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Silicon micromachining to tissue engineer branched vascular channels for liver fabrication.

To date, many approaches to engineering new tissue have emerged and they have all relied on vascularization from the host to provide permanent engraftment and mass transfer of oxygen and nutrients. Although this approach has been useful in many tissues, it has not been as successful in thick, complex tissues, particularly those comprising the large vital organs such as the liver, kidney, and heart. In this study, we report preliminary results using micromachining technologies on silicon and Pyrex surfaces to generate complete vascular systems that may be integrated with engineered tissue before implantation. Using standard photolithography techniques, trench patterns reminiscent of branched architecture of vascular and capillary networks were etched onto silicon and Pyrex surfaces to serve as templates. Hepatocytes and endothelial cells were cultured and subsequently lifted as single-cell monolayers from these two-dimensional molds. Both cell types were viable and proliferative on these surfaces. In addition, hepatocytes maintained albumin production. The lifted monolayers were then folded into compact three-dimensional tissues. Thus, with the use microfabrication technology in tissue engineering, it now seems feasible to consider lifting endothelial cells as branched vascular networks from two-dimensional templates that may ultimately be combined with layers of parenchymal tissue, such as hepatocytes, to form three-dimensional conformations of living vascularized tissue for implantation.

Animals↗

Synthesis and characterization of polypyrrole-hyaluronic acid composite biomaterials for tissue engineering applications.

New tissue engineering technologies will rely on biomaterials that physically support tissue growth and stimulate specific cell functions. The goal of this study was to create a biomaterial that combines inherent biological properties which can specifically trigger desired cellular responses (e.g., angiogenesis) with electrical properties which have been shown to improve the regeneration of several tissues including bone and nerve. To this end, composites of the biologically active polysaccharide hyaluronic acid (HA) and the electrically conducting polymer polypyrrole (PP) were synthesized and characterized. Electrical conductivity of the composite biomaterial (PP/HA) was measured by a four-point probe technique, scanning electron microscopy was used to characterize surface topography, X-ray photoelectron spectroscopy and reflectance infrared spectroscopy were used to evaluate surface and bulk chemistry, and an assay with biotinylated hyaluronic acid binding protein was used to determine surface HA content. PP/HA materials were also evaluated for in vitro cell compatibility and tissue response in rats. Smooth, conductive, HA-containing PP films were produced; these films retained HA on their surfaces for several days in vitro and promoted vascularization in vivo. PP/HA composite biomaterials are promising candidates for tissue engineering and wound-healing applications that may benefit from both electrical stimulation and enhanced vascularization.

Animals↗

Cardiovascular tissue engineering I. Perfusion bioreactors: a review.

Tissue engineering is a fast-evolving field of biomedical science and technology with future promise to manufacture living tissues and organs for replacement, repair, and regeneration of diseased organs. Owing to the specific role of hemodynamics in the development, maintenance, and functioning of the cardiovascular system, bioreactors are a fundamental of cardiovascular tissue engineering. The development of perfusion bioreactor technology for cardiovascular tissue engineering is a direct sequence of previous historic successes in extracorporeal circulation techniques. Bioreactors provide a fluidic environment for tissue engineered tissue and organs, and guarantee their viability, maturation, biomonitoring, testing, storage, and transportation. There are different types of bioreactors and they vary greatly in their size, complexity, and functional capabilities. Although progress in design and functional properties of perfusion bioreactors for tissue engineered blood vessels, heart valves, and myocardial patches is obvious, there are some challenges and insufficiently addressed issues, and room for bioreactor design improvement and performance optimization. These challenges include creating a triple perfusion bioreactor for vascularized tubular tissue engineered cardiac construct; designing and manufacturing fluidics-based perfused minibioreactors; incorporation of systematic mathematical modeling and computer simulation based on computational fluid dynamics into the bioreactor designing process; and development of automatic systems of hydrodynamic regime control. Designing and engineering of built-in noninvasive biomonitoring systems is another important challenge. The optimal and most efficient perfusion and conditioning regime, which accelerates tissue maturation of tissue-engineered constructs also remains to be determined. This is a first article in a series of reviews on critical elements of cardiovascular tissue engineering technology describing the current status, unsolved problems, and challenges of bioreactor technology in cardiovascular tissue engineering and outlining future trends and developments.

