Enhanced Tissue Engineering Research Articles

Inkjet printing of polyurethane micro/nanofibers using TiO2 and Ag NPs on as‐spun mats for enhancing tissue engineering applications

Inkjet printing of polyurethane micro/nanofibers using <scp>TiO₂</scp> and Ag <scp>NPs</scp> on as‐spun mats for enhancing tissue engineering applications

Advanced Hydrogels: Enhancing Tissue Bioengineering with RGD Peptides and Carbon Nanomaterials.

The advancement of tissue engineering (TE) is driven by the development of scaffolds that mimic the mechanical, spatial, and biological environment of the extracellular matrix (ECM), crucial for regulating cell behavior and tissue repair. Hydrogels, 3D networks of polymer chains, are particularly suited for TE due to their high biocompatibility, ability to mimic tissue water content, facilitate cell migration, sustain growth factor release, and offer controllable physical properties. However, hydrogels mimicking the ECM often face challenges related to cell adhesion due to the absence of specific receptors. This issue can be addressed by incorporating ECM components into the polymer matrix, such as the peptide sequence arginine-glycine-aspartic acid (RGD), known for its role in cell adhesion. Additionally, carbon nanomaterials (CNMs) offer unique physicochemical properties that can improve scaffold-cell interactions. Despite the potential benefits, there are limited reports on their combination. RGD-CNM hydrogels enable a more accurate emulation of the natural cellular environment, enhancing tissue engineering applications. This hybrid approach may promote robust cell adhesion along with exceptional mechanical and electrical properties. This review outlines the potential benefits of these hybrid scaffolds and their synergistic potential, aiming to inspire new research directions in this innovative field.

Impact of Fibrin Gel Architecture on Hepatocyte Growth Factor Release and Its Role in Modulating Cell Behavior for Tissue Regeneration.

A novel scaffold design has been created to enhance tissue engineering and regenerative medicine by optimizing the controlled, prolonged release of Hepatocyte Growth Factor (HGF), a powerful chemoattractant for endogenous mesenchymal stem cells. We present a new stacked scaffold that is made up of three different fibrin gel layers, each of which has HGF integrated into the matrix. The design attempts to preserve HGF's regenerative properties for long periods of time, which is necessary for complex tissue regeneration. These multi-layered fibrin gels have been mechanically evaluated using rheometry, and their degradation behavior has been studied using D-Dimer ELISA. Understanding the kinetics of HGF release from this novel scaffold configuration is essential for understanding HGF's long-term sustained bioactivity. A range of cell-based tests were carried out to verify the functionality of HGF following extended incorporation. These tests included 2-photon microscopy using phalloidin staining to examine cellular morphology, SEM analysis for scaffold-cell interactions, and scratch and scatter assays to assess migration and motility. The analyses show that the novel stacking scaffold promotes vital cellular processes for tissue regeneration in addition to supporting HGF's bioactivity. This scaffold design was developed for in situ tissue engineering. Using the body as a bioreactor, the scaffold should recruit mesenchymal stem cells from their niche, thus combining the regenerative abilities of HGF and MSCs to promote tissue remodeling and wound repair.

Plant-Derived Exosome-Like Nanoparticles: Emerging Nanosystems for Enhanced Tissue Engineering.

Tissue engineering holds great potential for tissue repair and rejuvenation. Plant-derived exosome-like nanoparticles (ELNs) have recently emerged as a promising avenue in tissue engineering. However, there is an urgent need to understand how plant ELNs can be therapeutically applied in clinical disease management, especially for tissue regeneration. In this review, we comprehensively examine the properties, characteristics, and isolation techniques of plant ELNs. We also discuss their impact on the immune system, compatibility with the human body, and their role in tissue regeneration. To ensure the suitability of plant ELNs for tissue engineering, we explore various engineering and modification strategies. Additionally, we provide insights into the progress of commercialization and industrial perspectives on plant ELNs. This review aims to highlight the potential of plant ELNs in regenerative medicine by exploring the current research landscape and key findings.

