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Fernández-Villa, Daniel, Mirta Jiménez Gómez-Lavín, Cristina Abradelo, Julio San Román, and Luis Rojo. "Tissue Engineering Therapies Based on Folic Acid and Other Vitamin B Derivatives. Functional Mechanisms and Current Applications in Regenerative Medicine." International Journal of Molecular Sciences 19, no. 12 (December 16, 2018): 4068. http://dx.doi.org/10.3390/ijms19124068.
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B-vitamins are a group of soluble vitamins which are cofactors of some of the enzymes involved in the metabolic pathways of carbohydrates, fats and proteins. These compounds participate in a number of functions as cardiovascular, brain or nervous systems. Folic acid is described as an accessible and multifunctional niche component that can be used safely, even combined with other compounds, which gives it high versatility. Also, due to its non-toxicity and great stability, folic acid has attracted much attention from researchers in the biomedical and bioengineering area, with an increasing number of works directed at using folic acid and its derivatives in tissue engineering therapies as well as regenerative medicine. Thus, this review provides an updated discussion about the most relevant advances achieved during the last five years, where folic acid and other vitamins B have been used as key bioactive compounds for enhancing the effectiveness of biomaterials’ performance and biological functions for the regeneration of tissues and organs.
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Nag, Kakon, and Toshihiro Akaike. "E-Cadherin – Fc Chimeric Protein-Based Biomaterial: Breaking the Barriers in Stem Cell Technology and Regenerative Medicine." Advanced Materials Research 810 (September 2013): 41–76. http://dx.doi.org/10.4028/www.scientific.net/amr.810.41.
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Chimeric proteins have been used for years for various purposes ranging from biomaterials to candidate drug molecules, and from bench to bulk. Regenerative medicine needs various kinds of proteins for providing essential factors for maintaining starting cells, like induced pluripotent stem cells (iPSC), and renewal, proliferation, targeted differentiation of these cells, and as extracellular matrix for the experimental cells. However, there are several challenges associated with making functional chimeric proteins for effective application as biomaterial in this field. Fc-chimeric protein technology could be an effective solution to overcome many of them. These tailored proteins are recently becoming superior choice of biomaterials in stem cell technology and regenerative medicine due to their specific advantageous biophysical and biochemical properties over other chimeric forms of same proteins. Recent advances in recombinant protein-related science and technology also expedited the popularity of this kind of engineered protein. Over the last decade our lab has been pioneering this field, and we and others have been successfully applied Fc-chimeric proteins to overcome many critical issues in stem cell technologies targeting regenerative medicine and tissue engineering. Fc-chimeric protein-based biomaterials, specifically, E-cad-Fc have been preferentially applied for coating of cell culture plates for establishing xenogeneic-agent free monolayer stem cell culture and their maintenance, enhanced directed differentiation of stem cells to specific lineages, and non-enzymatic on-site one-step purification of target cells. Here the technology, recent discoveries, and future direction related with the E-cad-Fc-chimeric protein in connection with regenerative medicine are described.
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Jahangirian, Azizi, Rafiee-Moghaddam, Baratvand, and Webster. "Status of Plant Protein-Based Green Scaffolds for Regenerative Medicine Applications." Biomolecules 9, no. 10 (October 17, 2019): 619. http://dx.doi.org/10.3390/biom9100619.
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In recent decades, regenerative medicine has merited substantial attention from scientific and research communities. One of the essential requirements for this new strategy in medicine is the production of biocompatible and biodegradable scaffolds with desirable geometric structures and mechanical properties. Despite such promise, it appears that regenerative medicine is the last field to embrace green, or environmentally-friendly, processes, as many traditional tissue engineering materials employ toxic solvents and polymers that are clearly not environmentally friendly. Scaffolds fabricated from plant proteins (for example, zein, soy protein, and wheat gluten), possess proper mechanical properties, remarkable biocompatibility and aqueous stability which make them appropriate green biomaterials for regenerative medicine applications. The use of plant-derived proteins in regenerative medicine has been especially inspired by green medicine, which is the use of environmentally friendly materials in medicine. In the current review paper, the literature is reviewed and summarized for the applicability of plant proteins as biopolymer materials for several green regenerative medicine and tissue engineering applications.
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Torres-Huerta, Ana Laura, Aurora Antonio-Pérez, Yolanda García-Huante, Nayelhi Julieta Alcázar-Ramírez, and Juan Carlos Rueda-Silva. "Biomolecule-Based Optical Metamaterials: Design and Applications." Biosensors 12, no. 11 (November 2, 2022): 962. http://dx.doi.org/10.3390/bios12110962.
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Metamaterials are broadly defined as artificial, electromagnetically homogeneous structures that exhibit unusual physical properties that are not present in nature. They possess extraordinary capabilities to bend electromagnetic waves. Their size, shape and composition can be engineered to modify their characteristics, such as iridescence, color shift, absorbance at different wavelengths, etc., and harness them as biosensors. Metamaterial construction from biological sources such as carbohydrates, proteins and nucleic acids represents a low-cost alternative, rendering high quantities and yields. In addition, the malleability of these biomaterials makes it possible to fabricate an endless number of structured materials such as composited nanoparticles, biofilms, nanofibers, quantum dots, and many others, with very specific, invaluable and tremendously useful optical characteristics. The intrinsic characteristics observed in biomaterials make them suitable for biomedical applications. This review addresses the optical characteristics of metamaterials obtained from the major macromolecules found in nature: carbohydrates, proteins and DNA, highlighting their biosensor field use, and pointing out their physical properties and production paths.
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Filipczak, Nina, Satya Siva Kishan Yalamarty, Xiang Li, Muhammad Muzamil Khan, Farzana Parveen, and Vladimir Torchilin. "Lipid-Based Drug Delivery Systems in Regenerative Medicine." Materials 14, no. 18 (September 17, 2021): 5371. http://dx.doi.org/10.3390/ma14185371.
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The most important goal of regenerative medicine is to repair, restore, and regenerate tissues and organs that have been damaged as a result of an injury, congenital defect or disease, as well as reversing the aging process of the body by utilizing its natural healing potential. Regenerative medicine utilizes products of cell therapy, as well as biomedical or tissue engineering, and is a huge field for development. In regenerative medicine, stem cells and growth factor are mainly used; thus, innovative drug delivery technologies are being studied for improved delivery. Drug delivery systems offer the protection of therapeutic proteins and peptides against proteolytic degradation where controlled delivery is achievable. Similarly, the delivery systems in combination with stem cells offer improvement of cell survival, differentiation, and engraftment. The present review summarizes the significance of biomaterials in tissue engineering and the importance of colloidal drug delivery systems in providing cells with a local environment that enables them to proliferate and differentiate efficiently, resulting in successful tissue regeneration.
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Kishan Shetty, Ashmitha, Serene Joy, Manasa Latha Biligowda, and Siddique Sha Muhammed Hussain. "Biomarkers of Pulpal Regeneration: Overview on Immunohistochemistry Analysis." ECS Transactions 107, no. 1 (April 24, 2022): 17193–99. http://dx.doi.org/10.1149/10701.17193ecst.
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Immunocytochemistry (IHC) is a method that uses monoclonal and polyclonal antibodies to determine the tissue distribution of an antigen of interest in health and disease. The method of recognizing a tissue component in situ utilizing unique antibody-antigen interactions in which the antibody is precisely labelled is referred to as IHC. It can be used to identify and localize well-known cellular structures and also extracellular matrix components. This method similarly provides information on the temporospatial distribution of newly discovered carbohydrates and proteins in development, illness and health. Endodontics can use IHC as a diagnostic tool and a potential marker for odontogenic tissues. Although histopathologic examination of periapical lesions reveals the true nature of regenerative tissue, immunohistochemical markers can be utilized to refine the tissue's composition.
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Khosropanah, Mohammad Hossein, Mahdi Alizadeh Vaghasloo, Mehdi Shakibaei, Anna‐Lena Mueller, Abdol‐Mohammad Kajbafzadeh, Leila Amani, Ismaeil Haririan, Ashkan Azimzadeh, Zahra Hassannejad, and Masoumeh Majidi Zolbin. "Biomedical applications of silkworm ( Bombyx Mori ) proteins in regenerative medicine (a narrative review)." Journal of Tissue Engineering and Regenerative Medicine 16, no. 2 (December 7, 2021): 91–109. http://dx.doi.org/10.1002/term.3267.
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Lee, Jung-Hwan, Ji-Young Yoon, Jun Hee Lee, Hae-Hyoung Lee, Jonathan C. Knowles, and Hae-Won Kim. "Emerging biogenesis technologies of extracellular vesicles for tissue regenerative therapeutics." Journal of Tissue Engineering 12 (January 2021): 204173142110190. http://dx.doi.org/10.1177/20417314211019015.
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Extracellular vesicles (EVs), including exosomes, carry the genetic packages of RNA, DNA, and proteins and are heavily involved in cell-cell communications and intracellular signalings. Therefore, EVs are spotlighted as therapeutic mediators for the treatment of injured and dysfunctional tissues as well as biomarkers for the detection of disease status and progress. Several key issues in EVs, including payload content and bioactivity, targeting and bio-imaging ability, and mass-production, need to be improved to enable effective therapeutics and clinical translation. For this, significant efforts have been made recently, including genetic modification, biomolecular and chemical treatment, application of physical/mechanical cues, and 3D cultures. Here we communicate those recent technological advances made mainly in the biogenesis process of EVs or at post-collection stages, which ultimately aimed to improve the therapeutic efficacy in tissue healing and disease curing and the possibility of clinical translation. This communication will help tissue engineers and biomaterial scientists design and produce EVs optimally for tissue regenerative therapeutics.
