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Development of biomaterials for gene therapy
Han, S., R. I. Mahato, et al. (2000), Mol Ther 2(4): 302-17.
Abstract: Novel biocompatible polymeric gene carriers have been examined for their potential in treating various genetic and acquired diseases. The use of polymeric gene carriers may overcome the current problems associated with viral vectors in safety, immunogenicity, and mutagenesis. However, effective polymer-based gene therapy requires the control of cellular access and uptake, intracellular trafficking, and nuclear retention of plasmid DNA. Inefficient endosomal release, cytoplasmic transport, and nuclear entry of plasmids are currently limiting factors in the use of polymers for effective plasmid-based gene therapy. Therefore, several different polymeric gene carriers have been designed recently in an attempt to overcome these problems. This review explores the conceptual and experimental aspects of polymer-based gene delivery and presents an overview on the recent use of polymers to enhance the effectiveness of plasmid-based systems. Despite their current limitations, polymeric carriers have significant potential as commercially viable gene medicines.

Development of FRP composite structural biomaterials: fatigue strength of the fiber/matrix interfacial bond in simulated in vivo environments
Latour, R. A., Jr. and J. Black (1993), J Biomed Mater Res 27(10): 1281-91.
Abstract: Fiber/matrix interfacial bonding in fiber reinforced polymer (FRP) composite materials is potentially sensitive to degradation in aqueous environments. Ultimate bond strength (UBS) in carbon fiber/polysulfone (CF/PSF) and polyaramid/polysulfone (K49/PSF) was previously reported to be significantly decreased in two simulated in vivo environments. While UBS is a useful parameter, for orthopedic implant applications the fatigue behavior of the interface is probably a more relevant indicator of long-term composite material performance. In this article, the effects of simulated in vivo environments (saline, exudate) upon the fatigue behavior of the interface of CF/PSF and K49/PSF are reported. The fatigue behavior of both material combinations was linearly dependent on the logarithm of fatigue life in the dry (control), saline, and exudate environments. Testing either material in saline and exudate resulted in significantly lower fatigue strength than in the dry environment; however, results in the two wet environments were indistinguishable. The CF/PSF interface experienced fatigue failure at approximately 10(5) load cycles at a maximum applied load level of only 15% of its ultimate dry bond strength without indication of an endurance limit being reached. These results raise some important questions regarding the durability of CF/PSF composite in load bearing orthopedic applications.

Development of FRP composite structural biomaterials: ultimate strength of the fiber/matrix interfacial bond in in vivo simulated environments
Latour, R. A., Jr. and J. Black (1992), J Biomed Mater Res 26(5): 593-606.
Abstract: Fiber reinforced polymer (FRP) composites are being developed as alternatives to metals for structural orthopedic implant applications. FRP composite fracture behavior and environmental interactions are distinctly different from those which occur in metals. These differences must be accounted for in the design and evaluation of implant performance. Fiber/matrix interfacial bond strength in a FRP composite is known to strongly influence fracture behavior. The interfacial bond strength of four candidate fiber/matrix combinations (carbon fiber/polycarbonate, carbon fiber/polysulfone, polyaramid fiber/polycarbonate, polyaramid fiber/polysulfone) were investigated at 37 degrees C in dry and in vivo simulated (saline, exudate) environments. Ultimate bond strength was measured by a single fiber-microdroplet pull-out test. Dry bond strengths were significantly decreased following exposure to either saline or exudate with bond strength loss being approximately equal in both the saline and exudate. Bond strength loss is attributed to the diffusion of water and/or salt ions into the sample and their interaction with interfacial bonding. Because bond degradation is dependent upon diffusion, diffusional equilibrium must be obtained in composite test samples before the full effect of the test environment upon composite mechanical behavior can be determined.

