Development, preclinical evaluation and validation of a novel quick vascular closure device for transluminal, cardiac and radiological arterial catheterization.

Following percutaneous coronary intervention, vascular closure devices (VCDs) are increasingly used to reduce time to ambulation, enhance patient comfort, and reduce potential complications compared with traditional manual compression. Newer techniques include complicated, more or less automated suture devices, local application of pads or the use of metal clips and staples. These techniques often have the disadvantage of being time consuming, expensive or not efficient enough. The VCD failure rate in association with vascular complications of 2.0-9.5%, depending on the type of VCD, is still not acceptable. Therefore, the aim of this study is to develop a self-expanding quick vascular closure device (QVCD) made from a bioabsorbable elastic polymer that can be easily applied through the placed introducer sheath. Bioabsorbable block-co-polymers were synthesized and the chemical and mechanical degradation were determined by in vitro tests. The best fitting polymer was selected for further investigation and for microinjection moulding. After comprehensive haemocompatibility analyses in vitro, QVCDs were implanted in arterial vessels following arteriotomy for different time points in sheep to investigate the healing process. The in vivo tests proved that the new QVCD can be safely placed in the arteriotomy hole through the existing sheath instantly sealing the vessel. The degradation time of 14 days found in vitro was sufficient for vessel healing. After 4 weeks, the remaining QVCD material was covered by neointima. Overall, our experiments showed the safety and feasibility of applying this novel QVCD through an existing arterial sheath and hence encourage future work with larger calibers.

Suprathel-an innovative, resorbable skin substitute for the treatment of burn victims.

Autologous split skin grafts are the most reliable method for closing third degree burns. Under this scheme, donor sites as well as second degree burns under conservative treatment, however, would benefit from rapid wound closure. For this treatment, biological as well as synthetic materials are available. For the improvement of these materials, primary goals are pain reduction and easy handling in the absence of biological risk. From a synthetic copolymer mainly based on DL-lactic acid a new skin substitute was developed, marketed as Suprathel. Within the scope of a bicentric study Suprathel was compared versus paraffin gauze intraindividually applied on split skin donor sites. Wound pain was measured on the Visual Pain Analog Scale over a period of 10 days as the critical criterion. Accordingly Suprathel versus Omiderm were compared on second degree burns (degree 2a, partial thickness burns). In both study parts, Suprathel significantly reduced pain. Its easy handling was superior compared to other materials. The Suprathel membrane adhered rapidly to the wound thus protecting against infections and promoting wound healing. No allergic reactions were observed. The ability of the material to resorb ensured pain-free removal after complete healing of the wound.

Acoustomechanical properties of open TTP titanium middle ear prostheses.

OBJECTIVE: The purpose of the study was to identify acoustcomechanical properties of various biostable and biocompatible materials to create a middle ear prosthesis with the following properties: (i) improved handling including a good view of the head of the stapes or footplate and adjustable length, (ii) improved acoustical characteristics that are adequate for ossiculoplastic. The identified material should serve to build CE and FDA approved prostheses for clinical use in patients. METHODS: Test models made of Teflon, polyetheretherketone, polyethylenterephtalate, polysulfone, gold, Al2O3 ceramics, carbon and titanium were investigated for their potential to fulfill the requirements. Acoustical properties were investigated by laser Doppler velocimetry (LDV) in mechanical middle ear models (MMM). Measured data were fed in to a recently created computer model of the middle ear (multibody systems approach, MBS). Using computer-aided design (CAD) measured and computed data allowed creation and fine precision of titanium prostheses (Tübingen Titanium Protheses, TTP). Their handling was tested in temporal bones. Acoustomechanical properties were investigated using the MBS and mechanical middle ear models. MAIN OUTCOME MEASURES: Input impedance, mass, stiffness, and geometry of test models and prostheses were determined. Furthermore, their influence on the intraprosthetic transfer functions and on coupling to either tympanic membrane or stapes was investigated. RESULTS: Final results were FDA- and CE-approved filigreed titanium prostheses with an open head that fulfilled the four requirements detailed above. The prostheses (TTP) were developed in defined lengths of between 1.75 and 3.5 mm (partial) and 3.0 and 6.5 mm (total) as well as in adjustable lengths (TTP-Vario). CONCLUSIONS: The results suggest acoustomechanical advantages of TTPs because they combine a significantly low mass with high stiffness. In contrast to closed prostheses, the open head and filigreed design allow an excellent view of the prosthesis foot during coupling to the head or footplate of stapes, contributing to an improved intraoperative reliability of prosthesis coupling.

