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From stem cells to insulin-producing cells: towards a bioartificial endocrine pancreas.

The total absence or low production of insulin by beta-cells avoids a proper control of glycemia forcing diabetic people to daily insulin injection for survival. Islet transplantation represents a hallmark in the cure of diabetes and has been successfully applied to more than 400 patients, resulting in insulin independency for periods longer than 4 years. However, transplantation trials for diabetes have to face the scarcity of islets from cadaveric donors. Therefore, the finding of renewable sources of cells could circumvent this problem. In this respect, embryonic or adult stem cells are representing an interesting alternative. Stem cells display robust proliferation and the plasticity to differentiate to other cell types, including insulin-containing cells. The current therapeutical use in the future of bioengineered insulin-secreting cells derived from stem cells needs at present to fulfill several criteria. These criteria concern to the type of stem cell to be used as starting biomaterial (embryonic or adult), the in vitro differentiation protocol applied, the functional phenotype reached for the final cell product and the transplantation associated problems (likely immune rejection and tumor formation). This review will try to focus on these different aspects in order to emphasize in the key points to consider for designing unified strategies for diabetes cell therapy.

Bioartificial Organs↗

Skin cell culture on an ear-shaped scaffold created by fused deposition modelling.

Tissue engineering, where cells attach and grow on a scaffold, has the potential to produce replacement ears made from natural tissues and replace the need for rubber prosthetic ears. This study investigated the feasibility of using the rapid prototyping technique of Fused Deposition Modelling (FDM) to produce an ear-shaped scaffold. A three-dimensional image of the ear was used to manufacture ear-shaped scaffolds from ABS (acrylonitrile/butadiene/styrene) plastic using FDM. Human dermal fibroblasts were seeded on the scaffold (coated with fibronectin) to attach and grow in culture medium in an incubator for two weeks. Human keratinocytes were then seeded on to the fibroblast layer to attempt to produce a more realistic skin covering. The morphology of the cells were observed using scanning electron microscopy. The results show that a realistic ear-shaped scaffold can be made using FDM. Human fibroblasts were found to attach and grow. Human keratinocytes were successfully attached and grown on top of the fibroblasts and this resulted in a skin covering over the scaffold. This study shows that FDM has great potential as a manufacturing technique for ear-shaped scaffolds for tissue engineering.

Bioartificial Organs↗

[Primary experimental study on the construction of tissue engineering blood vessel].

OBJECTIVE: To explore the primary experimental methods to construct tissue engineering blood vessel. METHODS: Using the collagen-chitosan to prefabricate the scaffolds with 3-dimensional structure, the proliferated human endothelial cells (ECs), smooth muscle cells (SMCs) and fibroblasts act as the seed cells. The cells were seeded to scaffolds in two-step method, and engineering tissue were matured by static culture or bioreactor culture. Extracellular matrix contents and the platelet aggregation were examined in engineering tissue, tissue engineering blood vessels were taken as patches to repair the man-made defaults on the rats aorta. RESULTS: The proliferated human ECs, SMCs and fibroblast can hold activity and act as seed cells. The prefabricated scaffolds, with excellently cell and tissue biocompatibility, can facilitated cells adherence and upgrowth, the cells quantities and extracellular matrix contents in engineered tissue are time dependent increase (P < 0.05). Platelet aggregation tests confirm the tissue engineering blood vessel have some anti-coagulability. Using the engineering tissue patch to repair the default, 6 aortas in 8 animal were patency till 10 days post-operation. CONCLUSIONS: The seeding cells can be seeded on the 3-dimensional collagen-chitosan scaffolds and matured, the tissue engineering blood vessel can be constructed primarily.

Animals↗

Stem cells are big news, but are they big business?

Future health care will involve treatment with stem cells and tissue engineering. This article, the first in a series reporting on the scientific breakthroughs in this area and relating them to the medical device industry, describes the science.

Animals↗

[Research advances of small-diameter vascular grafts].

This review is a summary of some useful methods and advances about improving clinical applications to small-diameter vascular grafts in recent years, and it points out the developing orientation of small-diameter vascular grafts in the future.

Bioartificial Organs↗

Stem cells: the next therapeutic frontier.