Development of infection resistant polyurethane biomaterials using textile dyeing technology
Phaneuf, M. D., M. J. Bide, et al. (2001), Asaio J 47(6): 634-40.
Abstract: Infection is a major complication when using biomaterials such as polyurethane in the clinical setting. The purpose of this study was to develop a novel infection resistant polyurethane biomaterial using textile dyeing technology. This procedure results in incorporation of the antibiotic into the polymer, resulting in a slow, sustained release of antibiotic from the material over time, without the use of exogenous binder agents. Polycarbonate based urethanes were synthesized that contained either a non-ionic (bdPU) or anionic (cPU) chain extender within the polymer backbone and cast into films. The fluoroquinolone antibiotic ciprofloxacin (Cipro) was applied to bdPU and cPU using textile dyeing technology, with Cipro uptake determined by absorbance reduction of the "dyebath." These dyed bdPU/cPU samples were then evaluated for prolonged Cipro release and antimicrobial activity by means of spectrophotometric and zone of inhibition assays, respectively. Cipro release and antimicrobial activity by dyed cPU segments that were aggressively washed persisted over 9 days, compared with dyed bdPU and dipped cPU control segments that lasted < 24 hours. Dyed cPU segments, which remained in a static wash solution, maintained antimicrobial activity for 11 days (length of study), whereas controls again lost antimicrobial activity within 24 hours. Thus, application of Cipro to the cPU polymer by means of dyeing technology results in a slow sustained release of antibiotic with persistent bacteriocidal properties over extended periods of time.

Development of mammalian cell-enclosing subsieve-size agarose capsules (<100 microm) for cell therapy
Sakai, S., K. Kawabata, et al. (2005), Biomaterials 26(23): 4786-92.
Abstract: Agarose capsules were prepared using a droplet breakup method in a coflowing stream. Subsieve-size capsules 76+/-9 microm in diameter were obtained by extruding 4 wt% agarose solution from a needle (300 microm inner diameter) at a velocity of 1.2 cm/s into an ambient liquid paraffin flow of 20.8 cm/s. Increasing the flow rate of the liquid paraffin and decreasing that of the agarose solution resulted in a decreased resultant capsule diameter. Reduction in diameter from several hundred micrometers to subsieve-size (<100 microm) enhanced molecular exchange and mechanical stability. Measurements based on the percentage of intact mitochondria in the cells demonstrated that the viability of the enclosed cells was independent of capsule diameter. No significant difference was observed between the viabilities of cells enclosed in capsules with diameters of 79+/-8 and 351+/-41 microm (p=0.43). Compared with cells seeded in a tissue culture dish, the cells enclosed in the subsieve-size capsules showed 89.2% viability.

Development of microporous covered stents: geometrical design of the luminal surface
Nakayama, Y., S. Nishi, et al. (2005), Int J Artif Organs 28(6): 600-8.
Abstract: To reduce in-stent restenosis rates we have developed newly designed covered stents, in which a stent strut is buried into a microporous elastomeric cover film to provide a physical barrier against tissue ingrowth and a pharmacological reservoir for drug-eluting. The covered stents were prepared by dip-coating balloon expandable stents mounted on a stainless steel rod in a segmented polyurethane (SPU) solution, and were subsequently subjected to laser-processed microporing (pore diameter, 100 microm; interpore distance, 200 microm). The covered stents, which possessed flat luminal surfaces and micropores that were homogeneously arranged on the whole surface of the covering film, were deployed into the bilateral common carotid arteries of normal New Zealand white rabbits. Angiography after one month of implantation showed all stents were patent with little thrombus formation. The mean thickness of the formed neointimal layers was 292 +/- 177 microm (n=8), which was close to the size in non-covered bare stent (231 +/- 58 microm, n=7), but markedly decreased (about 2/3) from that in the previously developed wrapping-type covered stents (415 +/- 173 microm, P<0.01, n=8).

Development of multifunctional polymer-mineral composite materials for bone tissue engineering
Vlakh, E. G., E. F. Panarin, et al. (2005), J Biomed Mater Res A 75(2): 333-41.
Abstract: The main goal of this article is the development of a novel approach to construct multifunctional composite scaffolds for bone tissue engineering. For this purpose, different kinds of mineral macroporous supports, water-soluble aldehyde-containing copolymers of N-vinylpyrrolidone, as well as different nonspecific and biospecific ligands governing cell adhesion and growth have been used. The composite materials were tested initially for cytotoxicity in cell culture experiments using a model cell line.

