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PubMedDate not supplied

In vivo response of polylactic acid-alginate scaffolds and bone marrow-derived cells for cartilage tissue engineering.

Successful application of tissue-engineering techniques to damaged biological structures is determined by functional performance in vivo. This study evaluated the in vivo response of a tissue-engineered construct composed of a polylactic acid-alginate amalgam seeded with bone marrow-derived mesenchymal stem cells and stimulated in vitro with transforming growth factor beta for cartilage tissue engineering. Constructs were placed in cylindrical osteochondral defects in the canine femoral condyle and examined 6 weeks postoperatively by gross, histological, immunohistochemical, and biomechanical analyses. In the course of 6 weeks in vivo, the defects filled with a cartilaginous tissue regardless of whether cell-seeded (experimental) or cell-free (control) constructs were implanted; however, the quality of the tissue differed between the experimental and control defects. Cell-seeded experimental defects showed more cartilage-like matrix quality, cell distribution, and proteoglycan staining. Biomechanically, experimental and control specimens exhibited similar behavior; however, both tissues were still immature compared with normal cartilage. The evidence accumulated in this study showed a modest acceleration of the in vivo healing of cell-seeded constructs but also demonstrated a reparative response of cell-free constructs. This finding suggests that the constructs prepared from the PLA-alginate amalgam may serve as a means for host cell attachment.

Alginates↗

PubMed2006

Stem cells and tissue engineering: past, present, and future.

Tissue engineering is an interdisciplinary field that brings together the principles of the life sciences and medicine with those of engineering. The increase in its development over the past decade has resulted from a variety of factors; advances in genomics and proteomics, the advent of new biomaterials as potential templates for tissue growth, improvements in bioreactor design, and increased understanding of healing processes. Possibly the greatest contribution has come from our increased knowledge and understanding of stem cell biology, which is paving the way for the generation of unlimited cells of specific phenotypes for incorporation into engineered tissue constructs. Thus, tissue engineering approaches for expanding and engrafting the differentiated progeny of embryonic, fetal, or adult stem cells have major potential for tissue repair and will make a major contribution to medicine in the 21st century.

Animals↗

PubMed2004

Potential tissue-engineering applications for neonatal surgery.

Tissue engineering attempts to build neotissue from its cellular building blocks. This neotissue can then be used for reconstructive surgical applications such as replacement of a congenitally abnormal heart valve or repair of a craniofacial abnormality. Since its inception in the late 1980s, tissue engineering has sparked the interests of physicians and scientists alike because of its great potential. Significant progress has been made in this burgeoning branch of science. This article reviews some of the ongoing preclinical and clinical tissue engineering research as it applies to neonatology.

Animals↗

PubMed2005

Effects of initial cell seeding density for the tissue engineering of the temporomandibular joint disc.

Tissue engineering may provide a better treatment modality for postoperative discectomy patients. The TMJ disc is an ideal candidate for tissue engineering approaches because of its lack of an intrinsic regenerative ability. Unfortunately, basic knowledge related to TMJ disc tissue engineering is still at an infancy level and not on par to that related to articular cartilage tissue engineering. The objective of this study was to examine the effects of initial cell density of TMJ disc cells seeded in nonwoven poly-glycolic acid (PGA) scaffolds on the biochemical and biomechanical properties of constructs examined at 0, 3, and 6 weeks after seeding. Low, medium, and high seeding densities were chosen to be 15, 30, and 120 million cells per ml of scaffold, which were seeded using a spinner flask. Significant differences were found temporally and as a function of seeding density in morphology, total collagen, GAG content, and permeability of the constructs, but not in aggregate modulus. The high seeding density group outperformed the low and medium groups in collagen and GAG content at all time points measured. The high-density group produced a total of 55.37 +/- 3.56 microg of collagen per construct, maintained 15.77 +/- 1.86 microg of GAG per construct, and only shrunk to 50% of the original scaffold size. Permeability of the constructs at 6 weeks was decreased by 70% compared to 0 weeks.

Animals↗

PubMed2004

Creating artificial perichondrium by polymer complex membrane macroencapsulation: immune protection and stabilization of subcutaneously transplanted tissue-engineered cartilage.

