Tissue Repair Applications Research Articles

Comprehensive metabolomic analysis of Mangifera indica leaves using UPLC-ESI-Q-TOF-MSE for cell differentiation: An in vitro and in vivo study

The comprehensive metabolic profiling was performed in the leaf extracts of Mangifera indica and assessed for their significant therapeutic application in tissue engineering and regenerative medicine in both in vitro and in vivo studies. About 147 compounds were identified in the ethyl acetate and methanol extracts of M. indica using MS/MS fragmentation analysis and the selected compounds were quantified using LC-QqQ-MS analysis. The in vitro cytotoxic activity showed that the M. indica extracts enhance the proliferation of mouse myoblast cells in concentration-dependent manner. As well, the extracts of M. indica induce the myotube formation by generating oxidative stress in the C2C12 cells was confirmed. The western blot analysis clearly showed that the M. indica induce myogenic differentiation by upregulating the myogenic marker proteins such as PI3K, Akt, mTOR, MyoG, and MyoD. The in vivo studies showed that the extracts expedites the acute wound repair by formation of crust, wound closure and improves the blood perfusion towards the wound area. Together, the leaves of M. indica can be used as excellent therapeutic agent for tissue repair and wound healing applications.

Collagen-mesenchymal stem cell spheroids in suspension promote high adipogenic capacity

Human mesenchymal stem cells (hMSC) represent a unique and promising platform because of their ability to promote soft tissue regeneration, particularly their ability to differentiate into adipocytes, which are important for adipose tissue regeneration. In this context, type I collagen is the most abundant extracellular matrix component of adipose tissue and can act as a natural spheroid source to support the differentiation process of stem cells. However, spheroids based on collagen and hMSCs without numerous pro-adipogenic factors that can induce adipogenesis have not yet been investigated. In this study, we focused on developing collagen-hMSC spheroids capable of differentiating into adipocyte-like cells in a short time (eight culture days) without adipogenic factors, with potential applications in adipose tissue repair. The physical and chemical properties of the spheroids indicated successful cross-linking of collagen. Upon spheroid development, stability, cell viability, and metabolic activity of the constructs were maintained. During adipogenesis, cell morphology shows significant changes, in which cells change from a fibroblast-like shape to an adipocyte-like shape, and adipogenic gene expression after eight days of cell culture. These results support the utility of collagen-hMSC 3 mg ml−1 collagen concentration spheroids to differentiate into adipocyte-like cells in a short time without adverse effects on biocompatibility, metabolic activity, or cell morphology, suggesting that this construct may be used in soft tissue engineering.

Composite beads of alginate and biological hydroxyapatite from poultry and mariculture for hard tissue repair

Synthesizing materials that stimulate the natural growth of living tissues and restore damaged parts of the body is one of the most challenging problems in regenerative medicine. Despite being the most commonly used biomaterial, synthetic hydroxyapatite is a calcium phosphate with a relatively low rate of bioresorption related to new tissue growth. For specific applications, the speed of resorption is essential, and synergy between polymer and hydroxyapatite composite materials from natural sources can be developed. Therefore, this study attempts to synthesize hydroxyapatite from poultry and mariculture by-products and produce spheres with alginate for use as biomaterials for tissue repair. Shells different shellfish and eggs were used as sources of CaCO3 and added to a phosphoric acid solution as precursors in wet synthesis. The powders were dried, thermally treated and characterized. Structural analysis revealed hydroxyapatite in nanometric crystallites (61–72 nm) with high crystallinity (86–89%). The calcium phosphate obtained from Mozambique shellfish acquired the best characteristics. Beads with various sizes and porosities were produced through changes in the process parameters, including the type and size of the dripper, speed of agitation of the solution, and type of drying. The results show that the type of dripper strongly influences the size of the beads and that the rotation speed influences the sphericity. The styling directly influenced the fluid absorption, demonstrating that the spheres dried by lyophilization can absorb up to 223% of their weight. In comparison, samples dried in a desiccator could absorb only 112% of their weight in body fluids. The porosity of the optimized beads was up to 90%, which is similar to that of human bone, and they did not show cytotoxicity. Therefore, the beads composed of alginate and hydroxyapatite produced here have the potential for application in tissue repair.

