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PubMed2004

The application of recombinant human collagen in tissue engineering.

Collagen is the main structural protein in vertebrates. It plays an essential role in providing a scaffold for cellular support and thereby affecting cell attachment, migration, proliferation, differentiation, and survival. As such, it also plays an important role in numerous approaches to the engineering of human tissues for medical applications related to tissue, bone, and skin repair and reconstruction. Currently, the collagen used in tissue engineering applications is derived from animal tissues, creating concerns related to the quality, purity, and predictability of its performance. It also carries the risk of transmission of infectious agents and precipitating immunological reactions. The recent development of recombinant sources of human collagen provides a reliable, predictable and chemically defined source of purified human collagens that is free of animal components. The triple-helical collagens made by recombinant technology have the same amino acid sequence as human tissue-derived collagen. Furthermore, by achieving the equivalent extent of proline hydroxylation via coexpression of genes encoding prolyl hydroxylase with the collagen genes, one can produce collagens with a similar degree of stability as naturally occurring material. The recombinant production process of collagen involves the generation of single triple-helical molecules that are then used to construct more complex three-dimensional structures. If one loosely defines tissue engineering as the use of a biocompatible scaffold combined with a biologically active agent (be it a gene or gene construct, growth factor or other biologically active agent) to induce tissue regeneration, then the production of recombinant human collagen enables the engineering of human tissue based on a human matrix or scaffold. Recombinant human collagens are an efficient scaffold for bone repair when combined with a recombinant bone morphogenetic protein in a porous, sponge-like format, and when presented as a membrane, sponge or gel can serve as a basis for the engineering of skin, cartilage and periodontal ligament, depending on the specific requirements of the chosen application.

Animals↗

PubMed2005

Vascular patches tissue-engineered with autologous bone marrow-derived cells and decellularized tissue matrices.

Synthetic polymer vascular patches used in cardiovascular surgery have shortcomings such as thrombosis, intimal hyperplasia, calcification, infection, and no growth potential. Tissue-engineered vascular patches using autologous vascular cells may solve these problems. In this study, we developed a tissue-engineered vascular patch using autologous bone marrow-derived cells (BMCs) and decellularized tissue matrices. Vascular smooth muscle cells and endothelial cells were differentiated from bone marrow mononuclear cells in vitro. Tissue-engineered vascular patches were fabricated by seeding these cells onto decellularized canine inferior vena cava matrices and implanted into the inferior vena cava of dogs. Three weeks after implantation, the tissue-engineered vascular patches were patent with no sign of thrombus formation. Histological, immunohistochemical, and electron microscopic analyses of the vascular patches retrieved 3 weeks after implantation revealed regeneration of endothelium and smooth muscle and the presence of collagen and elastin. BMCs labeled with a fluorescent dye prior to implantation were detected in the retrieved vascular patches, indicating that the BMCs survived after implantation and contributed to the vascular tissue regeneration. This study demonstrates that vascular patches can be tissue-engineered with autologous BMCs and decellularized tissue matrices.

Animals↗

PubMed2005

Prediction of vascular tissue engineering results with artificial neural networks.

Tissue engineers are often confused on finding the most successful strategy for specific patient. In this study, we used artificial neural networks to predict the outcomes of different vascular tissue engineering strategies, thus providing advisory information for experimental designers. Over 30 variables were used as features of the tissue engineering strategies. Different architectures of artificial neural networks with back propagation algorithm were tested to obtain the best model configuration for the prediction of the tissue engineering strategies. In the computational experiments, the artificial neural networks with one and two hidden layers could, respectively, detect unsuccessful strategies with the highest predictive accuracy of 91.45 and 94.24%. In conclusion, artificial intelligence has great potential in tissue engineering decision support. It can provide accurate advisory information for tissue engineers, thus reducing failures and improving therapeutic effects.

Animals↗

PubMed2006

Treatment of rabbit growth plate injuries with an autologous tissue-engineered composite. An experimental study.