Bioreactors↗

Injectable cartilage tissue engineering.

Cartilage is the tissue that lines the surface of bones in articulating joints, allowing painless joint movement. Cartilage loss is an increasingly significant problem, particularly with the ageing of active baby boomers, with few efficacious treatments available at present. Tissue engineering is a field that has evolved over recent years to combat tissue loss by providing a living tissue equivalent or substitute that can mimic the properties of the lost tissue. The general strategy of tissue engineering is to place cells on a biomaterial scaffold that is designed to promote cell function and form new tissue. This review describes the status of materials that are available as injectable scaffolds for tissue engineering and the numerous cell types that can be applied to cartilage repair, including cells derived from cartilage and stem cells. The current state of injectable cartilage tissue engineering and the hurdles that remain for widespread clinical application are discussed.

Animals↗

Review. Hyaluronan: a powerful tissue engineering tool.

Hyaluronan (HA) is a versatile molecular tool with considerable potential for tissue engineering applications. The inclusion of HA has created biocompatible biomaterials and engineered tissues that can be crosslinked or degraded controllably and can facilitate angiogenesis, osteointegration, and cell phenotype preservation. The utility of HA in tissue engineering has been broadened further by the recently identified HA synthases, which can be manipulated to stimulate the endogenous production of HA by cells seeded within biomaterial scaffolds. Overall, HA shows great promise in the development of engineered tissues and biomaterials for a variety of biomedical needs including orthopedic, cardiovascular, pharmacologic, and oncologic applications.

Animals↗

Repair of sheep metatarsus defects by using tissue-engineering technique.

Tissue-engineering bone with porous ,betatricalcium phosphate (3-TCP) ceramic and autologous bone marrow mesenchymal stem cells (MSC) was constructed and the effect of this composite on healing of segmental bone defects was investigated. 10-15 ml bone marrow aspirates were harvested from the iliac crest of sheep, and enriched for MSC by density gradient centrifugation over a Percoll cushion (1. 073 g/ml). After cultured and proliferated, tissue-engineering bones were constructed with these,cellS seeded onto porous f-TCP, and then the constructs were implanted in 8 sheep left metatarsus defect (25 mm in length) as experimental group. Porous ,-TCP only were implanted to bridge same size and position defects in 8 sheep as control group, and 25 mm segmental bone defects of left metatarsus were left empty in 4 sheep as blank group. Sheep were sacrificed on the 6th, 12th, and 24th week postoperatively and the implants samples were examined by radiograph, histology, and biomechanical test. The 4 sheep in blank group were sacrificed on the 24th week postoperatively. The results showed that new bone tissues were observed either radiographic or histologically at the defects of experimental group as early as 6th week postoperatively, but not in control group, and osteoid tissue, woven bone and lamellar bone occurred earlier than in control group in which the bone defects were repaired in "creep substitution" way, because of the new bone formed in direct manner without progression through a cartilaginous intermediate. At the 24th week, radiographs and biomechanical test revealed an almost complete repair of the defect of experimental group, only partly in control group. The bone defects in blank group were non-healing at the 24th week. It was concluded that engineering bones constructed with porous -TCP and autologous MSC were capable of repairing segmental bone defects in sheep metatarsus beyond "creep substitution" way and making it healed earlier. Porous ,-TCP being constituted with autologous MSC may be a good option in healing critical segmental bone defects in clinical practice and provide insight for future clinical repair of segmental defect.

Animals↗

Informatics challenges in tissue engineering and biomaterials.