Chitosan 3D scaffolds with resolvin D1 for vertebral arthrodesis: a pilot study

PurposeOver the last years, the number of vertebral arthrodesis has been steadily increasing. The use of iliac crest bone autograft remains the “gold standard” for bone graft substitute in these procedures. However, this solution has some side effects, such as the problem of donor site morbidity indicating that there is a real need for adequate alternatives. This pilot study aimed to evaluate the usefulness of chitosan (Ch) porous 3D scaffolds incorporated with resolvin D1 (RvD1) as an alternative implant to iliac bone autograft.MethodsWe have performed bilateral posterolateral lumbar vertebral arthrodesis in a rat animal model. Three experimental groups were used: (i) non-operated animals; (ii) animals implanted with Ch scaffolds incorporated with RvD1 and (iii) animals implanted with iliac bone autograft.ResultsThe collagenous fibrous capsule formed around the Ch scaffolds with RvD1 is less dense when compared with the iliac bone autograft, suggesting an important anti-inflammatory effect of RvD1. Additionally, new bone formation was observed in the Ch scaffolds with RvD1.ConclusionThese results demonstrate the potential of these scaffolds for bone tissue repair applications.

An adhesive gelatin-coated small intestinal submucosa composite hydrogel dressing aids wound healing

It is a challenging clinical task to determine how to repair large-area skin defects better. Traditional wound dressings (e.g., cotton and gauze) can only be used as a dressing; consequently, there is an increasing demand for wound dressings with additional properties (i.e., antibacterial and pro-repair) in clinical practice. In this study, a composite hydrogel with o-nitrobenzene-modified gelatin-coated decellularized small intestinal submucosa (GelNB@SIS) was designed for the repair of skin injuries. SIS is a natural extracellular matrix with a 3D microporous structure and also contains high levels of growth factors and collagen. GelNB provides this material photo-triggering tissue adhesive property. The structure, tissue adhesion, cytotoxicity, and bioactivity to cells were investigated. Based on in vivo study and histological analysis, we found the combination of GelNB and SIS improved the healing process by promoting vascular renewal, dermal remodeling, and epidermal regeneration. Based on our findings, GelNB@SIS is a promising candidate for tissue repair applications.

Filter cake-derived calcium carbonate polymorphs from sugar refinery for hydroxyapatite production as a sustainable material for biomedical application

In this study, sugarcane filter cake (SFC) was refined to produce calcium carbonate (CaCO3) via the calcination method under a controlled environment. The obtained CaCO3 and orthophosphoric acid were used as calcium and phosphorus precursors respectively to synthesize hydroxyapatite (HA) through the wet precipitation method. Morphological investigation showed different morphologies of CaCO3 particles. The scanning electron microscopy (SEM) image showed high agglomeration of CaCO3 particles and the synthesized HA nanoparticles. Particle size analysis of CaCO3 and the HA nanoparticles revealed an average size of 0.9 μm and a crystallite size of 24.33 nm, respectively. X-ray diffraction (XRD) and Wide-angle x-ray scattering (WAXS) analysis confirmed the phase of calcium carbonate coexisting with both calcium hydroxide and calcium oxide, and a single phase of carbonated HA. The calcium to phosphorus (Ca/P) ratio of HA derived from calcinated filter cake was 1.66, which is close to the stoichiometric value of 1.67. The HA nanoparticles were biocompatible, with no evidence of toxicity to MC3T3-E1 cells. This study has demonstrated the feasibility of utilizing sugarcane filter cake as a sustainable raw material for CaCO3 and HA production. The as prepared HA is similar to biological apatite found in mammals and is potentially useful for hard tissue repair and other biomedical applications.

