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At least 631 records · Page 35Linked to original sources

PubMed2003

[Preliminary research for constructing tissue-engineered oral mucosa].

OBJECTIVE: To search for a method of constructing tissue-engineered oral mucosa. METHODS: Hard palate mucoperiosteum were excised and extracted from raw SD milk rat. Tissue-engineered oral mucosa was made with the cultured oral keratinocytes that had been digested by Dispase and cultured in the serum-free keratinocytes medium, the supportive membrane being made from sodium alginate. RESULTS: Rat oral mucosal epithelial cells could be obtained with Dispase digestion. It was found that the best time for oral mucosa membrane to be digested to independent cells by Dispase again is ten minutes after the mucosa membrane has been obtained by Dispase; the best density of rat oral mucosa cells cultured is 1.5 x 10(5)/cm2, and the mucosa cells will be difficult to form the colon if the density of cells is too low. The keratinocytes can be cultured in serum-free keratinocytes medium without fibroblast contamination; the mucosa cells grow well on the sodium alginate membrane. CONCLUSION: Tissue-engineered oral mucosa can be constructed with the cultured oral keratinocytes in the serum-free keratinocytes medium and the self-made sodium alginate membrane.

Alginates↗

PubMed2002

Tissue-engineered small intestine: ontogeny of the immune system.

BACKGROUND: Using tissue-engineering techniques, we have developed a that regenerates structural and transporter properties of native jejunum. The purpose of this study was to characterize the mucosal immune system of the engineered neointestine. We hypothesized that the neointestinal mucosa is capable of developing a mature immunocyte population and that exposure to luminal stimuli is critical to this development. METHODS: Neointestinal cysts were engineered by implanting polymer-organoid constructs into syngeneic adult recipients. Neointestine (cysts left nonanastomosed [NA] and cysts anastomosed to native bowel [AN]) and native jejunum were harvested serially (3-56 weeks postoperatively). Immune cell subsets were characterized by the immunohistochemical detection of cell-specific antigens (T cells [CD3], B cells [CD32], NK cells [CD56], and macrophages [CD68]) combined with computer-based morphometry. RESULTS: Intraepithelial and lamina propria immunocyte population densities and subset distributions were identical in AN cysts harvested 20 weeks postoperatively and in native jejunum. Mucosal immunocyte population densities were lower in AN cysts harvested 10 weeks postoperatively and only rudimentary in NA cysts, even those harvested 20 weeks postoperatively. CONCLUSIONS: These results suggest that tissue-engineered intestine has the capacity to develop a mucosal immune system with an immunocyte population similar to that of native small intestine. The development of this immune system is a function of both exposure to luminal stimuli and the duration of this exposure. Tissue-engineered intestine offers promise as a new therapeutic approach for patients who have intestinal insufficiency.

Anastomosis, Surgical↗

PubMed2006

Tissue engineering of endothelial cells and the immune response.

BACKGROUND: While tissue engineering offers promise for organ and tissue transplantation, it can also be used to examine transplant and immune biology. Endothelial cells engrafted within 3-dimensional matrices create stable units that produce all of the factors of a functional quiescent endothelium. Perivascular implantation of tissue engineered endothelial cell constructs provides long-term control of vascular repair after injury. This control is established without restoration of the natural luminal:mural endothelium, and most intriguingly, without engendering host allo- and xenogeneic immune responses. We examined how endothelial immunogenicity is controlled by interaction with 3-dimensional matrices. MATERIALS AND METHODS: Human aortic endothelial cells (HAE) were either grown to confluence on polystyrene tissue culture plates or within 3-dimensional collagen-based matrices. Major histocompatibility complex (MHC) class II, integrin, interferon (IFN)-gamma receptor expression, and signaling were analyzed via confocal microscopy, flow cytometry, reverse transcription polymerase chain reaction (RT-PCR), and microarray. Splenocyte proliferation was assayed by thymidine incorporation. RESULTS: Despite similar expression levels of IFN-gamma receptors, matrix-embedded HAE elicited far less STAT-1 phosphorylation upon IFN-gamma stimulation, and expressed 2-fold less MHC II than HAE grown to confluence on culture plates (P < .001). This effect correlated with reduced expression of integrin alpha(v) and beta(3) (P < .002), and muted proliferation of porcine splenocytes (P < .001). CONCLUSIONS: Matrix architecture is critical for modulation of endothelial immunogenicity. Embedding HAE within a physiologic 3-dimensional environment affects activity of intracellular signaling pathways, MHC II expression, and subsequent activation of immune cells. These findings might offer novel insights into our understanding of endothelial-mediated diseases and might enhance our ability to leverage the potential for cell-based therapies.

