Tissue Engineering Research Research Articles

Study of composition of hydrogels and bioactive zinc (II) MOFs on plant tissue growth

In this work, we report the synthesis of metal-organic frameworks (MOFs) using zinc (II) and the amino acids L-phenylalanine (Phe), L-tryptophan (Trp), and L-histidine (His) as ligands. They were incorporated in hydrogel matrixes comprised of collagen, starch, green and red tomato cells to obtain composite hydrogels. The cell migration into the polymer matrices, the resistance to degradation and in vitro biocompatibility of hydrogels was studied. The cell migration assay showed a high confluence of plant cells, indicating a favorable environment for cell migration and proliferation in the presence of MOF-Zn(His) and MOF-Zn(Phe). During a degradation kinetics in vegetable substrate for 10 days, a rapid loss of water was observed in the first two days, indicating a remarkable water absorption capacity in the hydrogels. The results of metabolic activity in red and green tomato cells revealed a significant increase, exceeding 164±12 %, suggesting promising cell development in this type of composite hydrogels. These results support the potential of hydrogels synthesized with MOFs in agricultural and biotechnological applications, opening perspectives for future research and applications in plant tissue engineering.

Decellularised extracellular matrix-based injectable hydrogels for tissue engineering applications.

Decellularised extracellular matrix (dECM) is a biomaterial derived from natural tissues that has attracted considerable attention from tissue engineering researchers due to its exceptional biocompatibility and malleability attributes. These advantageous properties often facilitate natural cell infiltration and tissue reconstruction for regenerative medicine. Due to their excellent fluidity, the injectable hydrogels can be administered in a liquid state and subsequently formed into a gel state in vivo, stabilising the target area and serving in a variety of ways, such as support, repair, and drug release functions. Thus, dECM-based injectable hydrogels have broad prospects for application in complex organ structures and various tissue injury models. This review focuses on exploring research advances in dECM-based injectable hydrogels, primarily focusing on the applications and prospects of dECM hydrogels in tissue engineering. Initially, the recent developments of the dECM-based injectable hydrogels are explained, summarising the different preparation methods with the evaluation of injectable hydrogel properties. Furthermore, some specific examples of the applicability of dECM-based injectable hydrogels are presented. Finally, we summarise the article with interesting prospects and challenges of dECM-based injectable hydrogels, providing insights into the development of these composites in tissue engineering and regenerative medicine.

Elastomer in vascular tissue engineering research progress of scaffold materials for constructing tissue engineered blood vessels

This paper first introduces the requirements of tissue engineering for scaffold materials, and then analyzes the available scaffold materials, hoping to provide help for the research of tissue engineering vascular scaffold materials, and then promote the rapid development of tissue engineering industry.

Salivary Gland Bioengineering.

Salivary gland dysfunction affects millions globally, and tissue engineering may provide a promising therapeutic avenue. This review delves into the current state of salivary gland tissue engineering research, starting with a study of normal salivary gland development and function. It discusses the impact of fibrosis and cellular senescence on salivary gland pathologies. A diverse range of cells suitable for tissue engineering including cell lines, primary salivary gland cells, and stem cells are examined. Moreover, the paper explores various supportive biomaterials and scaffold fabrication methodologies that enhance salivary gland cell survival, differentiation, and engraftment. Innovative engineering strategies for the improvement of vascularization, innervation, and engraftment of engineered salivary gland tissue, including bioprinting, microfluidic hydrogels, mesh electronics, and nanoparticles, are also evaluated. This review underscores the promising potential of this research field for the treatment of salivary gland dysfunction and suggests directions for future exploration.

Carbon nanotube nanocomposite scaffolds: advances in fabrication and applications for tissue regeneration and cancer therapy.

