Tissue engineering最新文献

The development of minimally invasive therapeutics for orthopedic clinical conditions has substantial benefits, especially for osteoporotic fragility fractures and vertebral compression fractures. Poly(ester urethane)urea (PEUUR) foams are potentially useful for addressing these conditions because they cure in situ upon injection to form porous scaffolds. In this study, the effects of water concentration and polyester triol composition on the physicochemical, mechanical, and biological properties of PEUUR foams were investigated. A liquid resin (lysine diisocyanate) and hardener (poly(epsilon-caprolactone-co-glycolide-co-DL-lactide) triol, tertiary amine catalyst, anionic stabilizer, and fatty acid-derived pore opener) were mixed, and the resulting reactive liquid mixture was injected into a mold to harden. By varying the water content over the range of 0.5 to 2.75 parts per hundred parts polyol, materials with porosities ranging from 89.1 to 95.8 vol-% were prepared. Cells permeated the PEUUR foams after 21 days post-seeding, implying that the pores are open and interconnected. In vitro, the materials yielded non-cytotoxic decomposition products, and differences in the half-life of the polyester triol component translated to differences in the PEUUR foam degradation rates. We anticipate that PEUUR foams will present compelling opportunities for the design of new tissue-engineered scaffolds and delivery systems because of their favorable biological and physical properties.

One of the principal challenges facing the field of tissue engineering over the past 2 decades has been the requirement for large-scale engineered constructs comprising precisely organized cellular microenvironments. For vital organ assist and replacement devices, microfluidic-based systems such as the microcirculation, biliary, or renal filtration and resorption systems and other functional elements containing multiple cell types must be generated to provide for viable engineered tissues and clinical benefit. Over the last several years, microfabrication technology has emerged as a versatile and powerful approach for generating precisely engineered scaffolds for engineered tissues. Fabrication process tools such as photolithography, etching, molding, and lamination have been established for applications involving a range of biocompatible and biodegradable polymeric scaffolding materials. Computational fluid dynamic designs have been used to generate scaffold designs suitable for microvasculature and a number of organ-specific constructs; these designs have been translated into 3-dimensional scaffolding using microfabrication processes. Here a brief overview of the fundamental microfabrication technologies used for tissue engineering will be presented, along with a summary of progress in a number of applications, including the liver and kidney.

An appropriate cellular response to implanted surfaces is essential for tissue regeneration and integration. In this study, we investigated how human bone marrow stromal cells (hBMSCs) respond to scaffold substrates. We prepared wettable polymer surfaces by exposing polymer sheets to radio frequency plasma discharge, which gradually oxidizes the polymer surface, increasing the roughness and greatly reducing the hydrophobicity. We found that hBMSCs adhered better to highly hydrophilic and rough surfaces than to hydrophobic and smooth surfaces. In addition, the cells flattened extensively on hydrophilic surfaces. Further, c-fos gene expression increased in parallel with the degree of hydrophilicity, whereas the expression of the c-myc gene was higher on hydrophobic than on hydrophilic surfaces. Finally, p53 gene expression was higher on more hydrophobic or hydrophilic surfaces than on moderately hydrophobic or hydrophilic surfaces. These results indicate that the biological signals induced by cell adhesion depend on the wettability of the surface to which the cells attach.

The burgeoning field of regenerative medicine promises significant progress in the treatment of cardiac ischemia, liver disease, and spinal cord injury. Key to its success will be the ability to engineer tissue safely and reliably. Tissue functionality must be recapitulated in the laboratory and then integrated into surrounding tissue upon transfer to the patient. Scaffolding materials must be chosen such that the microenvironment surrounding the cells is a close analog of the native environment. In the early days of tissue engineering, these materials were largely borrowed from other fields, with much of the focus on biocompatibility and biodegradation. However, attention has shifted recently to cell-cell and cell-surface interactions, largely because of enabling technologies at the nanoscale and microscale. Studies on cellular behavior in response to various stimuli are now easily realized by using microfabrication techniques and devices (e.g., biomedical microelectromechanical systems). These experiments are reproducible and moderate in cost, and often can be accomplished at high throughput, providing the fundamental knowledge required to design biomaterials that closely mimic the biological system. It is our opinion that these novel materials and technologies will bring engineered tissues one step closer to practical application in the clinic. This review discusses their application to cardiac, liver, and nerve tissue engineering.