Cultivation of porcine hepatocytes in polyurethane nonwovens as part of a biohybrid liver support system.

Many patients suffering from end-stage liver disease cannot be transplanted within reasonable time due to the shortage of donor organs. Bioartificial liver support systems may contribute to the liver regeneration or bridging the time until a liver graft for transplantation becomes available. Nonwovens with integrated oxygenation capacity have been developed and manufactured by melt blow technology using thermoplastic polyurethane. Capillary membranes for oxygenation were integrated into the nonwoven during the processing. The polyurethane nonwoven structures with adapted pore size and high pore volume allow high cell densities in the hepatocyte culture. The three-dimensional cell culture was housed by a flow bioreactor system and was integrated in a closed loop circulation with monitoring possibilities for pressure, pH, temperature, ammonia, and oxygen. Hepatocytes were isolated from rats or pigs by collagenase perfusion and infused into the medium-perfused circulation. Cells showed high viability and hepatocyte specific cytochrome P450-dependent metabolic function in culture (MEGX test).

Resorbable polymer fibers for ligament augmentation.

Resorbable augmentation devices for cruciate ligament surgery have been developed to temporarily protect healing tendon grafts or sutured ligaments against high tensile loads during the postoperative healing period. Materials available at present [e.g., polydioxanone (PDS)] show a half-life tensile strength of only 4-6 weeks, whereas the process of revitalization and recovering of the transplanted tendon graft can take up to 12 months. Therefore, a device that provides gradually decreasing mechanical properties with a half-time strength of at least 6 months would be desirable. In order to obtain a suitable material, we investigated the degradation kinetics of a variety of different resorbable fibers made of poly(L-lactide) and poly(L-lactide-co-glycolide). The fiber materials differed in processing and treatment parameters like thermal posttreatment, irradiation, and fiber diameter. The fibers were degraded in vitro and were tested for mechanical properties and molecular weight at various time points up to 72 weeks. The half-time strength of the materials ranged between 5 and 64 weeks, depending on their treatment parameters. In contrast, the stiffness did not decrease adequately. However, an augmentation stiffness that does not change much versus time could not provide a gradual increase in graft load, which is important to stimulate the orientation of the collagenous tissue. Therefore, design of an augmentation construct braided out of more than one quickly degrading fiber materials is suggested. After the breakdown of the faster-degrading fiber components the stiffness would automatically decrease by the diminution of the load-carrying fiber volume.

Macroencapsulation of rat islets without alteration of insulin secretion kinetics.

Transplantation of encapsulated islets may restore endogenous insulin secretion in type 1 diabetics with no need of lifetime immunosuppression of the recipient. A biomaterial should be developed which combined immunoisolation with rapid and efficient diffusion of glucose and insulin. Rat islets were macroencapsulated in capillaries (molecular cut off 50 kD) of differently modified polysulphone. Macroencapsulated islets were perifused to study the kinetics of glucose induced insulin secretion into the perifusion medium. Blending polysulphone (PSU) with poly vinyl pyrrolidone or sodium dodecyl sulphate was not suited for islet macroencapsulation since glucose induced insulin release was absent after encapsulation. Hydroxy methylation (CH2OH) of PSU improved the secretory behaviour of macroencapsulated islets depending on the degree of substitution (DS). At 0.8 DS glucose induced insulin secretion was delayed and inefficient. At maximal degrees of PSU-substitution (1.8) the kinetics of insulin release and the efficiency of insulin release were very similar to that observed of free floating islets. In conclusion, highly substituted hydroxy methylated polysulphone allows a rapid and efficient insulin release after macroencapsulation and is suited for the further development of a bioartificial pancreas.

High density culturing of porcine hepatocytes immobilized on nonwoven polyurethane-based biomatrices.