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Vacchini, Mattia, Rana Edwards, Roberto Guizzardi, Alessandro Palmioli, Carlotta Ciaramelli, Alice Paiotta, Cristina Airoldi, Barbara La Ferla, and Laura Cipolla. "Glycan Carriers As Glycotools for Medicinal Chemistry Applications." Current Medicinal Chemistry 26, no. 35 (December 13, 2019): 6349–98. http://dx.doi.org/10.2174/0929867326666190104164653.
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Carbohydrates are one of the most powerful and versatile classes of biomolecules that nature uses to regulate organisms’ biochemistry, modulating plenty of signaling events within cells, triggering a plethora of physiological and pathological cellular behaviors. In this framework, glycan carrier systems or carbohydrate-decorated materials constitute interesting and relevant tools for medicinal chemistry applications. In the last few decades, efforts have been focused, among others, on the development of multivalent glycoconjugates, biosensors, glycoarrays, carbohydrate-decorated biomaterials for regenerative medicine, and glyconanoparticles. This review aims to provide the reader with a general overview of the different carbohydrate carrier systems that have been developed as tools in different medicinal chemistry approaches relying on carbohydrate-protein interactions. Given the extent of this topic, the present review will focus on selected examples that highlight the advancements and potentialities offered by this specific area of research, rather than being an exhaustive literature survey of any specific glyco-functionalized system.
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Miyata, T., and K. Kurokawa. "Carbonyl Stress: Increased Carbonyl Modification of Proteins by Autoxidation Products of Carbohydrates and Lipids in Uremia." International Journal of Artificial Organs 22, no. 4 (April 1999): 195–98. http://dx.doi.org/10.1177/039139889902200402.
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Zhang, Hengtong, Xixi Wu, Liang Quan, and Qiang Ao. "Characteristics of Marine Biomaterials and Their Applications in Biomedicine." Marine Drugs 20, no. 6 (May 31, 2022): 372. http://dx.doi.org/10.3390/md20060372.
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Oceans have vast potential to develop high-value bioactive substances and biomaterials. In the past decades, many biomaterials have come from marine organisms, but due to the wide variety of organisms living in the oceans, the great diversity of marine-derived materials remains explored. The marine biomaterials that have been found and studied have excellent biological activity, unique chemical structure, good biocompatibility, low toxicity, and suitable degradation, and can be used as attractive tissue material engineering and regenerative medicine applications. In this review, we give an overview of the extraction and processing methods and chemical and biological characteristics of common marine polysaccharides and proteins. This review also briefly explains their important applications in anticancer, antiviral, drug delivery, tissue engineering, and other fields.
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Ortiz-Arrabal, Olimpia, Ramón Carmona, Óscar-Darío García-García, Jesús Chato-Astrain, David Sánchez-Porras, Alberto Domezain, Roke-Iñaki Oruezabal, Víctor Carriel, Antonio Campos, and Miguel Alaminos. "Generation and Evaluation of Novel Biomaterials Based on Decellularized Sturgeon Cartilage for Use in Tissue Engineering." Biomedicines 9, no. 7 (July 4, 2021): 775. http://dx.doi.org/10.3390/biomedicines9070775.
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Because cartilage has limited regenerative capability, a fully efficient advanced therapy medicinal product is needed to treat severe cartilage damage. We evaluated a novel biomaterial obtained by decellularizing sturgeon chondral endoskeleton tissue for use in cartilage tissue engineering. In silico analysis suggested high homology between human and sturgeon collagen proteins, and ultra-performance liquid chromatography confirmed that both types of cartilage consisted mainly of the same amino acids. Decellularized sturgeon cartilage was recellularized with human chondrocytes and four types of human mesenchymal stem cells (MSC) and their suitability for generating a cartilage substitute was assessed ex vivo and in vivo. The results supported the biocompatibility of the novel scaffold, as well as its ability to sustain cell adhesion, proliferation and differentiation. In vivo assays showed that the MSC cells in grafted cartilage disks were biosynthetically active and able to remodel the extracellular matrix of cartilage substitutes, with the production of type II collagen and other relevant components, especially when adipose tissue MSC were used. In addition, these cartilage substitutes triggered a pro-regenerative reaction mediated by CD206-positive M2 macrophages. These preliminary results warrant further research to characterize in greater detail the potential clinical translation of these novel cartilage substitutes.
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Davies, Rebecca, and Nicola Kuiper. "Regenerative Medicine: A Review of the Evolution of Autologous Chondrocyte Implantation (ACI) Therapy." Bioengineering 6, no. 1 (March 13, 2019): 22. http://dx.doi.org/10.3390/bioengineering6010022.
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Articular cartilage is composed of chondrons within a territorial matrix surrounded by a highly organized extracellular matrix comprising collagen II fibrils, proteoglycans, glycosaminoglycans, and non-collagenous proteins. Damaged articular cartilage has a limited potential for healing and untreated defects often progress to osteoarthritis. High hopes have been pinned on regenerative medicine strategies to meet the challenge of preventing progress to late osteoarthritis. One such strategy, autologous chondrocyte implantation (ACI), was first reported in 1994 as a treatment for deep focal articular cartilage defects. ACI has since evolved to become a worldwide well-established surgical technique. For ACI, chondrocytes are harvested from the lesser weight bearing edge of the joint by arthroscopy, their numbers expanded in monolayer culture for at least four weeks, and then re-implanted in the damaged region under a natural or synthetic membrane via an open joint procedure. We consider the evolution of ACI to become an established cell therapy, its current limitations, and on-going strategies to improve its efficacy. The most promising developments involving cells and natural or synthetic biomaterials will be highlighted.
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Batool, Fareeha, Hayriye Özçelik, Céline Stutz, Pierre-Yves Gegout, Nadia Benkirane-Jessel, Catherine Petit, and Olivier Huck. "Modulation of immune-inflammatory responses through surface modifications of biomaterials to promote bone healing and regeneration." Journal of Tissue Engineering 12 (January 2021): 204173142110414. http://dx.doi.org/10.1177/20417314211041428.
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Control of inflammation is indispensable for optimal oral wound healing and tissue regeneration. Several biomaterials have been used to enhance the regenerative outcomes; however, the biomaterial implantation can ensure an immune-inflammatory response. The interface between the cells and the biomaterial surface plays a critical role in determining the success of soft and hard tissue regeneration. The initial inflammatory response upon biomaterial implantation helps in tissue repair and regeneration, however, persistant inflammation impairs the wound healing response. The cells interact with the biomaterials through extracellular matrix proteins leading to protein adsorption followed by recruitment, attachment, migration, and proliferation of several immune-inflammatory cells. Physical nanotopography of biomaterials, such as surface proteins, roughness, and porosity, is crucial for driving cellular attachment and migration. Similarly, modification of scaffold surface chemistry by adapting hydrophilicity, surface charge, surface coatings, can down-regulate the initiation of pro-inflammatory cascades. Besides, functionalization of scaffold surfaces with active biological molecules can down-regulate pro-inflammatory and pro-resorptive mediators’ release as well as actively up-regulate anti-inflammatory markers. This review encompasses various strategies for the optimization of physical, chemical, and biological properties of biomaterial and the underlying mechanisms to modulate the immune-inflammatory response, thereby, promoting the tissue integration and subsequent soft and hard tissue regeneration potential of the administered biomaterial.
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Rubí-Sans, Gerard, Irene Cano-Torres, Soledad Pérez-Amodio, Barbara Blanco-Fernandez, Miguel A. Mateos-Timoneda, and Elisabeth Engel. "Development and Angiogenic Potential of Cell-Derived Microtissues Using Microcarrier-Template." Biomedicines 9, no. 3 (February 25, 2021): 232. http://dx.doi.org/10.3390/biomedicines9030232.
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Tissue engineering and regenerative medicine approaches use biomaterials in combination with cells to regenerate lost functions of tissues and organs to prevent organ transplantation. However, most of the current strategies fail in mimicking the tissue’s extracellular matrix properties. In order to mimic native tissue conditions, we developed cell-derived matrix (CDM) microtissues (MT). Our methodology uses poly-lactic acid (PLA) and Cultispher® S microcarriers’ (MCs’) as scaffold templates, which are seeded with rat bone marrow mesenchymal stem cells (rBM-MSCs). The scaffold template allows cells to generate an extracellular matrix, which is then extracted for downstream use. The newly formed CDM provides cells with a complex physical (MT architecture) and biochemical (deposited ECM proteins) environment, also showing spontaneous angiogenic potential. Our results suggest that MTs generated from the combination of these two MCs (mixed MTs) are excellent candidates for tissue vascularization. Overall, this study provides a methodology for in-house fabrication of microtissues with angiogenic potential for downstream use in various tissue regenerative strategies.
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Goker, Funda, Lena Larsson, Massimo Del Fabbro, and Farah Asa’ad. "Gene Delivery Therapeutics in the Treatment of Periodontitis and Peri-Implantitis: A State of the Art Review." International Journal of Molecular Sciences 20, no. 14 (July 20, 2019): 3551. http://dx.doi.org/10.3390/ijms20143551.