Cell therapy is one of the most exciting fields in translational medicine. It stands at the intersection of a variety of rapidly developing scientific disciplines: stem cell biology, immunology, tissue engineering, molecular biology, biomaterials, transplantation biology, regenerative medicine, and clinical research. Cell-based therapy may develop into a new therapeutic platform to treat a vast array of clinical disorders. Blood transfusions and bone marrow transplantation are prime examples of the successful application of cell-based therapeutics; but recent advances in cellular and molecular biology have expanded the potential applications of this approach. Although recombinant genetic engineering to produce a variety of therapeutics such as human erythropoietin and insulin has proven successful, these treatments are unable to completely correct or reverse disease states, because most common disease processes are not due to the deficiency of a single protein but develop due to alterations in the complex interactions of a variety of cell components. In these complex situations, cell-based therapy may be a more successful strategy by providing a dynamic, interactive, and individualized therapeutic approach that responds to the pathophysiological condition of the patient. In this regard, cells may provide innovative methods for drug delivery of biologics, immunotherapy, and tissue regenerative or replacement engineering (1,2). The translation of this discipline to medical practice has tremendous potential, but in many applications technological issues need to be overcome. Since many cell-based indications are already being evaluated in the clinic, the field appears to be on the threshold of a number of successes. This review will focus on our group's use of human stem/progenitor cells in the treatment of acute and chronic renal failure as extensions to the current successful renal substitution processes of hemodialysis and hemofiltration.

Animals↗

[Separation and purification of skeletal muscle satellite cells for tissue engineering applications by Percoll].

OBJECTIVE: To investigate separation and purification of skeletal muscle satellite cells with improved incontinuous density Percoll gradient centrifugation technique. METHODS: The primary skeletal muscle satellite cells of New Zealand white rabbits were cultured with different adhesion time method and incontinuous density Percoll gradient centrifugation technique. The cells were observed under invert microscope and scanning electron microscope. The degree of purification was examined by celluar immunochemical stain. The growth curve was tested by thiazolyl blue assay. RESULTS: Over 90% satellite cells were harvested by incontinuous density Percoll gradient centrifugation technique, in contrast to which, only 30%-40% cells were harvested by different adhesion time. Morphological observation accorded with satellite cells. The growth curve indicated that the cells grew in a good status. CONCLUSION: The high purification satellite cells can be obtained by incontinuous density Percoll gradient centrifugation technique. It is a good method to culture seed cells for tissue engineering applications.

Animals↗

[Eyelid conjunctiva can be a potential cells source for conjunctival sack reconstruction purpose].

UNLABELLED: There is no ideal culture establishing method for the conjunctival sac reconstruction. Two methods of primary conjunctival culture establishing were compared. MATERIALS AND METHODS: The internal surface of rat's eyelid was used. In the 1st method cells migrate from specimen and attached to the surface of culture plate. In the 2nd method tissue was digested and placed in the flask. Epithelial character of cultured cells was confirmed. RESULTS: Both methods allow to set up the culture of epithelial cells. Efficiency of enzymatic isolation was higher then growing from previously attached specimen. At the end of the experiment the cultures were similar. CONCLUSIONS: Both methods allow to establish the primary culture of conjunctival epithelial cells. Eyelid conjunctiva can be a cells source for the conjunctival sack reconstruction purpose.

Animals↗

Artificial organs as a bridge to transplantation.

Current organ shortage is estimated to keep outpacing demand for years to come. Among the advocated strategies, artificial and bioartificial devices may prove beneficial to a wide category of patients on transplant waiting lists. Bionic organ science allows to reproduce organ architecture and function through a complex interplay of cellular and mechanical elements. Some bioartificial organs may well be used to replace anatomical defects, while others allow to compensate for failing organ functions and to bridge patients to transplantation. Among these latter, bioartificial liver (BAL) systems bear the highest potential for clinical application, even if their use is raising several controversial issues. These latter regard the identification and stratification of patients fit for transplantation, timing and type of transplantation after recovery, appropriateness of double-blind, randomized clinical trials and safety of animal and/or human cell lines. Nonetheless, bionic organ science needs to be regarded as a useful adjunct in the armamentarium of organ replacement therapies for the third millennium.