Tissue engineering has become a promising way of treating growth plate injuries. In this study, we attempted investigating the role of the autologous tissue-engineered composite in the treatment of rabbit growth plate injuries. Growth plate chondrocytes from iliac crest epiphyseal cartilage of immature New Zealand rabbits were obtained by dissection and sequential digestion with 0.2% collagenase (type II). After proliferating in monolayer culture in vitro for 3 weeks, the cells were harvested and seeded onto the demineralized bone matrix (DBM) scaffold to construct the composite. The autologous tissue-engineered composites were finally implanted into the proximal right tibia defects of the growth plate created in 12 rabbits (group A underwent the operation after obtaining chondrocytes 3 weeks beforehand), another 12 rabbits were implanted with only the DBM scaffold (group B), and the defects in group C (12 rabbits) were not implanted. The left tibias of all animals were left undone as the normal control. Two weeks after the operation, severe shortness and angulation deformity of the right tibia evaluated by X-ray were gradually observed in groups B and C. However, there were no obvious changes in group A and there were significant differences between group A and groups B and C (p < 0.05) at the 4-, 8-, and 16-weeks time points. 16 weeks after operation, histological examination revealed that the defects of the right tibias in group A had restored to almost the normal columnar structure of the growth plate. The results demonstrate that tissue-engineered composite established by combination of autologous growth plate chondrocytes and DBM can prevent the formation of a bone bridge and restore the growth of damaged growth plate.

Animals↗

PubMed2005

Stem cells, tissue engineering and organogenesis in transplantation.

Tissue engineering is an attempt to generate living tissues for surgical transplantation. In vitro and in vivo approaches have led to the production of vascular and cardiovascular components, bones, cartilages and gastrointestinal tissues. Organogenesis has a different aim, which is to create transplantable organs from embryonic tissue implanted into the recipient's omentum. This approach has been successful in creating kidneys and pancreases in animals. The use of stem cells in organogenesis and in tissue engineering has vastly enlarged the potential for clinical applications. The technique of nuclear transfer offers the possibility of creating cells, which are genetically identical to the host. Tissue engineering and organogenesis represent the future of transplantation in medicine. The progress in this field is of tremendous importance because it can produce a new generation of morphologically complex tissues and organs. In this review, the most relevant experiences in this area are summarized, including its perspectives for therapeutical applications.

Animals↗

PubMedDate not supplied

ProteinChip system technology: a powerful tool to analyze expression differences in tissue-engineered blood vessels.

At the time of implantation, tissue-engineered constructs should resemble native tissues as closely as possible. At present, histology and biochemical methods are commonly used to compare tissue-engineered constructs with native tissue. A ProteinChip system based on surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI) has been developed that allows visualization of complex protein profiles from biological samples. The aim of this study was to determine whether the ProteinChip system is a suitable tool with which to compare the protein expression profiles of tissue-engineered aortic blood vessels with native tissues. Tissue-engineered blood vessel substitutes were fabricated with poly-4-hydroxybutyrate scaffolds, ovine vascular cell seeding, and dynamic tissue culture conditions. Engineered, ovine aortic, and carotid tissues were homogenized and total protein was extracted. Samples were analyzed on ProteinChip arrays. Analysis yielded reproducible protein profiles from all samples. About 150 distinct protein peaks were detected. Comparative analysis with ProteinChip software revealed that the protein profiles from native aorta and native carotid arteries were similar whereas early tissue-engineered samples displayed more distinct deviations. In conclusion, ProteinChip system technology is rapid, reproducible, and highly sensitive in highlighting differentially expressed proteins in tissue-engineered blood vessel substitutes.

Animals↗

PubMed2002

Tissue engineering and ENT surgery.

Tissue engineering is the development of biological substitutes for the repair and regeneration of damaged tissues. We explain the principles of this emerging field of biotechology. The present and potential applications of tissue engineering technologies in ENT surgery are then reviewed.

