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In vitro biocompatibility assessment of naturally derived and synthetic biomaterials using normal human urothelial cells
Pariente, J. L., B. S. Kim, et al. (2001), J Biomed Mater Res 55(1): 33-9.
Abstract: The reconstruction of urinary tissues often employs various types of biomaterials, and adequate material biocompatibility is essential for the successful reconstruction of urinary tissues. In this study we utilized a primary normal human urothelial cell culture system to evaluate the in vitro biocompatibility of a number of naturally derived biomaterials [i.e., bladder submucosa, small intestinal submucosa, collagen, and alginate] and polymeric biomaterials [i.e., poly(glycolic acid), poly(L-lactic acid), poly(lactic-co-glycolic acid), and silicone] that are either experimentally or clinically used in urinary reconstructive surgery. To determine the cytotoxic and bioactive effects of these biomaterials, the cell viability, metabolic activity, apoptotic properties, and DNA-synthesis activity were measured with four types of assays [Neutral Red, 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide, apoptotic activity, and tritiated thymidine incorporation assays] using extract and direct contact methods. Most of the biomaterials tested did not induce significant cytotoxic effects and exhibited normal metabolic function and cell growth in vitro. This normal primary human urothelial cell culture model is suitable for in vitro biocompatibility assessments and is able to provide information on the cell-biomaterial interactions and the ability of biomaterials to support bioactive cell functions.

In vitro biocompatibility evaluation of naturally derived and synthetic biomaterials using normal human bladder smooth muscle cells
Pariente, J. L., B. S. Kim, et al. (2002), J Urol 167(4): 1867-71.
Abstract: PURPOSE: Tissue engineering of the urinary tract often requires the use of various biomaterials. Adequate biomaterial biocompatibility is necessary for successful urinary reconstruction. In this study using a primary normal human bladder smooth muscle cell culture system we evaluated the in vitro biocompatibility of a number of naturally derived biomaterials, including bladder submucosa, small intestinal submucosa, collagen and alginate, and polymeric biomaterials, including polyglycolic acid, poly(L-lactic acid) and poly(lactic-co-glycolic acid, which have been used for urinary reconstruction experimentally or clinically. MATERIALS AND METHODS: To determine the cytotoxic and bioactive effects of bladder submucosa, small intestinal submucosa, collagen, alginate, polyglycolic acid, poly(L-lactic acid) and poly(lactic-co-glycolic acid) we measured cell viability, metabolic activity, apoptotic properties and DNA synthesis activity with 4 types of assays, namely Neutral Red (Sigma Chemical Co., St. Louis, Missouri), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (Sigma Chemical Co.), apoptotic activity and tritiated thymidine incorporation (Dupont NEN, Boston, Massachusetts) assays. Normal human bladder smooth muscle cells were cultured with the extracts of the biomaterials or cultured in direct contact with the biomaterials. RESULTS: All naturally derived and synthetic biomaterials tested in this study except alginate exhibited nontoxic and bioactive effects on human bladder smooth muscle cells (HBSMCs) in vitro, as indicated by the 4 types of biocompatibility assays using the extract and direct contact methods. Cell viability, apoptotic properties, metabolic activity and DNA synthesis activity of HBSMCs cultured with the extracts of the biomaterials or cultured in direct contact with the biomaterials were not significantly different from those of negative controls (fresh medium with no extracts or tissue culture plates without biomaterials). CONCLUSIONS: All naturally derived and synthetic biomaterials tested in this study except alginate exhibited nontoxic and bioactive effects on HBSMCs in vitro. This normal primary human bladder smooth muscle cell culture model is suitable for in vitro biocompatibility assessment. It provides information on cell-biomaterial interactions and on the ability of biomaterials to support bioactive cell functions.