Development of new injectable bulking agents: biocompatibility of radiopaque polymeric microspheres studied in a mouse model
Emans, P. J., K. Saralidze, et al. (2005), J Biomed Mater Res A 73(4): 430-6.
Abstract: Radiopaque polymeric microspheres have a potential as new bulking agents for treatment of stress urinary incontinence (SUI). The advantage over existing bulking agents lies in their X-ray visibility in situ; other polymeric bulking agents (e.g., PTFE or silicone rubbers) are practically radiolucent (i.e., incapable of absorbing X-radiation). Radiopacity is useful in practice because of the high spatial accuracy of X-ray imaging. For instance, X-ray fluoroscopy can be used to assess possible migration of the bulking agent over time or to provide guidance in cases in which a second injection of a bulking agent is necessary (repeated treatment of SUI). Biocompatibility of injected radiopaque microspheres was investigated in vivo by using the mouse as a model. Microspheres were injected subcutaneously (9 animals) or intramuscularly (9 animals), and follow-up was 8 days or 3 months. X-ray fluoroscopy gave clear images of the microspheres as an ensemble, and it was found that no migration occurred during 3 months. Histopathology confirmed that all microspheres stayed close to the site of the injection. The microspheres appeared to be well tolerated; only a few giant cells, manifesting a mild inflammatory reaction, were encountered. At 3 months, capillary blood vessels were observed throughout the microsphere beds, and macrophages and fibroblast cells were seen in between the microspheres. This is encouraging with respect to the intended application, although it must be acknowledged that the data refer merely to a mouse model. Further experiments with larger, more representative models (rabbit and goat) are in progress.

Development of new reverse micellar microencapsulation technique to load water-soluble drug into PLGA microspheres
Kim, H., M. Cho, et al. (2005), Arch Pharm Res 28(3): 370-5.
Abstract: The objective of this study was to develop a new reverse micelle-based microencapsulation technique to load tetracycline hydrochloride into PLGA microspheres. To do so, a reverse micellar system was formulated to dissolve tetracycline hydrochloride and water in ethyl formate with the aid of cetyltrimethylammonium bromide. The resultant micellar solution was used to dissolve 0.3 to 0.75 g of PLGA, and microspheres were prepared following a modified solvent quenching technique. As a control experiment, the drug was encapsulated into PLGA microspheres via a conventional methylene chloride-based emulsion procedure. The microspheres were then characterized with regard to drug loading efficiency, their size distribution and morphology. The reverse micellar procedure led to the formation of free-flowing, spherical microspheres with the size mode of 88 microm. When PLGA microspheres were prepared following the conventional methylene chloride-based procedure, most of tetracycline hydrochloride leached to the aqueous external phase: A maximal loading efficiency observed our experimental conditions was below 5%. Their surfaces had numerous pores, while their internal architecture was honey-combed. In sharp contrast, the new reverse micellar encapsulation technique permitted the attainment of a maximal loading efficiency of 63.19 +/- 0.64%. Also, the microspheres had smooth and pore-free surfaces, and hollow cavities were absent from their internal matrices. The results of this study demonstrated that PLGA microspheres could be successfully prepared following the new reverse micellar encapsulation technique.

Development of polymer membranes with improved haemocompatibility for biohybrid organ technology
Groth, T., B. Seifert, et al. (2005), Clin Hemorheol Microcirc 32(2): 129-43.
Abstract: Biomedical technology has opened up possibilities of treating the failure of internal organs like kidney and liver by artificial organ therapy. Most of these techniques are based on polymer membranes, which allow the removal of excess of water, salts and toxins from the circulation. However, haemodialysis for the replacement of kidney function results in an increased morbidity and mortality of patients after long-term application. Conventional therapy, such as haemofiltration for the treatment of acute liver failure does not improve significantly the survival rate of patients. Biohybrid organ support as a combination of the artificial organ therapy with the functional activity of immobilised cells seems to be a solution of the problem. Membranes applied in these devices have to face both tissue cells and blood. Organ cells in biohybrid organs have to make intimate contact with the surface of membrane but must also develop close cell-cell-connections as a prerequisite for their survival and high functional activity. Blood to be detoxified will contact the other side of membrane and may not become activated by the synthetic material. New polymer membranes based on acrylonitrile were developed to address these requirements by tailoring the composition of copolymers and to be applied in a specific hollow fibre bioreactor with an outer fibre for blood contact, and an inner fibre for tissue contact or vice versa.