Animals↗

PubMed2005

Combined chondrocyte-copolymer implantation with slow release of basic fibroblast growth factor for tissue engineering an auricular cartilage construct.

Basic fibroblast growth factor (b-FGF) may have a role in tissue-engineered chondrogenesis. However, when applied in solution, b-FGF rapidly diffuses from the implant site. In another approach for tissue engineering, poly-lactide-based copolymers have shown promise as scaffolds for chondrocytes used to tissue engineer auricular cartilage in the shape of an ear. This study evaluated the effectiveness of b-FGF impregnated in gelatin microspheres to achieve slow growth factor release for augmenting the in vivo chondrogenic response. Whereas 125I-labeled b-FGF injected in solution showed rapid in vivo clearance from the injection site (only 3% residual after 24 h), when incorporated into gelatin microspheres, 44% and 18% of the b-FGF remained at 3 and 14 days, respectively. Canine chondrocytes were isolated and grown in vitro onto ear-shaped poly-lactide/caprolactone copolymers for 1 week, then implanted into the dorsal subcutaneous tissue of nude mice; implants contained b-FGF either in free solution or in gelatin microspheres. A third group underwent preinjection of b-FGF in gelatin microspheres 4 days before chondrocyte-copolymer implantation. The implants with b-FGF-incorporated microspheres showed the greatest chondrogenic characteristics at 5 and 10 weeks postoperatively: good shape and biomechanical trait retention, strong (histologic) metachromasia, rich vascularization of surrounding tissues, and increased gene expression for type II collagen (cartilage marker) and factor VIII-related antigen (vascular marker). In the case of implant site preadministration with b-FGF-impregnated microspheres, the implant architecture was not maintained as well, and reduced vascularization and metachromasia was also apparent. In conclusion, these findings indicate that a sustained release of b-FGF augments neovascularization and chondrogenesis in a tissue-engineered cartilage construct.

Animals↗

PubMed2003

Fabrication of precise cylindrical three-dimensional tissue engineering scaffolds for in vitro and in vivo bone engineering applications.

It is sometimes necessary to form highly porous polymeric tissue engineering scaffolds into various shapes and sizes. Ideally, in these cases, the three-dimensional morphology should be maintained to the outer margins of the scaffold so as to provide optimum function. Many biodegradable polymeric scaffolds are soft and delicate, however, and their poor physical strength presents a challenge when cutting these materials into the required shapes. We describe a simple device that can be used quickly and accurately to cut cylindrical shapes from such delicate polymeric scaffold materials, which maintain their morphological features to the margins of the shapes produced. We demonstrate that the device can be used to create scaffolds with reproducible dimensions having an SD in mass of less then 6%. The in vitro utility of scaffolds cut with the device was established through demonstrating bone marrow-derived cell invasion into fibrin-filled scaffolds that fit precisely into the wells of 24-well plates. We also demonstrate the in vivo utility of precise cylindrically shaped scaffolds by observing rapid bone invasion into 2.4-mm diameter scaffolds that have been placed into drill hole defects in the distal femur of young rats. When scaffolds are filled with fibrin before implantation as part of a bone tissue engineering strategy, less blood fills the defect site and the fibrin is gradually remodeled and replaced by bone. The ability to cut precise cylindrical scaffolds in the millimeter size range has allowed for the creation of a new small animal model that may prove useful for screening tissue engineering scaffolds for further study.

Animals↗

PubMed2005

Galactose-carrying polymers as extracellular matrices for liver tissue engineering.

Extracellular matrix (ECM) plays important roles in tissue engineering because cellular growth and differentiation, in the two-dimensional cell culture as well as in the three-dimensional space of the developing organism, require ECM with which the cells can interact. Especially, the bioartificial liver-assist device or regeneration of the liver-tissue substitutes for liver tissue engineering requires a suitable ECM for hepatocyte culture because hepatocytes are anchorage-dependent cells and are highly sensitive to the ECM milieu for the maintenance of their viability and differentiated functions. Galactose-carrying synthetic ECMs derived from synthetic polymers and natural polymers bind hepatocytes through a receptor-mediated mechanism, resulting in enhanced hepatocyte functions. Attachment and functions of hepatocytes were affected by physico-chemical properties including ECM geometry as well as the type, density and orientation of galactose. Also, cellular environment, medium composition and dynamic culture system influenced liver-specific functions of hepatocytes beside ECM.