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Background: Periodontal disease is a chronic inflammatory condition that affects supporting tissues around teeth, resulting in periodontal tissue breakdown. If left untreated, periodontal disease could have serious consequences; this condition is in fact considered as the primary cause of tooth loss. Being highly prevalent among adults, periodontal disease treatment is receiving increased attention from researchers and clinicians. When this condition occurs around dental implants, the disease is termed peri-implantitis. Periodontal regeneration aims at restoring the destroyed attachment apparatus, in order to improve tooth stability and thus reduce disease progression and subsequent periodontal tissue breakdown. Although many biomaterials have been developed to promote periodontal regeneration, they still have their own set of disadvantages. As a result, regenerative medicine has been employed in the periodontal field, not only to overcome the drawbacks of the conventional biomaterials but also to ensure more predictable regenerative outcomes with minimal complications. Regenerative medicine is considered a part of the research field called tissue engineering/regenerative medicine (TE/RM), a translational field combining cell therapy, biomaterial, biomedical engineering and genetics all with the aim to replace and restore tissues or organs to their normal function using in vitro models for in vivo regeneration. In a tissue, cells are responding to different micro-environmental cues and signaling molecules, these biological factors influence cell differentiation, migration and cell responses. A central part of TE/RM therapy is introducing drugs, genetic materials or proteins to induce specific cellular responses in the cells at the site of tissue repair in order to enhance and improve tissue regeneration. In this review, we present the state of art of gene therapy in the applications of periodontal tissue and peri-implant regeneration. Purpose: We aim herein to review the currently available methods for gene therapy, which include the utilization of viral/non-viral vectors and how they might serve as therapeutic potentials in regenerative medicine for periodontal and peri-implant tissues.
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Lindholm, T. Sam, and Tom C. Lindholm. "The Skull Defect Model in Measuring Osteoinductivity." Journal of Musculoskeletal Research 02, no. 02 (June 1998): 123–39. http://dx.doi.org/10.1142/s0218957798000147.
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Research on regeneration and experimental or clinical repair of cranial defects commenced already one hundred years ago. Principally spontaneous regeneration, intramembraneous bone healing, starts from the edges, proceeding to the center of the defect. A skull defect exceeding some critical limitations lacks the capacity for spontaneous healing. This kind of situation is named a "critical size" defect. The skull defect model is eminently suitable for testing osteoconduction and induction following local implantation of biomaterials and growth factors. There are numerous reports especially on osteoinductive proteins, native and recombinant bone morphogenetic proteins (BMPs), showing satisfactory complete healing of critical size skull defects in a relatively short period of time, while untreated defects have not had the capacity for complete regeneration. This article is a review of skull defect healing in different animal species showing optimized osteoinductive regenerative capacity especially with the use of BMPs. The results, indeed, also have clinical implications.
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Narayanan, Raghuvaran, Chun-Chieh Huang, and Sriram Ravindran. "Hijacking the Cellular Mail: Exosome Mediated Differentiation of Mesenchymal Stem Cells." Stem Cells International 2016 (2016): 1–11. http://dx.doi.org/10.1155/2016/3808674.
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Bone transplantation is one of the most widely performed clinical procedures. Consequently, bone regeneration using mesenchymal stem cells and tissue engineering strategies is one of the most widely researched fields in regenerative medicine. Recent scientific consensus indicates that a biomimetic approach is required to achieve proper regeneration of any tissue. Exosomes are nanovesicles secreted by cells that act as messengers that influence cell fate. Although exosomal function has been studied with respect to cancer and immunology, the role of exosomes as inducers of stem cell differentiation has not been explored. We hypothesized that exosomes can be used as biomimetic tools for regenerative medicine. In this study we have explored the use of cell-generated exosomes as tools to induce lineage specific differentiation of stem cells. Our results indicate that proosteogenic exosomes isolated from cell cultures can induce lineage specific differentiation of naïve MSCsin vitroandin vivo. Additionally, exosomes can also bind to matrix proteins such as type I collagen and fibronectin enabling them to be tethered to biomaterials. Overall, the results from this study show the potential of cell derived exosomes in bone regenerative medicine and opens up new avenues for future research.
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Chen, Jianwei, Duchao Zhou, Zhenguo Nie, Liang Lu, Zhidong Lin, Dezhi Zhou, Yi Zhang, Xiaoyan Long, Siyang Fan, and Tao Xu. "A scalable coaxial bioprinting technology for mesenchymal stem cell microfiber fabrication and high extracellular vesicle yield." Biofabrication 14, no. 1 (December 3, 2021): 015012. http://dx.doi.org/10.1088/1758-5090/ac3b90.
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Abstract Mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) are promising candidates for regenerative medicine; however, the lack of scalable methods for high quantity EV production limits their application. In addition, signature EV-derived proteins shared in 3D environments and 2D surfaces, remain mostly unknown. Herein, we present a platform combining MSC microfiber culture with ultracentrifugation purification for high EV yield. Within this platform, a high quantity MSC solution (∼3 × 108 total cells) is encapsulated in a meter-long hollow hydrogel-microfiber via coaxial bioprinting technology. In this 3D core–shell microfiber environment, MSCs express higher levels of stemness markers (Oct4, Nanog, Sox2) than in 2D culture, and maintain their differentiation capacity. Moreover, this platform enriches particles by ∼1009-fold compared to conventional 2D culture, while preserving their pro-angiogenic properties. Liquid chromatography-mass spectrometry characterization results demonstrate that EVs derived from our platform and conventional 2D culturing have unique protein profiles with 3D-EVs having a greater variety of proteins (1023 vs 605), however, they also share certain proteins (536) and signature MSC-EV proteins (10). This platform, therefore, provides a new tool for EV production using microfibers in one culture dish, thereby reducing space, labor, time, and cost.
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Mano, J. F., G. A. Silva, H. S. Azevedo, P. B. Malafaya, R. A. Sousa, S. S. Silva, L. F. Boesel, et al. "Natural origin biodegradable systems in tissue engineering and regenerative medicine: present status and some moving trends." Journal of The Royal Society Interface 4, no. 17 (April 3, 2007): 999–1030. http://dx.doi.org/10.1098/rsif.2007.0220.
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The fields of tissue engineering and regenerative medicine aim at promoting the regeneration of tissues or replacing failing or malfunctioning organs, by means of combining a scaffold/support material, adequate cells and bioactive molecules. Different materials have been proposed to be used as both three-dimensional porous scaffolds and hydrogel matrices for distinct tissue engineering strategies. Among them, polymers of natural origin are one of the most attractive options, mainly due to their similarities with the extracellular matrix (ECM), chemical versatility as well as typically good biological performance. In this review, the most studied and promising and recently proposed naturally derived polymers that have been suggested for tissue engineering applications are described. Different classes of such type of polymers and their blends with synthetic polymers are analysed, with special focus on polysaccharides and proteins, the systems that are more inspired by the ECM. The adaptation of conventional methods or non-conventional processing techniques for processing scaffolds from natural origin based polymers is reviewed. The use of particles, membranes and injectable systems from such kind of materials is also overviewed, especially what concerns the present status of the research that should lead towards their final application. Finally, the biological performance of tissue engineering constructs based on natural-based polymers is discussed, using several examples for different clinically relevant applications.
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Xu, Zhenyuan, Jacob A. Orkwis, and Greg M. Harris. "Cell Shape and Matrix Stiffness Impact Schwann Cell Plasticity via YAP/TAZ and Rho GTPases." International Journal of Molecular Sciences 22, no. 9 (May 1, 2021): 4821. http://dx.doi.org/10.3390/ijms22094821.
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Schwann cells (SCs) are a highly plastic cell type capable of undergoing phenotypic changes following injury or disease. SCs are able to upregulate genes associated with nerve regeneration and ultimately achieve functional recovery. During the regeneration process, the extracellular matrix (ECM) and cell morphology play a cooperative, critical role in regulating SCs, and therefore highly impact nerve regeneration outcomes. However, the roles of the ECM and mechanotransduction relating to SC phenotype are largely unknown. Here, we describe the role that matrix stiffness and cell morphology play in SC phenotype specification via known mechanotransducers YAP/TAZ and RhoA. Using engineered microenvironments to precisely control ECM stiffness, cell shape, and cell spreading, we show that ECM stiffness and SC spreading downregulated SC regenerative associated proteins by the activation of RhoA and YAP/TAZ. Additionally, cell elongation promoted a distinct SC regenerative capacity by the upregulation of Rac1/MKK7/JNK, both necessary for the ECM and morphology changes found during nerve regeneration. These results confirm the role of ECM signaling in peripheral nerve regeneration as well as provide insight to the design of future biomaterials and cellular therapies for peripheral nerve regeneration.
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Cicuéndez, Mónica, Laura Casarrubios, María José Feito, Iratxe Madarieta, Nerea Garcia-Urkia, Olatz Murua, Beatriz Olalde, Nerea Briz, Rosalía Diez-Orejas, and María Teresa Portolés. "Effects of Human and Porcine Adipose Extracellular Matrices Decellularized by Enzymatic or Chemical Methods on Macrophage Polarization and Immunocompetence." International Journal of Molecular Sciences 22, no. 8 (April 8, 2021): 3847. http://dx.doi.org/10.3390/ijms22083847.
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The decellularized extracellular matrix (ECM) obtained from human and porcine adipose tissue (AT) is currently used to prepare regenerative medicine bio-scaffolds. However, the influence of these natural biomaterials on host immune response is not yet deeply understood. Since macrophages play a key role in the inflammation/healing processes due to their high functional plasticity between M1 and M2 phenotypes, the evaluation of their response to decellularized ECM is mandatory. It is also necessary to analyze the immunocompetence of macrophages after contact with decellularized ECM materials to assess their functional role in a possible infection scenario. In this work, we studied the effect of four decellularized adipose matrices (DAMs) obtained from human and porcine AT by enzymatic or chemical methods on macrophage phenotypes and fungal phagocytosis. First, a thorough biochemical characterization of these biomaterials by quantification of remnant DNA, lipids, and proteins was performed, thus indicating the efficiency and reliability of both methods. The proteomic analysis evidenced that some proteins are differentially preserved depending on both the AT origin and the decellularization method employed. After exposure to the four DAMs, specific markers of M1 proinflammatory and M2 anti-inflammatory macrophages were analyzed. Porcine DAMs favor the M2 phenotype, independently of the decellularization method employed. Finally, a sensitive fungal phagocytosis assay allowed us to relate the macrophage phagocytosis capability with specific proteins differentially preserved in certain DAMs. The results obtained in this study highlight the close relationship between the ECM biochemical composition and the macrophage’s functional role.