LNP-RNA-engineered adipose stem cells for accelerated diabetic wound healing

Adipose stem cells (ASCs) have attracted considerable attention as potential therapeutic agents due to their ability to promote tissue regeneration. However, their limited tissue repair capability has posed a challenge in achieving optimal therapeutic outcomes. Herein, we conceive a series of lipid nanoparticles to reprogram ASCs with durable protein secretion capacity for enhanced tissue engineering and regeneration. In vitro studies identify that the isomannide-derived lipid nanoparticles (DIM1T LNP) efficiently deliver RNAs to ASCs. Co-delivery of self-amplifying RNA (saRNA) and E3 mRNA complex (the combination of saRNA and E3 mRNA is named SEC) using DIM1T LNP modulates host immune responses against saRNAs and facilitates the durable production of proteins of interest in ASCs. The DIM1T LNP-SEC engineered ASCs (DS-ASCs) prolong expression of hepatocyte growth factor (HGF) and C-X-C motif chemokine ligand 12 (CXCL12), which show superior wound healing efficacy over their wild-type and DIM1T LNP-mRNA counterparts in the diabetic cutaneous wound model. Overall, this work suggests LNPs as an effective platform to engineer ASCs with enhanced protein generation ability, expediting the development of ASCs-based cell therapies.

Spider silk enhanced tissue engineering of cartilage tissue: Approach of a novel bioreactor model using adipose derived stromal cells.

Human cartilage tissue remains a challenge for the development of therapeutic options due to its poor vascularization and reduced regenerative capacities. There are a variety of research approaches dealing with cartilage tissue engineering. In addition to different biomaterials, numerous cell populations have been investigated in bioreactor-supported experimental setups to improve cartilage tissue engineering. The concept of the present study was to investigate spider silk cocoons as scaffold seeded with adipose-derived stromal cells (ASC) in a custom-made bioreactor model using cyclic axial compression to engineer cartilage-like tissue. For chemical induction of differentiation, BMP-7 and TGF-β2 were added and changes in cell morphology and de-novo tissue formation were investigated using histological staining to verify chondrogenic differentiation. By seeding spider silk cocoons with ASC, a high colonization density and cell proliferation could be achieved. Mechanical induction of differentiation using a newly established bioreactor model led to a more roundish cell phenotype and new extracellular matrix formation, indicating a chondrogenic differentiation. The addition of BMP-7 and TGF-β2 enhanced the expression of cartilage specific markers in immunohistochemical staining. Overall, the present study can be seen as pilot study and valuable complementation to the published literature.

Angiogenesis driven extracellular matrix remodeling of 3D bioprinted vascular networks

Angiogenesis plays a pivotal role in development and tissue growth, as well as in pathological conditions such as cancer. Being able to understand the basic mechanisms involved in the vascularization of tissues and angiogenic network formation provides a window to advance the development of in vitro tissue models and enhance tissue engineering applications. In this study, we leveraged a novel microfluidic-based three dimensional (3D) bioprinting technology and alginate-collagen type I (AGC) bioink, to develop a 3D bioprinting strategy to enable the biofabrication of complex angiogenic networks within the 3D structure. These networks were comprised of simian vacuolating virus 40 (SV40) transformed adult rat brain endothelial cell (SV-ARBEC)-laden hydrogel rings. With mechanical properties relevant for vascular tissue engineering applications, these bioprinted constructs formed spontaneous vascular networks, reminiscent of anisotropic tissue-like structures, while retaining high cellular viability. The vascular network formation was accompanied by extracellular matrix (ECM) remodeling, confirming sequential SV-ARBEC mediated collagen type I fiber deposition and reorganization. Treatment with broad spectrum matrix metalloproteinase (MMP) inhibitor supressed SV-ARBEC angiogenic sprouting, highlighting requirements of ECM remodeling in angiogenic network formation. This novel 3D microfluidic bioprinting technology and biocompatible AGC hydrogel fiber rings supported robust SV-ARBEC angiogenesis and corresponding ECM remodeling, allowing us to present a strategy suitable to advancing applications in vascular research and supporting the further development of disease models, novel testing beds for drug discovery and tissue engineering applications.

