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Potential of nanofiber matrix as tissue-engineering scaffolds.

Tissue-engineering scaffolds should be analogous to native extracellular matrix (ECM) in terms of both chemical composition and physical structure. Polymeric nanofiber matrix is similar, with its nanoscaled nonwoven fibrous ECM proteins, and thus is a candidate ECM-mimetic material. Techniques such as electrospinning to produce polymeric nanofibers have stimulated researchers to explore the application of nanofiber matrix as a tissue-engineering scaffold. This review covers the preparation and modification of polymeric nanofiber matrix in the development of future tissue-engineering scaffolds. Major emphasis is also given to the development and applications of aligned, core shell-structured, or surface-functionalized polymer nanofibers. The potential application of polymer nanofibers extends far beyond tissue engineering. Owing to their high surface area, functionalized polymer nanofibers will find broad applications as drug delivery carriers, biosensors, and molecular filtration membranes in future.

Animals↗

Tissue engineering: prospect for regenerating periodontal tissues.

New advancements in technological fields, continually has had a major impact on dental practice. The emergence of tissue engineering and biomimetic concepts has enhanced the predictability of existing therapy and also has radically recast approaches towards the dentoalveolar reconstruction. Tissue engineering in the simplest sense is a combination of material sciences and biology to repair tissues and organs which will unquestionably offer an exciting therapeutic alternative that have never been available before. This article is a brief introduction to the ever expanding field of tissue engineering and its possible implication in periodontal regeneration.

Cell Transplantation↗

Manufacture and evaluation of bioactive and biodegradable materials and scaffolds for tissue engineering.

For tissue regeneration and tissue engineering applications, a number of bioactive and biodegradable composites, either porous or non-porous, were fabricated. The newly developed materials included tricalcium phosphate reinforced polyhydroxybutyrate and its copolymer, poorly crystallized hydroxyapatite reinforced chitin, and plasma sprayed hydroxyapatite reinforced poly(L-lactic acid). It was shown that these new materials could be successfully produced using the manufacturing techniques adopted. In vitro experiments revealed that the incorporation of bioceramic particles in biodegradable polymers rendered the composites bioactive and significantly improved the ability of composites to induce the formation of bone-like apatite on their surfaces. Degradation of composite scaffolds in simulated body fluid was observed and could be due to the simultaneous degradation of polymer matrix and dissolution of bioceramic particles.

Journal Article↗

[Advances in research on calcium polyphosphate bioceramic for bone tissue engineering scaffold].

Bone tissue engineering is a novel, developing and challenging science which provides a new way to repair bone lost from injury and disease. Porous calcium polyphosphate bioceramic is one kind of absorptable bioceramic. Owing to its fine biocompatibility and degradability, more and more pieces of research wark have been carried out in bone tissue engineering, and because of its special characteristics, calcium polyphosphate bioceramic is regarded as a promising material for solving the problem of how to match the degradation velocity of scaffold with the velocity of cell growth. The recent research of using calcium polyphosphate bioceramic as the scaffold in bone tissue engineering is summarized, including the property, synthesis and advances.

Bone Substitutes↗

A useful marker for evaluating tissue-engineered products: gap-junctional communication for assessment of the tumor-promoting action and disruption of cell differentiation in tissue-engineered products.

An in vitro system for evaluating the safety of tissue-engineered products is a convenient because of its rapidity and low cost. On the basis of recent studies, intercellular channels called gap-junctions are considered to play an important role on the tumor-promotion stage during the tumorigenesis induced by polyurethanes. Further, we also demonstrate the significance of the intercellular communication during neuronal cell differentiation. From these results, we propose a survey of the function of the gap-junctional communication as a probable useful marker for evaluating the safety of tissue-engineered products.

Animals↗

Design and analysis of tissue engineering scaffolds that mimic soft tissue mechanical anisotropy.

Tissue engineered constructs must exhibit tissue-like functional properties, including mechanical behavior comparable to the native tissues they are intended to replace. Moreover, the ability to reversibly undergo large strains can help to promote and guide tissue growth. Electrospun poly (ester urethane) ureas (ES-PEUU) are elastomeric and allow for the control of fiber diameter, porosity, and degradation rate. ES-PEUU scaffolds can be fabricated to have a well-aligned fiber network, which is important for applications involving mechanically anisotropic soft tissues. We have developed ES-PEUU scaffolds under variable speed conditions and modeled the effects of fiber orientation on the macro-mechanical properties of the scaffold. To illustrate the ability to simulate native tissue mechanical behavior, we demonstrated that the high velocity spun scaffolds exhibited highly anisotropic mechanical properties closely resembling the native pulmonary heart valve leaflet. Moreover, use of the present fiber-level structural constitutive model allows for the determination of electrospinning conditions to tailor ES-PEUU scaffolds for specific soft tissue applications. The results of this study will help to provide the basis for rationally designed mechanically anisotropic soft tissue engineered implants.