In vitro biological performances of phosphorylcholine-grafted ePTFE prostheses through RFGD plasma techniques
Chevallier, P., R. Janvier, et al. (2005), Macromol Biosci 5(9): 829-39.
Abstract: Arterial prostheses made of microporous Teflon (ePTFE) are currently used in vascular surgery as bypasses for small and medium vessels. However, several clinical complications, such as thrombosis, frequently occur in these prostheses when implanted in humans. In this work, an original strategy was developed to improve the hemocompatibility of ePTFE prostheses, based on glow-discharge surface modification followed by chemical grafting of phosphorylcholine, known for its hemocompatible properties. This procedure leads to a covalent attachment of the molecules, therefore preventing their removal by shear stress induced by blood flow at the implant wall. The improvement of the blood compatibility properties of the modified ePTFE arterial prostheses have been investigated by in vitro tests such as thromboelastography, neutrophil adsorption, platelet aggregation, and cell cultures. These in vitro tests put in evidence that thrombogenicity index, platelet aggregation, and neutrophil adhesion were decreased by the molecule grafted on the prostheses. Moreover, the cell growth on the surface of the PRC-grafted prostheses was greatly enhanced in comparison to the virgin prosthesis. Based on these results, it could be concluded that PRC grafting on ePTFE prostheses permit to improve in vitro hemocompatibility and biocompatibility in comparison with their virgin counterpart.

In vitro blood compatibility of polymeric biomaterials through covalent immobilization of an amidine derivative
Gouzy, M. F., C. Sperling, et al. (2004), Biomaterials 25(17): 3493-501.
Abstract: We present a surface coating with anticoagulant characteristics showing significantly reduced coagulation activation. The synthesis of a monomeric conjugate containing a benzamidine moiety was carried out and its inhibitory activity against human thrombin, the key enzyme of the blood coagulation cascade, was determined using a chromogenic assay. Based on that, low-thrombogenic interfaces were prepared by covalent attachment of this low-molecular weight thrombin inhibitor on poly(octadecene-alt-maleic anhydride) copolymer thin films and characterized using ellipsometry, XPS and dynamic contact angle measurements. The in vitro hemocompatibility tests using freshly drawn human whole blood showed, in agreement with the SEM images, that a PO-MA film modified with a benzamidine moiety using a PEG spacer decreased the activation of coagulation, platelets and the complement system. The decreased protein adsorption, in addition to the specific inhibition of thrombin, effectively enhanced the short-term hemocompatibility characteristics.

In vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells
Wang, Y., U. J. Kim, et al. (2005), Biomaterials 26(34): 7082-94.
Abstract: Adult cartilage tissue has limited self-repair capacity, especially in the case of severe damages caused by developmental abnormalities, trauma, or aging-related degeneration like osteoarthritis. Adult mesenchymal stem cells (MSCs) have the potential to differentiate into cells of different lineages including bone, cartilage, and fat. In vitro cartilage tissue engineering using autologous MSCs and three-dimensional (3-D) porous scaffolds has the potential for the successful repair of severe cartilage damage. Ideally, scaffolds designed for cartilage tissue engineering should have optimal structural and mechanical properties, excellent biocompatibility, controlled degradation rate, and good handling characteristics. In the present work, a novel, highly porous silk scaffold was developed by an aqueous process according to these criteria and subsequently combined with MSCs for in vitro cartilage tissue engineering. Chondrogenesis of MSCs in the silk scaffold was evident by real-time RT-PCR analysis for cartilage-specific ECM gene markers, histological and immunohistochemical evaluations of cartilage-specific ECM components. Dexamethasone and TGF-beta3 were essential for the survival, proliferation and chondrogenesis of MSCs in the silk scaffolds. The attachment, proliferation, and differentiation of MSCs in the silk scaffold showed unique characteristics. After 3 weeks of cultivation, the spatial cell arrangement and the collagen type-II distribution in the MSCs-silk scaffold constructs resembles those in native articular cartilage tissue, suggesting promise for these novel 3-D degradable silk-based scaffolds in MSC-based cartilage repair. Further in vivo evaluation is necessary to fully recognize the clinical relevance of these observations.

In vitro characterization of a bone marrow stem cell-seeded collagen gel composite for soft tissue grafts: effects of fiber number and serum concentration
Lewus, K. E. and E. A. Nauman (2005), Tissue Eng 11(7-8): 1015-22.
Abstract: Cell-seeded collagen hydrogels have been used in the engineering of many tissue types, from skin and vasculature to spinal cord. One of the primary limitations of collagen-based hydrogels for use in tissue-engineered grafts is that cells seeded within the gel cause it to contract as much as 70%. By forming a composite gel by adding short collagen fibers, Gentleman et al. (Tissue Eng. 10, 421, 2004) determined that the contraction due to fibroblasts was decreased and permeability was increased. Before these composite hydrogels can be used to design soft tissue replacements, however, the effect of fiber number and serum concentration should be addressed. Consequently, short collagen fibers were included in adult rat bone marrow stem cell-seeded hydrogels for composite support. The mass of fibers was varied from 1.6 to 31.3 mg per gel, and the effect of serum concentration in the growth medium was examined. It was determined that increasing fiber mass and decreasing serum concentration significantly decreased contraction, which plateaued after day 10. Cell number increased throughout the experiment, demonstrating the compatibility of bone marrow stem cells with the collagen composite gels. By using short collagen fibers to create a collagen composite gel, preservation of the original dimensions can be achieved without compromising cellular viability.