Front Cover: Digital light processing (DLP)‐based (bio)printing strategies for tissue modeling and regeneration

Digital light processing (DLP)-based bioprinting technology has been exploited for a wide range of applications in tissue repair, tissue regeneration, disease modeling and drug screening. In article number e270, Li and co-workers comprehensively summarize works about DLP-based bioprinting strategy to deliver biomedical materials and/or specific cells to create sophisticated structures for various tissue modeling and regeneration.

A Short Review on Nanostructured Carbon Containing Biopolymer Derived Composites for Tissue Engineering Applications.

The development of new scaffolds and materials for tissue engineering is a wide and open realm of material science. Among solutions, the use of biopolymers represents a particularly interesting area of study due to their great chemical complexity that enables creation of specific molecular architectures. However, biopolymers do not exhibit the properties required for direct application in tissue repair-such as mechanical and electrical properties-but they do show very attractive chemical functionalities which are difficult to produce through in vitro synthesis. The combination of biopolymers with nanostructured carbon fillers could represent a robust solution to enhance composite properties, producing composites with new and unique features, particularly relating to electronic conduction. In this paper, we provide a review of the field of carbonaceous nanostructure-containing biopolymer composites, limiting our investigation to tissue-engineering applications, and providing a complete overview of the recent and most outstanding achievements.

Immunomodulatory effect of human dedifferentiated fat cells: comparison with adipose-derived stem cells.

Dedifferentiated fat cells (DFATs), which are originated by the dedifferentiation of adipocytes, display surface markers of mesenchymal stem cells and are able to differentiate into different cell types, thus, yielding a huge therapeutic potential in repairing damaged tissues and organs. The use of allogeneic stem cells from healthy donors constitutes the basis of a new strategy for cell therapy in the field of transplantation and the first requirement for allografts is determining their immunological properties. In this study, human DFATs and ADSCs were passaged as in vitro models to investigate their immunomodulatory effects. Phenotypic analysis of cell surface markers and three-line differentiation protocols were used to identify stem cells. The immunogenic phenotypes of DFATs and ADSCs were analyzed by flow cytometry and a mixed lymphocyte reaction was used to assess their immune function. The characteristics of stem cells were confirmed by phenotypic identification of cell surface markers and three-line differentiation. Flow cytometry analysis showed that P3 generation DFATs and ADSCs contained human leukocyte antigen (HLA) class I molecules, but did not express HLA class II molecules and costimulatory molecules CD40, CD80 and CD86. Moreover, allogeneic DFATs and ADSCs could not induce the proliferation of peripheral blood mononuclear cells (PBMCs). In addition, both populations were shown to inhibit the Concanavalin A-stimulated proliferation of PBMCs and act as third-party cells responsible for inhibiting the mixed lymphocyte response. DFATs have immunosuppressive properties similar to ADSCs. Based on this, allogeneic DFATs have potential applications in tissue repair or cell therapy.

Study of platelet-rich plasma application for skin and plastic surgery in recent 20 years: A bibliometric analysis.

In recent years, platelet-rich plasma (PRP) has been used in plastic surgery, dermatology, and other treatment procedures worldwide. Since the number of scientific writings has been significantly increasing, it is challenging to generate a manual compilation and systematic review of PRP's therapeutic applications in dermatology and plastic surgeries. This study aimed to make a bibliometric analysis of the literature in the field and evaluate research hotspots and frontiers in this field in the past 20 years. Using the Academic Search Premier and ScienceDirect defined search terms, we searched the Web of Science Core Collection (WoSCC) and Scopus databases. All data were analyzed using CiteSpace 5.8.R3 and VOSviewer, including countries, institutions, authors, keywords, cited authors, cited journals, cited references, discovered research hotspots, and frontiers. A total of 1931 studies were retrieved. The number of publications on PRP application in dermatology and plastic surgeries showed a yearly increase. The United States was the most significant contributor to this field, while Italy's contribution was noteworthy. The journal with the highest number of relevant articles in dermatology and plastic surgery included the Journal of Cosmetic Dermatology. However, the Wound Repair and Regeneration and International Journal of Molecular Sciences were the leading journals that should be paid attention to in the future. Author Anitua E from the Tor Vergata University of Rome published the most publications in this field. In the keyword co-occurrence analysis, all keywords were divided into six clusters, and the most common one in recent years was "PRP for facial beauty." Facial rejuvenation, scar, and alopecia were the main hotspots and research trends in this field. Based on the current global trends, the use of PRP in cosmetics and skin care is receiving increasing attention from researchers and clinicians. Recently, an increasing number of articles on PRP's application in skin tissue repair have been published in the United States and Italy. The number of studies on hair loss, facial rejuvenation, and scar management is increasing, suggesting that these subjects may become research hotspots for PRP in dermatology and cosmetic surgeries in recent years.

