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PubMedDate not supplied

Review: application of stem cells for vascular tissue engineering.

As the prevalence of vascular disease has continued to expand, the need for a suitable arterial replacement has prompted researchers to look beyond synthetic and autologous grafts toward the field of tissue engineering. Advances in vascular tissue engineering have utilized both mesenchymal and hematopoietic stem cells as a cell source in an attempt to create a fully engineered small-diameter graft. Stem cells offer enormous potential as a cell source because of their proliferative and growth potential, and the application of stem cell technology has far-reaching implications for future applications. The innovative use of stem cells for vascular tissue engineering has opened new possibilities for a fully engineered blood vessel. The purpose of this review is to summarize the current perspective on the use of stem cells for vascular tissue engineering. It focuses principally on the classes of stem cells used, techniques for differentiation scaffolding technology, and the successes and failures of models.

Animals↗

PubMed2002

Recent advances in tissue engineering: an invited review.

Tissue formation within the body, as part of a development or repair process, is a complex event in which cell populations self-assemble into functional units. There is intense academic, medical, and commercial interest in finding methods of replicating these events outside the body. This interest has accelerated with the demonstration of the engineering of skin and cartilage tissue in the laboratory and there is now worldwide activity in the in vitro regeneration of tissues including nerve, liver, bone, heart valves, blood vessels, bladder, and kidney. Approaches to tissue engineering center on the need to provide signals to cell populations to promote cell proliferation and differentiation. This review considers recent advances in methods of providing these signals to cells using examples of progress in the engineering of complex tissues.

Absorbable Implants↗

PubMed2006

Clinical application of tissue-engineered transplants. Part I: mucosa.

OBJECTIVES: The study series aims at testing the feasibility of the clinical application of tissue-engineered oral mucosa. The preliminary results were gathered over a period varying from 6 months to 12 years depending on the surgical method. METHODS: Tissue-engineered oral mucosa was used to cover defects in various surgical procedures like vestibuloplasty (n=42), freeing of the tongue (n=10), prelaminating the radial flap (n=5) and reconstruction of the urethra (n=16). In all interventions small samples of oral mucosa were harvested, cut into small pieces, resuspended in culture medium and seeded into a culture flask. Cultured keratinocytes were transferred onto membranes which then were used to cover mucosal defects in the oral cavity. RESULTS: To gain a graft of 15 cm(2) size a mucosa biopsy of 4-8 mm(2) and 40 ml autologous patients serum is needed. Tissue-engineered oral mucosa was applied successfully in all four surgical methods. Six months after transplantation a regular epithelial layering with a histological delimitation of the stratum, epithelial crest and a strong basal membrane appeared. According to the reception site the tissue engineered oral mucosa differentiated in several ways. CONCLUSION: Tissue-engineered oral mucosa fulfils the requirements for clinical routine. With view to healing time and outcome it does not appear to be superior to regular harvested oral mucosa transplants. Because of a smaller harvesting defect and primary wound closure at the actual operation site the patients' convenience is increased. Thus this method reduces morbidity and advances the quality of life.

Aged↗

PubMed2006

The outlook for implants and endodontics: a review of the tissue engineering strategies to create replacement teeth for patients.

Ideally, root canal therapy involves the removal of diseased pulp tissues and permanent replacement with healthy pulp to revitalize teeth. Rather than placing implants, the ideal solution is to grow new replacement teeth. Success rates of implants and endodontic treatments can exceed 90%, which presents a formidable challenge to tissue engineering researchers to ensure that future dental treatments are even more successful. The purpose of this article is to explain how tissue engineering can be used to create replacement teeth. The science of tissue engineering has evolved from growing simple tissues in cell culture incubators to a multistep process. Although the problems of introducing tissue engineering therapies as part of routine dental treatments are substantial, the potential benefits are equally ground breaking.

Biomimetic Materials↗

PubMed2003

Tissue engineering of the gastrointestinal tract for surgical replacement: a nutrition tool of the future?