Anisotropy↗

Electrospun degradable polyesterurethane membranes: potential scaffolds for skeletal muscle tissue engineering.

Skeletal muscle tissue engineering represents an attractive approach to overcome problems associated with autologous transfer of muscle tissue and provides a valid alternative in muscle regeneration enhancement. The aim of this study was to investigate the suitability, as scaffold for skeletal muscle tissue engineering, of a known biodegradable block copolymer (DegraPol) processed by electrospinning in the novel form of microfibrous membranes. Scaffolds were characterized with reference to their morphological, degradative and mechanical properties. Subsequently, cell viability, adhesion and differentiation on coated and uncoated DegraPol) slides were investigated using line cells (C2C12 and L6) and primary human satellite cells (HSCs). The membranes exhibited absence of toxic residuals and satisfactory mechanical properties (linear elastic behavior up to 10% deformation, E modulus in the order of magnitude of MPa). A promising cellular response was also found in preliminary experiments: both line cells and HSCs adhered, proliferated and fused on differently coated electrospun membranes. Positive staining for myosin heavy chain expression indicated that differentiation of C2C12 multinucleated cells occurred within the porous elastomeric substrate. Together the results of this study provide significant evidence of the suitability of electrospun DegraPol) membranes as scaffolds for skeletal muscle tissue engineering and that they represent a promising alternative to scaffolds currently used in this field.

Cell Adhesion↗

Microrobotics and MEMS-based fabrication techniques for scaffold-based tissue engineering.

Scaffold based tissue engineering strategies use cells, biomolecules and a scaffold to promote the repair and regeneration of tissues. Although scaffold-based tissue engineering approaches are being actively developed, most are still experimental, and it is not yet clear what defines an ideal scaffold/cell construct. Solid free form fabrication (SFF) techniques can precisely control matrix architecture (size, shape, interconnectivity, branching, geometry and orientation). The SFF methods enable the fabrication of scaffolds with various designs and material compositions, thus providing a control of mechanical properties, biological effects and degradation kinetics. This paper reviews the application of micro-robotics and MEMS-based fabrication techniques for scaffold design and fabrication. It also presents a novel robotic technique to fabricate scaffold/cell constructs for tissue engineering by the assembly of microscopic building blocks.

Biocompatible Materials↗

[Bone tissue engineering scaffolds].

Bone tissue engineering may provide an alternative to the repairs to skeletal defects resulting from disease, trauma or surgery. Scaffold has played an important role in bone tissue engineering, which functions as the architecture for bone in growth. In this paper, the authors gave a brief introduction about the requirement of bone tissue engineering scaffold, the key of the design of scaffolds and the current research on this subject.

Biodegradation, Environmental↗

Growth factor delivery for tissue engineering.

A tissue-engineered implant is a biologic-biomaterial combination in which some component of tissue has been combined with a biomaterial to create a device for the restoration or modification of tissue or organ function. Specific growth factors, released from a delivery device or from co-transplanted cells, would aid in the induction of host parenchymal cell infiltration and improve engraftment of co-delivered cells for more efficient tissue regeneration or ameliorate disease states. The characteristic properties of growth factors are described to provide a biological basis for their use in tissue engineered devices. The principles of polymeric device development for therapeutic growth factor delivery in the context of tissue engineering are outlined. A review of experimental evidence illustrates examples of growth factor delivery from devices such as microparticles, scaffolds, and encapsulated cells, for their use in the application areas of musculoskeletal tissue, neural tissue, and hepatic tissue.

Animals↗

Comparative study of cellular and extracellular matrix composition of native and tissue engineered heart valves.