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Dzobo, Kevin, Keolebogile Shirley Caroline M. Motaung, and Adetola Adesida. "Recent Trends in Decellularized Extracellular Matrix Bioinks for 3D Printing: An Updated Review." International Journal of Molecular Sciences 20, no. 18 (September 18, 2019): 4628. http://dx.doi.org/10.3390/ijms20184628.
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The promise of regenerative medicine and tissue engineering is founded on the ability to regenerate diseased or damaged tissues and organs into functional tissues and organs or the creation of new tissues and organs altogether. In theory, damaged and diseased tissues and organs can be regenerated or created using different configurations and combinations of extracellular matrix (ECM), cells, and inductive biomolecules. Regenerative medicine and tissue engineering can allow the improvement of patients’ quality of life through availing novel treatment options. The coupling of regenerative medicine and tissue engineering with 3D printing, big data, and computational algorithms is revolutionizing the treatment of patients in a huge way. 3D bioprinting allows the proper placement of cells and ECMs, allowing the recapitulation of native microenvironments of tissues and organs. 3D bioprinting utilizes different bioinks made up of different formulations of ECM/biomaterials, biomolecules, and even cells. The choice of the bioink used during 3D bioprinting is very important as properties such as printability, compatibility, and physical strength influence the final construct printed. The extracellular matrix (ECM) provides both physical and mechanical microenvironment needed by cells to survive and proliferate. Decellularized ECM bioink contains biochemical cues from the original native ECM and also the right proportions of ECM proteins. Different techniques and characterization methods are used to derive bioinks from several tissues and organs and to evaluate their quality. This review discusses the uses of decellularized ECM bioinks and argues that they represent the most biomimetic bioinks available. In addition, we briefly discuss some polymer-based bioinks utilized in 3D bioprinting.
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Gasperini, Luca, João F. Mano, and Rui L. Reis. "Natural polymers for the microencapsulation of cells." Journal of The Royal Society Interface 11, no. 100 (November 6, 2014): 20140817. http://dx.doi.org/10.1098/rsif.2014.0817.
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The encapsulation of living mammalian cells within a semi-permeable hydrogel matrix is an attractive procedure for many biomedical and biotechnological applications, such as xenotransplantation, maintenance of stem cell phenotype and bioprinting of three-dimensional scaffolds for tissue engineering and regenerative medicine. In this review, we focus on naturally derived polymers that can form hydrogels under mild conditions and that are thus capable of entrapping cells within controlled volumes. Our emphasis will be on polysaccharides and proteins, including agarose, alginate, carrageenan, chitosan, gellan gum, hyaluronic acid, collagen, elastin, gelatin, fibrin and silk fibroin. We also discuss the technologies commonly employed to encapsulate cells in these hydrogels, with particular attention on microencapsulation.
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Chun, So Young, Jun Nyung Lee, Yun-Sok Ha, Bo Hyun Yoon, Eun Hye Lee, Bo Mi Kim, Haejung Gil, et al. "Optimization of extracellular matrix extraction from human perirenal adipose tissue." Journal of Biomaterials Applications 35, no. 9 (January 12, 2021): 1180–91. http://dx.doi.org/10.1177/0885328220984594.
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Human adipose tissue includes useful substrates for regenerative medicine such as the extracellular matrix (ECM), but most perirenal fat tissue is wasted after kidney surgery. Since a lot of adipose tissue can be procured after a kidney, we extracted ECM from human perirenal adipose tissue and optimized the extraction process. To verify the efficacy for ECM extraction, we compared the products in several steps. Perirenal adipose tissue was either finely homogenized or underwent crude manual dissection. The amount of extracted ECM was quantified with ELISA for verification of the initial tissue downsizing effect. To validate the drying effect for fast and complete delipidation, tissues were prepared in a dry or wet phase, and residual lipids were visualized with Oil-Red-O staining. The extracted lipid was assayed at each time point to quantify the appropriate delipidation time. To select the optimal decellularization method, tissues were treated with physical, chemical, or enzymatic method, and the residual cell debris were identified with histological staining. The biochemical properties of the ECM extracted by the above methods were analyzed. The ECM extracted by fine homogenization showed a significantly enhanced amount of collagen, laminin and fibronectin compared to the crude dissection method. The dried tissue showed fast and complete lipid elimination compared to the wet tissue. Complete delipidation was achieved at 45 min after acetone treatment. Additionally, 1% triton X-100 chemical treatment showed complete decellularization with well-preserved collagen fibers. Biochemical analysis revealed preserved ECM proteins, a high cell proliferation rate and normal cell morphology without cell debris or lipids. The established process of homogenization, drying, delipidation with acetone, and decellularization with Triton X-100 treatment can be an optimal method for ECM extraction from human perirenal adipose tissue. Using this technique, human perirenal adipose tissue may be a valuable source for tissue engineering and regenerative medicine.
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Perez-Valle, Arantza, Cristina Del Amo, and Isabel Andia. "Overview of Current Advances in Extrusion Bioprinting for Skin Applications." International Journal of Molecular Sciences 21, no. 18 (September 12, 2020): 6679. http://dx.doi.org/10.3390/ijms21186679.
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Bioprinting technologies, which have the ability to combine various human cell phenotypes, signaling proteins, extracellular matrix components, and other scaffold-like biomaterials, are currently being exploited for the fabrication of human skin in regenerative medicine. We performed a systematic review to appraise the latest advances in 3D bioprinting for skin applications, describing the main cell phenotypes, signaling proteins, and bioinks used in extrusion platforms. To understand the current limitations of this technology for skin bioprinting, we briefly address the relevant aspects of skin biology. This field is in the early stage of development, and reported research on extrusion bioprinting for skin applications has shown moderate progress. We have identified two major trends. First, the biomimetic approach uses cell-laden natural polymers, including fibrinogen, decellularized extracellular matrix, and collagen. Second, the material engineering line of research, which is focused on the optimization of printable biomaterials that expedite the manufacturing process, mainly involves chemically functionalized polymers and reinforcement strategies through molecular blending and postprinting interventions, i.e., ionic, covalent, or light entanglement, to enhance the mechanical properties of the construct and facilitate layer-by-layer deposition. Skin constructs manufactured using the biomimetic approach have reached a higher level of complexity in biological terms, including up to five different cell phenotypes and mirroring the epidermis, dermis and hypodermis. The confluence of the two perspectives, representing interdisciplinary inputs, is required for further advancement toward the future translation of extrusion bioprinting and to meet the urgent clinical demand for skin equivalents.
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Yeo, Giselle C., Alexey Kondyurin, Elena Kosobrodova, Anthony S. Weiss, and Marcela M. M. Bilek. "A sterilizable, biocompatible, tropoelastin surface coating immobilized by energetic ion activation." Journal of The Royal Society Interface 14, no. 127 (February 2017): 20160837. http://dx.doi.org/10.1098/rsif.2016.0837.
Full textAbstract:
Biomimetic materials which integrate with surrounding tissues and regulate new tissue formation are attractive for tissue engineering and regenerative medicine. Plasma immersion ion-implanted (PIII) polyethersulfone (PES) provides an excellent platform for the irreversible immobilization of bioactive proteins and peptides. PIII treatment significantly improves PES wettability and results in the formation of acidic groups on the PES surface, with the highest concentration observed at 40–80 s of PIII treatment. The elastomeric protein tropoelastin can be stably adhered to PIII-treated PES in a cell-interactive conformation by tailoring the pH and salt levels of the protein–surface association conditions. Tropoelastin-coated PIII-treated PES surfaces are resistant to molecular fouling, and actively promote high levels of fibroblast adhesion and proliferation while maintaining cell morphology. Tropoelastin, unlike other extracellular matrix proteins such as fibronectin, uniquely retains full bioactivity even after medical-grade ethylene oxide sterilization. This dual approach of PIII treatment and tropoelastin cloaking allows for the stable, robust functionalization of clinically used polymer materials for directed cellular interactions.
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Toledo, Anna L. M. M., Talita N. da Silva, Arianne C. dos S. Vaucher, Arthur H. V. Miranda, Gabriela C. C. Silva, Maria E. R. Vaz, Luísa V. da Silva, Thais N. Barradas, and Paulo H. S. Picciani. "Polymer Nanofibers for Biomedical Applications: Advances in Electrospinning." Current Applied Polymer Science 4, no. 3 (December 2021): 190–209. http://dx.doi.org/10.2174/2452271604666211122122557.
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Background: The demand for novel biomaterials has been exponentially rising in the last years as well as the searching for new technologies able to produce more efficient products in both drug delivery systems and regenerative medicine. Objective: The technique that can pretty well encompass the needs for novel and high-end materials with a relatively low-cost and easy operation is the electrospinning of polymer solutions. Methods: Electrospinning usually produces ultrathin fibers that can be applied in a myriad of biomedical devices including sustained delivery systems for drugs, proteins, biomolecules, hormones, etc that can be applied in a broad spectrum of applications, from transdermal patches to cancer-related drugs. Results: Electrospun fibers can be produced to mimic certain tissues of the human body, being an option to create new scaffolds for implants with several advantages. Conclusions: In this review, we aimed to encompass the use of electrospun fibers in the field of biomedical devices, more specifically in the use of electrospun nanofibers applications toward the production of drug delivery systems and scaffolds for tissue regeneration.