Optimal nutrition depends on the multiple complex functions performed by the gastrointestinal tract, which range from basic functions such as storage, conduit and mechanical processing to more finely regulated capabilities such as vectorial transport, immune defence and cell signalling. Surgical strategies to supply lacking gastrointestinal tract tissues have relied on either replacement by proxy (surgical substitution) or the introduction of prostheses. Tissue engineering seeks to replace missing tissues with engineered tissues that more accurately reproduce the native physiological and anatomical milieu. It is now possible to engineer several areas of the gastrointestinal tract with high fidelity, and to employ tissue-engineered bowel in replacement in animal models. These replacement models have reflected excellent anatomical and physiological recapitulation of native bowel by the tissue-engineered constructs in vivo.

Animals↗

PubMed2005

Fabrication and characterization of DTBP-crosslinked chitosan scaffolds for skin tissue engineering.

Chitosan, the deacetylated derivative of chitin, is a promising scaffold material for skin tissue engineering applications. It is biocompatible and biodegradable, and the degradation products are resorbable. However, the rapid degradation of chitosan and its low mechanical strength are concerns that may limit its use. In this study, chitosan with 80%, 90% and 100% degree of deacetylation (DDA) was crosslinked with dimethyl 3-3, dithio bis' propionimidate (DTBP) and compared to uncrosslinked scaffolds. The scaffolds were characterized with respect to important tissue engineering properties. The tensile strength of scaffolds made from 100% DDA chitosan was significantly higher than for scaffolds made from 80% and 90% DDA chitosan. Crosslinking of scaffolds with DTBP increased the tensile strength. Crosslinking with DTBP had no significant effect on water vapour transmission rate (WVTR) or water absorption but had significant effect on the pore size and porosity of the samples. All samples showed a WVTR and pore size distribution suitable for skin tissue engineering; however, the water absorption and porosity were lower than the optimal values for skin tissue engineering. The biodegradation rate of scaffolds crosslinked with DTBP and glutaraldehyde (GTA) were reduced while no significant effect was observed in biodegradation of the samples made from 100% DDA chitosan whether crosslinked or uncrosslinked after 24 days of degradation.

Absorption↗

PubMedDate not supplied

Biodegradable nanomats produced by electrospinning: expanding multifunctionality and potential for tissue engineering.

With increasing interest in nanotechnology, development of nanofibers (n-fibers) by using the technique of electrospinning is gaining new momentum. Among important potential applications of n-fiber-based structures, scaffolds for tissue-engineering represent an advancing front. Nanoscaffolds (n-scaffolds) are closer to natural extracellular matrix (ECM) and its nanoscale fibrous structure. Although the technique of electrospinning is relatively old, various improvements have been made in the last decades to explore the spinning of submicron fibers from biodegradable polymers and to develop also multifunctional drug-releasing and bioactive scaffolds. Various factors can affect the properties of resulting nanostructures that can be classified into three main categories, namely: (1) Substrate related, (2) Apparatus related, and (3) Environment related factors. Developed n-scaffolds were tested for their cytocompatibility using different cell models and were seeded with cells for to develop tissue engineering constructs. Most importantly, studies have looked at the potential of using n-scaffolds for the development of blood vessels. There is a large area ahead for further applications and development of the field. For instance, multifunctional scaffolds that can be used as controlled delivery system do have a potential and have yet to be investigated for engineering of various tissues. So far, in vivo data on n-scaffolds are scarce, but in future reports are expected to emerge. With the convergence of the fields of nanotechnology, drug release and tissue engineering, new solutions could be found for the current limitations of tissue engineering scaffolds, which may enhance their functionality upon in vivo implantation. In this paper electrospinning process, factors affecting it, used polymers, developed n-scaffolds and their characterization are reviewed with focus on application in tissue engineering.

Absorbable Implants↗

PubMed2006

Polyhydroxyalkanoate (PHA)/inorganic phase composites for tissue engineering applications.