In vitro characterization of chitosan-gelatin scaffolds for tissue engineering
Huang, Y., S. Onyeri, et al. (2005), Biomaterials 26(36): 7616-27.
Abstract: Recently, chitosan-gelatin scaffolds have gained much attention in various tissue engineering applications. However, the underlying cell-matrix interactions remain unclear in addition to the scaffold degradation and mechanical characteristics. In this study, we evaluated (i) the degradation kinetics of chitosan and chitosan-gelatin scaffolds in the presence of 10mg/L of lysozyme for dimensional stability, weight loss, and pH changes for a period of 2 months, (ii) tensile and compressive properties of films and scaffolds in wet state at 37 degrees C, (iii) viability of fibroblasts and human umbilical vein endothelial cells (HUVECs) on scaffolds, and (iv) the alteration in cell spreading characteristics, cytoskeletal actin distribution, focal adhesion kinase (FAK) distribution and PECAM-1 expression of HUVECs under static and 4.5, 8.5, 13 and 18 dyn/cm2 shear stress conditions. Degradation results showed that gelatin-containing chitosan scaffolds had faster degradation rate and significant loss of material than chitosan. Mechanical properties of chitosan are affected by the addition of gelatin although there was no clear trend. Three-dimensional chitosan and chitosan-gelatin scaffolds supported fibroblast viability equally. However, chitosan membranes decreased cell-spreading area, disrupted F-actin and localized FAK in the nucleus of HUVECs. Importantly, the lowest shear stress tested (4.5 dyn/cm2) for 3 h washed away cells on chitosan suggesting weak cell adhesion. In the blends, effect of gelatin was dominant; actin and FAK distribution were comparable to gelatin in static culture. However, at higher shear stresses, presence of chitosan inhibited shear-induced increase in cell spreading and weakened cell adhesive strength. No significant differences were observed in PECAM-1 expression. In summary, these results showed significant influence of blending gelatin with chitosan on scaffold properties and cellular behavior.

In vitro characterization of leukocyte mimetic for targeting therapeutics to the endothelium using two receptors
Omolola Eniola, A. and D. A. Hammer (2005), Biomaterials 26(34): 7136-44.
Abstract: Selectins (E- and P-selectin) and other endothelial expressed leukocyte adhesion molecules (ELAMs) are potential targets for site-specific delivery of therapeutics to the vascular endothelium due to their specific and highly regulated expression in vascular disease. It was recently shown that degradable microspheres coated with antibodies against E-selectin or other ELAMs can target inflammation in vivo. However, targeting ELAMs alone cannot differentiate between normal and diseased state, as a basal level of these LAMs are expressed on endothelium in healthy tissues. Furthermore, leukocytes usually employ two separate adhesion molecules in parallel to home to diseased tissues, and we recently quantified the advantages of a two-receptor display for the targeting of leukocyte mimetics (Eniola AO, Willcox PJ, Hammer DA. Interplay between rolling and firm adhesion elucidated with a cell-free system engineered with two distinct receptor-ligand pairs. Biophys J 2003;85:2720-31). Here, we describe a leukocyte mimetic for targeting therapeutics to the vasculature in inflammatory diseases via two receptors, selectin and intercellular cell adhesion molecule-1 (ICAM-1), where biodegradable, polymer microspheres were co-functionalized with the selectin ligand, sialyl Lewis(X) (sLe(X)), and an antibody against ICAM-1, anti-ICAM-1 (aICAM-1). These two-receptor targeted particles, at given ratios of sLe(X)/aICAM-1, firmly adhere to substrate surface in flow only when both targeting ligands can interact with their respective receptors, mimicking the multi-step in vivo leukocyte adhesion in inflammation. Thus, we have faithfully recreated the specificity and extent of leukocyte adhesion in a platform that can allow for local delivery of therapeutics.