Carbon nanotube (CNT) nanocomposite scaffolds have emerged as highly promising frameworks for tissue engineering research. By leveraging their intrinsic electrical conductivity and valuable mechanical properties, CNTs are commonly dispersed into polymers to create robust, electrically conductive scaffolds that facilitate tissue regeneration and remodeling. This article explores the latest progress and challenges related to CNT dispersion, functionalization, and scaffold printing techniques, including electrospinning and 3D printing. Notably, these CNT scaffolds have demonstrated remarkable positive effects across various cell culture systems, stimulating neuronal growth, promoting cardiomyocyte maturation, and facilitating osteocyte differentiation. These encouraging results have sparked significant interest within the regenerative medicine field, including neural, cardiac, muscle, and bone regenerations. However, addressing the concern of CNT cytotoxicity in these scaffolds remains critical. Consequently, substantial efforts are focused on exploring strategies to minimize cytotoxicity associated with CNT-based scaffolds. Moreover, researchers have also explored the intriguing possibility of utilizing the natural cytotoxic properties of CNTs to selectively target cancer cells, opening up promising avenues for cancer therapy. More research should be conducted on cutting-edge applications of CNT-based scaffolds through phototherapy and electrothermal ablation. Unlike drug delivery systems, these novel methodologies can combine 3D additive manufacturing with the innate physical properties of CNT in response to electromagnetic stimuli to efficiently target localized tumors. Taken together, the unique properties of CNT-based nanocomposite scaffolds position them as promising candidates for revolutionary breakthroughs in both regenerative medicine and cancer treatment. Continued research and innovation in this area hold significant promise for improving healthcare outcomes.

Non-invasive continuous imaging approaches to evaluate tissue structure and development

As the field of tissue engineering advances toward fully functional organoids and artificially generated tissues, methods to evaluate the condition of the cultured tissue from the beginning to the end continuously grow more importance. The structure and developmental status of tissues in vivo reveal considerably more information than current approaches to discrete analysis like histology. Optical imaging modalities present a promising future regarding its ability to unveil morphological and biochemical changes of the tissue in a continuous manner without compromising its viability. Therefore, in this paper, several popular and emerging methods include Fluorescence Lifetime Imaging, Hyperspectral Imaging, Optical Coherence Microscopy, Light Sheet Fluorescence Microscopy, Near-infrared II Fluorescence Imaging, Mueller-matrix imaging are discussed with regards to their advantages and limitations in tissue engineering research. Although these approaches allow long term monitoring of developing tissue in vivo, they are not suitable in all conditions and lack spatial resolution and depth penetration. The potential of optical imaging on tissue structure and interactions can be further realized by improving upon fixed sample imaging techniques and utilizing multimodal systems.

Microfluidic-Based Reconstitution of Functional Lymphatic Microvasculature: Elucidating the Role of Lymphatics in Health and Disease.

The knowledge of the blood microvasculature and its functional role in health and disease has grown significantly attributable to decades of research and numerous advances in cell biology and tissue engineering; however, the lymphatics (the secondary vascular system) has not garnered similar attention, in part due to a lack of relevant in vitro models that mimic its pathophysiological functions. Here, a microfluidic-based approach is adopted to achieve precise control over the biological transport of growth factors and interstitial flow that drive the in vivo growth of lymphatic capillaries (lymphangiogenesis). The engineered on-chip lymphatics with in vivo-like morphology exhibit tissue-scale functionality with drainage rates of interstitial proteins and molecules comparable to in vivo standards. Computational and scaling analyses of the underlying transport phenomena elucidate the critical role of the three-dimensionalgeometry and lymphatic endothelium in recapitulating physiological drainage. Finally, the engineered on-chip lymphatics enabled studies of lymphatic-immune interactions that revealed inflammation-driven responses by the lymphatics to recruit immune cells via chemotactic signals similar to in vivo, pathological events. This on-chip lymphatics platform permits the interrogation of various lymphatic biological functions, as well as screening of lymphatic-based therapies such as interstitial absorption of protein therapeutics and lymphatic immunomodulation for cancer therapy.

Live Imaging of 3D Hanging Drop Arrays through Manipulation of Light-Responsive Pyroelectric Slippery Surface and Chip Adhesion.

Three-dimensional (3D) hanging drop cell culture is widely used in organoid culture because of its lack of selection pressure and rapid cell aggregation. However, current hanging drop technology has limitations, such as a dependence on complex microfluidic transport channels or specific capillary force templates for drop formation, which leads to unchangeable drop features. These methods also hinder live imaging because of space and complexity constraints. Here, we have developed a hanging drop construction method and created a flexible 3D hanging drop construction platform composed of a manipulation module and an adhesion module. Their harmonious operation allows for the easy construction of hanging drops of varying sizes, types, and patterns. Our platform produces a cell hanging drop chip with small sizes and clear fields of view, thereby making it compatible with live imaging. This platform has great potential for personalized medicine, cancer and drug discovery, tissue engineering, and stem cell research.