Polyhydroxyalkanoates are emerging as a class of biodegradable polymers for applications in tissue engineering. Members of the polyhydroxyalkanoates family encompass a wide variety of materials, from hard and brittle materials to soft and elastomeric. Over the years, efforts have been made to extend the group of polyhydroxyalkanoates and to investigate their use in numerous biomedical applications, such as sutures, cardiovascular patches, wound dressings, guided tissue repair/regeneration devices, and tissue engineering scaffolds. Along with the development of polyhydroxyalkanoates, researchers have looked into the possibility of designing composites in combination with inorganic phases to further improve the mechanical properties, rate of degradation, and also impart bioactivity. Poly(3-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyvalerate), and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) are some of the polymers which have been studied extensively to fabricate composites in combination with hydroxyapatite, bioactive glass, and glass-ceramic fillers or coatings. This paper reviews international research carried out toward development of polyhydroxyalkanoates/inorganic phase composites in terms of systems investigated, microstructures, properties achieved, and applications, with special focus on tissue engineering scaffolds. A comparison between different composite systems developed in the past few years is presented. The paper also addresses the prospect of potential further development of polyhydroxyalkanoates/inorganic phase composites with optimized microstructure and properties for improved tissue engineering scaffolds.

Absorbable Implants↗

PubMed2000

Fabrication of a trileaflet heart valve scaffold from a polyhydroxyalkanoate biopolyester for use in tissue engineering.

Previously, we reported the implantation of a single tissue engineered leaflet in the posterior position of the pulmonary valve in a lamb model. The major problems with this leaflet replacement were the scaffold's inherent stiffness, thickness, and nonpliability. We have now created a scaffold for a trileaflet heart valve using a thermoplastic polyester. In this experiment, we show the suitability of this material in the production of a biodegradable, biocompatible scaffold for tissue engineered heart valves. A heart valve scaffold was constructed from a thermoplastic elastomer. The elastomer belongs to a class of biodegradable, biocompatible polyesters known as polyhydroxyalkanoates (PHAs) and is produced by fermentation (Metabolix Inc., Cambridge, MA). It was modified by a salt leaching technique to create a porous, three-dimensional structure, suitable for tissue engineering. The trileaflet heart valve scaffold consisted of a cylindrical stent (1 mm X 15 mm X 20 mm I.D.) containing three valve leaflets. The leaflets were formed from a single piece of PHA (0.3 mm thick), and were attached to the outside of the stent by thermal processing techniques, which required no suturing. After fabrication, the heart valve construct was allowed to crystallize (4 degrees C for 24 h), and salt particles were leached into doubly distilled water over a period of 5 days to yield pore sizes ranging from 80 to 200 microns. Ten heart valve scaffolds were fabricated and seeded with vascular cells from an ovine carotid artery. After 4 days of incubation, the constructs were examined by scanning electron microscopy. The heart valve scaffold was tested in a pulsatile flow bioreactor and it was noted that the leaflets opened and closed. Cells attached to the polymer and formed a confluent layer after incubation. One advantage of this material is the ability to mold a complete trileaflet heart valve scaffold without the need for suturing leaflets to the conduit. Second advantage is the use of only one polymer material (PHA) as opposed to hybridized polymer scaffolds. Furthermore, the mechanical properties of PHA, such as elasticity and mechanical strength, exceed those of the previously utilized material. This experiment shows that PHAs can be used to fabricate a three-dimensional, biodegradable heart valve scaffold.

Animals↗

PubMed2006

Modeling the mechanics of tissue-engineered human heart valve leaflets.

Mathematical models can provide valuable information to assess and evaluate the mechanical behavior of tissue-engineered constructs. In this study, a structurally based model is applied to describe and analyze the mechanics of tissue-engineered human heart valve leaflets. The results from two orthogonal uniaxial tensile tests are used to determine the model parameters of the constructs after two, three and four weeks of culturing. Subsequently, finite element analyses are performed to simulate the mechanical response of the engineered leaflets to a pressure load. The stresses in the leaflets induced by the pressure load increase monotonically with culture time due to a decrease in the construct's thickness. The strains, on the other hand, eventually decrease as a result of an increase in the elastic modulus. Compared to native porcine leaflets, the mechanical response of the engineered tissues after four weeks of culturing is more linear, stiffer and less anisotropic.

Biomechanical Phenomena↗

PubMed2005

Repair of osteochondral defect with tissue-engineered chondral plug in a rabbit model.