Animals↗

PubMed2002

Tissue engineering: chondrocytes and cartilage.

Tissue engineering offers new strategies for developing treatments for the repair and regeneration of damaged and diseased tissues. These treatments, using living cells, will exploit new developments in understanding the principles in cell biology that control and direct cell function. Arthritic diseases that affect so many people and have a major impact on the quality of life provide an important target for tissue engineering. Initial approaches are in cartilage repair; in our own programme we are elucidating the signals required by chondrocytes to promote new matrix assembly. These principles will extend to other tissues of the musculoskeletal system, including the repair of bone, ligament and tendon.

Animals↗

PubMed2002

Advanced bioreactor with controlled application of multi-dimensional strain for tissue engineering.

Advanced bioreactors are essential for meeting the complex requirements of in vitro engineering functional skeletal tissues. To address this need, we have developed a computer controlled bench-top bioreactor system with capability to apply complex concurrent mechanical strains to three-dimensional matrices independently housed in 24 reactor vessels, in conjunction with enhanced environmental and fluidic control. We demonstrate the potential of this new system to address needs in tissue engineering, specifically toward the development of a tissue engineered anterior cruciate ligament from human bone-marrow stromal cells (hBMSC), where complex mechanical and biochemical environment control is essential to tissue function. Well-controlled mechanical strains (resolution of < 0.1 micron for translational and < 0.1 degree for rotational strain) and dissolved oxygen tension (between 0%-95% +/- 1%) could be applied to the developing tissue, while maintaining temperature at 37 +/- 0.2 degrees C about developing tissue over prolonged periods of operation. A total of 48 reactor vessels containing cell culture medium and silk fiber matrices were run for up to 21 days under 90 degrees rotational and 2 mm translational deformations at 0.0167 Hz with only one succumbing to contamination due to a leak at an medium outlet port. Twenty-four silk fiber matrices seeded with human bone marrow stromal cells (hBMSCs) housed within reactor vessels were maintained at constant temperature (37 +/- 0.2 degrees C), pH (7.4 +/- 0.02), and pO2 (20 +/- 0.5%) over 14 days in culture. The system supported cell spreading and growth on the silk fiber matrices based on SEM characterization, as well as the differentiation of the cells into ligament-like cells and tissue (Altman et al., 2001).

Biomimetics↗

PubMedDate not supplied

Osseous tissue engineering in oncologic surgery.

Tissue engineering is an interdisciplinary field that will yield new sources of tissue for clinical and research purposes in oncology. Bone is under intense investigation by this field. Relevant areas of progress are in advanced computing, biomaterials, cell technology, growth factor fabrication and delivery, and gene manipulation. Clinical techniques will emerge from continued investigation in each of these areas. Techniques that are developed must be scaled up to industry with products cleared by regulatory agencies and acceptable to clinicians and patients. The goals of tissue engineering in oncology are improved tissue models for basic cancer research and a change in clinical practice. Semin. Surg. Oncol. 19:294-301, 2000.

Biomedical Engineering↗

PubMed1999

Acute excisional wounds treated with a tissue-engineered skin (Apligraf).

BACKGROUND: Tissue-engineered products are usually composed of living cells and their supporting matrices that have been grown in vitro, using a combination of engineering and life sciences principles. Apligraf is a bilayered product composed of neonatal-derived dermal fibroblasts and keratinocytes, and Type I bovine collagen. OBJECTIVE: To evaluate in a prospective, multicentered open study, the effects of tissue therapy with a tissue-engineered skin (Apligraf) with partial or full-thickness excisional wounds. METHODS: One hundred and seven patients participated in this study. The tissue-engineered skin was applied once, immediately after excisional surgery, usually for skin cancer, and patients were followed for up to one year. RESULTS: The safety results were impressive, with no clinical or laboratory evidence of rejection. Clinically, graft persistence was good to excellent in 77 of 105 (73.3%) of patients at one week, falling to 56.6% and 53.6% at two weeks and one month respectively. CONCLUSION: To date, this is the largest experience with a tissue-engineered skin product in acute wounds, and this study suggests that tissue therapy may be safe and useful.