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Malcor, Jean-Daniel, Emma J. Hunter, Natalia Davidenko, Daniel V. Bax, Ruth Cameron, Serena Best, Sanjay Sinha, and Richard W. Farndale. "Collagen scaffolds functionalized with triple-helical peptides support 3D HUVEC culture." Regenerative Biomaterials 7, no. 5 (August 18, 2020): 471–82. http://dx.doi.org/10.1093/rb/rbaa025.
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Abstract Porous biomaterials which provide a structural and biological support for cells have immense potential in tissue engineering and cell-based therapies for tissue repair. Collagen biomaterials that can host endothelial cells represent promising tools for the vascularization of engineered tissues. Three-dimensional collagen scaffolds possessing controlled architecture and mechanical stiffness are obtained through freeze–drying of collagen suspensions, followed by chemical cross-linking which maintains their stability. However, cross-linking scaffolds renders their biological activity suboptimal for many cell types, including human umbilical vein endothelial cells (HUVECs), by inhibiting cell–collagen interactions. Here, we have improved crucial HUVEC interactions with such cross-linked collagen biomaterials by covalently coupling combinations of triple-helical peptides (THPs). These are ligands for collagen-binding cell-surface receptors (integrins or discoidin domain receptors) or secreted proteins (SPARC and von Willebrand factor). THPs enhanced HUVEC adhesion, spreading and proliferation on 2D collagen films. THPs grafted to 3D-cross-linked collagen scaffolds promoted cell survival over seven days. This study demonstrates that THP-functionalized collagen scaffolds are promising candidates for hosting endothelial cells with potential for the production of vascularized engineered tissues in regenerative medicine applications.
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Sánchez-Romero, Natalia, Laura Martínez-Gimeno, Pedro Caetano-Pinto, Berta Saez, José Manuel Sánchez-Zalabardo, Rosalinde Masereeuw, and Ignacio Giménez. "A simple method for the isolation and detailed characterization of primary human proximal tubule cells for renal replacement therapy." International Journal of Artificial Organs 43, no. 1 (August 6, 2019): 45–57. http://dx.doi.org/10.1177/0391398819866458.
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The main physiological functions of renal proximal tubule cells in vivo are reabsorption of essential nutrients from the glomerular filtrate and secretion of waste products and xenobiotics into urine. Currently, there are several established cell lines of human origin available as in vitro models of proximal tubule. However, these cells appeared to be limited in their biological relevance, because essential characteristics of the original tissue are lost once the cells are cultured. As a consequence of these limitations, primary human proximal tubule cells constitute a suitable and a biologically more relevant in vitro model to study this specific segment of the nephron and therefore, these cells can play an important role in renal regenerative medicine applications. Here, we describe a protocol to isolate proximal tubule cells from human nephrectomies. We explain the steps performed for an in-depth characterization of the cells, including the study of markers from others segments of the nephron, with the goal to determine the purity of the culture and the stability of proteins, enzymes, and transporters along time. The human proximal tubule cells isolated and used throughout this study showed many proximal tubule characteristics, including monolayer organization, cell polarization with the expression of tight junctions and primary cilia, expression of proximal tubule–specific proteins, such as megalin and sodium/glucose cotransporter 2, among others. The cells also expressed enzymatic activity for dipeptidyl peptidase IV, as well as for gamma glutamyl transferase 1, and expressed transporter activity for organic anion transporter 1, P-glycoprotein, multidrug resistance proteins, and breast cancer resistance protein. In conclusion, characterization of our cells confirmed presence of putative proximal tubule markers and the functional expression of multiple endogenous organic ion transporters mimicking renal reabsorption and excretion. These findings can constitute a valuable tool in the development of bioartificial kidney devices.
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Ávila-Salas, Fabián, and Esteban F. Durán-Lara. "An Overview of Injectable Thermo-Responsive Hydrogels and Advances in their Biomedical Applications." Current Medicinal Chemistry 27, no. 34 (October 12, 2020): 5773–89. http://dx.doi.org/10.2174/0929867325666190603110045.
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Background:: Injectable hydrogels are a thermo-responsive system based on biomaterials. Injectable hydrogels have been broadly investigated mainly as vehicles or scaffolds of therapeutic agents that include drugs, proteins, cells, and bioactive molecules among others, utilized in the treatment of diseases such as cancers and the repair and regeneration of tissues. Results: : There are several studies that have described the multiple features of hydrogels. However, the main aspect that breaks the paradigm in the application of hydrogels is the thermoresponsiveness that some of them have, which is an abrupt modification in their properties in response to small variations in temperature. For that reason, the thermo-responsive hydrogels with the unique property of sol-gel transition have received special attention over the past decades. These hydrogels show phase transition near physiological human body temperature. This feature is key for being applied in promising areas of human health-related research. Conclusion: : The purpose of this study is the overview of injectable hydrogels and their latest advances in medical applications including bioactive compound delivery, tissue engineering, and regenerative medicine.
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Vielreicher, M., S. Schürmann, R. Detsch, M. A. Schmidt, A. Buttgereit, A. Boccaccini, and O. Friedrich. "Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine." Journal of The Royal Society Interface 10, no. 86 (September 6, 2013): 20130263. http://dx.doi.org/10.1098/rsif.2013.0263.
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This review focuses on modern nonlinear optical microscopy (NLOM) methods that are increasingly being used in the field of tissue engineering (TE) to image tissue non-invasively and without labelling in depths unreached by conventional microscopy techniques. With NLOM techniques, biomaterial matrices, cultured cells and their produced extracellular matrix may be visualized with high resolution. After introducing classical imaging methodologies such as µCT, MRI, optical coherence tomography, electron microscopy and conventional microscopy two-photon fluorescence (2-PF) and second harmonic generation (SHG) imaging are described in detail (principle, power, limitations) together with their most widely used TE applications. Besides our own cell encapsulation, cell printing and collagen scaffolding systems and their NLOM imaging the most current research articles will be reviewed. These cover imaging of autofluorescence and fluorescence-labelled tissue and biomaterial structures, SHG-based quantitative morphometry of collagen I and other proteins, imaging of vascularization and online monitoring techniques in TE. Finally, some insight is given into state-of-the-art three-photon-based imaging methods (e.g. coherent anti-Stokes Raman scattering, third harmonic generation). This review provides an overview of the powerful and constantly evolving field of multiphoton microscopy, which is a powerful and indispensable tool for the development of artificial tissues in regenerative medicine and which is likely to gain importance also as a means for general diagnostic medical imaging.
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Salih, Mohamed, Bakiah Shaharuddin, and Samar Abdelrazeg. "A Concise Review on Mesenchymal Stem Cells for Tissue Engineering with a Perspective on Ocular Surface Regeneration." Current Stem Cell Research & Therapy 15, no. 3 (April 25, 2020): 211–18. http://dx.doi.org/10.2174/1574888x15666200129145251.
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Organ and tissue transplantation are limited by the scarcity of donated organs or tissue sources. The success of transplantation is limited by the risk of disease transmission and immunological- related rejection. There is a need for new strategies and innovative solutions to make transplantation readily available, safer and with less complications to increase the success rates. Accelerating progress in stem cell biology and biomaterials development have pushed tissue and organ engineering to a higher level. Among stem cells repertoire, Mesenchymal Stem Cells (MSC) are gaining interest and recognized as a cell population of choice. There is accumulating evidence that MSC growth factors, its soluble and insoluble proteins are involved in several key signaling pathways to promote tissue development, cellular differentiation and regeneration. MSC as multipotent non-hematopoietic cells with paracrine factors is advantageous for regenerative therapies. In this review, we discussed and summarized the important features of MSC including its immunomodulatory properties, mechanism of homing in the direction of tissue injury, licensing of MSC and the role of MSC soluble factors in cell-free therapy. Special consideration is highlighted on the rapidly growing research interest on the roles of MSC in ocular surface regeneration.
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Suhito, Intan Rosalina, Kyeong-Mo Koo, and Tae-Hyung Kim. "Recent Advances in Electrochemical Sensors for the Detection of Biomolecules and Whole Cells." Biomedicines 9, no. 1 (December 26, 2020): 15. http://dx.doi.org/10.3390/biomedicines9010015.
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Electrochemical sensors are considered an auspicious tool to detect biomolecules (e.g., DNA, proteins, and lipids), which are valuable sources for the early diagnosis of diseases and disorders. Advances in electrochemical sensing platforms have enabled the development of a new type of biosensor, enabling label-free, non-destructive detection of viability, function, and the genetic signature of whole cells. Numerous studies have attempted to enhance both the sensitivity and selectivity of electrochemical sensors, which are the most critical parameters for assessing sensor performance. Various nanomaterials, including metal nanoparticles, carbon nanotubes, graphene and its derivatives, and metal oxide nanoparticles, have been used to improve the electrical conductivity and electrocatalytic properties of working electrodes, increasing sensor sensitivity. Further modifications have been implemented to advance sensor platform selectivity and biocompatibility using biomaterials such as antibodies, aptamers, extracellular matrix (ECM) proteins, and peptide composites. This paper summarizes recent electrochemical sensors designed to detect target biomolecules and animal cells (cancer cells and stem cells). We hope that this review will inspire researchers to increase their efforts to accelerate biosensor progress—enabling a prosperous future in regenerative medicine and the biomedical industry.