Both tissue engineering and biomaterials have made tremendous strides recently, yet major questions remain unanswered. Tissue-engineered products have come to the market; others are in development. A fundamental issue that informatics could address for tissue engineering is to describe and to predict the cascade of biochemical and cellular reactions that occur as a function of time and implant material: surface texture, microporosity; pore size, density, and connectivity; and three-dimensional configuration. Behavior of ceramics, a subset of tissue-engineering scaffold materials and a mainstay of dental restorations, has been studied extensively for very thin layers and for thicknesses greater than 2 mm. Until recently, little has been known about dentally relevant thickness of 1-2 mm. Results have been surprising and are continuing to develop. Still, at least one fundamental question remains that could be addressed by informatics techniques: Where, along the spectrum of flat-polished material to 10-year clinical in vivo study, can we test to predict clinical performance of all-ceramic crowns accurately?

Biocompatible Materials↗

Directions in cardiovascular tissue engineering.

The generation of tissue replacements or supplements for diseased tissue within the cardiovascular system has been the target of recent tissue engineering efforts. While clinically applicable methodologies remain to be achieved, important foundational experimentation has been performed in recent years to begin the move toward engineered tissue replacement therapy. Inadequacies of current valve, vessel, and other heart prostheses are reviewed briefly, followed by the discussion of selected progress in the supplementation or replacement of each cardiovascular component with tissue constructs. Topics addressed include the endothelialization of bio-prosthetic valves and synthetic vascular grafts, the generation of tissue valve leaflets and vascular conduits, the genetic manipulation of endothelial cells with implications for graft endothelialization, and cardiomyoplasty achieved through cellular and genetic means.

Animals↗

Advances in tissue engineering of blood vessels and other tissues.

Tissue engineering is a new and rapidly expanding field, in which techniques are being developed for culturing a variety of tissues both in vitro and in vivo using polymer 'scaffolds' to support tissue growth. Polymer scaffolds used in tissue engineering are generally biodegradable, often involving compounds which are already approved for human implantation. In some cases, these polymers may be chemically modified to exhibit selective cell adhesion properties, which enhance cell attachment and subsequent tissue growth. Many cell types have been successfully cultured on these scaffolds, including smooth muscle cells, endothelial cells, hepatocytes and chondrocytes. Tissue engineering holds the potential for the in vitro development of autologous or allogeneic transplantable vascular conduits. Each year in the USA, there are approximately 1.4 million procedures performed which require arterial prostheses. Most of these procedures are in small calibre (< 6 mm) vessels, for which synthetic graft materials are not generally suitable. While autologous venous or arterial vessels are generally used, not all patients possess adequate conduit for revascularization. Tubular scaffolds have been specially designed for culturing small calibre arteries in vitro. Bovine aortic vascular cells were seeded and cultured on these polymer scaffolds, and grown under conditions of pulsatile pressure and intra-luminal flow. To minimize contamination during the weeks of tissue culture required to produce an arterial prosthesis, a sterile incubator system was developed. Preliminary studies have achieved good cell densities of both smooth muscle cells and endothelial cells on biodegradable polymer scaffolds.

Animals↗

[Advances in research and development of tissue engineering].

OBJECTIVE: From the point of view of material science, the methods of tissue repair and defect reconstruct were discussed, including mesenchymal stem cells (MSCs), growth factors, gene therapy and tissue engineered tissue. METHODS: The advances in tissue engineering technologies were introduced based on the recent literature. RESULTS: Tissue engineering should solve the design and preparation of molecular scaffold, tissue vascularization and dynamic culture of cell on the scaffolds in vitro. CONCLUSION: Biomaterials play an important role in the tissue engineering. They can be used as the matrices of MSCs, the delivery carrier of growth factor, the culture scaffold of cell in bioreactors and delivery carrier of gene encoding growth factors.

Biocompatible Materials↗

Evolving concepts in bone tissue engineering.

