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PubMed2003

[Tissue engineering: chances and challenges for application in rheumatic diseases].

Current technologies of tissue engineering offer new strategies for the treatment of cartilage and bone defects. Beyond implantation of cell suspensions, second generation products of biomaterial enforced with in vitro preformed tissues are clinically applied. Ongoing research and development focus on differentiation factors and tissue protection. In search for sources of autologous cells which are easier to collect and which may serve for more complex tissues like osteochondral implants, mesenchymal stem cells are investigated. The design of in vitro experiments, which are required for these investigations, has produced tissue engineering technologies, which may serve for pathophysiology research in inflammatory joint diseases and for exploration of treatment strategies. These together with the advances in biological therapies of rheumatic diseases are the basis of new concepts, which promise application of tissue engineering also in inflammatory joint diseases.

Animals↗
PubMed2005

Second generation of meniscus transplantation: in-vivo study with tissue engineered meniscus replacement.

INTRODUCTION: The options available after meniscus loss offer only limited chances for a long-term success. In the following experimental study, we investigated the effect of meniscus tissue engineering on properties of the collagen meniscus implant (CMI). METHODS: Autologous fibrochondrocytes, obtained per biopsy from adult Merino sheep (n=25), were released from the matrix, cultured in-vitro and seeded into CMI scaffolds (n=10, group 1). Following a 3-week in-vitro culture, the tissue engineered menisci were used for autologous transplantation. Macroscopical and histological evaluation were performed in comparison with non-seeded CMI controls (n=10, group 2) and with meniscus-resected controls (n=5, group 3) after 3 weeks (each 1 animal group 1 and 2) and 3 months. RESULTS: The lameness score did not show any difference between the groups. Meniscus tissue was found in seven knee joints (group 1), in five knee joints (group 2) and in two knee joints (group 3). The size of the transplants reduced from 25.9+/-4.5 to 20.1+/-10.8 mm (group 1) and from 25.9+/-1.5 to 14.4+/-12.5 mm (group 2). Histologically, enhanced vascularisation, accelerated scaffold re-modelling, higher content of extra-cellular matrix and lower cell number were noted in the pre-seeded menisci in comparison with non-seeded controls. Dense high-cellular fibrous scar tissue was found in two of five cases in the resection control group. CONCLUSION: Tissue engineering of meniscus with autologous fibrochondrocytes demonstrates a macroscopic and histological improvement of the transplants. However, further development of the methods, especially of the scaffold and of the cell-seeding procedure must prove the feasibility of this procedure for human applications.

Animals↗
PubMed2003

Immunomodulation of tissue-engineered transplants: in vivo bone generation from methylprednisolone-stimulated chondrocytes.

Subcutaneously implanted, in vitro engineered tissue is generally affected by the immune system of the host even in autogenous transplantation. The aim of this study was to investigate immunomodulation of subcutaneously implanted tissue-engineered cartilage transplants by intramuscular methylprednisolone application. Transplants consisted of auricular rabbit chondrocytes, polylactide-polyglycolide co-polymer fleeces and species-specific fibrin or agarose. The transplants were subcutaneously implanted in the ridge. Thereafter, animals were separated into two groups, one with and one without methylprednisolone treatment. The specimens were histologically investigated after 6 and 12 weeks. Fleece fiber degradation was complete after 12 weeks, and all transplants showed areas of calcification. The corticosteroid-treated group presented pronounced trabecular bone generation without fibrous tissue infiltration. The untreated group showed sporadic islets of calcification without coherent bone formation, and adjacent fibrous tissue had infiltrated the transplants. Native controls and corticoid-treated transplants did not exhibit bone generation or signs of fibrous tissue infiltration. This study found that immunomodulation by intramuscular methylprednisolone application protects tissue-engineered autogenous chondrocyte transplants from fibrous tissue infiltration and induces trabecular bone formation.

Animals↗
PubMed2000

Engineering pancreatic islets.