Functional organ or tissue failure is one of the most frequent, devastating and costly problems in modern health care. The field of tissue engineering has tremendous potential for developing new functional tissue. In reconstructive surgery, cartilage engineering could be a serious alternative to the established method of autologous cartilage transplantation. Recent studies demonstrate cartilage engineering by subcutaneous implantation of chondrocyte-seeded PGA/PLA-fibrin glue scaffolds in the backs of nude mice. In both autologous cartilage transplantation and cartilage engineering, the host immune response affects transplant integrity and cartilage morphology to an unforeseeable extent. To investigate whether polyelectrolyte complex (PEC) membranes can prevent rejection of cartilage transplants without neglecting tissue metabolism, tissue-engineered cartilage encapsulated with a PEC membrane was subcutaneously implanted in the backs of nude mice. Non-encapsulated tissue-engineered cartilage was used for the control group. Histochemistry and scanning electron microscopy were performed 4 and 12 weeks after implantation. There was no interaction between the host and the implant with an intact PEC membrane. With protection by PEC encapsulation, implanted tissue-engineered cartilage showed no signs of degeneration and had a significantly weaker cellular immune response than without it. Thus, PEC membrane encapsulation appears to be a novel approach for protecting cartilage implants from host immune response after autologous transplantation.

Animals↗

PubMed2006

Tissue engineering of blood vessels.

BACKGROUND: Tissue engineering techniques have been employed successfully in the management of wounds, burns and cartilage repair. Current prosthetic alternatives to autologous vascular bypass grafts remain poor in terms of patency and infection risk. Growing biological blood vessels has been proposed as an alternative. METHODS: This review is based on a literature search using Medline, PubMed, ISIS and CAS of original articles and reviews, and unpublished material and abstracts. RESULTS AND CONCLUSIONS: Complete incorporation into host tissues and the maintenance of a viable and self-renewing endothelial layer are the fundamental goals to be achieved when developing a tissue-engineered blood vessel. Sourcing of cells and modulating their interaction with extracellular matrix and supporting scaffold have been the focus of intense research. Although the use of tissue-engineered blood vessels in humans is so far limited, advances in our knowledge of stem cell precursors and the development of new biomaterials should enable this technology to reach routine clinical practice within a decade.

Biotechnology↗

PubMed2000

[Tissue engineering: possibilities and perspectives].

Successful tissue engineering requires intensive co-oporation between clinicians, biologists (cell culture, gene therapy), chemical engineers (biomaterials) and industrial partners. In case of wound healing tissue engineered constructs have already been applied successfully in burns and chronic wounds. In order to improve carrier and matrix function biomaterials still have to be optimized. The potential of such constructs might even be enhanced by gene therapeutical methods. The complex mammalian organism has to be considered as the gold standard and the model for perfect tissue engineering. The problem of vascularization of complex organs yet has to be solved. In general it seems to be more promising to substitute deficient components in vivo and to rely on modulating influences within the host organism rather than to create complex organs ex vivo.

Animals↗

PubMed2003

[Construction of tissue engineering skin containing melanocytes].

OBJECTIVE: To construct a tissue engineering skin containing melanocytes by employing tissue engineering method. METHODS: The keratinocytes, dermal fibroblasts and melanocytes were isolated and purified. Then the cells were used to construct a tissue engineering skin containing melanocytes. The location of melanocytes in the tissue engineering skin were detected by Dopa staining, transmission electron microscope (TEM) and S-100 immunohistochemical staining. RESULTS: Melanocytes can be detected in the basal layer of the constructed tissue engineering skin. The results of TEM showed that the melanocytes were in good conditions. CONCLUSION: The artificial skin containing melanocytes was successfully constructed in vitro and can be used to repair the full-thickness skin defects.

Adolescent↗

PubMed2000

Biomaterial developments for bone tissue engineering.

The development of bone tissue engineering is directly related to changes in materials technology. While the inclusion of materials requirements is standard in the design process of engineered bone substitutes, it is also critical to incorporate clinical requirements in order to engineer a clinically relevant device. This review presents the clinical need for bone tissue-engineered alternatives to the present materials used in bone grafting techniques, a status report on clinically available bone tissue-engineering devices, and recent advances in biomaterials research. The discussion of ongoing research includes the current state of osseoactive factors and the delivery of these factors using bioceramics and absorbable biopolymers. Suggestions are also presented as to the desirable design features that would make an engineered device clinically effective.

Biocompatible Materials↗

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↗

PubMed2006

Small molecule inducers of angiogenesis for tissue engineering.

Engineering of implantable tissues requires rapid induction of angiogenesis to meet the significant oxygen and nutrient demands of cells during tissue repair. To this end, our laboratories have utilized medicinal chemistry to synthesize non-peptide-based inducers of angiogenesis to aid tissue engineering. In this study, we describe the evaluation of SC-3-149, a small molecule compound with proliferative effects on vascular endothelial cells. Specifically, exogenous exposure of SC-3-149 induced an 18-fold increase in proliferation of human microvascular endothelial cells in vitro at low micromolar potency by day 14 in culture. Moreover, SC-3-149 significantly increased the formation of endothelial cord and tubelike structures in vitro, and improved endothelial scratch wound healing within 24 h. SC-3-149 also significantly inhibited vascular endothelial cell death owing to serum deprivation and high acidity (pH 6). Concurrent incubation of SC-3-149 with vascular endothelial growth factor increased cell survivability under serum-deprived conditions by an additional 7%. In addition, in vivo injection of SC-3-149 into the rat mesentery produced qualitative increases in microvessel length density. Taken together, our studies suggest that SC-3-149 and its analogs may serve as promising new angiogenic agents for targeted drug delivery and therapeutic angiogenesis in tissue engineering.