The field of tissue engineering integrates the latest advances in molecular biology, biochemistry, engineering, material science, and medical transplantation. Researchers in the developing field of regenerative medicine have identified bone tissue engineering as an attractive translational target. Clinical problems requiring bone regeneration are diverse, and no single regeneration approach will likely resolve all defects. Recent advances in the field of tissue engineering have included the use of sophisticated biocompatible scaffolds, new postnatal multipotent cell populations, and the appropriate cellular stimulation. In particular, synthetic polymer scaffolds allow for fast and reproducible construction, while still retaining biocompatible characteristics. These criteria relate to the immediate goal of determining the ideal implant. The search is becoming a reality with widespread availability of biocompatible scaffolds; however, the desired parameters have not been clearly defined. Currently, most research focuses on the use of bone morphogenetic proteins (BMPs), specifically BMP-2 and BMP-7. These proteins induce osteogenic differentiation in vitro, as well as bone defect healing in vivo. Protein-scaffold interactions that enhance BMP binding are of the utmost importance, since prolonged BMP release creates the most osteogenic microenvironment. Transition into clinical studies has had only mild success and relies on large doses of BMPs for bone formation. Advances within the field of bone tissue engineering will likely overcome these challenges and lead to more clinically relevant therapies.

Animals↗

Extracellular stimulation in tissue engineering.

The field of tissue engineering has created a need for biomaterials that are capable of providing biofunctional and structural support for living cells outside of the body. Most of the commonly used biomaterials in tissue engineering are designed based on their physicochemical properties, thus achieving precise control over mechanical strength, compliance, porosity, and degradation kinetics. Biofunctional signals are added to the scaffold by tethering, immobilizing, or supplementing biofunctional macromolecules, such as growth factors, directly to the scaffold material. The challenge in tissue engineering remains to find the correct balance between the biofunctional and the physical properties of the scaffold material for each application. Moreover, the ability to modulate communication between cells and the extracellular environment using the engineered scaffold as the actuator can provide a significant advantage in tissue engineering. In this study, a unique scaffold material is presented. The material interchangeably combines biofunctional and structural molecules by fusing the two into a single backbone macromolecule. This integration provides the basis for practical, effective, and high-resolution control of both the biofunctional and the physical properties of the scaffold material. This new scaffold material has proven effective with smooth muscle, cardiac, cartilage, and human embryonic stem cell cultures. The advantages of this approach as well as the potential applications of this unique scaffold material are discussed.

Animals↗

Functional tissue engineering: the role of biomechanics.

"Tissue engineering" uses implanted cells, scaffolds, DNA, protein, and/or protein fragments to replace or repair injured or diseased tissues and organs. Despite its early success, tissue engineers have faced challenges in repairing or replacing tissues that serve a predominantly biomechanical function. An evolving discipline called "functional tissue engineering" (FTE) seeks to address these challenges. In this paper, the authors present principles of functional tissue engineering that should be addressed when engineering repairs and replacements for load-bearing structures. First, in vivo stress/strain histories need to be measured for a variety of activities. These in vivo data provide mechanical thresholds that tissue repairs/replacements will likely encounter after surgery. Second, the mechanical properties of the native tissues must be established for subfailure and failure conditions. These "baseline data" provide parameters within the expected thresholds for different in vivo activities and beyond these levels if safety factors are to be incorporated. Third, a subset of these mechanical properties must be selected and prioritized. This subset is important, given that the mechanical properties of the designs are not expected to completely duplicate the properties of the native tissues. Fourth, standards must be set when evaluating the repairs/replacements after surgery so as to determine, "how good is good enough?" Some aspects of the repair outcome may be inferior, but other mechanical characteristics of the repairs and replacements might be suitable. New and improved methods must also be developed for assessing the function of engineered tissues. Fifth, the effects of physical factors on cellular activity must be determined in engineered tissues. Knowing these signals may shorten the iterations required to replace a tissue successfully and direct cellular activity and phenotype toward a desired end goal. Finally, to effect a better repair outcome, cell-matrix implants may benefit from being mechanically stimulated using in vitro "bioreactors" prior to implantation. Increasing evidence suggests that mechanical stress, as well as other physical factors, may significantly increase the biosynthetic activity of cells in bioartificial matrices. Incorporating each of these principles of functional tissue engineering should result in safer and more efficacious repairs and replacements for the surgeon and patient.

Biocompatible Materials↗

Tissue engineering of artificial organs.