Animals↗

Tissue engineering science: consequences of cell traction force.

Blood and tissue cells mechanically interact with soft tissues and tissue-equivalent reconstituted collagen gels in a variety of situations relevant to biomedicine and biotechnology. A key phenomenon in these interactions is the exertion of traction force by cells on local collagen fibers which typically constitute the solid network of these tissues and gels and impart gross mechanical integrity. Two important consequences of cells exerting traction on such collagen networks are first, when the cells co-ordinate their traction, resulting in cell migration, and second, when their traction is sufficient to deform the network. Such cell-collagen network interactions are coupled in a number of ways. Network deformation, for example, can result in net alignment of collagen fibers, eliciting contact guidance, wherein cells move with bidirectional bias along an axis of fiber alignment, potentially leading to a nonuniform cell distribution. This may govern cell accumulation in wounds and be exploited to control cell infiltration of bioartificial tissues and organs. Another consequence of cell traction is the resultant stress and strain in the network which modulate cell protein and DNA synthesis and differentiation. We summarize, here, relevant mathematical theories which we have used to describe the inherent coupling of cell dynamics and tissue mechanics in cell-populated collagen gels via traction. The development of appropriate models based on these theories, in an effort to understand how events in wound healing govern the rate and extent of wound contraction, and to measure cell traction forces in vitro, are described. Relevant observations and speculation from cell biology and medicine that motivate or serve to critique the assumptions made in the theories and models are also summarized.

Biomechanical Phenomena↗

Bioartificial kidney in the treatment of acute renal failure associated with sepsis.

Acute renal failure (ARF) associated with sepsis has a high rate of mortality. It is not merely a surrogate marker for severity of disease but also an independent predictor of mortality and a separate pathogenic entity, even when nearly physiological doses of fluid and small-molecule clearance are maintained with currently available renal replacement therapies (RRT). The techniques to remove cytokines, including high-volume haemofiltration, haemodialysis using high-cut-off haemofilters, and absorptive techniques, lead to some improvement in outcome but are still insufficient to reverse the complicated biological dysregulation resulting from ARF associated with sepsis. The novel and exciting technique of cell therapy, which is based on the principle of using functional cells to replace a greater range of renal functions, may add significant benefit to current RRT in dealing with this disease process. Because renal tubule cells appear to play critical roles in immunoregulation, renal tubule cell therapy during ARF associated with sepsis should alter the detrimental multiple-organ consequences of sepsis. The development of a bioartificial kidney consisting of a conventional haemofiltration cartridge in series with a renal tubule assist device containing renal proximal tubule cells represents a new therapeutic approach to this clinical disorder. The results to date of large animal studies and recent Phase I/II and Phase II clinical trials show that such a device replaces multiple kidney functions and modifies the sepsis condition to improve survival in ARF.

Acute Kidney Injury↗

The use of a pig liver xenograft for temporary support of a patient with fulminant hepatic failure.

A 26-year-old female patient with fulminant hepatic failure and a history of autoimmune hepatitis was heterotopically transplanted with a pig hepatic xenograft to provide temporary metabolic support prior to transplantation with a human donor organ. Circulating natural antipig antibodies were removed prior to transplantation by plasmapheresis and ex vivo en bloc perfusion of the donor pig kidneys. The liver xenograft functioned after transplantation as measured by active bile production, stabilization of prothrombin levels, and reduction in the circulating levels of lactic acid and the enzymes AST and ALT. Despite the removal of greater than 90% of the recipient's natural xenoantibodies prior to transplantation, the levels of antibody rapidly returned and were associated with antibody and complement-mediated rejection of the donor graft. Immunohistochemical evidence of graft rejection could be detected by the deposition of antibody, complement components including properdin, and endothelial swelling as early as 3 hr posttransplantation. These lesions progressed in severity and were accompanied by evidence of thrombosis and ischemic necrosis of the liver xenograft by 34 hrs posttransplantation. The main portal vein, hepatic artery, and vena cava were patent. The placement of the liver graft did not result in any improvement in the neurological status of the patient and she died 34 hr after xenografting due to irreversible brain damage. The information derived from this case has renewed interest in the clinical use of bioartificial devices and whole organ perfusion using xenogeneic tissue for temporary bridging of patients prior to allografting.