OBJECTIVE: Hepatocytes are increasingly used as functional units in bioartificial liver devices. The objective of the present study was to investigate the feasibility of culturing porcine hepatocytes in high density on a novel polyurethane-based nonwoven three-dimensional matrix. We investigated (1) the optimal cell density within this culture configuration, (2) the maintenance of liver-specific morphology and cell functions over long-term periods and (3) the necessity to apply an additional extracellular matrix component (collagen gel). METHODS: Nonwoven polyurethane matrices were manufactured by a specially developed fiber extrusion technology. Pig hepatocytes were cultured at various cell densities of 0.1, 0.25, 0.5, 0.75, 1 and 2 x 10(6) cells/cm(2) on three-dimensional networks of nonwoven polyurethane matrices and cell adhesion as well as functional parameters (DNA of nonattached/attached cells, lactate dehydrogenase release and cytochrome P450 activity) were determined. To assess the performance of cells within this configuration albumin and urea excretion was measured over 8 days. The potentially beneficial effect of an additional extracellular matrix configuration was evaluated by comparing the average albumin synthesis in groups of identical cell numbers. RESULTS: The optimal cell density in this three-dimensional culture configuration was 1 x 10(6) cells/cm(2). The functional capacity of hepatocytes was stable for 8 days at an average level of 53.7 +/- 5.6 ng/h/microg DNA and of 1.8 +/- 0.14 microg/h/microg DNA for albumin and urea excretion, respectively. The supplementation of an extracellular matrix configuration did not improve functional activity of cells. Average albumin synthesis was 35.6 ng/h/microg DNA (28.7, 42.8) and 32.7 ng/h/microg DNA (23.4, 49.2) for collagen-immobilized and control cultures, respectively. CONCLUSION: The results of the study indicate that nonwoven polyurethane sheets supply a biocompatible support structure for functionally active high density cultures. Thus, nonwoven polyurethane matrices should be further investigated on with respect to their role in the development, optimization and design of bioartificial liver systems.

In vitro test of new biomaterials for the development of a bioartificial pancreas.

The implantation of macroencapsulated islets has the potential to restore endogenous insulin secretion in type 1 diabetics, with no need for lifetime immunosuppression. To match the physiological fluctuations of blood glucose concentrations with appropriate insulin release, the macroencapsulation material must combine immunoprotection with optimal diffusion properties for glucose and insulin. The impact of chemical modifications of polysulphone (PSU) capillary polymers with a cutoff of 50 kD on glucose-induced insulin secretion of macroencapsulated rat islets was studied in perifusion experiments. The insulin release of free-floating islets showed the typical rapid response to glucose stimulation. Total insulin release (AUC between minute 30 and 120 of perifusion) reached 117+/-22 ng/ml. Blending PSU with polyvinylpyrrolidone or sodium-dodecyl-sulfate was not suitable for islet macroencapsulation, since glucose-induced insulin release was absent or disturbed. Hydroxy-methylation (CH2OH) of PSU improved the secretory behavior of macroencapsulated islets depending on the degree of PSU substitution (DS 0.8, AUC 62+/-15 ng/ml; DS 1.8, 111+/-24 ng/ml). In highly substituted PSU-capillaries the kinetics of glucose-induced insulin release was very similar to that observed in free-floating islets. Two consecutive glucose stimulations potentiated insulin release of free-floating islets during the second period of stimulation. Furthermore, freshly isolated macroencapsulated islets responded with more efficient insulin secretion after the initial priming. In conclusion, in vitro membrane screening identified highly substituted hydroxy-methylated PSU as the material of choice for islet encapsulation in a bioartificial pancreas.

Alveolar ridge repair using resorbable membranes and autogenous bone particles with simultaneous placement of implants: an experimental pilot study in dogs.

The aim of this experimental study was to evaluate the use of autogenous bone harvested during preparation of implant sites in combination with resorbable membranes for vertical ridge augmentation under 2 different defect site conditions. Combined vertical/horizontal alveolar bone defects were created by experimentally induced periodontal infections around all premolar teeth in the mandibles of 3 dogs (group 1). In another 3 dogs, fresh surgical defects were created after extraction of all premolar teeth in the mandibles (group 2). In all dogs, 2 implants were placed on each side of the mandible into the defect areas. One implant on each side of the mandible received augmentation with autogenous bone particles, and both implants on one side of the mandible were covered with polylactic acid membranes. After 5 months, the material was evaluated histologically. There was a small but significant increase in bone regeneration in the defects augmented with bone particles with and without membrane coverage in group 1. In group 2, no significant difference was seen between the controls and the augmented sites. The major limiting effect for bone regeneration appeared to be insufficient stability of the bone material to withstand the overlying soft tissue pressure. It was concluded that the placement of autogenous bone particles, either with or without membrane coverage, had little effect on the regeneration of peri-implant bone defects.

Guided bone regeneration around endosseous implants using a resorbable membrane vs a PTFE membrane.