Tissue engineering of heart valves utilizes biodegradable or metabolizable scaffolds for remodeling by seeded autologous cells. The aim of this study was to determine and compare extracellular matrix (ECM) formations, cellular phenotypes and cell location of native and tissue engineered (TE) valve leaflets. Ovine carotid arteries, ovine and porcine hearts were obtained from slaughterhouses. Cells were isolated from carotid arteries and dissected ovine, porcine and TE leaflets. TE constructs were fabricated from decellularized porcine pulmonary valves, seeded ovine arterial cells and subsequent 16 days dynamic in vitro culture using a pulsatile bioreactor. Native and TE valves were studied by histology (hematoxylin-eosin, resorcin-fuchsin, Movat pentachrome), NIR femtosecond multiphoton laser scanning microscopy and scanning electron microscopy (SEM). Cells of native and TE tissues were identified and localized by immunohistochemistry. Arterial, valvular and re-isolated TE-construct cells were processed for immunocytochemistry and Western blotting. ECM analysis and SEM revealed characteristical and comparable structures in native and TE leaflets. Most cells in native leaflets stained strongly positive for vimentin. Cells positive to alpha-smooth muscle actin (alpha-SMA), myosin and calponin were only found at the ventricular (inflow) side of ovine aortic and porcine pulmonary valve leaflets. Cells from TE constructs had a strong expression of vimentin, alpha-SMA, myosin, calponin and h-caldesmon throughout the entire leaflet. Comparable ECM formation and endothelial cell lining of native and TE leaflets could be demonstrated. However, immunostaining revealed significant differences between valvular cell phenotypes of native and TE leaflets. These results may be essential for further cardiovascular tissue engineering efforts.

Animals↗

Biomechanical and biochemical characterization of composite tissue-engineered intervertebral discs.

Composite tissue-engineered intervertebral tissue was assembled in the shape of cylindrical disks composed of an outer shell of PGA mesh seeded with annulus fibrosus cells with an inner core of nucleus pulposus cells seeded into an alginate gel. Samples were implanted subcutaneously in athymic mice and retrieved at time points up to 16 weeks. At all retrieval times, samples maintained shape and contained regions of distinct tissue formation. Histology revealed progressive tissue formation with distinct morphological differences in tissue formation in regions seeded with annulus fibrosus and nucleus pulposus cells. Biochemical analysis indicated that DNA, proteoglycan, and collagen content in tissue-engineered discs increased with time, reaching >50% of the levels of native tissue by 16 weeks. The exception to this was the collagen content of the nucleus pulposus portion of the implants with were approximately 15% of native values. The equilibrium modulus of tissue-engineered discs was 49.0+/-13.2 kPa at 16 weeks, which was between the measured values for the modulus of annulus fibrosus and nucleus pulposus. The hydraulic permeability of tissue-engineered discs was 5.1+/-1.7x10(-14) m2/Pa at 16 weeks, which was between the measured values for the hydraulic permeability of annulus fibrosus and nucleus pulposus. These studies document the feasibility of creating composite tissue-engineered intevertebral disc implants with similar composition and mechanical properties to native tissue.

Alginates↗

The potency of culture-expanded nasal septum chondrocytes for tissue engineering of cartilage.

Tissue engineering techniques to create extra autologous cartilage for reconstructive surgery receive more and more scientific and industrial attention. The objective of this experimental study was to assess the use of in vitro multiplied chondrocytes of the nasal septum for generation of cartilage grafts using tissue engineering techniques. Cells isolated from a biopsy of septal cartilage of rabbits and humans were expanded in culture to get a sufficient number of cells to engineer a cartilage graft. The drawback of the expansion procedure is that the cells lose their cartilaginous phenotype (dedifferentiation). We studied a method to reverse the dedifferentiation of expanded cells to stimulate them to produce cartilage matrix of good quality. Rabbit chondrocytes showed reversion of dedifferentiation (redifferentiation) when fetal calf serum was replaced by the growth factors IGF1 and TGFbeta2. This was expressed by increased glycosaminoglycan synthesis and increased numbers of collagen type II-producing cells. The redifferentiation capacity of septal cartilage cells of young rabbits was higher than that of adult rabbits. In human chondrocytes from the nasal septum redifferentiation could also be induced by replacement of serum with IGF1 and TGFbeta2. This method, however, was less efficient than in rabbits. Chondrocytes of older patients (>40 years old) were no longer sensitive to the growth factor treatment. In conclusion, our study demonstrates a method to regain cartilage phenotype in multiplied cells of nasal septum cartilage needed for tissue engineering of new cartilage. These results are promising for this technique to generate cartilage grafts for facial plastic surgery of the nasal septum.

Adult↗

[Primary experimental study on the construction of tissue engineering blood vessel].