Photo biomodulation of dental derived stem cells to ameliorate regenerative capacity: In vitro study

BackgroundDental regeneration benefits from improving the features of dental derived stem cells. Gallium-aluminum-arsenide laser had a significant role in modification of cell behavior in different cell lines and culture conditions. Hence, exploring its mechanism and effect on dental derived stem cells would benefit prospective regenerative dental therapies. ObjectivesTo assess the impact of photo biomodulation by Low-Level-Laser on isolated Dental Pulp derived Stem Cells and Periodontal Ligament derived Stem Cells regarding their proliferation and osteogenic differentiation. MethodsIsolated DPSCs and PDLSCs from impacted third molars were subjected to Gallium-aluminum-arsenide laser for 12 sec and 3.6 J/cm2. The proliferative capacity was evaluated via 3-(4,5-dimethylthiazol-2-yl),2,5-diphenyltetrazolium bromide (MTT) Assay and Trypan blue stain. Cell osteogenic differentiation potentials were assessed by alkaline phosphatase assay and alizarin red stain, polymerase chain reaction was performed to quantify Nuclear factor Kappa gene expression. ResultsDPSCs subjected to laser bio-stimulation showed the best results regarding cell viability (MTT) and osteogenic differentiation (ALP assay), and calcium deposition at 3 intervals (3, 7, 14 days), meanwhile, PDLSCs subjected to laser bio-stimulation showed better result than control but less than DPSCs. While NF-KB gene expression was proven to be approximately comparable for both groups. Generally, the Photo-bio modulated groups showed better results than their control groups. ConclusionLow-level laser bio-stimulation (LLL) therapy improves DPSC and PDLSC osteogenic differentiation and proliferation via the activation of the NF-KB pathway. Also, the DPSCs outperformed PDLSCs in terms of performance. Clinical significanceThese results can be beneficial information and a reference database for more research in tissue engineering, dental therapy, and regeneration.

Stretchable Sponge-Based Electrochemical Biosensor for Real-Time Sensing of Cells in Three-Dimensional Culture.

For the study of cell biology, real-time information on cell physiological processes will be more accurate and closer to the in vivo condition in a three-dimensional (3D) culture system. Although most reported 3D cell culture scaffolds can better mimic the in vivo dynamic microenvironment, the real-time analysis technique is deficient or lacking. Herein, a stretchable and conductive 3D scaffold is developed to construct an electrochemical biosensor for real-time monitoring of cell release in 3D culture under stimulation of drug stimulant and mechanical force. In our design, the polyurethane sponge (PU) dipped with conductive carbon ink (CC/PU) was used as a conductive scaffold, and gold nanoparticles (nano-Au) were electrodeposited on the CC/PU (nano-Au CC/PU) to improve the electrochemical sensing performance. The prepared nano-Au CC/PU scaffold exhibits a good electrocatalytic ability to H2O2 with a linear range from 20 nM to 43 μM. Due to the great biocompatibility, HeLa cells can be cultured directly on the nano-Au CC/PU and the in situ and real-time tracking of H2O2 secretion from cells was achieved. The results demonstrate that both the drug stimulant and mechanical force can rapidly activate the release of reactive oxygen species. This study indicates that the stretchable 3D sensing scaffold has good potential for cell biology research in an in vivo-like microenvironment and can be extensively used in the fields of tissue engineering, drug screening, and pathological research.

Three-Dimensional Bioprinting in Soft Tissue Engineering for Plastic and Reconstructive Surgery.