Animals↗

PubMed2005

A tissue-engineered suburethral sling in an animal model of stress urinary incontinence.

OBJECTIVE: To create and evaluate the functional effects of a tissue-engineered sling in an animal model of stress urinary incontinence (SUI). MATERIALS AND METHODS: Twenty female Sprague-Dawley rats were divided into four equal groups: a control group (C) had no intervention before the leak-point pressure (LPP) was measured; a denervated group (D) had bilateral proximal sciatic nerve transection (PSNT) and periurethral dissection with no sling placed; group S had concomitant bilateral PSNT and a suburethral sling of small intestinal submucosa (SIS) placed; and group (M) had concomitant bilateral PSNT with implantation of a tissue-engineered sling. The suburethral sling was placed via a transabdominal approach with the sling sutured to the pubic bone. Tissue-engineered slings were prepared with muscle-derived cells obtained via the pre-plate technique and subsequently seeded for 2 weeks on a SIS scaffold. Suburethral slings were implanted 2 weeks before LPP testing, using the vertical-tilt method. RESULTS: Surgically placing a suburethral sling is feasible in the female rat, with few complications. LPPs from both sling groups (S and M) were not significantly different from untreated controls (C). The S, M and C groups all had significantly higher LPPs than group D. Importantly, no rat from either sling group (S and M) had signs of urinary retention. CONCLUSIONS: Placing tissue-engineered slings in an animal model of SUI resulted in LPP values that were not significantly different from those in untreated control or SIS (S) groups. These data show that incorporating muscle stem cells into SIS slings does not adversely alter the advantageous mechanical properties of the SIS sling in a model of SUI, and provide the basis for future functional studies of tissue-engineered sling materials with long-term retention.

Animals↗

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↗

PubMedDate not supplied

Characterization of the response of bone marrow-derived progenitor cells to cyclic strain: implications for vascular tissue-engineering applications.

One of the major failings in vascular tissue engineering is the limited capacity of autologous differentiated cells to reconstitute tissues. A logical solution is to use multipotent progenitor cells, which in vascular treatments have been underutilized. Although biochemical stimulation has been explored to differentiate bone marrow-derived progenitor cells (BMPCs) to smooth muscle cells (SMCs), the use of biomechanical forces in differentiation remains unexplored. The purpose of this work was to explore the effects of cyclic strain alone on BMPC morphology, proliferation, and differentiation. BMPCs were isolated from rat bone marrow and, after 7 days in culture, the cells grew in distinct multilayered colonies. BMPCs were stimulated with 10% strain at 1 Hz for 7 days. Observations showed that cyclic strain inhibited proliferation (p < 0.05) and caused alignment of the cells (p < 0.05) and of the F-actin cytoskeleton perpendicular to the direction of strain. In addition, cyclic strain resulted in expression by the cells of vascular smooth muscle alpha-actin and h1-calponin. This work demonstrates the potential of physiologic biomechanical stimulation in the differentiation of BMPCs to SMCs, and this could have important implications for vascular tissue engineering and other therapies in which cell sourcing is a major concern.

Actins↗

PubMed2000

The living shunt: a tissue engineering approach in the treatment of hydrocephalus.

Tissue engineering is the use of cultured cells seeded into biodegradable polymers to create custom designed, living implantable devices. As a first approach to the use of this technique in the treatment of hydrocephalus, we have prepared chondrocyte-seeded polyglycolic acid (PGA) tubes coated with polylactic glycolic acid (PGLA), implanted initially with thin silastic stents removed four weeks after shunt insertion. The use of bovine xenograft cells in athymic (nude) rats resulted in more efficient seeding with chondrocytes, stiffer tube walls, and better patency. When implanted in 6-week-old rats made hydrocephalic by cisternal injection of kaolin at 4 weeks of age, six of eight 'living shunts' remained patent to radio-opaque contrast injection at two weeks after stent removal. At four weeks after stent removal, all four of the shunts had occluded at the ventricular end, three of the four apparently due to growth of the animal. We conclude that polymer type, cell type, and cell density will require considerable optimization, but a working tissue engineered shunt is feasible and may one day address some problems of interactions of living tissue and inert polymer.