Development of specific adsorbents for human tumor necrosis factor-alpha: influence of antibody immobilization on performance and biocompatibility
Weber, V., I. Linsberger, et al. (2005), Biomacromolecules 6(4): 1864-70.
Abstract: To develop adsorbents for the specific removal of tumor necrosis factor-alpha (TNF) in extracorporeal blood purification, cellulose microparticles were functionalized either with a monoclonal anti-TNF antibody (mAb) or with recombinant human antibody fragments (Fab). The TNF binding capacity of the adsorbents was determined with in vitro batch experiments using spiked human plasma (spike: 1200 pg TNF/mL; 1 mg particles in 250 muL plasma). Random immobilization of the full-sized monoclonal antibody to periodate-activated cellulose yielded particles with excellent adsorption capacity (258.1 +/- 48.6 pg TNF per mg adsorbent wet weight). No leaching of antibody was detectable, and the adsorbents retained their activity for at least 12 months at 4 degrees C. We found that the conditions used during immobilization of the antibody (pH, nature of the reducing agent) profoundly influenced the biocompatibility of the resulting adsorbents, especially with respect to activation of the complement system. Particles obtained by random immobilization of the monovalent Fab fragments on periodate-activated cellulose using the same conditions as for immobilization of the mAb exhibited only low adsorption capacity (44 +/- 7 pg/mg adsorbent wet weight). Oriented coupling of the Fab fragments on chelate-epoxy cellulose via a C-terminal histidine tag, however, increased the adsorption capacity to 178.3 +/- 8.6 pg TNF/mg adsorbent wet weight. Thus, in the case of small, monovalent ligands, the orientation on the carrier is critical to retain full binding activity.

Development of the human umbilical vein scaffold for cardiovascular tissue engineering applications
Daniel, J., K. Abe, et al. (2005), Asaio J 51(3): 252-61.
Abstract: Biologic function and the mechanical performance of vascular grafting materials are important predictors of graft patency. As such, "functional" materials that improve biologic integration and function have become increasingly sought after. An important alternative to synthetic materials is the use of biomaterials derived from ex vivo tissues that retain significant biologic and mechanical function. Unfortunately, inconsistent mechanical properties that result from tedious, time consuming, manual dissection methods have reduced the potential usefulness of many of these materials. We describe the preparation of the human umbilical vein (HUV) for use as an acellular, three-dimensional, vascular scaffold using a novel, automated dissection methodology. The goal of this investigation was to determine the effectiveness of the autodissection methodology to yield an ex vivo biomaterial with improved uniformity and reduced variance. Mechanical properties, including burst pressure, compliance, uniaxial tension testing, and suture holding capacity, were assessed to determine the suitability of the HUV scaffold for vascular tissue engineering applications. The automated methodology results in a tubular scaffold with significantly reduced sample to sample variation, requiring significantly less time to excise the vein from the umbilical cord than manual dissection methods. Short-term analysis of the interactions between primary human vascular smooth muscle cells and fibroblasts HUV scaffold have shown an excellent potential for cellular integration by native cellular remodeling processes. Our work has shown that the HUV scaffold is mechanically sound, uniform, and maintains its biphasic stress-strain relationship throughout tissue processing. By maintaining the mechanical properties of the native blood vessels, in concert with promising cellular interactions, the HUV scaffold may lead to improved grafts for vascular reconstructive surgeries.

Diagnosis and treatment of biomaterial-associated vascular infections
Bandyk, D. F. (1992), Infect Dis Clin North Am 6(3): 719-29.
Abstract: Management of biomaterial-associated vascular infections requires an understanding of pathogenetic mechanisms, risk factors, and microbiologic characteristics. Staphylococci sp., especially slime-producing strains of S. epidermidis are the prevalent pathogens. Experimental and clinical studies have indicated in situ replacement, particularly with an antibiotic-bonded prosthesis, as effective treatment for infections caused by coagulase-negative staphylococci. When sepsis is a presenting sign, prompt intervention, total excision of the prosthesis, and antibiotic administration are required.

