Tissue Engineering Technology Research Articles

Mesenchymal stem cell sheets: a new cell-based strategy for bone repair and regeneration

Mesenchymal stem cells (MSCs), a class of adult stem cells, are considered a promising source for bone regeneration. Although combining MSCs with biomaterial scaffolds offers an interesting clinical strategy for bone tissue engineering, the presence of the scaffolds could induce an undesirable effect on cell-cell interactions. Moreover, before the application of scaffold materials in bone tissue reconstruction, cells must be manipulated with proteolytic enzymes, such as trypsin or dispase that degrade extracellular matrix (ECM) molecules and cell surface proteins, which can result in the cell damage and loss of cellular activity. Therefore, the development of alternative strategies for bone regeneration is required to solve these problems. Recently, a novel tissue engineering technology named 'cell sheet' has been efficaciously utilized in the regeneration of bone, corneal, cardiac, tracheal and periodontal ligament-like tissues. The cell sheet is a layer of cells, which contains intact ECM and cell surface proteins such as growth factor receptors, ion channels and cell-to-cell junction proteins. MSC sheets can be easily fabricated by layering the recovered cell sheets without any scaffolds or complicated manipulation. This review summarizes the current state of the literature regarding the use of MSCs to produce cell sheets and assesses their applicability in bone tissue regeneration and repair.

Developing trend of wound dressing

The ideal wound dressing should have the functions of keeping wound moist and warm, preventing and treating wound infection, promoting wound healing, and so on; However there is no such ideal wound dressing in clinic. Dressings are likely to capable application to different kinds of wounds with multi-functions in the future. For the purpose of good tissue compatibility and permanent wound cover, auto- or allo- skin living cells should be integrated with biological dressings as real artificial skin by employing tissue engineering technology. Clinical application of smart dressings can enable wound management more personalized, effective, optimized, and convenient.

Immunomodulatory Functions of Mesenchymal Stem Cells in Tissue Engineering.

The inflammatory response to chronic injury affects tissue regeneration and has become an important factor influencing the prognosis of patients. In previous stem cell treatments, it was revealed that stem cells not only have the ability for direct differentiation or regeneration in chronic tissue damage but also have a regulatory effect on the immune microenvironment. Stem cells can regulate the immune microenvironment during tissue repair and provide a good “soil” for tissue regeneration. In the current study, the regulation of immune cells by mesenchymal stem cells (MSCs) in the local tissue microenvironment and the tissue damage repair mechanisms are revealed. The application of the concepts of “seed” and “soil” has opened up new research avenues for regenerative medicine. Tissue engineering (TE) technology has been used in multiple tissues and organs using its biomimetic and cellular cell abilities, and scaffolds are now seen as an important part of building seed cell microenvironments. The effect of tissue engineering techniques on stem cell immune regulation is related to the shape and structure of the scaffold, the preinflammatory microenvironment constructed by the implanted scaffold, and the material selection of the scaffold. In the application of scaffold, stem cell technology has important applications in cartilage, bone, heart, and liver and other research fields. In this review, we separately explore the mechanism of MSCs in different tissue and organs through immunoregulation for tissue regeneration and MSC combined with 3D scaffolds to promote MSC immunoregulation to repair damaged tissues.

Regeneration of a neoartery through a completely autologous acellular conduit in a minipig model: a pilot study

BackgroundVascular grafts are widely used as a treatment in coronary artery bypass surgery, hemodialysis, peripheral arterial bypass and congenital heart disease. Various types of synthetic and natural materials were experimented to produce tissue engineering vascular grafts. In this study, we investigated in vivo tissue engineering technology in miniature pigs to prepare decellularized autologous extracellular matrix-based grafts that could be used as vascular grafts for small-diameter vascular bypass surgery.MethodsAutologous tissue conduits (3.9 mm in diameter) were fabricated by embedding Teflon tubings in the subcutaneous pocket of female miniature pigs (n = 8, body weight 25–30 kg) for 4 weeks. They were then decellularized by CHAPS decellularization solution. Heparin was covalently-linked to decellularized tissue conduits by Sulfo-NHS/EDC. We implanted these decellularized, completely autologous extracellular matrix-based grafts into the carotid arteries of miniature pigs, then sacrificed the pigs at 1 or 2 months after implantation and evaluated the patency rate and explants histologically.ResultsAfter 1 month, the patency rate was 100% (5/5) while the inner diameter of the grafts was 3.43 ± 0.05 mm (n = 5). After 2 months, the patency rate was 67% (2/3) while the inner diameter of the grafts was 2.32 ± 0.14 mm (n = 3). Histological staining confirmed successful cell infiltration, and collagen and elastin deposition in 2-month samples. A monolayer of endothelial cells was observed along the inner lumen while smooth muscle cells were dominant in the graft wall.ConclusionA completely autologous acellular conduit with excellent performance in mechanical properties can be remodeled into a neoartery in a minipig model. This proof-of-concept study in the large animal model is very encouraging and indicates that this is a highly feasible idea worthy of further study in non-human primates before clinical translation.