Animals↗

PubMed2002

To fabricate artificial nerves with tissue engineering methods.

OBJECTIVE: To fabricate artificial nerves with tissue engineering methods in vitro. METHODS: Schwann cells (SCs) were cultured and seeded on polyglactin 910 fibers wrapped by biomembrane coated with rat tail glue and laminin for 2 weeks. The absorbability on the scaffolds, growth and migration of SCs were assessed with a light microscope, a scanning electron microscope and a transmission electron microscope. RESULTS: SCs could migrate and proliferate on polyglactin 910 fibers. They were well distributed between scaffolds and absorbed on surface of scaffolds and formed a bungner band, on which SCs produced more matrices. SCs seeded on the biomembrane could also grow well. Axon regeneration in the distal nerve stump was observed at 8 weeks. CONCLUSIONS: Adult SCs can be expanded on coated fibers and biomembrane. Three-dimensional scaffold of SCs has the basic characteristics of artificial nerves. These findings offer a novel method to fabricate artificial nerves with tissue engineering methods for repairing defected long nerves.

Animals↗

PubMed2006

[A comparative study on seeding methods of human mesenchymal stem cells in bone tissue engineering].

Seeding is the crucial step in bone tissue engineering. In current study, static and dynamic seeding methods for human bone marrow stem cells (hBMSCs) were compared. The methods for assay of DNA content in the constructs after seeding were adopted. The optic microscopy for histological apearance and the scanning electron microscopy (SEM) and fluorescent RT-PCR for osteogenic markers were performed. The maximal initial seeding concentration in static seeding is lower than that in dynamic seeding. Histology and SEM revealed the even distribution and spreading of cells in the dynamically seeded constructs, but showed cell aggregation in the statically seeded counterparts. Fluorescent RT-PCR again revealed stronger osteogenic potential of dynamically seeded constructs. Therefore, this initial study demonstrated that dynamic seeding of human bone marrow stem cells is a promising technique in bone tissue engineering.

Bone Marrow Cells↗

PubMed2003

[Functional reconstruction with tissue engineered myoblast in facial muscle of rat].

OBJECTIVE: The purpose of this article is to discuss the reconstruction of facial muscle defects with tissue engineered myoblast in SD rats. METHODS: Using purified, subcultured myoblast of neonatal rats and type I collagen gels as extracellular matrix (ECM) and scaffold, tissue engineered muscle was transplanted in the face of syngeneic nutured rats. RESULTS: Tissue engineered myoblast generated and differentiated in vitro were observed with microscope. Myoblast fused each other and formed myofibers in the face of rats, nerve fibers and vessels regeneration could also be found in some samples. Postoperative electromiographs showed the myofibers were active when stimulating the nerve trunk that innervates the engineered facial muscle. CONCLUSION: Tissue engineered method is hopeful to be used as a new technique to reconstruct defects of facial muscle.

Animals↗

PubMed1998

An economic survey of the emerging tissue engineering industry.

The contemporary scope of worldwide tissue engineering research and development was estimated by totaling the relevant annual spending and other economic parameters of firms involved the field. Operating expenses allocated to tissue engineering in 1997 exceed $450 million and fund the activities of nearly 2,500 scientists and support personnel. Growth rate is 22.5% per annum. Most activity is centered in the United States. Government spending in this field represents <10% of the total. The aggregate capital value of start-ups that have gone public was approximately $1.7 billion as of January 1, 1998; total capital value of all firms and business units in the field was estimated to be roughly $3.5 billion. The level of investment and valuation represents a remarkable act of faith in the future of a technology yet to produce its first significant revenue-generating product.

Artificial Organs↗

PubMed2005

Optimal segmentation of microcomputed tomographic images of porous tissue-engineering scaffolds.