In vitro characterization of TGF-beta1 release from genetically modified fibroblasts in Ca(2+)-alginate microcapsules
Paek, H. J., A. B. Campaner, et al. (2005), Asaio J 51(4): 379-84.
Abstract: This study was undertaken to develop an in situ source of transforming growth factor-beta1 (TGF-beta1), one of several molecules potentially useful for a tissue-engineered bioartificial cartilage. Primary human fibroblasts and murine NIH 3T3 cells were genetically modified via viral transfection to express human TGF-beta1. Two viral constructs were used, one expressing a gene encoding for the latent and the other for the constitutively active form of the growth factor. Unmodified cells served as controls. Four genetically modified cohorts and two controls were separately encapsulated in a 1.8% alginate solution using a vibrating nozzle and 0.15M calcium chloride crosslinking bath. Diameter of the spherical capsules was 410 +/- 87 microm. In vitro release rate measured over 168 hours varied with cell types and ranged from 2-17 pg/(milligram of capsules x 24 h) or 2-17 ng/(10(6) cells x 24 h). None of the formulations exhibited a large initial bolus release. Even when serum-supplemented medium was not replenished, cell viabilities remained over 55% after 1 week for all cell types. Microencapsulated genetically modified cells were capable of a constitutive synthesis and delivery of biologically significant quantity of TGF-beta1 for at least 168 hours and thus are of potential utility for artificial cartilage and other orthopedic tissue engineering applications.

In vitro comparison of three materials as apical sealants of equine premolar and molar teeth
Steenkamp, G., A. Olivier-Carstens, et al. (2005), Equine Vet J 37(2): 133-6.
Abstract: REASONS FOR PERFORMING STUDY: Surgical endodontic therapy is a conservative dental technique used in horses with some degree of clinical success. Failure of this procedure can partially be explained by inadequate sealing of the root apices with resultant microleakage in the periapical area. OBJECTIVES: To assess and compare in vitro sealing ability of 3 different dental restorative materials used as apical sealants during equine surgical endodontics. METHODS: Thirty extracted equine cheek teeth were divided randomly into 3 groups and subjected to apicoectomy and apical sealing using 3 materials: reinforced zinc oxide-eugenol cement; intermediate restorative material (IRM); a resin-modified glass ionomer; and amalgam. After apical sealing, the teeth were submerged in a solution of Procion Brilliant Cresyl Blue stain for a period of 7 days. The teeth were then washed, embedded in resin, sectioned and assessed microscopically for dye leakage around the apical restorations. RESULTS: Although the materials proved effective as apical sealants, some dye leakage was encountered in all 3 groups with no statistical difference (P = 0.114). CONCLUSIONS AND POTENTIAL RELEVANCE: IRM, a resin-modified glass ionomer and amalgam all showed comparative features as apical sealants when used in vitro in equine teeth. IRM is currently regarded as the superior material in clinical situations due to its ease of handling and lesser sensitivity to environmental moisture during placement compared to the other 2 materials.

In vitro cytotoxicity evaluation of biomaterials on human osteoblast cells CRL-1543; hydroxyapatite, natural coral and polyhydroxybutarate
Shamsuria, O., A. S. Fadilah, et al. (2004), Med J Malaysia 59 Suppl B: 174-5.
Abstract: The aim of this study was to evaluate the in vitro cytotoxicity of biomaterials; Hydroxyapatite (HA), Natural coral (NC) and Polyhydroxybutarate (PHB). Three different materials used in this study; HA (Ca10(PO4)6(OH)2), NC (CaCO3) and PHB (Polymer) were locally produced by the groups of researcher from Universiti Sains Malaysia. The materials were separately extracted in the complete culture medium (100mg/ml) for 72h and introduced to the osteoblast cells CRL-1543. The viability of osteoblast CRL-1543 cultivated with these extraction materials after 72h incubation period was compared to negative control with neutral red assay by using spectrophotometer at 540nm. The results showed the non-cytotoxicity of the materials. After 72h of incubation period, HA showed 123% viable cells, NC was 99.43% and PHB was 176.75%. In this study, cytotoxicity test dealt mainly with the substances that leached out from the biomaterial. The results obtained showed that the materials were not toxic and also promoted cells growth in the sense of biofunctionality.