Constructing artificial urinary conduits: current capabilities and future potential

ABSTRACTIntroduction: Intestinal segments are currently used in reconstructive urology to create urinary diversion after cystectomy. Ileal conduit (IC) is the dominant type of urinary diversion. Nevertheless, IC is not an ideal solution as the procedure still requires entero-enterostomy to restore the bowel continuity. This step is a source of relevant complications that might prolong recovery time. Fabrication of artificial urinary conduit is a tempting idea to introduce an alternative form of urinary diversion which might improve cystectomy outcomes.Area covered: The aim of this review is to discuss available research data about artificial urinary conduit and identify major challenges for future studies.Expert opinion: Fabrication of artificial urinary conduit is in range of current tissue engineering technology but there are still many challenges to overcome. There is an urgent need for studies to be conducted on large animal models with long follow up to expose the limitation of experimental strategies and to gather data for translational research.

Bioprinting Technologies in Tissue Engineering.

Bioprinting technology is a strong tool in producing living functional tissues and organs from cells, biomaterial-based bioinks, and growth factors in computer-controlled platform. The aim of this chapter is to present recent progresses in bioprinting of nerve, skin, cardiac, bone, cartilage, skeletal muscle, and other soft tissues and highlight the challenges in these applications. Various composite bioinks with bioactive ceramic-based scaffolds having patient-specific design and controlled micro-architectures were used at clinical and preclinical applications successfully for regeneration of bone. In nerve tissue engineering, bioprinting of alginate- and gelatin-based gel bioinks by extrusion presented a controllable 3D microstructures and showed satisfactory cytocompatibility and axonal regeneration. Bioprinting of cardiac progenitors in biopolymers resulted in limited success, while the use of bioinks from extracellular matrix induced satisfactory results in cardiac regeneration. Osteochondral scaffold bioprinting is challenging due to the complex hierarchical structure and limited chondral regeneration. Therefore, current approaches focused on osteochondral scaffold with vascular network and mimicking hierarchical structures. The applications of bioprinting in other types of tissues were also studied, and results showed significant potentials in regeneration of tissues such as cornea, liver, and urinary bladder.

The Application Prospect Autologous Bone Marrow Extract Buffy Coat in Cartilage Tissue Engineering: A Mini-Review

Cartilage injury is an important cause of osteoarthritis; however, there is no effective method of cartilage regeneration at present. Therefore, it would be of great value to explore and develop new methods of cartilage regeneration. In recent years, it has become a hot topic on repairing damaged cartilage by using tissue engineered cartilage domestic and abroad. It’s the key factor to success for getting cell source which is effective, applicable and easy to obtain in the technology of cartilage tissue engineering filed. This article summarizes the research progress of bone marrow extract buffy coat in cartilage tissue engineering.

Organ-on-chip in development: Towards a roadmap for organs-on-chip.

Organ-on-Chip is considered a potentially game-changing technology born from the convergence of tissue engineering and microfluidic technology. Organ-on-Chip devices (OoCs) are expected to offer effective solutions to persisting problems in drug development and personalized disease treatments. This opinion paper surveys the current landscape in research, development, application and market opportunities for OoCs towards establishing a global and multi-stakeholder OoC eco-system. Based on panel discussions held at the Vision Workshop (Stuttgart, 23 May 2018) organized by the EU ORCHID consortium, as well as on additional bibliometric study, market analysis and expert interviews conducted within the EU ORCHID project, we outline perceived unmet needs, key challenges, barriers and perspectives of the field. We finally propose recommendations to-wards the definition of a comprehensive roadmap that could render OoCs realistic models of hu-man (patho)physiology in the near future.