Animals↗

PubMed2005

Perfusion bioreactor system for human mesenchymal stem cell tissue engineering: dynamic cell seeding and construct development.

Human mesenchymal stem cells (hMSCs) have great potential for therapeutic applications. A bioreactor system that supports long-term hMSCs growth and three-dimensional (3-D) tissue formation is an important technology for hMSC tissue engineering. A 3-D perfusion bioreactor system was designed using non-woven poly (ethylene terepthalate) (PET) fibrous matrices as scaffolds. The main features of the perfusion bioreactor system are its modular design and integrated seeding operation. Modular design of the bioreactor system allows the growth of multiple engineered tissue constructs and provides flexibility in harvesting the constructs at different time points. In this study, four chambers with three matrices in each were utilized for hMSC construct development. The dynamic depth filtration seeding operation is incorporated in the system by perfusing cell suspensions perpendicularly through the PET matrices, achieving a maximum seeding efficiency of 68%, and the operation effectively reduced the complexity of operation and the risk of contamination. Statistical analyses suggest that the cells are uniformly distributed in the matrices. After seeding, long-term construct cultivation was conducted by perfusing the media around the constructs from both sides of the matrices. Compared to the static cultures, a significantly higher cell density of 4.22 x 10(7) cell/mL was reached over a 40-day culture period. Cellular constructs at different positions in the flow chamber have statistically identical cell densities over the culture period. After expansion, the cells in the construct maintained the potential to differentiate into osteoblastic and adipogenic lineages at high cell density. The perfusion bioreactor system is amenable to multiple tissue engineered construct production, uniform tissue development, and yet is simple to operate and can be scaled up for potential clinical use. The results also demonstrate that the multi-lineage differentiation potential of hMSCs are preserved even after extensive expansion, thus indicating the potential of hMSCs for functional tissue construct development. The system has important applications in stem cell tissue engineering.

Adipocytes↗

PubMed2006

Cellular to tissue informatics: approaches to optimizing cellular function of engineered tissue.

Tissue engineering is a rapidly expanding, multi-disciplinary field in biomedicine. It provides the ability to manipulate living cells and biomaterials for the purpose of restoring, maintaining, and enhancing tissue and organ function. Scientists have engineered various tissues in the body, from skin substitutes to artificial nerves to heart tissues, with varying degrees of success. Although the field of tissue engineering has come a long way since its first successful demonstration by Bisceglie in the 1930s, methods of coaxing them into functional tissues have been predominantly empirical to date. To successfully develop tissue-engineered organs, it is important to understand how to maintain the cells under conditions that maximize their ability to perform their physiological roles, regardless of their environment. In that context, a methodology that combines empirical data with mathematical and statistical techniques, such as metabolic engineering and cellular informatics, to systematically determine the optimal (1) type of cell to use, (2) scaffold properties and the corresponding processing conditions to achieve those properties, and (3) the required types and levels of environmental factors and the operating conditions needed in the bioreactor, will enable the design of viable and functional tissues tailored to the specific requirements of individual situations.

Animals↗

PubMed2004

Rapid prototyping in tissue engineering: challenges and potential.

Tissue engineering aims to produce patient-specific biological substitutes in an attempt to circumvent the limitations of existing clinical treatments for damaged tissue or organs. The main regenerative tissue engineering approach involves transplantation of cells onto scaffolds. The scaffold attempts to mimic the function of the natural extracellular matrix, providing a temporary template for the growth of target tissues. Scaffolds should have suitable architecture and strength to serve their intended function. This paper presents a comprehensive review of the fabrication methods, including conventional, mainly manual, techniques and advanced processing methods such as rapid prototyping (RP) techniques. The potential and challenges of scaffold-based technology are discussed from the perspective of RP technology.