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Dolganyuk, V. F., O. O. Babich, S. A. Sukhikh, E. V. Ulrikh, E. V. Kashirskikh, and A. P. Andreeva. "In vitro study of biologically active properties of complexes isolated from biomass of microscopic algae." Food systems 5, no. 3 (October 7, 2022): 212–22. http://dx.doi.org/10.21323/2618-9771-2022-5-3-212-222.
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Microalgae are rich in biologically active substances: proteins, carbohydrates, lipids, polyunsaturated fatty acids, vitamins, pigments, phycobiliproteins, enzymes, which are able to provide antioxidant, immunostimulating, antibacterial, antiviral, antitumor, antihypertensive, regenerative and neuroprotective effects on a body. The aim of this study is to run in vitro study of the antioxidant, antibacterial, fungicidal, antihypertensive and prebiotic properties of protein concentrate (PC), lipid-pigment complex (LPC) and carbohydrate-mineral complexes (CMC) obtained from the biomass of microscopic algae. To determine in vitro the antioxidant, antibacterial, fungicidal, antihypertensive and prebiotic properties of protein concentrates, lipid-pigment complexes and carbohydratemineral complexes obtained from the biomass of microscopic algae, the following methods were used: method of diffusion (on a solid nutritional medium); optical density method (in a liquid nutritional medium); the method for determining the antioxidant activity of the researched samples by their ability to restore free radicals; the method of inhibition of the angiotensin-converting enzyme. It was shown in this study that among the studied samples the carbohydrate-mineral complex provided the most pronounced antioxidant effect. It was found that both protein concentrates, and lipid-pigment complexes and carbohydrate-mineral complexes feature antibacterial and fungicidal properties. It was proven that among the studied samples the carbohydrate-mineral complex provided the least pronounced antihypertensive effect. It was found that the lipid-pigment complexes and carbohydrate-mineral complexes practically have no prebiotic properties. The biological activity (antimicrobial, fungicidal, antioxidant and prebiotic activities), as well as the antihypertensive properties, were confirmed in the protein concentrates, lipid-pigment complexes and carbohydrate-mineral complexes obtained from the biomass of microscopic algae (Chlorella vulgaris, Arthrospira platensis, Nostoc sp., Dunaliella salina, Pleurochrysis carterae). All these factors open up promising prospects for the practical application of protein concentrates, as well as lipid-pigment complexes and carbohydrate-mineral complexes.
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Muzio, Giuliana, Germana Martinasso, Francesco Baino, Roberto Frairia, Chiara Vitale-Brovarone, and Rosa A. Canuto. "Key role of the expression of bone morphogenetic proteins in increasing the osteogenic activity of osteoblast-like cells exposed to shock waves and seeded on bioactive glass-ceramic scaffolds for bone tissue engineering." Journal of Biomaterials Applications 29, no. 5 (July 2, 2014): 728–36. http://dx.doi.org/10.1177/0885328214541974.
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In this work, the role of shock wave-induced increase of bone morphogenetic proteins in modulating the osteogenic properties of osteoblast-like cells seeded on a bioactive scaffold was investigated using gremlin as a bone morphogenetic protein antagonist. Bone-like glass-ceramic scaffolds, based on a silicate experimental bioactive glass developed at the Politecnico di Torino, were produced by the sponge replication method and used as porous substrates for cell culture. Human MG-63 cells, exposed to shock waves and seeded on the scaffolds, were treated with gremlin every two days and analysed after 20 days for the expression of osteoblast differentiation markers. Shock waves have been shown to induce osteogenic activity mediated by increased expression of alkaline phosphatase, osteocalcin, type I collagen, BMP-4 and BMP-7. Cells exposed to shock waves plus gremlin showed increased growth in comparison with cells treated with shock waves alone and, conversely, mRNA contents of alkaline phosphatase and osteocalcin were significantly lower. Therefore, the shock wave-mediated increased expression of bone morphogenetic protein in MG-63 cells seeded on the scaffolds is essential in improving osteogenic activity; blocking bone morphogenetic protein via gremlin completely prevents the increase of alkaline phosphatase and osteocalcin. The results confirmed that the combination of glass-ceramic scaffolds and shock waves exposure could be used to significantly improve osteogenesis opening new perspectives for bone regenerative medicine.
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Khabibrakhmanova, Venera Ravilevna, Anna Evgenievna Rassabina, Aisylu Faridovna Khayrullina, and Farida Vilevna Minibayeva. "PHYSICO-CHEMICAL CHARACTERISTICS AND ANTIOXIDANT PROPERTIES OF MELANINS EXTRACTED FROM LEPTOGIUM FURFURACEUM (HARM.)." chemistry of plant raw material, no. 4 (December 15, 2022): 115–25. http://dx.doi.org/10.14258/jcprm.20220411774.
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Melanins are universal biopolymers, the main biological function of which is to protect living organisms from adverse factors. Interest in these dark pigments arises due to the prospects for their use in medicine, cosmeceuticals, bioremediation, bioelectronics, and other fields. In this work, a complex scheme was developed to isolate melanins from the lichen Leptogium furfuraceum (Harm.), which is a symbiotic extremophilic organism. Screening of the conditions for the extraction of melanin, its subsequent precipitation and purification made it possible to isolate three samples of melanin with a total yield of 7.5% of the dry matter of the lichen. It was established that all isolated melanins have a polyaromatic structure, they contain carbohydrates and proteins, the content of which, depending on the conditions for melanin isolation, is 7.3–9.9% and 13.5–32.7%, respectively. Among the isolated melanins, melanin, which is a water-soluble fraction of acid-precipitated melanin, displays the highest antioxidant activity. This melanin, unlike other melanins, is almost fully soluble in distilled water, 95% ethanol, 99% dimethyl sulfoxide, and phosphate buffer (pH 7.4). The differences in solubility, antioxidant activity, and chelating capacity of isolated lichen melanins make it possible to determine potential areas for their practical use.
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Veiga, Anabela, Filipa Castro, Fernando Rocha, Beatriz Bernardes, Marta M. Duarte, and Ana Leite Oliveira. "Opening New Avenues for Bioceramics: Oscillatory Flow Reactors and Upcoming Technologies in Skin-Tissue Engineering." Solid State Phenomena 339 (December 19, 2022): 79–87. http://dx.doi.org/10.4028/p-1f087p.
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An aging population and lifestyle-related practices increase the incidence of chronic diseases and consequently its costs. The increasing requests for efficient chronic wound care constitute an opportunity for the field of regenerative medicine but, at the same time, it represents a challenge due to the need to limit treatment costs. Calcium-based materials have enormous potential for skin applications, as calcium has an established role in the normal homeostasis of wounded skin and serves as a modulator in keratinocyte proliferation and differentiation. On the other hand, several natural biopolymers, as silk proteins are known for their antioxidant and moisturizing properties as well as a mitogenic influence on mammalian cells. In the present work, a cost-effective method using an oscillatory flow reactor to produce a calcium phosphate/sericin composite system with controlled properties is presented, to be applied in skin wound healing and regeneration. Future perspectives for the produced biomaterials are also addressed.
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Andrée, Lea, Rik Oude Egberink, Josephine Dodemont, Negar Hassani Besheli, Fang Yang, Roland Brock, and Sander C. G. Leeuwenburgh. "Gelatin Nanoparticles for Complexation and Enhanced Cellular Delivery of mRNA." Nanomaterials 12, no. 19 (September 29, 2022): 3423. http://dx.doi.org/10.3390/nano12193423.
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Messenger RNA (mRNA) is increasingly gaining interest as a modality in vaccination and protein replacement therapy. In regenerative medicine, the mRNA-mediated expression of growth factors has shown promising results. In contrast to protein delivery, successful mRNA delivery requires a vector to induce cellular uptake and subsequent endosomal escape to reach its end destination, the ribosome. Current non-viral vectors such as lipid- or polymer-based nanoparticles have been successfully used to express mRNA-encoded proteins. However, to advance the use of mRNA in regenerative medicine, it is required to assess the compatibility of mRNA with biomaterials that are typically applied in this field. Herein, we investigated the complexation, cellular uptake and maintenance of the integrity of mRNA complexed with gelatin nanoparticles (GNPs). To this end, GNPs with positive, neutral or negative surface charge were synthesized to assess their ability to bind and transport mRNA into cells. Positively charged GNPs exhibited the highest binding affinity and transported substantial amounts of mRNA into pre-osteoblastic cells, as assessed by confocal microscopy using fluorescently labeled mRNA. Furthermore, the GNP-bound mRNA remained stable. However, no expression of mRNA-encoded protein was detected, which is likely related to insufficient endosomal escape and/or mRNA release from the GNPs. Our results indicate that gelatin-based nanomaterials interact with mRNA in a charge-dependent manner and also mediate cellular uptake. These results create the basis for the incorporation of further functionality to yield endosomal release.
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Oprita, Elena Iulia, Andreea Iosageanu, and Oana Craciunescu. "Progress in Composite Hydrogels and Scaffolds Enriched with Icariin for Osteochondral Defect Healing." Gels 8, no. 10 (October 12, 2022): 648. http://dx.doi.org/10.3390/gels8100648.