Extracellular vesicle-loaded hydrogels for tissue repair and regeneration.

Extracellular vesicles (EVs) are a collective term for nanoscale or microscale vesicles secreted by cells that play important biological roles. Mesenchymal stem cells are a class of cells with the potential for self-healing and multidirectional differentiation. In recent years, numerous studies have shown that EVs, especially those secreted by mesenchymal stem cells, can promote the repair and regeneration of various tissues and, thus, have significant potential in regenerative medicine. However, due to the rapid clearance capacity of the circulatory system, EVs are barely able to act persistently at specific sites for repair of target tissues. Hydrogels have good biocompatibility and loose and porous structural properties that allow them to serve as EV carriers, thereby prolonging the retention in certain specific areas and slowing the release of EVs. When EVs are needed to function at specific sites, the EV-loaded hydrogels can stand as an excellent approach. In this review, we first introduce the sources, roles, and extraction and characterization methods of EVs and describe their current application status. We then review the different types of hydrogels and discuss factors influencing their abilities to carry and release EVs. We summarize several strategies for loading EVs into hydrogels and characterizing EV-loaded hydrogels. Furthermore, we discuss application strategies for EV-loaded hydrogels and review their specific applications in tissue regeneration and repair. This article concludes with a summary of the current state of research on EV-loaded hydrogels and an outlook on future research directions, which we hope will provide promising ideas for researchers.

Integrated data-driven modeling and experimental optimization of granular hydrogel matrices

Integrated data-driven modeling and experimental optimization of granular hydrogel matrices

Advances in polymer-based cell encapsulation and its applications in tissue repair.

Cell microencapsulation is a more widely accepted area of biological encapsulation. In most cases, it involves fixing cells in polymer scaffolds or semi-permeable hydrogel capsules, providing the environment for protecting cells, allowing the exchange of nutrients and oxygen, and protecting cells against the attack of the host immune system by preventing the entry of antibodies and cytotoxic immune cells. Hydrogel encapsulation provides a three-dimensional (3D) environment similar to that experienced in vivo, so it can maintain normal cellular functions to produce tissues similar to those in vivo. Embedded cells can be genetically modified to release specific therapeutic products directly at the target site, thereby eliminating the side effects of systemic treatments. Cellular microcarriers need to meet many extremely high standards regarding their biocompatibility, cytocompatibility, immunoseparation capacity, transport, mechanical, and chemical properties. In this article, we discuss the biopolymer gels used in tissue engineering applications and the brief introduction of cell encapsulation for therapeutic protein production. Also, we review polymer biomaterials and methods for preparing cell microcarriers for biomedical applications. At the same time, in order to improve the application performance of cell microcarriers in vivo, we also summarize the main limitations and improvement strategies of cell encapsulation. Finally, the main applications of polymer cell microcarriers in regenerative medicine are summarized.

Living Chinese Herbal Scaffolds from Microfluidic Bioprinting for Wound Healing.