PURPOSE: The purpose of this study was to evaluate the macroscopic and histologic results of transplanting a tissue-engineered chondral plug made of atelocollagen sponge and PLLA mesh to treat osteochondral defects. TYPE OF STUDY: Controlled experimental study. METHODS: Twelve-week-old male Japanese white rabbits were used. Fresh articular cartilage slices were taken from the humeral head, and isolated chondrocytes were embedded in atelocollagen gel which does not have antigenic portions of collagen (2.0 x 10(6) cells/mL). They were seeded on the top of the atelocollagen sponge/PLLA mesh composite and cultured for 2 weeks. The culture medium was changed every 3 days and L-ascorbic acid (50 microg/mL) was added every 2 days. Culturing the composites for 2 weeks produced tissue-engineered chondral plugs. These tissue-engineered chondral plugs (4-mm diameter, 4-mm thick) were transplanted into the osteochondral defects (4 mm diameter, 4 mm deep) in the patellar grooves of the same rabbits from which the chondrocytes had been harvested (the experimental group). In the control group, the defects were treated with the plugs without chondrocytes. The rabbits were killed 4 and 12 weeks after transplantation. The repaired tissues were evaluated macroscopically and histologically, and analyzed immunohistochemically for expression of type II collagen. RESULTS: Four weeks after transplantation in the experimental group, the defects were partially repaired with cartilage-like tissue with good subchondral bone formation. Twelve weeks after transplantation, the defects were repaired with hyaline cartilage-like tissue densely stained by Safranin O. Well-organized subchondral bone formation was also observed. In the control group, the defects were covered with only soft fibrous tissue at 4 and 12 weeks macroscopically. Immunohistochemically, type II collagen was detected in about 90% of the repaired area. Histologic scores in the experimental group were significantly higher than those in the control group at both 4 and 12 weeks after transplantation. CONCLUSIONS: This study shows that the defects treated with tissue engineered chondral plug developed type II collagen in about 90% of the repaired area. CLINICAL RELEVANCE: The transplantation of a tissue-engineered chondral plug will be one option for treating osteochondral defects. The next step in testing our hypothesis is to evaluate the repaired tissue biomechanically and biochemically over a longer period of time.

Animals↗

PubMed2005

[Experimental study on biomechanics characteristics of combined collagen tissue engineering tendon].

OBJECTIVE: To investigate the influence of collagen on the biomechanics strength of tissue engineering tendon. METHODS: All of 75 nude mice were made the defect models of achilles tendons, and were divided into 5 groups randomly. Five different materials including human hair, carbon fibre (CF), polyglycolic acid (PGA), human hair and PGA, and CF and PGA with exogenous collagen were co-cultured with exogenous tendon cells to construct the tissue engineering tendons. These tendons were implanted to repair defect of achilles tendons of right hind limb in nude mice as experimental groups, while the materials without collagen were implanted to repair the contralateral achilles tendons as control groups. In the 2nd, 4th, 6th, 8th and 12th weeks after implantation, the biomechanical characteristics of the tissue engineering tendon was measured, meanwhile, the changes of the biomechanics strength were observed and compared. RESULTS: From the 2nd week to the 4th week after implantation, the experimental groups were stronger than the control groups in biomechanics, there was statistically significant difference (P < 0.05). From the 6th to 12th weeks, there was no statistically significant difference between the experiment and control groups (P > 0.05). Positive correlation existed between time and intensity, there was statistically significant difference (P < 0.05). The strength of materials was good in human hair, followed by CF, and PGA was poor. CONCLUSION: Exogenous collagen can enhance the mechanics strength of tissue engineering tendon, and is of a certain effect on affected limb

Animals↗

PubMed2004

Electrospinning collagen and elastin: preliminary vascular tissue engineering.