Four-Dimensional Bioprinting: Current Developments and Applications in Bone Tissue Engineering

Four-dimensional (4D) bioprinting, in which the concept of time is integrated with three-dimensional (3D) bioprinting as the fourth dimension, is widely proposed as the next-generation of tissue engineering technology as it presents the possibility of constructing complex, functional structures. 4D bioprinting can be used to fabricate dynamic 3D patterned biological architectures that change their shapes or functions under various stimuli by employing stimuli-responsive materials or novel 3D bioprinting strategies, providing unprecedented potential for bone tissue engineering. The shape memory properties of printed structures cater to the need for personal bone defect repair and biomimetic microenvironments that promote the osteogenic differentiation of stem cells. In this review, we introduce the application of different stimuli-responsive biomaterials in tissue engineering and a series of novel 4D bioprinting strategies based on functional transformation of printed structures through the establishment of biomimetic constructs. Furthermore, we discuss the application of 4D bioprinting in bone tissue engineering, as well as future challenges and perspectives.

Biofunctionalization of decellularized porcine aortic valve with OPG-loaded PCL nanoparticles for anti-calcification.

Decellularized valve stents are widely used in tissue-engineered heart valves because they maintain the morphological structure of natural valves, have good histocompatibility and low immunogenicity. However, the surface of the cell valve loses the original endothelial cell coverage, exposing collagen and causing calcification and decay of the valve in advance. In this study, poly ε-caprolactone (PCL) nanoparticles loaded with osteoprotegerin (OPG) were bridged to a decellularized valve using a nanoparticle drug delivery system and tissue engineering technology to construct a new anti-calcification composite valve with sustained release function. The PCL nanoparticles loaded with OPG were prepared via an emulsion solvent evaporation method, which had a particle size of 133 nm and zeta potential of −27.8 mV. Transmission electron microscopy demonstrated that the prepared nanoparticles were round in shape, regular in size, and uniformly distributed, with an encapsulation efficiency of 75%, slow release in vitro, no burst release, no cytotoxicity to BMSCs, and contained OPG nanoparticles in vitro. There was a delay in the differentiation of BMSCs into osteoblasts. The decellularized valve modified by nanoparticles remained intact and its collagen fibers were continuous. After 8 weeks of subcutaneous implantation in rats, the morphological structure of the valve was almost complete, and the composite valve showed anti-calcification ability to a certain extent.

Gingiva-Derived Mesenchymal Stem Cell-Extracellular Vesicles Activate Schwann Cell Repair Phenotype and Promote Nerve Regeneration.

Peripheral nerve injuries (PNIs) are common and debilitating, usually resulting in considerable long-term disability and remaining an unmet clinical need. Even though the combination of mesenchymal stem cells (MSCs) and the state-of-the-art tissue engineering technologies has shown promising therapeutic potentials for PNI, there is still not a single licensed stem cell-based product for peripheral nerve repair/regeneration. Emerging evidence indicates that MSC-derived extracellular vesicles (EVs) are comparably effective as MSCs in the therapy of a variety of disease models or pathological conditions. This report shows that local delivery of gingiva-derived mesenchymal stem cell (GMSC)-derived EVs could obviously promote axonal regeneration and functional recovery of injured mice sciatic nerves. Importantly, the findings suggest that GMSC-derived EVs promoted the expression of Schwann cell dedifferentiation/repair phenotype-related genes in vitro, particularly c-JUN, a key transcription factor that drives the activation of repair phenotype of Schwann cells during PNI and regeneration.

Evaluation of Poly (Carbonate-Urethane) Urea (PCUU) Scaffolds for Urinary Bladder Tissue Engineering.

Although the previous success of bladder tissue engineering demonstrated the feasibility of this technology, most polyester based scaffolds used in previous studies possess inadequate mechanical properties for organs that exhibit large deformation. The present study explored the use of various biodegradable elastomers as scaffolds for bladder tissue engineering and poly (carbonate-urethane) urea (PCUU) scaffolds mimicked urinary bladder mechanics more closely than polyglycerol sebacate-polycaprolactone (PGS-PCL) and poly (ether-urethane) urea (PEUU). The PCUU scaffolds also showed cyto-compatibility as well as increased porosity with increasing stretch indicating its ability to aid in infiltration of smooth muscle cells. Moreover, a bladder outlet obstruction (BOO) rat model was used to test the safety and efficacy of the PCUU scaffolds in treating a voiding dysfunction. Bladder augmentation with PCUU scaffolds led to enhanced survival of the rats and an increase in the bladder capacity and voiding volume over a 3week period, indicating that the high-pressure bladder symptom common to BOO was alleviated. The histological analysis of the explanted scaffold demonstrated smooth muscle cell and connective tissue infiltration. The knowledge gained in the present study should contribute towards future improvement of bladder tissue engineering technology to ultimately aide in the treatment of bladder disorders.