The morphometric properties of the porous tissue-engineering scaffolds play a dominant role in the initial cell attachment and subsequent tissue regeneration. These properties can be derived nondestructively with the use of quantitative analysis of high-resolution microcomputed tomography (microCT) imaging of scaffolds. Accurate segmentation of these acquired images into solid and porous subspaces is critical to the integrity of morphometric analysis. The absence of a single image-processing technique to provide such accurate separability immune to all the intricacies of the acquired data makes this seemingly simple task significantly error prone. Consequently, an optimal segmentation has to be selected by ranking the segmentations produced by a multiplicity of methods. This article proposes a robust, easy-to-implement, unambiguous, signal-processing-based, ground-truth-free, segmentation rating metric that correlates with visual acuity. With the use of this metric it is possible, for the first time, to threshold the data with a wide range of techniques and select automatically the technique that best delineates the acquired image. The proposed solution has been extensively tested on microCT images of scaffolds fabricated with biodegradable poly (propylene fumarate) (PPF) with the use of a solvent casting particulate leaching process. The approaches proposed and the results obtained may have profound implications for accurate image-based characterization of tissue-engineering scaffolds.

Biocompatible Materials↗

PubMed2005

Porous scaffold design for tissue engineering.

A paradigm shift is taking place in medicine from using synthetic implants and tissue grafts to a tissue engineering approach that uses degradable porous material scaffolds integrated with biological cells or molecules to regenerate tissues. This new paradigm requires scaffolds that balance temporary mechanical function with mass transport to aid biological delivery and tissue regeneration. Little is known quantitatively about this balance as early scaffolds were not fabricated with precise porous architecture. Recent advances in both computational topology design (CTD) and solid free-form fabrication (SFF) have made it possible to create scaffolds with controlled architecture. This paper reviews the integration of CTD with SFF to build designer tissue-engineering scaffolds. It also details the mechanical properties and tissue regeneration achieved using designer scaffolds. Finally, future directions are suggested for using designer scaffolds with in vivo experimentation to optimize tissue-engineering treatments, and coupling designer scaffolds with cell printing to create designer material/biofactor hybrids.

Animals↗

PubMed2005

Characterization of knitted polymeric scaffolds for potential use in ligament tissue engineering.

Different scaffolds have been designed for ligament tissue engineering. Knitted scaffolds of poly-L-lactic acid (PLLA) yarns and co-polymeric yarns of PLLA and poly(glycolic acid) (PLGA) were characterized in the current study. The knitted scaffolds were immersed in medium for 20 weeks, before mass loss, molecular weight, pH value change in medium were tested; changes in mechanical properties were evaluated at different time points. Results showed that the knitted scaffolds had 44% porosity. There was no significant pH value change during degradation, while there was obvious mass loss at initial 4 week, as well as smooth molecular weight drop of PLLA. PLGA degraded more quickly, while PLLA kept its integrity for at least 20 weeks. Young's modulus increased while tensile strength and strain at break decreased with degradation time; however, all of them could maintain the basic requirements for ACL reconstruction. It showed that the knitted polymeric structures could serve as potential scaffolds for tissue-engineered ligaments.

Biodegradation, Environmental↗

PubMed1998

A completely biological tissue-engineered human blood vessel.

Mechanically challenged tissue-engineered organs, such as blood vessels, traditionally relied on synthetic or modified biological materials for structural support. In this report, we present a novel approach to tissue-engineered blood vessel (TEBV) production that is based exclusively on the use of cultured human cells, i.e., without any synthetic or exogenous biomaterials. Human vascular smooth muscle cells (SMC) cultured with ascorbic acid produced a cohesive cellular sheet. This sheet was placed around a tubular support to produce the media of the vessel. A similar sheet of human fibroblasts was wrapped around the media to provide the adventitia. After maturation, the tubular support was removed and endothelial cells were seeded in the lumen. This TEBV featured a well-defined, three-layered organization and numerous extracellular matrix proteins, including elastin. In this environment, SMC reexpressed desmin, a differentiation marker known to be lost under standard culture conditions. The endothelium expressed von Willebrand factor, incorporated acetylated LDL, produced PGI2, and strongly inhibited platelet adhesion in vitro. The complete vessel had a burst strength over 2000 mmHg. This is the first completely biological TEBV to display a burst strength comparable to that of human vessels. Short-term grafting experiment in a canine model demonstrated good handling and suturability characteristics. Taken together, these results suggest that this novel technique can produce completely biological vessels fulfilling the fundamental requirements for grafting: high burst strength, positive surgical handling, and a functional endothelium.

Animals↗