Significant challenges must be overcome before the true benefit and economic impact of vascular tissue engineering can be fully realized. Toward that end, we have pioneered the electrospinning of micro- and nano-fibrous scaffoldings from the natural polymers collagen and elastin and applied these to development of biomimicking vascular tissue engineered constructs. The vascular wall composition and structure is highly intricate and imparts unique biomechanical properties that challenge the development of a living tissue engineered vascular replacement that can withstand the high pressure and pulsatile environment of the bloodstream. The potential of the novel scaffold presented here for the development of a viable vascular prosthetic meets these stringent requirements in that it can replicate the complex architecture of the blood vessel wall. This replication potential creates an "ideal" environment for subsequent in vitro development of a vascular replacement. The research presented herein provides preliminary data toward the development of electrospun collagen and elastin tissue engineering scaffolds for the development of a three layer vascular construct.

Blood Vessel Prosthesis↗

PubMed2005

Effect of passage number and matrix characteristics on differentiation of endothelial cells cultured for tissue engineering.

Cells can sense the physical and chemical properties of artificial materials used as scaffolds for tissue engineering and regulate their behavior. Therefore, biomimetic and biospecific molecules are coated on materials to regulate function of cells on the tissue-engineered product. These bioactive molecules can be attached in a defined spectrum, concentration and spatial distribution in order to control adhesion, growth, viability, differentiation, and function of the cells. When autologous cells are used for tissue engineering, initially limited cells obtained may often need an amplification of cell number by passage in tissue culture before they are seeded on a biomaterial or scaffold. We have conducted this study to understand how the characteristics of bioactive molecule coating might affect proliferation, apoptosis and differentiation when endothelial cell (EC) is serially passaged. Proliferation was assessed by proliferating cell nuclear antigen (PCNA) staining along with counting of cells harvested from confluent monolayer. Apoptosis was assessed by Annexin V staining and differentiation by semi quantitative reverse transcriptase polymerase chain reaction (RT-PCR) for von Willebrand factor (vWF) expression and quantification of its release using enzyme linked immunosorbant assay (ELISA), and thrombogenicity by comparing platelet adhesion to EC monolayer Dacron grafts (DG) with specific protein coating. The results indicate that ECs easily lose its proliferation potential when they are cultured repeatedly on gelatin, turn apoptotic and over express the prothrombotic protein- vWF. Whereas, when it is grown on a matrix composed of fibrin, fibronectin, gelatin and vascular EC growth factor (VEGF), the cells retained their ability to proliferate, remained viable and were relatively less thrombogenic, even when passage number progressed. It is concluded that if ECs are grown on the composite matrix that mimics natural vessel scaffold, the cell number can be amplified without affecting its normal physiological function and may be used to generate effective tissue-engineered cardiovascular constructs.

Cell Adhesion↗

PubMedDate not supplied

Poly(lactic-co-glycolic acid) microspheres as an injectable scaffold for cartilage tissue engineering.

Injectable scaffold has raised great interest for tissue regeneration in vivo, because it allows easy filling of irregularly shaped defects and the implantation of cells through minimally invasive surgical procedures. In this study, we evaluated poly(lactic-co-glycolic acid) (PLGA) microsphere as an injectable scaffold for in vivo cartilage tissue engineering. PLGA microspheres (30-80 microm in diameter) were injectable through various gauges of needles, as the microspheres did not obstruct the needles and microsphere size exclusion was not observed at injection. The culture of chondrocytes on PLGA microspheres in vitro showed that the microspheres were permissive for chondrocyte adhesion to the microsphere surface. Rabbit chondrocytes were mixed with PLGA microspheres and injected immediately into athymic mouse subcutaneous sites. Chondrocyte transplantation without PLGA microspheres and PLGA microsphere implantation without chondrocytes served as controls. Four and 9 weeks after implantation, chondrocytes implanted with PLGA microspheres formed solid, white cartilaginous tissues, whereas no gross evidence of cartilage tissue formation was noted in the control groups. Histological analysis of the implants by hematoxylin and eosin staining showed mature and well-formed cartilage. Alcian blue/safranin O staining and Masson's trichrome staining indicated the presence of highly sulfated glycosaminoglycans and collagen, respectively, both of which are the major extracellular matrices of cartilage. Immunohistochemical analysis showed that the collagen was mainly type II, the major collagen type in cartilage. This study demonstrates the feasibility of using PLGA microspheres as an injectable scaffold for in vivo cartilage tissue engineering. This scaffold may be useful to regenerate cartilaginous tissues through minimally invasive surgical procedures in orthopedic, maxillofacial, and urologic applications.