The aim of the present experimental pilot study was to assess bone regeneration underneath resorbable barrier membranes vs non-resorbable extended polytetrafluoroethylene (ePTFE) membranes in peri-implant defects. Two implants were inserted into surgically created defects on each side of the mandibles of 6 adult beagle dogs 3 months after extraction of all premolar teeth. One implant on each side was covered with a porous polylactic acid membrane or a ePTFE membrane, respectively, while the second implant served as control. Fluorochrome labelling was administered during the 1st, 5th, 12th and the 18th week. Three animals each were evaluated after 3 and 6 months. Bone regeneration was assessed by measuring the distance from the first fluorochrome label to the level of the regenerated bone immediately adjacent to the implant surface and to the top of the newly formed alveolar contour both on the lingual and buccal side. The increase in bone height was significantly higher compared to the controls under both barrier membranes after 3 months at the top of the alveolar crest but not immediately adjacent to the implant surface. After 6 months, bone height was significantly increased only at the top of the alveolar contour underneath the ePTFE membranes, while bone underneath the polylactic acid membranes showed signs of superficial resorption. It is concluded that guided bone regeneration underneath barrier membranes can restore alveolar bone contour but is not necessarily associated with a higher bone/implant contact. The use of resorbable membranes may be associated with untoward biological effects at later stages, when membrane degradation starts due to degradation products of the polymer material or decreasing membrane stability. Future efforts have to refine the relation between degradation kinetics, membrane porosity and mechanical properties of degradable barrier membranes to improve membrane performance.

New non-woven polyurethane-based biomaterials for the cultivation of hepatocytes: expression of differentiated functions.

A new non-woven polyetherurethane support suitable to host cultured hepatocytes has been developed. Prior to its use in bioreactors and artificial liver devices, the biocompatibility of this new material was investigated. The experiments have shown that the survival and functionality of hepatocytes entrapped in the non-woven polymer were longer than that of monolayer cultured hepatocytes, under serum-free culture conditions. Hepatic specific metabolic functions, namely, synthesis of urea and synthesis and secretion of plasma proteins, were well maintained by hepatocytes entrapped in non-woven polyetherurethane sheets. Cells also retained the expression of biotransformation activities of 7-ethoxycoumarin-O-deethylase as well as CYP2A1, CYP2B1 and CYP3A1. The results presented in this paper point to non-woven polyetherurethane sheets as a suitable biocompatible support for functional, three-dimensional hepatocyte cultures.

Biohybride nerve guide for regeneration: degradable polylactide fibers coated with rat Schwann cells.

The restricted capacity of the nervous system to regenerate calls for novel therapeutic concepts. We have tested biocompatible polylactide fibers as potential nerve guides that could bridge proximal nerve stumps and synaptic target regions after nerve lesion. Polylactides have the great advantage that they degrade and resorb after completion of regeneration. Material surface properties were optimized three-fold by oxygen plasma treatment, polyanion coating and the seeding of Schwann cells from rat sciatic nerve. Immunocytochemistry and scanning electron microscopy revealed that in vitro axonal outgrowth of dorsal root ganglia on two specifically synthesized lactide polymers can be greatly improved by these surface treatments. The approach aims to develop an 'intelligent neuroprosthesis' that in vivo facilitates directed axonal regrowth in the first place and disappears thereafter.

Cartilage reconstruction in head and neck surgery: comparison of resorbable polymer scaffolds for tissue engineering of human septal cartilage.

New cell culture techniques raise the possibility of creating cartilage in vitro with the help of tissue engineering. In this study, we compared two resorbable nonwoven cell scaffolds, a polyglycolic acid/poly-L-lactic acid (PGA/PLLA) (90/10) copolymer (Ethisorb) and pure PLLA (V 7-2), with different degradation characteristics in their aptitude for cartilage reconstruction. Chondrocytes were isolated enzymatically from human septal cartilage. The single cells were resuspended in agarose and transferred into the polymer scaffolds to create mechanical stability and retain the chondrocyte-specific phenotype. The cell-polymer constructs were then kept in perfusion culture for 1 week prior to subcutaneous transplantation into thymusaplastic nude mice. After 6, 12, and 24 weeks, the specimens were explanted and analyzed histochemically on the presence of collagen (azan staining), proteoglycans (Alcian blue staining), and calcification areas (von Kossa staining). Furthermore, different collagen types (collagen type I, which is found in most tissues, but not in hyaline cartilage matrix; and collagen type II, which is cartilage specific) were differentiated immunohistochemically by the indirect immunoperoxidase technique. Vascular ingrowth was investigated by a factor VIII antibody, which is a endothelial marker. Quantification of several matrix components was performed using the software Photoshop. Significant differences were found between both nonwoven structures concerning matrix synthesis and matrix quality as well as vascular ingrowth. Ethisorb, with a degradation time of approximately 3 weeks in vitro, showed no significant differences from normal human septal cartilage in the amount of collagen types I and II 24 weeks after transplantation. Thin fibrous tissue layers containing blood vessels encapsulated the transplants. V 7-2 constructs, which did not show strong signs of degradation even 24 weeks after transplantation, contained remarkably smaller amounts of cartilage-specific matrix components. At the same time, there was vascular ingrowth even in central parts of the transplants. In conclusion, polymer scaffolds with a short degradation time are suitable materials for the development of cartilage matrix products, while longer stability seems to inhibit matrix synthesis. Thus, in vitro engineering of human cartilage can result in a cartilage-like tissue when appropriate nonwovens are used. Therefore, this method could be the ideal cartilage replacement method without the risk of infection and with the possibility of reconstructing large defects with different configurations.