Angiogenesis Inducing Agents↗

PubMed2002

[Clinical application of tissue-engineered autologous oral mucosa transplants].

BACKGROUND: Tissue engineering of autogenous oral mucosa is a significant enrichment for various indications in oral and maxillofacial surgery. Besides preprosthetic surgery and prelamination of microvascular fasciocutaneous flaps, interdisciplinary work such as reconstruction procedures in cases of hypospadias and epispadias are the main indications for tissue-engineered oral mucosa. RESULTS: One main advantage of this technique is the reduction of second surgical procedures. The morbidity is decreased and the quality of life may be significantly increased. The manufacture of tissue-engineered autogenous oral mucosa up to 15 cm(2) requires a small biopsy of oral mucosa (4-8 mm(3)) and 40 ml of autogenous serum. DISCUSSION: The clinical and morphological long-term follow-up after different surgical procedures with tissue-engineered oral mucosa establishes this technique as an excellent additional tool in oral and maxillofacial surgery.

Female↗

PubMed2006

Testing the utility of rationally engineered recombinant collagen-like proteins for applications in tissue engineering.

Collagens are attractive proteins as materials for tissue engineering. Over the last decade, significant progress has been made in developing technologies for large-scale production of native-like human recombinant collagens. Yet, the rational design of customized collagen-like proteins for smart biomaterials to enhance the quality of engineered tissues has not been explored. We mapped the D4 domain of human collagen II as most critical for supporting migration of chondrocytes and used this information to genetically engineer a collagen-like protein consisting of tandem repeats of the D4 domain (mD4 collagen). This novel collagen has been utilized to fabricate a scaffold for support of chondrocytes. We determined superior qualities of cartilaginous constructs created by chondrocytes cultured in scaffolds containing the mD4 collagen in comparison to those formed by chondrocytes cultured in bare scaffolds or those coated with wild-type collagen II. Our results are a first attempt to rationally engineer collagen-like proteins with characteristics tailored for specific needs of cartilage engineering and provide a basis for rational engineering of similar proteins for a variety of biomedical applications.

Cartilage↗

PubMed2001

Construction and biological characterization of an HB-GAM/FGF-1 chimera for vascular tissue engineering.

OBJECTIVE: Cardiovascular tissue engineering approaches to vessel wall restoration have focused on the potent but relatively nonspecific and heparin-dependent mesenchymal cell mitogen fibroblast growth factor 1 (FGF-1). We hypothesized that linking FGF-1 to a sequence likely to bind to cell surface receptors relatively more abundant on endothelial cells (ECs) might induce a relative greater EC bioavailability of the FGF-1. We constructed a heparin-binding growth-associated molecule (HB-GAM)/FGF-1 chimera by linking full-length human HB-GAM to the amino-terminus of human FGF-1beta (21-154) and tested its activities on smooth muscle cells (SMCs) and ECs. METHODS: Primary canine carotid SMCs and jugular vein ECs were plated in 96-well plates in media containing 10% fetal bovine serum and grown to approximately 80% confluence. After being growth arrested in serum-free media for 24 hours, the cells were exposed to concentration ranges of cytokines and heparin, and proliferation was measured with tritiated-thymidine incorporation. Twenty percent fetal bovine serum was used as positive control, and phosphate-buffered saline was used as negative control. RESULTS: In the presence of heparin the HB-GAM/FGF-1 chimera stimulated less SMC proliferation than did the wild-type FGF-1 with a median effective dose of approximately 0.3 nmol versus approximately 0.1 nmol (P <.001). By contrast, the chimera retained full stimulating activity on EC proliferation with a median effective dose of 0.06 nmol for both cytokines. Unlike the wild-type protein, the chimera possessed heparin-independent activity. In the absence of heparin, the chimera induced dose-dependent EC and SMC proliferation at 0.06 nmol or more compared with the wild-type FGF-1, which stimulated minimal DNA synthesis at 6.0-nmol concentrations. CONCLUSIONS: The HB-GAM/FGF-1 chimera displays significantly greater and uniquely heparin-independent mitogenic activity for both cell types, and in the presence of heparin it displays a significantly greater EC specificity.