Skeletal muscle tissue engineering (TE) and adipose tissue engineering have undergone significant progress in recent years. This review focuses on the key findings in these areas, particularly highlighting the integration of 3D bioprinting techniques to overcome challenges and enhance tissue regeneration. In skeletal muscle TE, 3D bioprinting enables the precise replication of muscle architecture. This addresses the need for the parallel alignment of cells and proper innervation. Satellite cells (SCs) and mesenchymal stem cells (MSCs) have been utilized, along with co-cultivation strategies for vascularization and innervation. Therefore, various printing methods and materials, including decellularized extracellular matrix (dECM), have been explored. Similarly, in adipose tissue engineering, 3D bioprinting has been employed to overcome the challenge of vascularization; addressing this challenge is vital for graft survival. Decellularized adipose tissue and biomimetic scaffolds have been used as biological inks, along with adipose-derived stem cells (ADSCs), to enhance graft survival. The integration of dECM and alginate bioinks has demonstrated improved adipocyte maturation and differentiation. These findings highlight the potential of 3D bioprinting techniques in skeletal muscle and adipose tissue engineering. By integrating specific cell types, biomaterials, and printing methods, significant progress has been made in tissue regeneration. However, challenges such as fabricating larger constructs, translating findings to human models, and obtaining regulatory approvals for cellular therapies remain to be addressed. Nonetheless, these advancements underscore the transformative impact of 3D bioprinting in tissue engineering research and its potential for future clinical applications.

Oriented Porous Polymer Scaffolds in Tissue Engineering: A Comprehensive Review of Preparation Strategies and Applications

AbstractThe pursuit of effective therapeutic strategies for tissue damage has prompted extensive scholarly investigations worldwide. Tissue engineering has emerged as a prominent approach, particularly through the utilization of artificial scaffolds that closely resemble the natural extracellular matrix (ECM). These scaffolds exhibit multi‐scale topological structures and surface physicochemical properties, which significantly influence cellular behavior, thereby attracting considerable attention from numerous researchers. This comprehensive review is concentrated on the primary techniques employed in the fabrication of biodegradable polymer scaffolds possessing oriented porous structures, and the most recent advancements in tissue engineering research are presented. Significantly, the profound influence of scaffold surface characteristics is underscored on cellular behavior, elucidating the superiority of oriented pore structures over disordered ones in mimicking the distinctive attributes of the ECM. Enhanced cell adhesion, proliferation, and tissue differentiation represent notable advantages associated with oriented porous scaffolds. Additionally, the critical interplay between scaffold structure, performance, and functionalization is emphasized, highlighting the imperative to optimize the clinical application of tissue engineering scaffolds.

An overview of the production of tissue extracellular matrix and decellularization process.

Thousands of patients need an organ transplant yearly, while only a tiny percentage have this chance to receive a tissue/organ transplant. Nowadays, decellularized animal tissue is one of the most widely used methods to produce engineered scaffolds for transplantation. Decellularization is defined as physically or chemically removing cellular components from tissues while retaining structural and functional extracellular matrix (ECM) components and creating an ECM-derived scaffold. Then, decellularized scaffolds could be reseeded with different cells to fabricate an autologous graft. Effective decellularization methods preserve ECM structure and bioactivity through the application of the agents and techniques used throughout the process. The most valuable agents for the decellularization process depend on biological properties, cellular density, and the thickness of the desired tissue. ECM-derived scaffolds from various mammalian tissues have been recently used in research and preclinical applications in tissue engineering. Many studies have shown that decellularized ECM-derived scaffolds could be obtained from tissues and organs such as the liver, cartilage, bone, kidney, lung, and skin. This review addresses the significance of ECM in organisms and various decellularization agents utilized to prepare the ECM. Also, we describe the current knowledge of the decellularization of different tissues and their applications.

Global Bibliometric and Visualized Analysis of Tracheal Tissue Engineering Research.

The development of tracheal tissue engineering (TTE) has seen a rapid growth in recent years. The purpose of this study was to investigate the global status, trends, and hotspots of TTE research based on bibliometrics and visualization analysis. Publications related to TTE were retrieved and included in the Web of Science Core Collection. VOSviewer and CiteSpace were used to generate knowledge maps. Six hundred fifty-five publications were identified, and the quantity of the annual publications worldwide was on the increase. International collaboration is a widespread reality. The United States led the world in the field of trachea tissue engineering, whereas University College London was the institution with the greatest contribution. In addition, Biomaterials had a great influence in this field, attracting the largest number of papers. Moreover, the topics of TTE research largely concentrated on the biomechanical scaffold preparation, the vascularization and epithelialization of scaffold, the tracheal cartilage regeneration, and the tissue-engineered tracheal transplantation. And the research on the application of decellularization and 3D printing for the construction of a tissue-engineered trachea was likely to receive more widespread attention in the future. Impact statement In recent years, tracheal tissue engineering (TTE) has experienced rapid growth. In this study, we investigated the worldwide status and trends of TTE research, and revealed the countries, institutions, journals, and authors that had made significant contributions to the field of TTE. Moreover, the possible research hotspots in the future were predicted. According to our research, researchers can gain a better understanding of the trends in this field, and stay informed of the most current research by tracking key journals, institutions, and authors.