Graphene-Lined Porous Gelatin Glycidyl Methacrylate Hydrogels: Implications for Tissue Engineering

Despite rigorous research, inferior mechanical properties and structural homogeneity are the main challenges constraining hydrogel's suturability to host tissue and limiting its clinical applications. To tackle those, we developed a reverse solvent interface trapping method, in which organized, graphene-coated microspherical cavities were introduced into a hydrogel to create heterogeneity and make it suturable. To generate those cavities, (i) graphite exfoliates to graphene sheets, which spread at the water/ heptane interfaces of the microemulsion, (ii) heptane fills the microspheres coated by graphene, and (iii) a cross-linkable hydrogel dissolved in water fills the voids. Cross-linking solidifies such microemulsion to a strong, suturable, permanent hybrid architecture, which has better mechanical properties, yet it is biocompatible and supports cell adhesion and proliferation. These properties along with the ease and biosafety of fabrication suggest the potential of this strategy to enhance tissue engineering outcomes by generating various suturable scaffolds for biomedical applications, such as donor cornea carriers for Boston keratoprosthesis (BK).

Nanoparticles: Promising Tools for the Treatment and Prevention of Myocardial Infarction.

Despite several recent advances, current therapy and prevention strategies for myocardial infarction are far from satisfactory, owing to limitations in their applicability and treatment effects. Nanoparticles (NPs) enable the targeted and stable delivery of therapeutic compounds, enhance tissue engineering processes, and regulate the behaviour of transplants such as stem cells. Thus, NPs may be more effective than other mechanisms, and may minimize potential adverse effects. This review provides evidence for the view that function-oriented systems are more practical than traditional material-based systems; it also summarizes the latest advances in NP-based strategies for the treatment and prevention of myocardial infarction.

Advances in Drug Delivery and Theranostics

Advances in Drug Delivery and Theranostics

Stem Cells Enhance Tissue Engineering in Head and Neck Surgery

Stem Cells Enhance Tissue Engineering in Head and Neck Surgery

Ectopic chondrogenesis of nude mouse induced by nano gene delivery enhanced tissue engineering technology.

Background: Many techniques and methods have been used clinically to relieve pain from cartilage repair, but the long-term effect is still unsatisfactory.Purpose: The objective of this study was to form an artificial chondroid tissue gene enhanced tissue engineering system to repair cartilage defects via nanosized liposomes.Methods: Cationic nanosized liposomes were prepared and characterized using transmission electron microscope (TEM) and dynamic laser light scattering (DLS). The rat mesenchymal stem cells (rMSCs) were isolated, cultivated, and induced by SRY (Sex-Determining Region Y)-Box 9 (Sox9) via cationic nanosized liposomes. The induced rMSCs were mixed with a thermo-sensitive chitosan hydrogel and subcutaneously injected into the nude mice. Finally, the newly-formed chondroid tissue obtained in the injection parts, and the transparent parts were detected by HE, collagen II, and safranin O.Results: It was found that the presently prepared cationic nanosized liposomes had the diameter of 85.76±3.48 nm and the zeta potential of 15.76±2.1 mV. The isolated rMSCs proliferation was fibroblast-like, with a cultivated confluence of 90% confluence in 5–8 days, and stained positive for CD29 and CD44 while negative for CD34 and CD45. After transfection with cationic nanosized liposomes, we observed changes of cellular morphology and a higher expression of SOX9 compared with control groups, which indicated that rMSCs could differentiate into chondrocyte in vitro. By mixing transfected rMSCs with the thermo-sensitive hydrogel of chitosan in nude mice, chondroid tissue was successfully obtained, demonstrating that rMSCs can differentiate into chondrogenic cells in vivo. Conclusion: This study explored new ways to improve the quality of tissue engineered cartilage, thus accelerating clinical transformation and reducing patient pain.

Constructing Gene-Enhanced Tissue Engineering for Regeneration and Repair of Osteochondral Defects.

In situ sustained release of endogenous growth factors from cells is a challenge for repair and regeneration of tissue. Although recombinant adenovirus vectors are an effective delivery system that can prolong the release of growth factors and is very suitable for the therapy of growth factors, these recombinant adenovirus vectors that are widely used at present have low safety and stability in terms of long-term expression. In this study, the above problems are solved by knocking out both E1 and E3 genes at the same time and directly inserting the gene fragments encoding target proteins after the inverted terminal repeats. Finally, the combination of gene therapy with tissue engineering in regeneration and repair of full-thickness defects of osteochondral tissue are applied as an example. The results show that this strategy can achieve complete repair of articular osteochondral defects and recovery of their function, and meanwhile solve the problems of low safety and expression instability of recombinant adenovirus vectors. This method provides a bright prospect for the application of gene enhanced tissue engineering in the regeneration and repair of joint tissue, and also provides a reference for the repair and regeneration of other tissues.