Resorbable continuous-fibre reinforced polymers for osteosynthesis.

Four institutes from three countries in the European Union have collaborated under the BRITE-EURAM framework programme for the development of processing technologies for resorbable osteosynthesis devices. The devices should be continuous-fibre reinforced, and the technology should offer the possibility of orienting the fibres in the main trajectories. Poly-L-lactide and poly-L-DL-lactides have been synthesized for reinforcement fibres and matrix material, respectively. Melt-spun P-L-LA fibres of a strength of 800 MPa have been embedded in an amorphous P-L-DL-LA 70 : 30 matrix by compression moulding. Ethyleneoxide sterilized samples have been tested in vitro and in vivo. A satisfying bending modulus has been reached (6 GPa). Yet with 50% strength retention after ten weeks, fast degradation occurred that could be related to residual monomers. By this fast degradation 70% resorption after one year could be observed in the non-functional animal studies in rabbits. There was only a mild inflammatory reaction, which confirmed the good biocompatibility of the materials even during the resorption period. Further effort has to concentrate on the reduction of initial monomer content. The great advantage of the processing method to orient fibres in the device will be utilized in prototype samples, e.g. an osteosynthesis plate with fixation holes.

Experimental in vitro and in vivo studies of epithelium formation on biomaterials seeded with isolated respiratory cells.

Extensive tracheal defects after intensive care medicine, trauma, or large resections in tumor surgery remain a major challenge in plastic and reconstructive surgery. Defects that cannot be satisfactorily treated by complicated and costly reconstructive techniques reveal a need for an alloplastic tracheal replacement. Recent experimental and clinical studies in the development of alloplastic tracheal prostheses proved that the lack of an epithelial lining on the luminal surfaces and inadequate biophysical properties and shapes of the prostheses were the main causes for failure of these prostheses. In this study a cell-seeding technique has been used. Adhesion, spreading, and differentiation of seeded mucosa cells on biomaterials in vitro were observed by scanning electron microscopy (SEM). Chemical properties and surface structure of the material influenced the differentiation process. Epithelium formation of incorporated tracheal prostheses was tested in animal experiments. Isolated respiratory cells were seeded into implanted tubular prostheses of porous polyurethane or expanded polytetrafluorethylene. Light microscopy and SEM showed the tendency of epithelium formation on the surface of the lumen. Vigorous cell layers, predominantly as multiple cell layers of squamous epithelium, were observed. Ciliated or mucus cells were not detected. It can be stated that the epithelium formation on incorporated porous implants is possible. Further studies of the stability and the differentiation process of the epithelium on such implants is needed before an introduction of tracheal replacements into the clinical practice can be considered.

Resorbable polyesters in cartilage engineering: affinity and biocompatibility of polymer fiber structures to chondrocytes.

The resorbable polymers polyglycolic acid (PGA) and polylactic acid (PLA) are gaining increasing importance in tissue engineering and cell transplantation. The present investigation was focused on the biocompatibility and cell retaining behavior of PGA/poly-L-lactide (PLLA) (90/10) and PLLA nonwoven structures for the in vitro development of chondrocyte-polymer constructs. The effect of the relevant monomers to chondrocytes was analyzed. Type II collagen and poly-L-lysine were compared to improve loading of PGA/PLLA and PLLA polymer nonwovens with chondrocytes. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra-zoliumbrom ide (MTT) test was applied for quantification. At concentrations above 2 mg/mL, glycolic acid was more cytotoxic than lactic acid. As shown by pH equilibration, the cytotoxic effect is not due merely to the acidity of the alpha-hydroxy acids. Regarding the degradation products, glycolic acid, and L(+) lactic acid, nonwovens of PLLA are more biocompatible with chondrocytes than nonwovens of polyglycolide. Collagen type II and poly-L-lysine generally improved cell seeding on resorbable polymers in tissue engineering; however, their efficiency varies depending on the type of fiber structure.