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Osteochondral structure reconstruction by tissue engineering, a challenge in regenerative medicine, requires a scaffold that ensures both articular cartilage and subchondral bone remodeling. Functional hydrogels and scaffolds present a strategy for the controlled delivery of signaling molecules (growth factors and therapeutic drugs) and are considered a promising therapeutic approach. Icariin is a pharmacologically-active small molecule of prenylated flavonol glycoside and the main bioactive flavonoid isolated from Epimedium spp. The in vitro and in vivo testing of icariin showed chondrogenic and ostseoinductive effects, comparable to bone morphogenetic proteins, and suggested its use as an alternative to growth factors, representing a low-cost, promising approach for osteochondral regeneration. This paper reviews the complex structure of the osteochondral tissue, underlining the main aspects of osteochondral defects and those specifically occurring in osteoarthritis. The significance of icariin’s structure and the extraction methods were emphasized. Studies revealing the valuable chondrogenic and osteogenic effects of icariin for osteochondral restoration were also reviewed. The review highlighted th recent state-of-the-art related to hydrogels and scaffolds enriched with icariin developed as biocompatible materials for osteochondral regeneration strategies.
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Khan, Mohammad R., Nicola Mordan, Mohamed Parkar, Vehid Salih, Nikolaos Donos, and Peter M. Brett. "Atypical Mesenchymal Stromal Cell Responses to Topographic Modifications of Titanium Biomaterials Indicate Cytoskeletal- and Genetic Plasticity-Based Heterogeneity of Cells." Stem Cells International 2019 (July 1, 2019): 1–16. http://dx.doi.org/10.1155/2019/5214501.
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Titanium (Ti) is widely used as a biomaterial for endosseous implants due to its relatively inert surface oxide layer that enables implanted devices the ability of assembling tissue reparative components that culminate in osseointegration. Topographic modifications in the form of micro- and nanoscaled structures significantly promote osseointegration and enhance the osteogenic differentiation of adult mesenchymal stromal cells (MSCs). While the biological mechanisms central to the differential responses of tissues and cells to Ti surface modifications remain unknown, adhesion and morphological adaptation are amongst the earliest events at the cell-biomaterial interface that are highly influenced by surface topography and profoundly impact the regulation of stem cell fate determination. This study correlated the effects of Ti topographic modifications on adhesion and morphological adaptation of human MSCs with phenotypic change. The results showed that modified Ti topographies precluded the adhesion of a subset of MSCs while incurring distinct morphological constraints on adherent cells. These effects anomalously corresponded with a differential expression of stem cell pluripotency and Wnt signalling-associated markers on both modified surfaces while additionally differing between hydrophobic and hydrophilic surface modifications—though extent of osteogenic differentiation induced by both modified topographies yielded similarly significant higher levels of cellular mineralisation in contrast to polished Ti. These results suggest that in the absence of deposited proteins and soluble factors, both modified topographies incur the selective adhesion of a subpopulation of progenitors with relatively higher cytoskeletal plasticity. While the presence of deposited proteins and soluble factors does not significantly affect adherence of cells, nanotopographic modifications enhance expression of pluripotency markers in proliferative conditions, which are conversely overridden by both modified topographies in osteogenic inductive conditions. Further deciphering the mechanisms underlying cellular selectivity and Ti topographic responsiveness will improve our understanding of stem cell heterogeneity and advance the potential of MSCs in regenerative medicine.
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Shanbhag, Siddharth, Carina Kampleitner, Niyaz Al-Sharabi, Samih Mohamed-Ahmed, Karol Ali Apaza Alccayhuaman, Patrick Heimel, Stefan Tangl, et al. "Functionalizing Collagen Membranes with MSC-Conditioned Media Promotes Guided Bone Regeneration in Rat Calvarial Defects." Cells 12, no. 5 (February 28, 2023): 767. http://dx.doi.org/10.3390/cells12050767.
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Functionalizing biomaterials with conditioned media (CM) from mesenchymal stromal cells (MSC) is a promising strategy for enhancing the outcomes of guided bone regeneration (GBR). This study aimed to evaluate the bone regenerative potential of collagen membranes (MEM) functionalized with CM from human bone marrow MSC (MEM-CM) in critical size rat calvarial defects. MEM-CM prepared via soaking (CM-SOAK) or soaking followed by lyophilization (CM-LYO) were applied to critical size rat calvarial defects. Control treatments included native MEM, MEM with rat MSC (CEL) and no treatment. New bone formation was analyzed via micro-CT (2 and 4 weeks) and histology (4 weeks). Greater radiographic new bone formation occurred at 2 weeks in the CM-LYO group vs. all other groups. After 4 weeks, only the CM-LYO group was superior to the untreated control group, whereas the CM-SOAK, CEL and native MEM groups were similar. Histologically, the regenerated tissues showed a combination of regular new bone and hybrid new bone, which formed within the membrane compartment and was characterized by the incorporation of mineralized MEM fibers. Areas of new bone formation and MEM mineralization were greatest in the CM-LYO group. Proteomic analysis of lyophilized CM revealed the enrichment of several proteins and biological processes related to bone formation. In summary, lyophilized MEM-CM enhanced new bone formation in rat calvarial defects, thus representing a novel ‘off-the-shelf’ strategy for GBR.
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Ben Abla, Amina, Guilhem Boeuf, Ahmed Elmarjou, Cyrine Dridi, Florence Poirier, Sylvie Changotade, Didier Lutomski, and Abdellatif Elm’selmi. "Engineering of Bio-Adhesive Ligand Containing Recombinant RGD and PHSRN Fibronectin Cell-Binding Domains in Fusion with a Colored Multi Affinity Tag: Simple Approach for Fragment Study from Expression to Adsorption." International Journal of Molecular Sciences 22, no. 14 (July 8, 2021): 7362. http://dx.doi.org/10.3390/ijms22147362.
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Engineering of biomimetic motives have emerged as promising approaches to improving cells’ binding properties of biomaterials for tissue engineering and regenerative medicine. In this study, a bio-adhesive ligand including cell-binding domains of human fibronectin (FN) was engineered using recombinant protein technology, a major extracellular matrix (ECM) protein that interacts with a variety of integrins cell-surface’s receptors and other ECM proteins through specific binding domains. 9th and 10th fibronectin type III repeat containing Arginine-Glycine-Aspartic acid (RGD) and Pro-His-Ser-Arg-Asn (PHSRN) synergic site (FNIII9-10) were expressed in fusion with a Colored Multi Affinity Tag (CMAT) to develop a simplified production and characterization process. A recombinant fragment was produced in the bacterial system using E. coli with high yield purified protein by double affinity chromatography. Bio-adhesive surfaces were developed by passive coating of produced fragment onto non adhesive surfaces model. The recombinant fusion protein (CMAT-FNIII9/10) demonstrated an accurate monitoring capability during expression purification and adsorption assay. Finally, biological activity of recombinant FNIII9/10 was validated by cellular adhesion assay. Binding to α5β1 integrins were successfully validated using a produced fragment as a ligand. These results are robust supports to the rational development of bioactivation strategies for biomedical and biotechnological applications.
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Nyambat, Batzaya, Yankuba B. Manga, Chih-Hwa Chen, Uuganbayar Gankhuyag, Andi Pratomo WP, Mantosh Kumar Satapathy, and Er-Yuan Chuang. "New Insight into Natural Extracellular Matrix: Genipin Cross-Linked Adipose-Derived Stem Cell Extracellular Matrix Gel for Tissue Engineering." International Journal of Molecular Sciences 21, no. 14 (July 9, 2020): 4864. http://dx.doi.org/10.3390/ijms21144864.
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The cell-derived extracellular matrix (ECM) is associated with a lower risk of pathogen transfer, and it possesses an ideal niche with growth factors and complex fibrillar proteins for cell attachment and growth. However, the cell-derived ECM is found to have poor biomechanical properties, and processing of cell-derived ECM into gels is scarcely studied. The gel provides platforms for three-dimensional cell culture, as well as injectable biomaterials, which could be delivered via a minimally invasive procedure. Thus, in this study, an adipose-derived stem cell (ADSC)-derived ECM gel was developed and cross-linked by genipin to address the aforementioned issue. The genipin cross-linked ADSC ECM gel was fabricated via several steps, including rabbit ADSC culture, cell sheets, decellularization, freeze–thawing, enzymatic digestion, neutralization of pH, and cross-linking. The physicochemical characteristics and cytocompatibility of the gel were evaluated. The results demonstrated that the genipin cross-linking could significantly enhance the mechanical properties of the ADSC ECM gel. Furthermore, the ADSC ECM was found to contain collagen, fibronectin, biglycan, and transforming growth factor (TGF)-β1, which could substantially maintain ADSC, skin, and ligament fibroblast cell proliferation. This cell-derived natural material could be suitable for future regenerative medicine and tissue engineering application.
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Medda, Rebecca, Arne Helth, Patrick Herre, Darius Pohl, Bernd Rellinghaus, Nadine Perschmann, Stefanie Neubauer, et al. "Investigation of early cell–surface interactions of human mesenchymal stem cells on nanopatterned β-type titanium–niobium alloy surfaces." Interface Focus 4, no. 1 (February 6, 2014): 20130046. http://dx.doi.org/10.1098/rsfs.2013.0046.
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Multi-potent adult mesenchymal stem cells (MSCs) derived from bone marrow have therapeutic potential for bone diseases and regenerative medicine. However, an intrinsic heterogeneity in their phenotype, which in turn results in various differentiation potentials, makes it difficult to predict the response of these cells. The aim of this study is to investigate initial cell–surface interactions of human MSCs on modified titanium alloys. Gold nanoparticles deposited on β-type Ti–40Nb alloys by block copolymer micelle nanolithography served as nanotopographical cues as well as specific binding sites for the immobilization of thiolated peptides present in several extracellular matrix proteins. MSC heterogeneity persists on polished and nanopatterned Ti–40Nb samples. However, cell heterogeneity and donor variability decreased upon functionalization of the gold nanoparticles with cyclic RGD peptides. In particular, the number of large cells significantly decreased after 24 h owing to the arrangement of cell anchorage sites, rather than peptide specificity. However, the size and number of integrin-mediated adhesion clusters increased in the presence of the integrin-binding peptide (cRGDfK) compared with the control peptide (cRADfK). These results suggest that the use of integrin ligands in defined patterns could improve MSC-material interactions, not only by regulating cell adhesion locally, but also by reducing population heterogeneity.