Pancreatic islets are neuroendocrine organs that control blood glucose homeostasis. The precise interplay of a heterogeneous group of cell populations (beta, alpha, delta and PP cells) results in the fine-tuned release of counterbalanced hormones (insulin, glucagon, somatostatin and pancreatic polypeptide respectively). Under the premises of detailed knowledge of the physiological basis underlying this behaviour, two lines of investigation might be inferred: generating computational and operational models to explain and predict this behaviour and engineering islet cells to reconstruct pancreatic endocrine function. Whilst the former is being fuelled by new computational strategies, giving biophysicists the possibility of modelling a system in which new "emergent" properties appear, the latter is benefiting from the useful tools and strategic knowledge achieved by molecular, cell and developmental biologists. This includes using tumour cell lines, engineering islet cell precursors, knowledge of the mechanisms of differentiation, regeneration and growth and, finally, therapeutic cloning of human tissues. Gaining deep physiological understanding of the basis governing these processes is instrumental for engineering new pancreatic islets.

Animals↗
PubMed1998

Tissue engineering: generation of differentiated artificial tissues for biomedical applications.

A new field in biomedical science has been established. Cell biologists, engineers, and surgeons now work within a team. Artificial connective, epithelial, or neuronal tissues are being constructed using living cells and different kinds of biomaterials. Numerous companies and laboratories are presenting dynamic developments in this field. Prognoses predict that, at the beginning of the coming century, the industry of tissue engineering will reach the importance of the present genetic technology. An enormous demand for organ and tissue transplants motivates research activities and drives the acquisition of innovative techniques and creative solutions. At the front of this development is the creation of artificial skin for severely burned patients and the generation of artificial cartilage for implantation in articular joint diseases. Future challenges are the construction of liver organoids and the development of an artificial kidney on the basis of cultured cells. In this paper we show strategies, needs, tools, and equipment for tissue engineering. The presupposition for all projects is the induction, development, and maintenance of differentiation within the tissue under in vitro conditions. As experiments in conventional culture dishes continued to fail, new cell and tissue culture methods had to be developed. Tissues are cultured under conditions as close as possible to their natural environment. To optimize adherence or embedding, cells are grown on novel tissue carriers and on individually selected biomatrices or scaffolds. The tissues are subsequently transferred into different types of containers for permanent perfusion with fresh culture medium. This guarantees constant nutrition of the developing tissue and prevents the accumulation of harmful metabolites. An organo-typical environment for epithelial cells, for example, is obtained in gradient containers, which are permanently superfused at the apical and basal sides with different media. Long term experiments result in cultured tissues in a quality thus far unreached.

Biomedical Engineering↗
PubMed2005

Bone tissue engineering with porous hydroxyapatite ceramics.

The main principle of bone tissue engineering strategy is to use an osteoconductive porous scaffold in combination with osteoinductive molecules or osteogenic cells. The requirements for a scaffold in bone regeneration are: (1) biocompatibility, (2) osteoconductivity, (3) interconnected porous structure, (4) appropriate mechanical strength, and (5) biodegradability. We recently developed a fully interconnected porous hydroxyapatite (IP-CHA) by adopting the "form-gel" technique. IP-CHA has a three-dimensional structure with spherical pores of uniform size that are interconnected by window-like holes; the material also demonstrated adequate compression strength. In animal experiments, IP-CHA showed superior osteoconduction, with the majority of pores filled with newly formed bone. The interconnected porous structure facilitates bone tissue engineering by allowing the introduction of bone cells, osteotropic agents, or vasculature into the pores. In this article, we review the accumulated data on bone tissue engineering using the novel scaffold, focusing especially on new techniques in combination with bone morphogenetic protein (BMP) or mesenchymal stem cells.

Animals↗
PubMed2005

Evaluation of three growth factors for TMJ disc tissue engineering.