Diagnostic Raman spectroscopy for the forensic detection of biomaterials and the preservation of cultural heritage
Edwards, H. G. and T. Munshi (2005), Anal Bioanal Chem 382(6): 1398-406.
Abstract: This paper reviews the contributions of analytical Raman spectroscopy to the non-destructive characterisation of biological materials of relevance to forensic science investigations, including the sourcing of resins and the identification of the biodegradation of art and archaeological artefacts. The advantages of Raman spectroscopy for non-destructive analysis are well-appreciated; however, the ability to record molecular information about organic and inorganic species present in a heterogeneous specimen at the same time, the insensitivity of the Raman scattering process to water and hydroxyl groups, which removes the necessity for sample desiccation, and the ease of illumination for samples of very small and very large sizes and unusual shapes are also apparent. Several examples are used to illustrate the application of Raman spectroscopic techniques to the characterisation of forensic biomaterials and for the preservation of cultural heritage through case studies in the following areas: wall-paintings and rock art, human and animal tissues and skeletal remains, fabrics, resins and ivories.

Dialysis needle puncture of Wallgrafts placed in polytetrafluoroethylene hemodialysis grafts
Rhodes, E. S. and A. M. Silas (2005), J Vasc Interv Radiol 16(8): 1129-34.
Abstract: This study evaluates the effect of routine venipuncture at hemodialysis on the durability of Wallgraft covered stents. Thirteen covered stents were placed in six aging, failing polytetrafluoroethylene grafts for treatment of pseudoaneurysms and recurrent stenoses. Four patients did not experience significant graft complications. One graft was ligated for an infected overlying skin ulcer. After surgical revision, the graft remains functional. Another patient experienced fraying of the stent edges and recurrence of a small pseudoaneurysm. Our experience suggests that the Wallgraft covered stent can withstand routine venipuncture at dialysis without flow-limiting stent distortion.

Diameter-selective encapsulation of metallocenes in single-walled carbon nanotubes
Li, L. J., A. N. Khlobystov, et al. (2005), Nat Mater 4(6): 481-5.
Abstract: Encapsulation of organic molecules in carbon nanotubes has opened a new route for the fabrication of hybrid nanostructures. Here we show that diameter-selective encapsulation of two metallocene compounds bis(cyclopentadienyl) cobalt and bis(ethylcyclopentadienyl) cobalt has been observed in single-walled carbon nanotubes. In particular, bis(cyclopentadienyl) cobalt is observed to fill only nanotubes of one specific diameter. Electron transfer from the cobalt ions to the nanotubes has been directly observed through a change in the charge state of the encapsulated molecules. The filling of the tubes is found to induce a red-shift of the photoluminescence emission, which is attributed to the formation of localized impurity states below the conduction band of the nanotubes.