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Norris, Emma G., Diane Dalecki, and Denise C. Hocking. "Acoustic Fabrication of Collagen–Fibronectin Composite Gels Accelerates Microtissue Formation." Applied Sciences 10, no. 8 (April 23, 2020): 2907. http://dx.doi.org/10.3390/app10082907.
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Ultrasound can influence biological systems through several distinct acoustic mechanisms that can be manipulated by varying reaction conditions and acoustic exposure parameters. We recently reported a new ultrasound-based fabrication technology that exploits the ability of ultrasound to generate localized mechanical forces and thermal effects to control collagen fiber microstructure non-invasively. Exposing solutions of type I collagen to ultrasound during the period of microfibril assembly produced changes in collagen fiber structure and alignment, and increased the biological activity of the resultant collagen hydrogels. In the extracellular matrix, interactions between fibronectin and collagen fibrils influence the biological activity of both proteins. Thus, in the present study, we examined how addition of fibronectin to collagen solutions prior to ultrasound exposure affects protein organization and the biological activity of the composite hydrogels. Results indicate that ultrasound can alter the distribution of fibronectin within 3D hydrogels via thermal and non-thermal mechanisms to produce composite hydrogels that support accelerated microtissue formation. The use of acoustic energy to drive changes in protein conformation to functionalize biomaterials has much potential as a unique, non-invasive technology for tissue engineering and regenerative medicine.
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Oude Egberink, Rik, Helen M. Zegelaar, Najoua El Boujnouni, Elly M. M. Versteeg, Willeke F. Daamen, and Roland Brock. "Biomaterial-Mediated Protein Expression Induced by Peptide-mRNA Nanoparticles Embedded in Lyophilized Collagen Scaffolds." Pharmaceutics 14, no. 8 (August 2, 2022): 1619. http://dx.doi.org/10.3390/pharmaceutics14081619.
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In our aging society, the number of patients suffering from poorly healing bone defects increases. Bone morphogenetic proteins (BMPs) are used in the clinic to promote bone regeneration. However, poor control of BMP delivery and thus activity necessitates high doses, resulting in adverse effects and increased costs. It has been demonstrated that messenger RNA (mRNA) provides a superior alternative to protein delivery due to local uptake and prolonged expression restricted to the site of action. Here, we present the development of porous collagen scaffolds incorporating peptide-mRNA nanoparticles (NPs). Nanoparticles were generated by simply mixing aqueous solutions of the cationic cell-penetrating peptide PepFect14 (PF14) and mRNA. Peptide-mRNA complexes were uniformly distributed throughout the scaffolds, and matrices fully preserved cell attachment and viability. There was a clear dependence of protein expression on the incorporated amount of mRNA. Importantly, after lyophilization, the mRNA formulation in the collagen scaffolds retained activity also at 4 °C over two weeks. Overall, our results demonstrate that collagen scaffolds incorporating peptide-mRNA complexes hold promise as off-the-shelf functional biomaterials for applications in regenerative medicine and constitute a viable alternative to lipid-based mRNA formulations.
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Chandra, Prafulla, and Sang Jin Lee. "Synthetic Extracellular Microenvironment for Modulating Stem Cell Behaviors." Biomarker Insights 10s1 (January 2015): BMI.S20057. http://dx.doi.org/10.4137/bmi.s20057.
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The innate ability of stem cells to self-renew and differentiate into multiple cell types makes them a promising source for tissue engineering and regenerative medicine applications. Their capacity for self-renewal and differentiation is largely influenced by the combination of physical, chemical, and biological signals found in the stem cell niche, both temporally and spatially. Embryonic and adult stem cells are potentially useful for cell-based approaches; however, regulating stem cell behavior remains a major challenge in their clinical use. Most of the current approaches for controlling stem cell fate do not fully address all of the complex signaling pathways that drive stem cell behaviors in their natural microenvironments. To overcome this limitation, a new generation of biomaterials is being developed for use as three-dimensional synthetic microenvironments that can mimic the regulatory characteristics of natural extracellular matrix (ECM) proteins and ECM-bound growth factors. These synthetic microenvironments are currently being investigated as a substrate with surface immobilization and controlled release of bioactive molecules to direct the stem cell fate in vitro, as a tissue template to guide and improve the neo-tissue formation both in vitro and in vivo, and as a delivery vehicle for cell therapy in vivo. The continued advancement of such an intelligent biomaterial system as the synthetic extracellular microenvironment holds the promise of improved therapies for numerous debilitating medical conditions for which no satisfactory cure exists today.
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Tran, Thuoc Linh, Fuyu Tamanoi, Mong-Hong Lee, and Phuc Van Pham. "Welcome to CRRM2017." Biomedical Research and Therapy 4, S (September 2, 2017): 1. http://dx.doi.org/10.15419/bmrat.v4is.364.
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On behalf of the entire the international conference Innovations in Cancer Research and Regenerative Medicine 2017 (CRRM2017) Organizing Committee, we would like to extend to you a warm welcome to Ho Chi Minh city, Vietnam to join the 3rd international conference of CRRM. We are very glad to announce that this time, CRRM has made some exciting changes with more plenary sessions, more sessions, more oral invited speakers and especially a day of pre-conference. With two main topics of cancer research and regenerative medicine, this conference has many sessions organized into some parallel sessions with different topics of cancer research and regenerative medicine. By doing so, the audience can arrange their time for networking and speaking to our commercial partners. From the basic science to clinical trials, all sessions at the conference will cover from gene to human body. Therefore, we believe that every session will provide exciting presentations for attendees at all levels from students to scientists/researchers. With this conference, all authors not only share their latest results, but also can publish their works in the reputed journals. That is a reason why the keywords for this conference is Share & Publish. Like many developing countries, Vietnam has no shortage of health challenges, from infectious diseases such as annual flu epidemics to rising rates of unmet and chronic illnesses such as cancers and diabetes. Vietnam also has to contend with increasing drug resistance for killer diseases including AIDS, tuberculosis, and malaria. There is, however, increasing evidence that Vietnamese researchers are tackling the challenges of harnessing biotechnology to improve health care for healthier society as well as advancing the sciences. The biomedicine progresses in Cancer Research, and Regenerative Medicine in Vietnam include gene diagnostics for genetic disorders of human diseases, infectious pathogens; in-vitro production of therapeutic proteins including insulin, IFNs or biosimilar forms of monoclonal antibodies for targeted therapies; biomaterials; cell therapy; stem cell therapy and tissue engineering. In this conference, with 14 sessions of 100 oral presentations and 100 poster presentations, all latest topics of cancer research and regenerative medicine will be covered. New biomarkers of cancers, immunotherapy strategy for cancers and stem cells as well as tissue engineering are hot topics of CRRM2017. In addition to the conference program, there is an extensive set of pre-conference and lunch-on innovation showcases for on 10th and 11th Sept, respectively. We are very grateful for the support from sponsors for the CRRM2017. More vents will be taking place in the exhibition area, and we encourage attendees to take advantage of the opportunities to visit the exhibit booths and corporate symposia. We wish to thank everyone who helped so enthusiastically in the organization of the conference. Our thanks also go out to all of the speakers who have generously agreed to share their research results and experiences at this conference. We hope that you will enjoy this meeting and have a wonderful time here in Ho Chi Minh city, Vietnam.
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Pizzicannella, Jacopo, Francesca Diomede, Agnese Gugliandolo, Luigi Chiricosta, Placido Bramanti, Ilaria Merciaro, Tiziana Orsini, Emanuela Mazzon, and Oriana Trubiani. "3D Printing PLA/Gingival Stem Cells/ EVs Upregulate miR-2861 and -210 during Osteoangiogenesis Commitment." International Journal of Molecular Sciences 20, no. 13 (July 2, 2019): 3256. http://dx.doi.org/10.3390/ijms20133256.
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Bone tissue regeneration strategies require approaches that provide an osteogenic and angiogenic microenvironment able to drive the bone growth. Recently, the development of 3D printing biomaterials, including poly(lactide) (3D-PLA), enriched with mesenchymal stem cells (MSCs) and/or their derivatives, such as extracellular vesicles (EVs) has been achieving promising results. In this study, in vitro results showed an increased expression of osteogenic and angiogenic markers, as RUNX2, VEGFA, OPN and COL1A1 in the living construct 3D-PLA/human Gingival MSCs (hGMSCs)/EVs. Considering that EVs carry and transfer proteins, mRNA and microRNA into target cells, we evaluated miR-2861 and miR-210 expression related to osteoangiogenesis commitment. Histological examination of rats implanted with 3D-PLA/hGMSCs/EVs evidenced the activation of bone regeneration and of the vascularization process, confirmed also by MicroCT. In synthesis, an upregulation of miR-2861 and -210 other than RUNX2, VEGFA, OPN and COL1A1 was evident in cells cultured in the presence of the biomaterial and EVs. Then, these results evidenced that EVs may enhance bone regeneration in calvaria defects, in association with an enhanced vascularization offering a novel regulatory system in the osteoangiogenesis evolution. The application of new strategies to improve biomaterial engraftment is of great interest in the regenerative medicine and can represent a way to promote bone regeneration.