Arguably one of the most complex joints in the body, the temporomandibular joint (TMJ) presents one of the most difficult problems in modem medicine. Tissue engineering, for the TMJ disc in particular, has been proposed as a potential breakthrough treatment strategy for TMJ disorders. Central to tissue engineering is understanding growth factor effects on TMJ disc cells, and to the best of our knowledge, this is the first 3D growth factor study for these cells. The purpose was to examine the effects of high and low concentrations of basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF), and transforming growth factor-beta1 (TGF-beta) on porcine TMJ disc cells. Cells were seeded onto non-woven PGA scaffolds (95% porosity) in spinner flasks, then cultured with a growth factor for 6 weeks. Constructs were analyzed for mechanical and structural integrity, cell number, and matrix biosynthesis. All growth factors improved mechanical and structural integrity compared to the control. IGF and TGF-beta were most effective at promoting collagen synthesis, although there were no significant differences in glycosaminoglycan synthesis or cell number between any groups. After considering the economic advantage of IGF over TGF-beta, the conclusion of this study is to use IGF in future TMJ disc tissue engineering experiments.

Animals↗
PubMed2005

Ex vivo adipose tissue engineering by human marrow stromal cell seeded gelatin sponge.

The limitation of current clinical treatment for restoration extended defects of soft tissue associated with trauma, tumor resections, and congenital deformities are well known. This study demonstrates that human bone marrow stromal cells (MSCs) can be utilized to tissue engineer adipose tissue for therapeutic purposes. Adipogenic potentials of monolayer-cultured human MSCs were evaluated by biochemical measurement of an adipogenic differentiation marker (glycerol-3-phosphate dehydrogenase, G-3-PDH) and cellular morphology. After preparation by seeding human MSCs on a 3-dimensional gelatin sponge and exposure to adipogenic differentiation medium, the ex vivo tissue-engineered adipose constructs were assessed histomorphologically and biochemically. Lipid droplets accumulated and expanded within the constructs accompanied by a significant increase of G-3-PDH activity. The present study indicates that bone MSCs could be a cell resource in tissue engineering adipose tissue, while gelatin sponge could be a good scaffold in this approach to improve the outcome of clinical treatment.

Adipose Tissue↗
PubMed2005

Design of ligand binding to an engineered protein cavity using virtual screening and thermal up-shift evaluation.

Proteins could be used to carry and deliver small compounds. As a tool for designing ligand binding sites in protein cores, a three-step virtual screening method is presented that has been optimised using existing data on T4 lysozyme complexes and tested in a newly engineered cavity in flavodoxin. The method can pinpoint, in large databases, ligands of specific protein cavities. In the first step, physico-chemical filters are used to screen the library and discard a majority of compounds. In the second step, a flexible, fast docking procedure is used to score and select a smaller number of compounds as potential binders. In the third step, a finer method is used to dock promising molecules of the hit list into the protein cavity, and an optimised free energy function allows discarding the few false positives by calculating the affinity of the modelled complexes. To demonstrate the portability of the method, several cavities have been designed and engineered in the flavodoxin from Anabaena PCC 7119, and the W66F/L44A double mutant has been selected as a suitable host protein. The NCI database has then been screened for potential binders, and the binding to the engineered cavity of five promising compounds and three tentative non-binders has been experimentally tested by thermal up-shift assays and spectroscopic titrations. The five tentative binders (some apolar and some polar), unlike the three tentative non-binders, are shown to bind to the host mutant and, importantly, not to bind to the wild type protein. The three-step virtual screening method developed can thus be used to identify ligands of buried protein cavities. We anticipate that the method could also be used, in a reverse manner, to identify natural or engineerable protein cavities for the hosting of ligands of interest.

Bacteriophage T4↗
PubMed2006

Historic and current strategies in bone tissue engineering: do we have a hope in Hench?