CAM Model: Intriguing Natural Bioreactor for Sustainable Research and Reliable/Versatile Testing.

We are witnessing the revival of the CAM model, which has already used been in the past by several researchers studying angiogenesis and anti-cancer drugs and now offers a refined model to fill, in the translational meaning, the gap between in vitro and in vivo studies. It can be used for a wide range of purposes, from testing cytotoxicity, pharmacokinetics, tumorigenesis, and invasion to the action mechanisms of molecules and validation of new materials from tissue engineering research. The CAM model is easy to use, with a fast outcome, and makes experimental research more sustainable since it allows us to replace, reduce, and refine pre-clinical experimentation ("3Rs" rules). This review aims to highlight some unique potential that the CAM-assay presents; in particular, the authors intend to use the CAM model in the future to verify, in a microenvironment comparable to in vivo conditions, albeit simplified, the angiogenic ability of functionalized 3D constructs to be used in regenerative medicine strategies in the recovery of skeletal injuries of critical size (CSD) that do not repair spontaneously. For this purpose, organotypic cultures will be planned on several CAMs set up in temporal sequences, and a sort of organ model for assessing CSD will be utilized in the CAM bioreactor rather than in vivo.

Analysis of cognitive framework and biomedical translation of tissue engineering in otolaryngology

Tissue engineering is a relatively recent research area aimed at developing artificial tissues that can restore, maintain, or even improve the anatomical and/or functional integrity of injured tissues. Otolaryngology, as a leading surgical specialty in head and neck surgery, is a candidate for the use of these advanced therapies and medicinal products developed. Nevertheless, a knowledge-based analysis of both areas together is still needed. The dataset was retrieved from the Web of Science database from 1900 to 2020. SciMAT software was used to perform the science mapping analysis and the data for the biomedical translation identification was obtained from the iCite platform. Regarding the analysis of the cognitive structure, we find consolidated research lines, such as the generation of cartilage for use as a graft in reconstructive surgery, reconstruction of microtia, or the closure of perforations of the tympanic membrane. This last research area occupies the most relevant clinical translation with the rest of the areas presenting a lower translational level. In conclusion, Tissue engineering is still in an early translational stage in otolaryngology, otology being the field where most advances have been achieved. Therefore, although otolaryngologists should play an active role in translational research in tissue engineering, greater multidisciplinary efforts are required to promote and encourage the translation of potential clinical applications of tissue engineering for routine clinical use.

The promoter effect of laminin-derived IKVAV peptide on three dimensional HUVEC microtissue

Tissue engineering research is recently a popular field but the vascularization process of existing methods limits the study area. Human Umbilical Vein Endothelial Cells (HUVEC) are essential cell models for vascularization study in vitro. Although studies about vascular biomaterial are mostly performed in traditional 2 Dimensional (D) cell culture, the system has some disadvantages. However, 3D scaffold-free microtissue can be used to overcome these disadvantages for the identification of the optimum concentration of biomaterials. IKVAV is an active unit of laminin which is an effective protein in the extracellular matrix. IKVAV may increase cell adhesion, proliferation, migration, and cellular differentiation. Since IKVAV directly affects endothelial cells, the definition of the optimum concentration of IKVAV is critically important for HUVEC growth and viability during vascularization. Thus, the study aimed identification of the optimal IKVAV peptide concentration for the production and viability of 3D HUVEC SFM. After peptide synthesis, 3D SFM was fabricated. 0.5 mM and 1 mM concentrations of IKVAV peptide were treated with SFM. The control group was incubated without any peptide concentration. Diameters and viabilities of SFMs were evaluated. 1 mM concentration showed the highest diameter and viability. The increasing concentrations may support HUVEC growth and viability so it may induce vascularization in vivo conditions.

