co-registration of 3D-bioluminescent imagingand µCT reconstructions enable the assessment of scaffold material and cell co-localization. The trackable microspheres are also compatible with minimally-invasive delivery by ultrasound-guided transthoracic intramyocardial injections in rats. These findings suggest that BaSO4-loaded microspheres can be used as a novel tool for optimizing delivery techniques and tracking persistence and distribution of implanted scaffold materials. Additionally, the microspheres can be cellularized and have the potential to be developed into an injectable tissue-engineered combination product for cardiac regeneration.

Dimensional and Structural Instability of Electrospun Polylactic Acid Membranes in Liquid Environments: Role of Water, Ethanol, and Temperature

Electrospun nanofibres of polylactic acid (PLA) are suggested for a variety of uses, including scaffolds for tissue engineering, components of drug delivery devices, sustainable packaging materials and membranes for liquid filtration/purification. For all these applications, it is critical to consider the stability of the PLA electrospun materials once in operation. Exposure to certain liquids and temperatures can modify their dimensions, shape, surface topography and mechanical response and compromise their performance. In this study, electrospun PLA mats were exposed to water and ethanol solutions, at different temperatures and for defined time periods, and changes in their properties were analysed. It was found that the impact of water on area shrinkage and fibre arrangement strongly depended on temperature, particularly if the treatment was performed at the glass transition temperature of PLA. Ethanol, instead, induced significant alterations in the size, morphology, and elastic modulus of the electrospun mats, even at room temperature and determined the formation of crimped structures. This work provides insights into the conditions that can critically affect the properties of PLA electrospun fibres and, hence, impact on their usage.

Physical cues of scaffolds promote peripheral nerve regeneration

The effective treatment of long-gap peripheral nerve injury (PNI) remains a challenge in clinical settings. The autograft, the gold standard for the long-gap PNI therapy, has several limitations, including a limited supply of donor nerve, size mismatch between the donor and recipient sites, functional loss at the donor site, neuroma formation, and the requirement for two operations. With the increasing abundance of biocompatible materials with adjustable structures and properties, tissue engineering provides a promising avenue for bridging peripheral nerve gaps and addressing the above issues of autograft. The physical cues provided by tissue engineering scaffolds, essential for regulating the neural cell fate and microenvironments, have received considerable research attention. This review elaborates on three major physical cues of tissue engineering scaffolds for peripheral nerve regeneration: topological structure, mechanical support, and electrical stimulation. These three aspects are analogs to Lego bricks, wherein different combinations result in diverse functions. Innovative and more effective bricks, along with multi-level and all-around integration, are expected to provide new advances in tissue engineering for peripheral nerve generation.

Stem Cell‐Based Therapies and Tissue Engineering Innovations for Tendinopathy: A Comprehensive Review of Current Strategies and Future Directions

AbstractTendon diseases commonly lead to physical disability, exerting a profound impact on the routine of affected patients. These conditions respond poorly to existing treatments, presenting a substantial challenge for orthopedic scientists. Research into clinical translational therapy has yet to yield highly versatile interventions capable of effectively addressing tendon diseases, including tendinopathy. Stem cell‐based therapies have emerged as a promising avenue for modifying the biological milieu through the secretion of regenerative and immunomodulatory factors. The current review provides an overview of the intricate tendon microenvironment, encompassing various tendon stem progenitor cells within distinct tendon sublocations, gene regulation, and pathways pertinent to tendon development, and the pathology of tendon diseases. Subsequently, the advantages of stem cell‐based therapies are discussed that utilize distinct types of autologous and allogeneic stem cells for tendon regeneration at the translational level. Moreover, this review outlines the challenges, gaps, and future innovations to propose a consolidated stem cell‐based therapy to treat tendinopathy. Finally, regenerative soluble therapies, insoluble bio‐active therapies, along with insoluble bio‐active therapies, and implantable 3D scaffolds for tendon tissue engineering are discussed, thereby presenting a pathway toward enhanced tissue regeneration and engineering.