Professors Larry Hench and Julia Polak formed the Tissue Engineering and Regenerative Medicine Centre (TERM) at Imperial College London to foster collaborations between biologists and materials scientists. Early work at the center elucidated the biomolecular interactions between primary human osteoblasts and 45S5 Bioglass . As research efforts expanded, the team discovered that the dissolution products of both 45S5 Bioglass and 58S sol-gel bioactive glasses had osteoblastic stimulatory properties. To address the shortage of appropriate cells for bone tissue engineering applications, TERM scientists also demonstrated the differentiation of embryonic stem (ES) cells to osteoblasts when treated with the dissolution products of bioactive glasses. They also found that the soluble factors ascorbic acid, beta -glycerophosphate, and dexamethasone preferentially differentiated ES cells to osteoblasts, and their combination with the dissolution products of bioactive glasses stimulated differentiation even further. Taken together, these results demonstrate the suitability of bioactive glasses as scaffolds for bone tissue engineering as they not only provide an osteoconductive and osteoproductive substrate, but also actively stimulate cells to express appropriate osteoblastic phenotypes. Professor Hench's vision to pioneer regenerative medicine research continues with the aim of developing novel therapeutics to treat musculoskeletal disability.

Animals↗
PubMed2006

Characterisation of a collagen membrane for its potential use in cardiovascular tissue engineering applications.

In this study, Biomend, a collagen membrane conventionally used in the regeneration of periodontal tissue, is investigated for its possible use in the field of cardiovascular tissue engineering. A key requirement of most potential tissue engineering scaffolds is that degradation occurs in tandem with tissue regeneration and extra cellular matrix remodelling. To this end, it is crucial to understand the degradation mechanics and mechanisms of the material and to investigate the practicability of using Biomend as a possible scaffold material. With this in mind, methodologies for the initial characterisation of the scaffold material were determined. The mechanical properties of Biomend samples, subjected to various degrees of hydration and enzymatic degradation, were examined primarily through tensile testing experiments. The effects of enzymatic degradation were monitored quantitatively, by observing weight loss, and visually, by studying micrographs. Cell adhesion and viability were of primary concern. Confocal laser scanning microscopy was employed to illustrate endothelialisation on the surface of this collagen membrane. Fluorescence microscopy was used to visualise cell viability on the membrane surface. These images, coupled with assays to measure cell activity, suggest that Biomend is not a suitable substrate to allow endothelialisation. In summary, this collagen membrane has suitable mechanical properties with the potential to control its degradation rate. However, since poor endothelial cell viability was observed on the membrane, it may not be suitable for use in cardiovascular tissue engineering applications.

Biocompatible Materials↗
PubMed2006

Micro-CT-based screening of biomechanical and structural properties of bone tissue engineering scaffolds.

The development of successful scaffolds for bone tissue engineering requires a concurrent engineering approach that combines different research fields. In order to limit in vivo experiments and reduce trial and error research, a scaffold screening technique has been developed. In this protocol seven structural and three biomechanical properties of potential scaffold materials are quantified and compared to the desired values. The property assessment is done on computer models of the scaffolds, and these models are based on micro-CT images. As a proof of principle, three porous scaffolds were evaluated with this protocol: stainless steel, hydroxyapatite, and titanium. These examples demonstrate that the modelling technique is able to quantify important scaffold properties. Thus, a powerful technique for automated screening of bone tissue engineering scaffolds has been developed that in a later stage may be used to tailor the scaffold properties to specific requirements.

Biocompatible Materials↗
PubMed2004

Protein engineering and applications of Candida rugosa lipase isoforms.