Biological scaffolds have been widely employed in wound healing applications, while their practical efficiency is compromised by insufficient oxygen delivery to the 3-dimensional constructs and inadequate nutrient supply for the long-term healing process. Here, we present an innovative living Chinese herbal scaffold to provide a sustainable oxygen and nutrient supply for promoting wound healing. Through a facile microfluidic bioprinting strategy, a traditional Chinese herbal medicine (Panax notoginseng saponins [PNS]) and a living autotrophic microorganism (microalgae Chlorella pyrenoidosa [MA]) were successfully encapsulated into the scaffolds. The encapsulated PNS could be gradually released from the scaffolds, which promoted cell adhesion, proliferation, migration, and tube formation invitro. In addition, benefiting from the photosynthetic oxygenation of the alive MA, the obtained scaffolds would produce sustainable oxygen under light illumination, exerting a protective effect against hypoxia-induced cell death. Based on these features, we have demonstrated through invivo experiments that these living Chinese herbal scaffolds could efficiently alleviate local hypoxia, enhance angiogenesis, and thereby accelerate wound closure in diabetic mice, indicating their great potential in wound healing and other tissue repair applications.

Recent advances in adhesive materials used in the biomedical field: adhesive properties, mechanism, and applications.

Adhesive materials are natural or synthetic polymers with the ability to adhere to the surface of luminal mucus or epithelial cells. They are widely used in the biomedical field due to their unique adhesion, biocompatibility, and excellent surface properties. When used in the human body, they can adhere to an accessible target and remain at the focal site for a longer period, improving the therapeutic effect on local disease. An adhesive material with bacteriostatic properties can play an antibacterial role at the focal site and the adhesive properties of the material can prevent the focal site from being infected by bacteria for a period. In addition, some adhesive materials can promote cell growth and tissue repair. In this review, the properties and mechanism of natural adhesive materials, organic adhesive materials, composite adhesive materials, and underwater adhesive materials have been introduced systematically. The applications of these adhesive materials in drug delivery, antibacterials, tissue repair, and other applications are described in detail. Finally, we have discussed the prospects and challenges of using adhesive materials in the field of biomedicine.

The fabrication of conductive material-decorated hydrogels for tissue repair

Conductive hydrogels have recently attracted considerable attention as a class of soft medical materials with high water content to mimic the electrophysiological environment of biological tissues for tissue repair applications.

Three-dimensional biofabrication of nanosecond laser micromachined nanofibre meshes for tissue engineered scaffolds.

There is a high demand for bespoke grafts to replace damaged or malformed bone and cartilage tissue. Three-dimensional (3D) printing offers a method of fabricating complex anatomical features of clinically relevant sizes. However, the construction of a scaffold to replicate the complex hierarchical structure of natural tissues remains challenging. This paper reports a novel biofabrication method that is capable of creating intricately designed structures of anatomically relevant dimensions. The beneficial properties of the electrospun fibre meshes can finally be realised in 3D rather than the current promising breakthroughs in two-dimensional (2D). The 3D model was created from commercially available computer-aided design software packages in order to slice the model down into many layers of slices, which were arrayed. These 2D slices with each layer of a defined pattern were laser cut, and then successfully assembled with varying thicknesses of 100 μm or 200 μm. It is demonstrated in this study that this new biofabrication technique can be used to reproduce very complex computer-aided design models into hierarchical constructs with micro and nano resolutions, where the clinically relevant sizes ranging from a simple cube of 20 mm dimension, to a more complex, 50 mm-tall human ears were created. In-vitro cell-contact studies were also carried out to investigate the biocompatibility of this hierarchal structure. The cell viability on a micromachined electrospun polylactic-co-glycolic acid fibre mesh slice, where a range of hole diameters from 200 μm to 500 μm were laser cut in an array where cell confluence values of at least 85% were found at three weeks. Cells were also seeded onto a simpler stacked construct, albeit made with micromachined poly fibre mesh, where cells can be found to migrate through the stack better with collagen as bioadhesives. This new method for biofabricating hierarchical constructs can be further developed for tissue repair applications such as maxillofacial bone injury or nose/ear cartilage replacement in the future.