Animals↗

PubMed2005

[Experimental study of tissue engineered bone with cryopreservation on healing of bone defects].

OBJECTIVE: To investigate the effect of tissue engineered bone with cryopreservation on healing of bone defects and to explore feasibility of cryopreservation for tissue engineered bone. METHODS: Tissue engineered bones were constructed with osteoblasts being seeded onto bio-derived materials made from fresh human bones,and they were preserved at 4 degrees C and - 196 degrees C for 3 months and 6 months respectively. They were applied to repair segmental bone defects of rabbit's radius while the tissue engineered bone without cryopreservation and bio-derived materials were brought into control groups. The experiment was divided into groups A3, A6, B3, B6, C and D (group A3: tissue engineered bones were preserved at 4 degrees C for 3 months; group A6: tissue engineered bones were preserved at 4 degrees C for 6 months; group B3:tissue engineered bones were preserved at --196 degrees C for 3 months; group B6:tissue engineered bones were preserved at - 196 degrees C for 6 months; group C: tissue engineered bones without cryopreservation; group D: bio-derived materials). Macroscopical and histological examination were done at the 2nd, 4th, 6th, 12th weeks, X-ray examination was done at the 6th, 12th weeks and biomechanics were determined at 12th weeks after operation respectively. RESULTS: Macroscopical observation showed no significant differences among group A3, A6, B3, B6 and C, but less new bone formation and more obvious boundary in group D were observed. Histological observation showed more collagen and new bone around the edge of implant of group A3, A6, B3, B6 and C than group D, and histological evaluation showed significant differences between group D and other groups (P <0.05). Radiographic observation showed no absorbability of the implant cortex and less new bone formation in group D, but the unity between implant and host bone, medullary cavity reopened, disappearance of fracture line and fine bone modelling were observed in other groups at 12 weeks after operation. Biomechanics between group D and other groups showed significant differences (P< 0. 05). CONCLUSION: Cryopreservation (4 degrees C and - 196 degrees C) were capable of preserving tissue engineered bone for long time, and tissue engineered bone with cryopreservation has significant effect on healing of bone defects. The methods fit clinical application.

Animals↗

PubMed1996

Tissue engineering and autologous transplant formation: practical approaches with resorbable biomaterials and new cell culture techniques.

The engineering of living tissues in vivo requires new concepts in cell culture technology. In contrast to conventional cell cultures, the development of tissues depends on a three-dimensional arrangement of cells and the formation or synthesis of an appropriate extracellular matrix. Special emphasis is given to the major role of the extracellular matrix and cell differentiation in an artificial tissue. New technical approaches of in vitro tissue engineering are compared to the natural development of tissues in vivo. Current methods using resorbable biomaterials, tissue encapsulation and perfusion culture are discussed. Major consideration is given to scaffold structures of biomaterials that define a three-dimensional shape of a tissue or guide matrix formation. The different goals of tissue engineering such as in vitro models and transplant production are taken into account in the described techniques. Practical concepts comprising cell multiplication and differentiation in subsequent steps for future clinical applications are outlined.

Animals↗

PubMed1999

Bioreactor development for tissue-engineered cartilage.

The development of tissue engineered cartilage is emerging as a potential treatment for the repair of cartilage defects. By seeding chondrocytes onto poly-glycolic acid (PGA) biodegradable scaffolds within a convective-flow bioreactor, the synthesis of tissue-engineered articular cartilage has been recently demonstrated. The ability to cultivate and manipulate this cell-polymer construct to possess specific dimensions, as well as biochemical and biomechanical properties is critical for potential application as an in vivo therapy of damaged articular surfaces. Bioreactor design requirements for stages from research to development to commercialization are discussed. Advantages and limitations to various bioreactor designs are critiqued. These studies illustrate the ability to synthesize tissue-engineered cartilage under convective-flow conditions for potential human tissue repair.

Bioreactors↗

PubMed1992

Tissue engineering in the USA.

Tissue engineering is the application of the principles and methods of engineering and the life sciences towards the development of biological substitutes to restore, maintain or improve functions. It is an area which is emerging in importance worldwide. In the USA it has been actively fostered by the National Science Foundation, both through research grants and the sponsorship of a series of workshops starting in 1988. This brief review of activities in the USA focuses on cell culture technology as a foundation for tissue engineering and then discusses examples of applications. These include artificial skin and the use of encapsulated cells in the development of bioartificial organs. Also discussed is the reconstitution of a blood vessel in culture, both for use in basic research and for implantation as an artificial blood vessel in bypass surgery. In conclusion, other potential applications are mentioned as well as generic areas of technology for future development.

Artificial Organs↗