Commercial preparations of Candida rugosa lipase (CRL) are mixtures of lipase isoforms used for the hydrolysis and synthesis of various esters. The presence of variable isoforms and the amount of lipolytic protein in the crude lipase preparations lead to a lack of reproducibility of biocatalytic reactions. Purification of crude CRL improve their substrate specificity, enantioselectivity, stability, and specific activities. The expression of the isoforms is governed by culture or fermentation conditions. Unfortunately, the nonsporogenic yeast C. rugosa does not utilize the universal codon CTG for leucine; therefore, most of the CTG codons were converted to universal serine triplets by site-directed mutagenesis to gain expression of functional lipase in heterologous hosts. Recombinant expressions by multiple-site mutagenesis or complete synthesis of the lipase gene are other possible ways of obtaining pure and different CRL isoforms, in addition to culture engineering. Protein engineering of purified CRL isoforms allows the tailoring of enzyme function. This involves computer modeling based on available 3-D structures of lipase isoforms. Lid swapping and DNA shuffling techniques can be used to improve the enantioselectivity, thermostability, and substrate specificity of CRL isoforms and increase their biotechnological applications. Lid swapping can result in chimera proteins with new functions. The sequence of the lid can affect the activity and specificity of recombinant CRL isoforms. Candida rugosa lipase is toxicologically safe for food applications. Protein engineering through lid swapping and rationally designed site-directed mutagenesis will continue to lead to the production of CRL isoforms with improved catalytic power, thermostability, enantioselectivity, and substrate specificity, while providing evidence for the mechanisms of actions of the various isoforms.

Candida↗
PubMed2001

Investigating ethical issues in engineering design.

This paper aims at contributing to a research agenda in engineering ethics by exploring the ethical aspects of engineering design processes. A number of ethically relevant topics with respect to design processes are identified. These topics could be a subject for further research in the field of engineering ethics. In addition, it is argued that the way design processes are now organised and should be organised from a normative point of view is an important topic for research.

Engineering↗
PubMed2002

Personal meaning and ethics in engineering.

The study of engineering ethics tends to emphasize professional codes of ethics and, to lesser degrees, business ethics and technology studies. These are all important vantage points, but they neglect personal moral commitments, as well as personal aesthetic, religious, and other values that are not mandatory for all members of engineering. This paper illustrates how personal moral commitments motivate, guide, and give meaning to the work of engineers, contributing to both self-fulfillment and public goods. It also explores some general frameworks for thinking about these commitments and calls for further exploration of them.

Attitude↗
PubMed2004

Internationalizing professional codes in engineering.

Professional engineering societies which are based in the United States, such as the American Society of Mechanical Engineers (ASME, now ASME International) are recognizing that their codes of ethics must apply to engineers working throughout the world. An examination of the ethical code of the ASME International shows that its provisions pose many problems of application, especially in societies outside the United States. In applying the codes effectively in the international environment, two principal issues must be addressed. First, some Culture Transcending Guidelines must be identified and justified. Nine such guidelines are identified Second, some methods for applying the codes to particular situations must be identified Three such methods are specification, balancing, and finding a creative middle way.

Codes of Ethics↗
PubMed2004

The dilemma of ethics in engineering education.

This paper briefly summarizes current thinking in engineering ethics education, argues that much of that ethical instruction runs the risk of being only superficially effective, and explores some of the underlying systemic barriers within academia that contribute to this result. This is not to criticize or discourage efforts to improve ethics instruction. Rather it is to point to some more fundamental problems that still must be addressed in order to realize the full potential of enhanced ethics instruction. Issues discussed will include: intellectual engagement versus emotional engagement; the gravitational pull of curricular structures; the nature of engineering faculty; and the "engineer-ization" of ethics.

Curriculum↗
PubMed2004

Esssential ethics--embedding ethics into an engineering curriculum.

Ethical decision-making is essential to professionalism in engineering. For that reason, ethics is a required topic in an ABET approved engineering curriculum and it must be a foundational strand that runs throughout the entire curriculum. In this paper the curriculum approach that is under development at the Padnos School of Engineering (PSE) at Grand Valley State University will be described. The design of this program draws heavily from the successful approach used at the service academies--in particular West Point and the United States Naval Academy. As is the case for the service academies, all students are introduced to the "Honor Concept" (which includes an Honor Code) as freshmen. As an element of professionalism the PSE program requires 1500 hours of co-op experience which is normally divided into three semesters of full-time work alternated with academic semesters during the last two years of the program. This offers the faculty an opportunity to teach ethics as a natural aspect of professionalism through the academic requirements for co-op. In addition to required elements throughout the program, the students are offered opportunities to participate in service projects which highlight responsible citizenship. These elements and other parts of the approach will be described.

Curriculum↗