Bio Mimicking of Extracellular Matrix.

Biomaterials play a critical role in regenerative strategies such as stem cell-based therapies and tissue engineering, aiming to replace, remodel, regenerate, or support damaged tissues and organs. The design of appropriate three-dimensional (3D) scaffolds is crucial for generating bio-inspired replacement tissues. These scaffolds are primarily composed of degradable or non-degradable biomaterials and can be employed as cells, growth factors, or drug carriers. Naturally derived and synthetic biomaterials have been widely used for these purposes, but the ideal biomaterial remains to be found. Researchers from diversified fields have attempted to design and fabricate novel biomaterials, aiming to find novel theranostic approaches for tissue engineering and regenerative medicine. Since no single biomaterial has been found to possess all the necessary characteristics for an ideal performance, over the years scientists have tried to develop composite biomaterials that complement and combine the beneficial properties of multiple materials into a superior matrix. Herein, we highlight the structural features and performance of various biomaterials and their application in regenerative medicine and for enhanced tissue engineering approaches.

Cluster-assembled zirconia substrates promote long-term differentiation and functioning of human islets of Langerhans

Ex vivo expansion and differentiation of human pancreatic β-cell are enabling steps of paramount importance for accelerating the development of therapies for diabetes. The success of regenerative strategies depends on their ability to reproduce the chemical and biophysical properties of the microenvironment in which β-cells develop, proliferate and function. In this paper we focus on the biophysical properties of the extracellular environment and exploit the cluster-assembled zirconia substrates with tailored roughness to mimic the nanotopography of the extracellular matrix. We demonstrate that β-cells can perceive nanoscale features of the substrate and can convert these stimuli into mechanotransductive processes which promote long-term in vitro human islet culture, thus preserving β-cell differentiation and function. Proteomic and quantitative immunofluorescence analyses demonstrate that the process is driven by nanoscale topography, via remodelling of the actin cytoskeleton and nuclear architecture. These modifications activate a transcriptional program which stimulates an adaptive metabolic glucose response. Engineered cluster-assembled substrates coupled with proteomic approaches may provide a useful strategy for identifying novel molecular targets for treating diabetes mellitus and for enhancing tissue engineering in order to improve the efficacy of islet cell transplantation therapies.

Matrix Stiffness Modulates Mesenchymal Stem Cell Sensitivity to Geometric Asymmetry Signals.

Human stem cells hold significant potential for the treatment of various diseases. However, their use as a therapy is hampered because of limited understanding of the mechanisms by which they respond to environmental stimuli. Efforts to understand extracellular biophysical cues have demonstrated the critical roles of geometrical and mechanical signals in determining the fate of stem cells. The goal of this study was to explore the interplay between cell polarity and matrix stiffness in stem cell lineage specification. We hypothesize that confining cells to asymmetric extracellular matrix islands will impart polarity at a single-celllevel and will interact with mechanical signals to define the lineage of stem cells. To test these hypotheses, we employed microcontact printing to create patterned symmetric and asymmetric hydrogel islands of soft and hard surface stiffness. Human mesenchymal stem cells (hMSCs) were confined to these islands at the single-cell level and given the ability to differentiate along adipogenic or osteogenic routes. Our results demonstrated that cell polarity defines the lineage specification of hMSCs only on islands with low stiffness. Insight gained from this study provides a rational basis for designing stem cell cultures to enhance tissue engineering and regenerative medicine strategies.

VEGF heparinized-decellularized adipose tissue scaffolds enhance tissue engineering vascularization in vitro.