Diamine-extended glutaraldehyde- and carbodiimide crosslinks act synergistically in mitigating bioprosthetic aortic wall calcification
Zilla, P., D. Bezuidenhout, et al. (2005), J Heart Valve Dis 14(4): 538-45.
Abstract: BACKGROUND AND AIM OF THE STUDY: The extension of glutaraldehyde (GA) crosslinks with diamine bridges was shown previously to reduce bioprosthetic heart valve calcification to a significant degree. The aim of the present study was to investigate whether the additional crosslinking of functional carboxyl groups could augment this anticalcific effect at the low glutaraldehyde concentrations typically used in commercial heart valve production. METHODS: Entire aortic roots of medium-sized pigs were fixed after 48 h of cold storage. Crosslinking of amino-functional groups was achieved either by GA fixation alone (0.2% or 0.7%) or with an interim treatment with the diamine L-lysine (25, 50 or 100 mM; 37 degrees C; 2 days). Carboxyl groups were activated with carbodiimide (N'-{3-dimethylaminopropyl}-N-ethyl carbodiimide hydrochloride (EDC), 240 mM) and crosslinked with an oligomeric diamine (polypropylene glycol-bis-aminopropyl ether (Jeffamine), 60 mM, 230D). By permutation of treatments and combinations thereof, a total of 17 groups was compared. Aortic wall discs (12 mm diameter) were implanted subcutaneously into seven-week-old Long-Evans rats for 60 days. Tissue calcification was determined by histology and atomic absorption spectrophotometry. RESULTS: There was no significant difference in tissue calcification if either GA or carbodiimide fixation was used alone. Equally, the combined crosslinking with GA and EDC/Jeffamine did not achieve a mitigation of tissue calcification below levels seen in at least one of the two treatments alone. When commercial GA fixation was mildly diamine-enhanced with L-lysine (25 mM), additional EDC/Jeffamine crosslinking of carboxyl groups resulted in a distinct additive effect in both 0.2% (-31%; p < 0.0002) and 0.7% (-36%; p = 0.0073) GA-fixed tissue. Relative to conventional GA fixation, this combination mitigated aortic wall calcification by 43% (p < 0.0001) and 34% (p = 0.0014) in 0.2% and 0.7% GA-fixed tissue, respectively. An increase in L-lysine concentration to 100 mM further reduced calcification of 0.7% GA-fixed tissue (18.5%; p = 0.016), but had no additional effect on 0.2% GA-fixed tissue (0.6%; p = 0.463). CONCLUSION: A distinct reduction in bioprosthetic aortic wall calcification can be achieved by combining diamine-extended conventional GA fixation with a diamine-extended carbodiimide based crosslinking step.

Diamond: the biomaterial of the 21st century?
Dion, I., C. Baquey, et al. (1993), Int J Artif Organs 16(9): 623-7.

Diethyl methyl chitosan as an intestinal paracellular enhancer: ex vivo and in vivo studies
Avadi, M. R., A. Jalali, et al. (2005), Int J Pharm 293(1-2): 83-9.
Abstract: Chitosan exhibits favorable biological properties such as no toxicity, biocompatibility and biodegradability; therefore, it has attracted great attention in both pharmaceutical and biomedical fields. Chitosan exhibits poor solubility at pH values above 6 that prevents enhancing effects at the sites of absorption of drugs. In the present work, N-diethyl methyl chitosan (DEMC) was prepared and the enhancing effect of this polymer was investigated. Ex vivo studies have shown a significant increase in absorption of brilliant blue in the presence of diethyl methyl chitosan in comparison with chitosan. DEMC with positive charges is able to interact with tight junctions of colon epithelial cells and hence increases permeability of brilliant blue across the tight junctions. In vivo investigations have exhibited the absorption enhancer effects of DEMC on the colon absorption of insulin in normal and diabetic rats. The insulin absorption from the rat's colon was evaluated by its hypoglycemic effect. A significant decrease in blood glucose was observed, when mixture of insulin and DEMC was introduced in ascending colon of rats.

Differences in stability and bone remodeling between a customized uncemented hydroxyapatite coated and a standard cemented femoral stem A randomized study with use of radiostereometry and bone densitometry
Grant, P., A. Aamodt, et al. (2005), J Orthop Res 23(6): 1280-5.
Abstract: The custom made Unique stem is designed to fit closely to the metaphyseal region of the femur in order to obtain maximum mechanical stability and optimal load transfer. Thirty-seven patients (38 hips) with non-inflammatory arthritis were randomized to the uncemented custom made Unique stem or the Elite Plus stem inserted with cement. The patients have been followed clinically as well as with radiostereometry (RSA) and Dual-energy X-ray Absorptiometry (DXA) for 2 years. After 2 years the RSA result showed minimal translation and rotation for the Unique stem while the Elite Plus rotated slightly (mean 1.05 degrees) into retroversion. Compared to previous studies the Elite Plus was as stable as the Charnley prosthesis. The DXA results showed a significantly higher proximal and total (10% for the Unique versus 5% for Elite) bone loss for the Unique stem compared to the Elite Plus. Thus the optimal proximal press-fit of the custom made stem did secure a stable fixation, but did not decrease the proximal bone loss.

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Last Modified: 8 February 2006