Preparation, Characterization, and Drug Delivery of Hexagonal Boron Nitride-Borate Bioactive Glass Biomimetic Scaffolds for Bone Tissue Engineering

In this study, biomimetic borate-based bioactive glass scaffolds containing hexagonal boron nitride hBN nanoparticles (0.1, 0.2, 0.5, 1, and 2% by weight) were manufactured with the polymer foam replication technique to be used in hard tissue engineering and drug delivery applications. To create three-dimensional cylindrical-shaped scaffolds, polyurethane foams were used as templates and covered using a suspension of glass and hBN powder mixture. Then, a heat treatment was applied at 570 °C in an air atmosphere to remove the polymer foam from the structure and to sinter the glass structures. The structural, morphological, and mechanical properties of the fabricated composites were examined in detail. The in vitro bioactivity of the prepared composites was tested in simulated body fluid, and the release behavior of gentamicin sulfate and 5-fluorouracil from glass scaffolds were analyzed separately as a function of time. The cytotoxicity was investigated using osteoblastic MC3T3-E1 cells. The findings indicated that the hBN nanoparticles, up to a certain concentration in the glass matrix, improved the mechanical strength of the glass scaffolds, which mimic the cancellous bone. Additionally, the inclusion of hBN nanoparticles enhanced the in vitro hydroxyapatite-forming ability of bioactive glass composites. The presence of hBN nanoparticles accelerated the drug release rates of the system. It was concluded that bioactive glass/hBN composite scaffolds mimicking native bone tissue could be used for bone tissue repair and regeneration applications.

Rapid fabrication and screening of tailored functional 3D biomaterials: Validation in bone tissue repair – Part II

Regenerative medicine strategies place increasingly sophisticated demands on 3D biomaterials to promote tissue formation at sites where tissue would otherwise not form. Ideally, the discovery/fabrication of the 3D scaffolds needs to be high-throughput and uniform to ensure quick and in-depth analysis in order to pinpoint appropriate chemical and mechanical properties of a biomaterial. Herein we present a versatile technique to screen new potential biocompatible acrylate-based 3D scaffolds with the ultimate aim of application in tissue repair. As part of this process, we identified an acrylate-based 3D porous scaffold that promoted cell proliferation followed by accelerated tissue formation, pre-requisites for tissue repair. Scaffolds were fabricated by a facile freeze-casting and an in-situ photo-polymerization route, embracing a high-throughput synthesis, screening and characterization protocol. The current studies demonstrate the dependence of cellular growth and vascularization on the porosity and intrinsic chemical nature of the scaffolds, with tuneable 3D scaffolds generated with large, interconnected pores suitable for cellular growth applied to skeletal reparation. Our studies showed increased cell proliferation, collagen and ALP expression, while chorioallantoic membrane assays indicated biocompatibility and demonstrated the angiogenic nature of the scaffolds. VEGRF2 expression in vivo observed throughout the 3D scaffolds in the absence of growth factor supplementation demonstrates a potential for angiogenesis. This novel platform provides an innovative approach to 3D scanning of synthetic biomaterials for tissue regeneration.

Processing and Characterization of a New Quaternary Alloy Ti10Mo8Nb6Zr for Potential Biomedical Applications

The study of new metallic biomaterials for application in bone tissue repair has improved due to the increase in life expectancy and the aging of the world population. Titanium alloys are one of the main groups of biomaterials for these applications, and beta-type titanium alloys are more suitable for long-term bone implants. The objective of this work was to process and characterize a new Ti10Mo8Nb6Zr beta alloy. Alloy processing involves arc melting, heat treatment, and cold forging. The characterization techniques used in this study were X-ray fluorescence spectroscopy, X-ray diffraction, differential scanning calorimetry, optical microscopy, microhardness measurements, and pulse excitation technique. In vitro studies using adipose-derived stem cells (ADSC) were performed to evaluate the cytotoxicity and cell viability after 1, 4, and 7 days. The results showed that the main phase during the processing route was the beta phase. At the end of processing, the alloy showed beta phase, equiaxed grains with an average size of 228.7 µm, and low Young's modulus (83 GPa). In vitro studies revealed non-cytotoxicity and superior cell viability compared to CP Ti. The addition of zirconium led to a decrease in the beta-transus temperature and Young's modulus and improved the biocompatibility of the alloy. Therefore, the Ti10Mo8Nb6Zr alloy is a promising candidate for application in the biomedical field.