Assessing non-synthetic crosslinkers in biomaterial inks based on polymers of marine origin to increase the shape fidelity in 3D extrusion printing

In the past decade, there has been significant progress in 3D printing research for tissue engineering (TE) using biomaterial inks made from natural and synthetic compounds. These constructs can aid in the regeneration process after tissue loss or injury, but achieving high shape fidelity is a challenge as it affects the construct’s physical and biological performance with cells. In parallel with the growth of 3D bioprinting approaches, some marine-origin polymers have been studied due to their biocompatibility, biodegradability, low immunogenicity, and similarities to human extracellular matrix components, making them an excellent alternative to land mammal-origin polymers with reduced disease transmission risk and ethical concerns. In this research, collagen from shark skin, chitosan from squid pens, and fucoidan from brown algae were effectively blended for the manufacturing of an adequate biomaterial ink to achieve a printable, reproducible material with a high shape fidelity and reticulated using four different approaches (phosphate-buffered saline, cell culture medium, 6% CaCl2, and 5 mM Genipin). Materials characterization was composed by filament collapse, fusion behavior, swelling behavior, and rheological and compressive tests, which demonstrated favorable shape fidelity resulting in a stable structure without deformations, and interesting shear recovery properties around the 80% mark. Additionally, live/dead assays were conducted in order to assess the cell viability of an immortalized human mesenchymal stem cell line, seeded directly on the 3D printed constructs, which showed over 90% viable cells. Overall, the Roswell Park Memorial Institute cell culture medium promoted the adequate crosslinking of this biopolymer blend to serve the TE approach, taking advantage of its capacity to hamper pH decrease coming from the acidic biomaterial ink. While the crosslinking occurs, the pH can be easily monitored by the presence of the indicator phenol red in the cell culture medium, which reduces costs and time.

Injectable and Cell-Laden Hydrogel in the Contained Bone Defect Animal Model: A Systematic Review.

Due to its high water content and biomimetic properties simulating extracellular matrix (ECM), hydrogels have been used as preferred cell culture and delivery systems. Similarly, cell-loaded hydrogels can be easily injected into target areas in a minimally invasive manner, minimizing surgical trauma, adapting to irregular shaped defects, and benefiting patients. In this study, we systematically reviewed multiple studies on hydrogel-based bone defect research and briefly summarized the progress of injectable and cell-loaded hydrogels in bone defect repair. A systematic search was conducted in the PubMed and Web of Science databases using selected search terms. Initially, 185 articles were retrieved from the databases. After full-text screening based on inclusion and exclusion criteria, 26 articles were included in this systematic review. Data collected from each study included culture model, seed cell type and origin, cell concentration, scaffold material, scaffold shape, experimental animal and site, bioactive agents, and binding method. This injectable and cell-loaded hydrogel shows certain feasibility in bone tissue engineering applications. Injectable and cell-loaded hydrogels have been widely applied in bone tissue engineering research. The future direction of bone tissue engineering for bone defect treatment involves the use of new hydrogel materials and biochemical stimulation.

2D Collagen Membranes from Marine Demosponge Chondrosia reniformis (Nardo, 1847) for Skin-Regenerative Medicine Applications: An In Vitro Evaluation.

Research in tissue engineering and regenerative medicine has an ever-increasing need for innovative biomaterials suitable for the production of wound-dressing devices and artificial skin-like substitutes. Marine collagen is one of the most promising biomaterials for the production of such devices. In this study, for the first time, 2D collagen membranes (2D-CMs) created from the extracellular matrix extract of the marine demosponge Chondrosia reniformis have been evaluated in vitro as possible tools for wound healing. Fibrillar collagen was extracted from a pool of fresh animals and used for the creation of 2D-CMs, in which permeability to water, proteins, and bacteria, and cellular response in the L929 fibroblast cell line were evaluated. The biodegradability of the 2D-CMs was also assessed by following their degradation in PBS and collagenase solutions for up to 21 days. Results showed that C. reniformis-derived membranes avoided liquid and protein loss in the regeneration region and also functioned as a strong barrier against bacteria infiltration into a wound. Gene expression analyses on fibroblasts stated that their interaction with 2D-CMs is able to improve fibronectin production without interfering with the regular extracellular matrix remodeling processes. These findings, combined with the high extraction yield of fibrillar collagen obtained from C. reniformis with a solvent-free approach, underline how important further studies on the aquaculture of this sponge could be for the sustainable production and biotechnological exploitation of this potentially promising and peculiar biopolymer of marine origin.