Adult↗

What should we expect from a bioartificial liver in fulminant hepatic failure?

Fulminant hepatic failure is associated with high mortality. Liver transplantation is an effective therapy that improves survival, and because of donor organ shortage and urgency, a bioartificial liver could act as an effective bridge to liver transplantation in patients with fulminant hepatic failure. We discuss the place of the bioartificial liver in the treatment of other causes of liver failure.

Europe↗

Nandrolone decanoate and load increase remodeling and strength in human supraspinatus bioartificial tendons.

BACKGROUND: To date, no studies document the effect of anabolic steroids on rotator cuff tendons. STUDY DESIGN: Controlled laboratory study. HYPOTHESIS: Anabolic steroids enhance remodeling and improve the biomechanical properties of bioartificially engineered human supraspinatus tendons. METHODS: Bioartificial tendons were treated with either nandrolone decanoate (nonload, steroid, n = 18), loading (load, nonsteroid, n = 18), or both (load, steroid, n = 18). A control group received no treatment (nonload, nonsteroid [NLNS], n = 18). Bioartificial tendons' remodeling was assessed by daily scanning, cytoskeletal organization by staining, matrix metalloproteinase-3 levels by ELISA assay, and biomechanical properties by load-to-failure testing. RESULTS: The load, steroid group showed the greatest remodeling and the best organized actin cytoskeleton. Matrix metallo-proteinase-3 levels in the load, steroid group were greater than those of the nonload, nonsteroid group (P <.05). Ultimate stress and ultimate strain in the load, steroid group were greater than those of the nonload, nonsteroid and nonload, steroid groups (P <.05). The strain energy density in the load, steroid group was greater when compared to other groups (P <.05). CONCLUSIONS: Nandrolone decanoate and load acted synergistically to increase matrix remodeling and biomechanical properties of bioartificial tendons. CLINICAL RELEVANCE: Data suggest anabolic steroids may enhance production of bioartificial tendons and rotator cuff tendon healing in vitro. More research is necessary before such clinical use is recommended.

Adult↗

Early clinical experience with a hybrid bioartificial liver.

BACKGROUND: Severe liver failure is associated with high mortality. Orthotopic liver transplantation (OLT) is the only effective therapeutic modality; there is a need for a 'bridge' system to support patients until an organ becomes available. METHODS: A bioartificial liver (BAL) was used to treat 10 patients with severe liver failure. A plasmapheresis system was used to pump patient plasma through a module with porcine hepatocytes. Each treatment lasted 6-7 h. RESULTS: All patients tolerated the procedure(s) well. Eight patients underwent OLT following BAL treatment(s). There were two late deaths after recovery from liver failure. Five patients with increased intracranial pressure (ICP) and decerebration had ICP normalization, increased cerebral perfusion pressure and full neurologic recovery after OLT. There was improvement in the level of encephalopathy and a significant decrease in serum ammonia after BAL treatment(s). CONCLUSIONS: BAL treatment is safe and beneficial and can be successfully used as a 'bridge' to transplantation.

Adult↗

[Fulminant liver failure: intensive care, extracorporeal treatment and liver transplantation].

Fulminant liver failure is characterized by massive acute Hepatocyte dysfunction associated with severe coagulopathy, acute hyperdynamic circulatory failure and hepatic encephalopathy. According to the more recent classification, which takes into account the interval between the onset of jaundice and the hepatic encephalopathy, three are the main forms of ALF hyperacute, acute or subacute. Despite recent and relevant advances in intensive care management and organ support techniques (both artificial and bioartificial), mortality remains extremely high, early deaths being related to cerebral oedema and circulatory failure, whereas late deaths are associated with sepsis and multiple organ failure. Orthotopic liver transplantation has proven to be the only treatment modality able to change radically the ALF natural course. he experiences with artificial and bioartificial devices, in spite of being interesting and sometimes very promising, are far from giving a real impact on survival and remain, so far, important interim measures for patients eventually candidate to liver transplantation.

Artificial Organs↗