Animals↗

PubMedDate not supplied

Tissue-engineered bone for lateral alveolar ridge augmentation: a case report.

Bone matrix derived from mandibular periosteal cells and cultivated by tissue engineering on a polymer fleece has recently been used for sinus floor elevation and augmentation. This case report focuses on clinical and histologic results after lateral ridge augmentation of a localized non-space maintaining defect in the right posterior area of the mandible using tissue-engineered bone. Implant supported prosthetic rehabilitation of a partially edentulous 32-year-old woman was planned involving a fixed partial denture. Preoperative investigations revealed a transversely reduced alveolar ridge width on the right side of the posterior mandible. Lateral augmentation was performed using tissue-engineered bone obtained by autogenous periosteum cells from the same area. Six months after augmentation 2 implants were placed and a bone biopsy was obtained from the augmented area. Transverse ridge dimensions were found to be enhanced. Histologic examination of the biopsy revealed dense lamellar bone. Wound healing was uneventful after all surgical interventions. This case report demonstrates the successful clinical application of tissue-engineered bone for lateral augmentation of the transversely reduced alveolar ridge. The results suggest that periosteum-derived tissue-engineered bone can be used to create a sufficient implant site not only for the sinus floor elevation and augmentation procedure for vertical bone enhancement but also for lateral augmentation.

Adult↗

PubMed2006

Tissue engineering of articular cartilage with autologous cultured adipose tissue-derived stromal cells using atelocollagen honeycomb-shaped scaffold with a membrane sealing in rabbits.

Adipose tissue derived stromal cells (ATSCs), which were isolated from adipose tissue of rabbit, have shown to possess multipotential, that is, they differentiate into osteoblasts and adipocytes in plate-culturing and into chondrocytes in an established aggregate culture using defined differentiation-inductive medium. The aim of this study was to evaluate the utility of ATSCs in tissue engineering procedures for repair of articular cartilage-defects using the atelocollagen honeycomb-shaped scaffold with a membrane sealing (ACHMS-scaffold). We intended to repair full-thickness articular cartilage defects in rabbit knees using autologously cultured ATSCs embedded in the ACHMS-scaffold. ATSCs were incubated within the ACHMS-scaffold to allow a high density and three-dimensional culture with control medium. An articular cartilage defect was created on the patellar groove of the femur, and the defect was filled with the ATSCs-containing ACHMS-scaffold, ACHMS-scaffold alone, or empty (control). Twelve weeks after the operation, the histological analyses showed that only the defects treated with the ATSCs-containing ACHMS-scaffold were filled with reparative hyaline cartilage, highly expressed Type II collagen. These results indicate that transplantation of autologous ATSCs-containing ACHMS-scaffold is effective in repairing articular cartilage defects.

Adipose Tissue↗

PubMed2001

Tissue engineering in urology.

Congenital abnormalities, cancer, trauma, infection, inflammation, iatrogenic injuries, and other conditions may lead to genitourinary organ damage or loss, requiring eventual reconstruction. Tissue engineering follows the principles of cell transplantation, materials science, and engineering toward the development of biological substitutes that would restore and maintain normal function. Tissue engineering may involve matrices alone, wherein the body's natural ability to regenerate is used to orient or direct new tissue growth, or the use of matrices with cells. Both synthetic (polyglycolic acid polymer scaffolds alone and with co-polymers of poly-1-lactic acid and poly-DL-lactide-coglycolide) and natural biodegradable materials (processed collagen derived from allogeneic donor bladder submucosa and intestinal submucosa) have been used, either alone or as cell delivery vehicles. Tissue engineering has been applied experimentally for the reconstitution of several urologic tissues and organs, including bladder, ureter, urethra, kidney, testis, and genitalia. Fetal applications have also been explored. Recently, several tissue engineering technologies have been used clinically, including the use of cells as bulking agents for the treatment of vesicoureteral reflux and incontinence, urethral replacement, and bladder reconstruction. Recent progress suggests that engineered urologic tissues may have clinical applicability in the future.

Humans↗