Tissue engineering efforts are currently being undertaken for every type of tissue and organ within the urinary system. Most of the effort expended to engineer genitourinary tissues has occurred within the last decade. Tissue engineering techniques require expertise in growth factor biology, a cell culture facility designed for human application, and personnel who have mastered the techniques of cell harvest, culture, and expansion. Polymer scaffold design and manufacturing resources are essential for the successful application of this technology. In order to apply these engineering techniques to humans, further studies need to be performed with many of the tissues described. The first human application of cell-based tissue engineering technology for urologic applications took place at our institution, with the injection of autologous cells for the correction of vesicoureteral reflux in children. The same technology has been expanded to treat adult patients with urinary incontinence. Trials of urethral tissue replacement with processed collagen matrices are in progress, and bladder replacement using tissue engineering techniques are currently being arranged. Recent progress suggests that engineered urologic tissues may have clinical applicability in the future.

Artificial Organs↗

Tissue engineering scheming by artificial intelligence.

Tissue engineers are often confused when seeking the most effective, economical and secure scheme for tissue engineering. The aim of this study is to generate tissue engineering schemes with artificial intelligence instead of human intelligence. The experimental data of tissue engineered cartilage were integrated and standardized with a centralized database, and a scheme engine was developed using artificial intelligent methods (artificial neural networks and decision trees). The scheme engine was trained with existing cases in the database, and then was used to generate tissue engineering schemes for new experimental animals. Following the schemes generated by the artificial intelligent system, we cured 18 of the 20 experimental animals. In conclusion, artificial intelligence is a powerful method for decision making in the tissue engineering realm.

Algorithms↗

Viral vectors for gene delivery in tissue engineering.

The goal of tissue engineering is the production of functional, biocompatible tissues by seeding cells within biological or synthetic scaffolds. One tissue engineering approach involves the genetic modification of cells that are seeded onto (or into) scaffolds prior to implantation. The genetic modification is achieved through gene delivery, with can utilize viral transduction or non-viral transfection systems. Although novel non-viral systems have continued to emerge as innovative vehicles for controlled gene delivery, viruses remain the most efficient means by which exogenous genes can be introduced into and expressed by mammalian cells. Retrovirus, adenovirus, adeno-associated virus and herpes virus are widely studied viral gene transfer systems and have attracted the most attention in the field of transduction. This review thoroughly discusses the genomic structures of each virus type, along with the advantages and disadvantages of their use in tissue engineering applications.

Adenoviridae↗

Tissue engineering: implications in the treatment of organ and tissue defects.

The loss or failure of organs and tissues due to injury/trauma or ageing is one of the major causes for concern in human health care. Tissue engineering, a new emerging field in modern biomedical sciences, promises a new ray of hope for transplantation biology by creating viable substitutes for such failing organs or tissues. Tissue engineering is rapidly growing in both scope and importance within biomedical engineering. It represents the marriage of rapid developments in cellular and molecular biology on the one hand and materials, chemical and mechanical engineering on the other. The ability to manipulate and reconstitute tissue function has tremendous clinical implications and is likely to play a key role in cell and gene therapies in the coming years. This article discusses the basic principles of tissue engineering and some of the attempts made to provide solutions by engineering tissues in vitro.

Aged↗

Adipose tissue engineering: the future of breast and soft tissue reconstruction following tumor resection.

Reconstructive surgeons have always been at the forefront of medical technology. The history of reconstructive surgery began with ablative surgery, which was followed by tissue and organ transplantation, leading to contemporary tissue reconstruction. The field of reconstructive surgery is poised at the next stage of its evolution, namely tissue regeneration. The field of tissue engineering has largely defined this evolutionary leap. One active area of investigation is the development of tissue engineering strategies for adipose tissue. Bioengineers, life scientists, and reconstructive surgeons are synergistically coupling expertise in areas such as cell culture technology, tissue transfer, cell differentiation, angiogenesis, computer modeling, and polymer chemistry to regenerate adipose tissue de novo for breast replacement and soft-tissue augmentation following tumor resection. This work presents the current state of the art in adipose tissue engineering, as well the clinically translatable strategies currently under development. Semin. Surg. Oncol. 19:302-311, 2000.

Adipose Tissue↗