Scaffolds based on decellularized adipose tissue (DAT) are gaining popularity in the adipose tissue engineering field due to their high biocompatibility and vascularizing properties. Previous studies involving decellularized adipose tissue (DAT) scaffolds have not fully demonstrated their ability to induce in vitro vascularization of engineered tissue. With the aim of developing a more effective adipose tissue engineering vascularization technique based on DAT, we investigated the vascularizing potential of a vascular endothelial growth factor (VEGF) delivery system utilizing a heparinized DAT (Hep-DAT) scaffold in vitro. To generate this system, heparins were cross-linked to DATs with 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide and N-hydroxysuccinimide. Encapsulated Hep-DATs were able to control the release of a significantly higher amount of VEGF in vitro than non-heparinized DATs. Human bone marrow stromal cells (hBMSCs), when seeded on these VEGF–Hep-DATs, differentiated into endothelial cells which expressed vascular endothelial markers CD34 and VWF, thus resulting in accelerated vascularization of transplanted tissue as compared to the control DAT-only scaffold. In conclusion, these studies demonstrate that VEGF–Hep-DATs promoted greater tissue vascularization as compared to the DAT control scaffold and that VEGF–Hep-DATs are an effective and biocompatible angiogenesis system.

Heparin Augmentation Enhances Bioactive Properties of Acellular Extracellular Matrix Scaffold.

Extracellular matrix (ECM) maintains a reservoir of bioactive growth factors and matricellular proteins that provide bioinductive effects on local cells that influence phenotype and behaviors. Bioactive acellular ECM scaffolds can be used therapeutically to stimulate adaptive tissue repair. Fibroblast growth factor-2 (FGF-2) attenuates transforming growth factor-β1 (TGF-β1)-mediated cardiac fibrosis. Heparin glycosaminoglycan can influence FGF-2 bioactivity and could be leveraged to enhance tissue engineering strategies. We explored the effects of heparin on FGF-2 enhancement of bioactive ECM scaffold biomaterials for its antifibrotic effect on attenuating human cardiac myofibroblast activation. Increasing heparin concentration at a fixed concentration of FGF-2 markedly increased the amount of FGF-2 retained and eluted by ECM scaffolds. To explore synergistic bioinductive effects of heparin and FGF-2, collagen gel contraction assay using human cardiac myofibroblasts was performed in vitro. Myofibroblast activation was induced by profibrotic cytokine, TGF-β1. FGF-2 and heparin in combination reduced human cardiac myofibroblast-mediated collagen gel contraction to a greater extent than FGF-2 alone. These observations were confirmed for both human atrial and human ventricular cardiac fibroblasts. Cell death was not different between groups. In summary, heparin is an effective adjuvant to enhance FGF-2 loading and elution of acellular ECM scaffold biomaterials. Heparin increases the bioactive effects of FGF-2 in attenuating human cardiac myofibroblast activation in response to profibrotic TGF-β1. These data may inform tissue engineering strategies for myocardial repair to prevent fibrosis.

Restore a 9 mm diameter osteochondral defect with gene enhanced tissue engineering followed mosaicplasty in a goat model

ObjectiveThe aim of this study was to evaluate the efficacy of gene enhanced tissue engineering followed mosaicplasty in a goat model. MethodsAn acute cylindrical defect 9 mm in diameter was created in the weight bearing area of the medial femoral condyle in a goat model. Thirty-six medial femoral condyles were divided into 6 groups using different proportion of gene enhanced tissue engineering and mosaicplasty to restore the defects. The specimen received gross and histology observation, which was evaluated by the histological grading scale of O'Driscoll, Keeley and Salter. Transmission electron microscope observation was also performed. Two factors analysis of variance and Student-Newman-Kewls test were used to compare the specimen. ResultsThe gross and histology observation revealed that each defects of six groups had different restoration. The scores of the reparative tissue of three groups with gene enhancement were significantly higher than those in other three groups without gene enhancement (p > 0.05). ConclusionGene enhanced tissue engineering followed mosaicplasty could restore a 9 mm diameter osteochondral defects in a goat model effectively. With the reduction of covering area of the graft, the advantages of the combined gene enhanced tissue engineering method can be better reflected.

Gene therapy for bone tissue engineering.

Gene therapy holds a great promise and has been extensively investigated to improve bone formation and regeneration therapies in bone tissue engineering. A variety of osteogenic genes can be delivered by combining different vectors (viral or non-viral), scaffolds and delivery methodologies. Ex vivo & in vivo gene enhanced tissue engineering approaches have led to successful osteogenic differentiation and bone formation. In this article, we review recent advances of gene therapy-based bone tissue engineering discussing strengths and weaknesses of various strategies as well as general overview of gene therapy.