Biomimetic 3D Printing of Hierarchical and Interconnected Porous Hydroxyapatite Structures with High Mechanical Strength for Bone Cell Culture

Human bone demonstrates superior mechanical properties due to its sophisticated hierarchical architecture spanning from the nano/microscopic level to the macroscopic. Bone grafts are in high demand due to the rising number of surgeries because of increasing incidence of orthopedic disorders, non‐union fractures, and injuries in the geriatric population. The bone scaffolds need to provide porous matrix with interconnected porosity for tissue growth as well as sufficient strength to withstand physiological loads, and be compatible with physiological remodeling by osteoclasts/osteoblasts. The‐state‐of‐art additive manufacturing (AM) technologies for bone tissue engineering enable the manipulation of gross geometries, for example, they rely on the gaps between printed materials to create interconnected pores in 3D scaffolds. Herein, the authors firstly print hierarchical and porous hydroxyapatite (HAP) structures with interconnected pores to mimic human bones from microscopic (below 10 µm) to macroscopic (submillimeter to millimeter level) by combining freeze casting and extrusion‐based 3D printing. The compression test of 3D printed scaffold demonstrates superior compressive stress (22 MPa) and strain (4.4%). The human mesenchymal stromal cells (MSCs) tests demonstrate the biocompatibility of printed scaffold.

Engineering a biomimetic integrated scaffold for intervertebral disc replacement

Tissue engineering technology provides a promising alternative to restore physiological functionality of damaged intervertebral disc (IVD). Advanced tissue engineering strategies for IVD have increasingly focused on engineering IVD regions combined the inner nucleus pulposus (NP) and surrounding annulus fibrosus (AF) tissue. However, simulating the cellular and matrix structures and function of the complex structure of IVD is still a critical challenge. Toward this goal, this study engineered a biomimetic AF-NP composite with circumferentially oriented poly(ε-caprolactone) microfibers seeded with AF cells, with an alginate hydrogel encapsulating NP cells as a core. Fluorescent imaging and histological analysis showed that AF cells spread along the circumferentially oriented PCL microfibers and NP cells colonized in the alginate hydrogel similar to native IVD, without obvious migration and mixing between the AF and NP region. Engineered IVD implants showed progressive tissue formation over time after subcutaneous implantation in nude mice, which were indicated by deposition and organization of extracellular matrix and enhanced mechanical properties. In terms of form and function of IVD-like tissue, our engineered biomimetic AF-NP composites have potential application for IVD replacement.

Human breast cancer decellularized scaffolds promote epithelial-to-mesenchymal transitions and stemness of breast cancer cells in vitro.

Breast cancer, with unsatisfactory survival rates, is the leading cause of cancer-related death in women worldwide. Recent advances in the genetic basis of breast cancer have benefitted the development of gene-based medicines and therapies. Tissue engineering technologies, including tissue decellularizations and reconstructions, are potential therapeutic alternatives for cancer research and tissue regeneration. In our study, human breast cancer biopsies were decellularized by a detergent technique, with sodium lauryl ether sulfate (SLES) solution, for the first time. And the decellularization process was optimized to maximally maintain tissue microarchitectures and extracellular matrix (ECM) components with minimal DNA compounds preserved. Histology analysis and DNA quantification results confirmed the decellularization effect with maximal genetic compounds removal. Quantification, immunofluorescence, and histology analyses demonstrated better preservation of ECM components in 0.5% SLES-treated scaffolds. Scaffolds seeded with MCF-7 cells demonstrated the process of cell recellularization in vitro, with increased cell migration, proliferation, and epithelial-to-mesenchymal transition (EMT) process. When treated with 5-fluorouracil, the expressions of stem cell markers, including Oct4, Sox2, and CD49F, were maximally maintained in the recellularized scaffold with decreased apoptosis rates compared with monolayer cells. These results showed that the decellularized breast scaffold model with SLES treatments would help to simulate the pathogenesis of breast cancer in vitro. And we hope that this model could further accelerate the development of effective therapies for breast cancer and benefit drug screenings.

Toughness enhancers for bone scaffold materials based on biocompatible photopolymers

ABSTRACTProviding access to the benefits of additive manufacturing technologies in tissue engineering, vinyl esters recently came into view as appropriate replacements for (meth)acrylates as precursors for photopolymers. Their low cytotoxicity and good biocompatibility as well as favorable degradation behavior are their main assets. Suffering from rather poor mechanical properties, particularly in terms of toughness, several improvements have been made over the last years. Especially, thiol–ene chemistry has been investigated to overcome those shortcomings. In this study, we focused on additional means to further improve the toughness of an already established biocompatible vinyl ester‐thiol formulation, eligible for digital light processing‐based stereolithography. All molecules were based on poly(ε‐caprolactone) as building block and the formulations were tested regarding their reactivity and the resulting mechanical properties. They all performed well as toughness enhancer, ultimately doubling the impact resistance of the reference system. © 2018 The Authors. Journal of Polymer Science Part A: Polymer Chemistry published by Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2019, 57, 110–119

Autologous platelet-rich plasma (PRP) effect on intervertebral disc restoration: an experimental rabbit model.