OBJECTIVE: To explore the primary experimental methods to construct tissue engineering blood vessel. METHODS: Using the collagen-chitosan to prefabricate the scaffolds with 3-dimensional structure, the proliferated human endothelial cells (ECs), smooth muscle cells (SMCs) and fibroblasts act as the seed cells. The cells were seeded to scaffolds in two-step method, and engineering tissue were matured by static culture or bioreactor culture. Extracellular matrix contents and the platelet aggregation were examined in engineering tissue, tissue engineering blood vessels were taken as patches to repair the man-made defaults on the rats aorta. RESULTS: The proliferated human ECs, SMCs and fibroblast can hold activity and act as seed cells. The prefabricated scaffolds, with excellently cell and tissue biocompatibility, can facilitated cells adherence and upgrowth, the cells quantities and extracellular matrix contents in engineered tissue are time dependent increase (P < 0.05). Platelet aggregation tests confirm the tissue engineering blood vessel have some anti-coagulability. Using the engineering tissue patch to repair the default, 6 aortas in 8 animal were patency till 10 days post-operation. CONCLUSIONS: The seeding cells can be seeded on the 3-dimensional collagen-chitosan scaffolds and matured, the tissue engineering blood vessel can be constructed primarily.

Animals↗

Interface tissue engineering and the formulation of multiple-tissue systems.

Interface tissue engineering is an exciting field which focuses on the development of tissue engineered grafts capable of promoting integration between different types of tissue and between the implant and surrounding tissue. Focusing on interface tissue engineering, and using the insertion site between the anterior cruciate ligament and bone as an example, this chapter discusses strategies in soft tissue to bone integration as well as current tissue engineering efforts in this area. This review begins with the clinical significance of this problem, followed by a review of existing fixation methods, and tissue engineering efforts aimed at addressing this critical issue. The development of multiphased scaffolds designed for the replacement of more than one type of tissue, as well as novel in vitro co-culture systems will be introduced. Future directions in the field of interface tissue engineering will also be discussed.

Animals↗

Angiogenesis in tissue engineering: breathing life into constructed tissue substitutes.

Long-term function of three-dimensional (3D) tissue constructs depends on adequate vascularization after implantation. Accordingly, research in tissue engineering has focused on the analysis of angiogenesis. For this purpose, 2 sophisticated in vivo models (the chorioallantoic membrane and the dorsal skinfold chamber) have recently been introduced in tissue engineering research, allowing a more detailed analysis of angiogenic dysfunction and engraftment failure. To achieve vascularization of tissue constructs, several approaches are currently under investigation. These include the modification of biomaterial properties of scaffolds and the stimulation of blood vessel development and maturation by different growth factors using slow-release devices through pre-encapsulated microspheres. Moreover, new microvascular networks in tissue substitutes can be engineered by using endothelial cells and stem cells or by creating arteriovenous shunt loops. Nonetheless, the currently used techniques are not sufficient to induce the rapid vascularization necessary for an adequate cellular oxygen supply. Thus, future directions of research should focus on the creation of microvascular networks within 3D tissue constructs in vitro before implantation or by co-stimulation of angiogenesis and parenchymal cell proliferation to engineer the vascularized tissue substitute in situ.

Animals↗

Fibrin gel as a three dimensional matrix in cardiovascular tissue engineering.

OBJECTIVE: In tissue engineering, three-dimensional biodegradable scaffolds are generally used as a basic structure for cell anchorage, cell proliferation and cell differentiation. The currently used biodegradable scaffolds in cardiovascular tissue engineering are potentially immunogenic, they show toxic degradation and inflammatory reactions. The aim of this study is to establish a new three-dimensional cell culture system within cells achieve uniform distribution and quick tissue development and with no toxic degradation or inflammatory reactions. METHODS: Human aortic tissue is harvested from the ascending aorta in the operation room and worked up to pure human myofibroblasts cultures. These human myofibroblasts cultures are suspended in fibrinogen solution and seeded into 6-well culture plates for cell development for 4 weeks and supplemented with different concentrations of aprotinin. Hydroxyproline assay and histological studies were performed to evaluate the tissue development in these fibrin gel structures. RESULTS: The light microscopy and the transmission electron microscopy studies for tissue development based on the three-dimensional fibrin gel structures showed homogenous cell growth and confluent collagen production. No toxic degradation or inflammatory reactions could be detected. Furthermore, fibrin gel myofibroblasts structures dissolved within 2 days in medium without aprotinin, but medium supplemented with higher concentration of aprotinin retained the three-dimensional structure and had a higher collagen content (P<0.005) and a better tissue development. CONCLUSIONS: A three-dimensional fibrin gel structure can serve as a useful scaffold for tissue engineering with controlled degradation, excellent seeding effects and good tissue development.

Aorta↗