Engineering a robust and anisotropic cardiac-specific extracellular matrix scaffold for cardiac patch tissue engineering

Extracellular matrix (ECM) fabricated using human induced pluripotent stem cells (hiPSCs)-derived cardiac fibroblasts (hiPSC-CFs) could serve as a completely biological scaffold for an engineered cardiac patch, leveraging the unlimited source and outstanding reproducibility of hiPSC-CFs. Additionally, hiPSC-CF-derived ECM (hiPSC-CF-ECM) holds the potential to enhance maturation of exogenous cardiomyocytes, such as hiPSC-derived cardiomyocytes (hiPSC-CMs), by providing a microenvironment rich in cardiac-specific biochemical and signaling cues. However, achieving sufficient robustness of hiPSC-CF-ECM is challenging. This study aims to achieve appropriate ECM deposition, scaffold thickness, and mechanical strength of an aligned hiPSC-CF-ECM by optimizing the culture period, ranging from 2 to 10 weeks, of hiPSC-CFs grown on micro-grated substrates, which can direct the alignment of both hiPSC-CFs and their secreted ECM. The hiPSC-CFs demonstrated a production rate of 13.5 µg ECM per day per 20,000 cells seeded. An anisotropic nanofibrous hiPSC-CF-ECM scaffold with a thickness of 20.0 ± 2.1 µm was achieved after 6 weeks of culture, followed by decellularization. Compositional analysis through liquid chromatography-mass spectrometry (LC-MS) revealed the presence of cardiac-specific fibrillar collagens, non-fibrillar collagens, and matricellular proteins. Uniaxial tensile stretching of the hiPSC-CF-ECM scaffold indicated robust tensile resilience. Finally, hiPSCs-CMs cultured on the hiPSC-CF-ECM exhibited alignment following the guidance of ECM nanofibers and demonstrated mature organization of key structural proteins. The culture duration of the anisotropic hiPSC-CF-ECM was successfully refined to achieve a robust scaffold containing structural proteins that resembles cardiac microenvironment. This completely biological, anisotropic, and cardiac-specific ECM holds great potential for cardiac patch engineering.

Silk-Based 3D Porous Scaffolds for Tissue Engineering.

Silk fibroin, extracted from the silk of the Bombyx mori silkworm, stands out as a biomaterial due to its nontoxic nature, excellent biocompatibility, and adjustable biodegradability. Porous scaffolds, a type of biomaterial, are crucial for creating an optimal microenvironment that supports cell adhesion and proliferation, thereby playing an essential role in tissue remodeling and repair. Therefore, this review focuses on 3D porous silk fibroin-based scaffolds, first summarizing their preparation methods and then detailing their regenerative effects on bone, cartilage, tendon, vascular, neural, skin, hepatic, and tracheal epithelial tissue engineering in recent years.

Potential use of a bone tissue engineering scaffold based on electrospun poly (ɛ-caprolactone) - Poly (vinyl alcohol) hybrid nanofibers containing modified cockle shell nanopowder

Today, the construction of scaffolds promoting the differentiation of stem cells is an intelligent innovation that accelerates the differentiation toward the target tissue. The use of calcium and phosphate compounds is capable of elevating the precision and efficiency of the osteogenic differentiation of stem cells. In this research, osteoconductive electrospun poly (ɛ-caprolactone) (PCL) - poly (vinyl alcohol) (PVA) hybrid nanofibrous scaffolds containing modified cockle shell (CS) nanopowder were prepared and investigated. In this regard, the modified CS nanopowder was prepared by grinding and modifying with phosphoric acid, and it was then added to PVA nanofibers at different weight percentages. Based on the SEM images, the optimum content of the modified CS nanopowder was set at 7 wt %, since reaching the threshold of agglomeration restricted this incorporation. In the second step, the PVA-CS7 nanofibrous sample was hybridized with different PCL ratios. Concerning the hydrophilicity and mechanical strength, the sample named PCL50-PVA50-CS7 was ultimately selected as the optimized and suitable candidate scaffold for bone tissue application. The accelerated hydrolytic degradation of the sample was also studied by FTIR and SEM analyses, and the results confirmed that the mineral deposits of CS are available approximately 7 days for mesenchymal stem cells. Moreover, Alizarin red staining illustrated that the presence of CS in the PCL50-PVA50-CS7 hybrid nanofibrous scaffold may potentially lead to an increase in calcium deposits with high precipitates, authenticating the differentiation of stem cells towards osteogenic cells.