Platelet-rich plasma (PRP) treatment for intervertebral disc (IVD) repair and tissue engineering technologies have been the target of intense research with promising results. The purpose of this study was to investigate the effect of only one intradiscal injection of PRP in the degenerated rabbit IVD and to assess the restoration process over a 6-week follow-up period. The L3-L4 and L4-L5 discs of 18 adult female rabbits were injured, according to an established degenerative model, with an 18-gauge needle, and classified into two groups: In the discs of group A rabbits, after needle puncture, an intradiscal injection of autologous PRP growth factors was performed, using a 27-gauge needle, and in the discs of the control group (group B), the same procedure was followed by intradiscal injection of normal saline. The PRP preparation was carried out aseptically, after blood collection from the same rabbit. During the 6weeks, there was a noteworthy progression of degeneration process in group B, whereas the grade of degeneration was significantly lower in group A, both for annulus fibrosus (AF) and for nucleus pulposus (NP). The intervertebral disc regeneration and reversal process of the lesions are obvious on 45days after the injury, in group A. The hematoxylin and eosin histology grading score and the expression of collagen type II in NP and inner layer of AF were the markers better mirroring the degeneration and restoration process. PRP intradiscal treatment in degenerative disc disease provokes the maintenance of the disc's basic morphological characteristics with restoration being evident early after injury.

A 3D tumor microenvironment regulates cell proliferation, peritoneal growth and expression patterns

Peritoneal invasion through the mesothelial cell layer is a hallmark of ovarian cancer metastasis. Using tissue engineering technologies, we recreated an ovarian tumor microenvironment replicating this aspect of disease progression. Ovarian cancer cell-laden hydrogels were combined with mesothelial cell-layered melt electrospun written scaffolds and characterized with proliferation and transcriptomic analyses and used as intraperitoneal xenografts. Here we show increased cancer cell proliferation in these 3D co-cultures, which we validated using patient-derived cells and linked to peritoneal tumor growth in vivo. Transcriptome-wide expression analysis identified IGFBP7, PTGS2, VEGFC and FGF2 as bidirectional factors deregulated in 3D co-cultures compared to 3D mono-cultures, which we confirmed by immunohistochemistry of xenograft and patient-derived tumor tissues and correlated with overall and progression-free survival. These factors were further increased upon expression of kallikrein-related proteases. This clinically predictive model allows us to mimic the complexity and processes of the metastatic disease that may lead to therapies that protect from peritoneal invasion or delay the development of metastasis.

The application and research progress of in-situ tissue engineering technology in bone and cartilage repair

To review the application and research progress of in-situ tissue engineering technology in bone and cartilage repair. The original articles about in-situ tissue engineering technology in bone and cartilage repair were extensively reviewed and analyzed. In-situ tissue engineering have been shown to be effective in repairing bone defects and cartilage defects, but biological mechanisms are inadequate. At present, most of researches are mainly focused on animal experiments, and the effect of clinical repair need to be further studied. In-situ tissue engineering technology has wide application prospects in bone and cartilage tissue engineering. However, further study on the mechanism of related cytokines need to be conducted.

Porous decellularized trachea scaffold prepared by a laser micropore technique

Rapid development of tissue engineering technology provides new methods for tracheal cartilage regeneration. However, the current lack of an ideal scaffold makes engineering of trachea cartilage tissue into a three-dimensional (3-D) tubular structure a great challenge. Although a decellularized trachea matrix (DTM) has become a recognized scaffold for trachea cartilage regeneration, it is difficult for cells to detach from or penetrate the matrix because of its non-porous structure. To tackle these problems, a laser micropore technique (LMT) was applied in the current study to enhance trachea sample porosity, and facilitate decellularizing treatment and cell ingrowth. Furthermore, after optimizing LMT and decellularizing treatment parameters, LMT-treated DTM (LDTM) retained its natural tubular structure with only minor extracellular matrix damage. Moreover, compared with DTM, the current study showed that LDTM significantly improved the adherence rate of cells with perfect cell biocompatibility. Moreover, the optimal implantation cell density for chondrogenesis with LDTM was determined to be 1 × 108 cells/ml. Collectively, the results suggest that the novel LDTM is an ideal scaffold for trachea tissue engineering.