Animals↗

PubMed2000

Bioabsorbable polymer scaffolds for tissue engineering capable of sustained growth factor delivery.

Engineering new tissues utilizing cell transplantation on biodegradable polymer matrices is an attractive approach to treat patients suffering from the loss or dysfunction of a number of tissues and organs. The matrices must maintain structural integrity during the process of tissue formation, and promote the vascularization of the developing tissue. A number of molecules (angiogenic factors) have been identified that promote the formation of new vascular beds from endothelial cells present within tissues, and the localized, controlled delivery of these factors from a matrix may allow an enhanced vascularization of engineered tissues. We have developed a gas foaming polymer processing approach that allows the fabrication of three-dimensional porous matrices from bioabsorbable materials (e.g., copolymers of lactide and glycolide [PLG]) without the use of organic solvents or high temperatures. The effects of several processing parameters (e.g., gas type, polymer composition and molecular weight) on the process were studied. Several gases (CO(2), N(2), He) were utilized in the fabrication process, but only CO(2) resulted in the formation of highly porous, structurally intact matrices. Crystalline polymers (polylactide and polyglycolide) did not form porous matrices, while amorphous copolymers (50:50, 75:25, and 85:15 ratio of lactide:glycolide) foamed to yield matrices with porosity up to 95%. The mechanical properties of matrices were also regulated by the choice of PLG composition and molecular weight. Angiogenic factors (e.g., vascular endothelial growth factor) were subsequently incorporated into matrices during the fabrication process, and released in a controlled manner. Importantly, the released growth factor retains over 90% of its bioactivity. In summary, a promising system for the incorporation and delivery of angiogenic factors from three-dimensional, biodegradable polymer matrices has been developed, and the fabrication process allows incorporation under mild conditions.

Absorption↗

PubMed2004

Tissue engineering: current state and perspectives.

Tissue engineering is an interdisciplinary field that involves cell biology, materials science, reactor engineering, and clinical research with the goal of creating new tissues and organs. Significant advances in tissue engineering have been made through improving singular aspects within the overall approach, e.g., materials design, reactor design, or cell source. Increasingly, however, advances are being made by combining several areas to create environments which promote the development of new tissues whose properties more closely match their native counterparts. This approach does not seek to reproduce all the complexities involved in development, but rather seeks to promote an environment which permits the native capacity of cells to integrate, differentiate, and develop new tissues. Progenitors and stem cells will play a critical role in understanding and developing new engineered tissues as part of this approach.

Animals↗

PubMed2002

The history and current status of tissue engineering: The future of pediatric surgery.

Progress in the pediatric surgical treatment of numerous tissue deficits has been achieved through the relatively new field of tissue engineering. Tissue engineering has distinct advantages over native tissue or prosthetic substitution including self-repair and growth with the patient, avoiding multiple surgeries. The application of microfabrication has allowed more precision in the control of cell interactions and resulting tissue architecture.

Animals↗

PubMed2005

Fibrin as a cell carrier in cardiovascular tissue engineering applications.

In cardiovascular tissue engineering approaches, efficient seeding methods are essential. To achieve this and to save time, cells can be encapsulated in gels. Combining the advantages of a gel as a cell carrier with the advantages of a fiber-based scaffold, providing structural integrity to the developing tissue, might offer several advantages. In this study, seeding by using fibrin as a cell carrier is compared to the conventional static seeding method with regard to tissue development. Seeding with fibrin resulted in less loss of soluble collagen into the medium and a more mature extracellular matrix in a shorter period of time. The use of fibrin degradation inhibitors was shown to inhibit extracellular matrix formation, although it did not hamper cell proliferation. The use of fibrin as a cell carrier to seed cells into a fiber-based scaffold may represent a promising, timesaving approach in cardiovascular tissue engineering applications.

Biocompatible Materials↗