Tissue engineering is an emerging area of applied research with a goal of repairing or regenerating the functions of damaged tissue that fails to heal spontaneously by using cells and synthetic functional components called scaffolds. Scaffolds made of nanofibers (herein called "nano-fibrous scaffolds") play a key role in the success of tissue engineering by providing a structural support for the cells to accommodate and guiding their growth in the three-dimensional space into a specific tissue. The orientation of these fibers is considered as one of the important features of a perfect tissue scaffold, because the fiber orientation greatly influences cell growth and related functions. Therefore, engineering scaffolds with a control over fiber orientation is essential and a prerequisite for controlling cell orientation and tissue growth. Recent advances in electrospinning have made it possible to create nano-featured scaffolds with controlled fiber orientation. Electrospinning is a straightforward, cost-effective, and versatile method, which is recently applied in engineering well-defined nano-fibrous scaffolds that hold promise in serving as a synthetic extra-cellular matrix (ECM). This article reviews the current trends in electrospinning nano-fibrous scaffolds with fiber orientation. A detailed mechanism involved in the spinning process is discussed, followed by experimental examples that show how the fiber orientation influences cellular growth behavior. This review is expected to be useful for readers to gain knowledge on the state-of-the-art of scaffold engineering by electrospinning.

In this paper, we review the progress toward developing strategies to engineer improved structural grafting of bone. Three strategies are typically used to augment massive bone defect repair. The first is to engraft mesenchymal stem cells (MSCs) onto a graft or a biosynthetic matrix to provide a viable osteoinductive scaffold material for segmental defect repair. The second strategy is to introduce critical factor(s), for example, bone morphogenetic proteins (BMPs), in the form of bone-derived or recombinant proteins onto the graft or matrix directly. The third strategy uses targeted delivery of therapeutic genes (using viral and nonviral vectors) that either transduce host cells in vivo or stably transduce cells in vitro for subsequent implantation in vivo. We developed a murine femoral model in which allografts can be revitalized via recombinant adeno-associated virus (rAAV) gene transfer. Specifically, allografts coated with rAAV expressing either the constitutively active BMP type I receptor Alk2 (caAlk2), or the angiogenic factor vascular endothelial growth factor (VEGF) combined with the osteoclastogenic factor receptor activator of NF-kappa B ligand (RANKL) have remarkable osteogenic, angiogenic, and remodeling effects that have not been previously documented in healing allografts. Using histomorphometric and micro computed tomography (muCT) imaging we show that rAAV-mediated delivery of caAlk2 induces significant osteoinduction manifested by a mineralized callus on the surface of the allograft, which resembles the healing response of an autograft. We also demonstrate that the rAAV-mediated gene transfer of the combination of VEGF and RANKL can induce significant vascularization and remodeling of processed structural allografts. By contrast, rAAV-LacZ coated allograft controls appeared similar to necrotic allografts and lacked significant mineralized callus, neovascularization, and remodeling. Therefore, innovations in gene delivery offer promising therapeutic approaches for tissue engineering of structural bone substitutes that can potentially have clinical applications in challenging indications.

The process by which cells self-assemble to form three-dimensional (3D) structures is central to morphogenesis and development of living tissues and hence is of growing interest to the field of tissue engineering. Using rapid prototyping technology we made micromolded nonadhesive hydrogels to study the dynamics of self-assembly in a low-shear environment with simple spherical geometries as well as more complex geometries such as a toroid. Aggregate size, shape, and composition were easily controlled; aggregates were easily retrieved; and the dynamics of the assembly process were readily observed by time-lapse microscopy. When two cell types, normal human fibroblasts (NHFs) and human umbilical vein endothelial cells (HUVECs), were seeded together, they self-segregated into multilayered spherical microtissues with a core of NHFs enveloped by a layer of HUVECs. Surprisingly, when a single cell suspension of NHFs was added to 7-day-old HUVEC spheroids, the HUVEC spheroid reorganized such that NHFs occupied the center and HUVECs coated the outside, demonstrating that self-assembly is not terminal and that spheroids are fluid structures that retain the ability to reassemble. We also showed that cells can self-assemble to form a complex toroid shape, and we observed several phenomena indicating that cellular contraction and tension play a significant role in the assembly process of complex shapes.

Stromal-derived factor 1alpha (SDF-1alpha) is a key stem cell homing factor that is crucial for mobilization of stem cells from bone marrow to peripheral blood and subsequent engraftment to the tissue of diseased organs. It has been reported that SDF-1alpha is transiently over-expressed in ischemic myocardium. Therefore, there may be a limited time window after acute myocardial infarction (AMI) during which stem cells are recruited to injured myocardium for repair. This study aimed at investigating whether controlled release of SDF-1alpha via a novel conjugated poly(ethylene glycol) (PEG) (PEGylated) fibrin patch at the infarct site would increase the rate of stem cell recruitment and offer potential therapeutic benefits. Recombinant mouse SDF-1alpha was covalently bound to the PEGylated fibrinogen as evidenced by immunoprecipitation and western blotting. The PEGylated fibrinogen, bound with recombinant mouse SDF-1alpha, was mixed with thrombin to form the PEGylated fibrin patch. The release kinetics of SDF-1alpha were detected in vitro using enzyme-linked immunosorbent assay. Using a mouse AMI model produced by a ligature on the left anterior descending coronary artery, a PEGylated fibrin patch bound with SDF-1alpha (100 ng) was placed on the surface of the infarct area of the left ventricle. Infarct size, left ventricular (LV) function, and the percentage of sca-1(+)/c-kit(+) cells within the infarct area were measured at days 7, 14, and 28 after AMI. In vitro results showed that SDF-1alpha was successfully bound to the PEGylated fibrin patch and can be released from the patch constantly for up to 10 days. Two weeks after infarction, the myocardial recruitment of c-kit(+) cells was significantly higher in the group treated with the SDF-1alpha PEGylated fibrin patch (n = 9) than in the AMI control group (n = 10) (p < 0.05; 11.20 +/- 1.71% vs. 4.22 +/- 0.96%, respectively). At day 28 post-AMI, unlike the control group, the group with the SDF-1alpha-releasing patch maintained stable release of SDF-1alpha concurrent with additional stem cell homing. Moreover, LV function was significantly better than in the control group. These data demonstrate that the PEGylated fibrin patch based SDF-1alpha delivery can improve the rate of c-kit(+) cell homing and improve LV function in hearts with postinfarction LV remodeling.

The surface modification of islets using poly(ethylene glycol) (PEG) is being studied as a means of preventing host immune responses against transplanted islets. In this study, to completely shield islets with PEG molecules, we increased the amount of PEG conjugated to islet surfaces, by multiple PEGylation or amplified PEGylation using poly-L-lysine, poly(allylamine), or poly(ethyleneimine), respectively. Amplified PEGylation was associated with islet cytotoxicity and functional impairment, but multiple PEGylation affected neither islet viability nor functionality. In addition, when triply PEGylated islets were allotransplanted into diabetic recipients, these islets survived in 3 of the 7 recipients for more than 100 days without any immunosuppressive treatment. Moreover, the blood glucose levels of these 3 recipients were stable and in the normal range. Immunohistochemical analysis showed that 3 of 7 triply PEGylated islets transplants survived for 100 days and that 4 that were rejected before day 20 were all immunologically protected from immune cells. However, unmodified islets were completely destroyed within 1 week. Consequently, we suggest that multiple PEGylation offers an effective means of reducing the immunogenicity of transplanted islets by increasing the amount of surface-bound PEG.

The field of cardiovascular tissue engineering has experienced tremendous advances in the past several decades, but the clinical reality of engineered heart tissue and vascular conduits remains immature. Stem cells and progenitor cells are promising cell sources for engineering functional cardiovascular tissues. To realize the therapeutic potential of stem cells and progenitor cells, we need to understand how microenvironmental cues modulate and guide stem cell differentiation and organization. This review describes the current understanding of the chemical and physical regulation of embryonic and adult stem cells for potential applications in cardiovascular repair, focusing on cardiac therapies after myocardial infarction and the engineering of vascular conduits.