In vitro cytotoxicity of melanized poly(2-hydroxyethyl methacrylate) hydrogels, a novel class of ocular biomaterials
Chirila, T. V., D. E. Thompson, et al. (1992), J Biomater Sci Polym Ed 3(6): 481-98.
Abstract: Due to their ability to absorb ultraviolet and visible radiation, we have proposed the melanized poly(2-hydroxyethyl methacrylate) hydrogels as biomaterials suitable for the manufacture of soft artificial intraocular lenses. Their biocompatibility has not been evaluated so far. In this study, poly(2-hydroxyethyl methacrylate) containing various amounts of adrenochrome-melanin were synthesized and the cytotoxicity of their aqueous extracts was assessed by using four in vitro testing techniques (trypan blue dye exclusion, inhibition of DNA synthesis, lactate dehydrogenase release, and inhibition of cell growth). Assays were based on incubation with human choroidal fibroblasts. By the first three techniques, no cytotoxicity was found in the extracts. The inhibition of cell growth test showed a slight cellular protein loss, however only in the extracts of polymers with high melanin content. The results suggest that the release of potentially toxic agents from melanized hydrogels into an aqueous medium is not significant. However, when an assay in collagen gel was carried out in the presence of specimens of melanized hydrogels, a toxic reaction was clearly revealed. This can be caused by a delayed release of toxic molecules from melanin, or by some other mechanism. The use of melanin-containing polymers as implant materials becomes questionable and further research is necessary.

In vitro degradation and cytocompatibility evaluation of novel soy and sodium caseinate-based membrane biomaterials
Silva, G. A., C. M. Vaz, et al. (2003), J Mater Sci Mater Med 14(12): 1055-66.
Abstract: Soy- and casein-based membranes are newly proposed materials disclosing a combination of properties that might allow for their use in a range of biomedical applications. Two of the most promising applications are drug delivery carrier systems and wound dressing membranes. As for all newly proposed biomaterials, a cytotoxic scanning must be performed as a preliminary step in the process of the determination of the compatibility with biological systems (biocompatibility). In this study, the cytotoxicity of both soy- and casein-based protein biomaterials has been evaluated and correlated with the materials degradation behavior. It was possible to show, through morphological and biochemical tests that these natural origin materials do not exert any cytotoxic effect over cells, and in some cases can in fact enhance cell proliferation. The different treatments to which the membranes were subjected during their processing (that include crosslinking with glyoxal and tannic acid, and physical modification by thermal treatment) seemed to have a clear effect both on the materials mechanical properties and on their in vitro biological behavior.

In vitro echogenicity characterization of poly[lactide-coglycolide] (plga) microparticles and preliminary in vivo ultrasound enhancement study for ultrasound contrast agent application
Lavisse, S., A. Paci, et al. (2005), Invest Radiol 40(8): 536-44.
Abstract: OBJECTIVES: This work includes (1) the characterization of a reproducible poly[lactide-coglycolide] (PLGA) microparticle preparation with an optimial mean diameter and size distribution and (2) the preliminary in vivo ultrasonographic investigation of PLGA microparticles. METHODS: A first series of PLGA microparticle preparations (1 to 15 mum) was acoustically characterized on a hydrodynamic device to select the most appropriate for ultrasound contrast agent application. Preparations of 3-microm microparticles were selected, characterized at different doses, and then injected into 20 melanoma grafted mice for contrast-enhanced power Doppler ultrasonography evaluation. RESULTS: The 3-microm microparticles (3.26-microm mean diameter with 0.41-microm standard deviation) led to in vitro enhancement of 18.3 dB at 0.62 mg/mL. In vivo experiments showed 47% enhancement of intratumoral vascularization detection after PLGA injection, significantly correlated (P < 0.0001) with preinjection intravascularization and tumoral volume. No toxicity was histologically observed. CONCLUSION: The 3-microm PLGA microparticles provided significant enhancement in vitro and in vivo without any toxicity.

In vitro effects of alloplastic biomaterials on the functional features of human erythrocytes and platelets
Di Massimo, C., M. G. Tozzi-Ciancarelli, et al. (1993), Riv Eur Sci Med Farmacol 15(5-6): 199-203.
Abstract: In vitro experiments were performed to evaluate the biocompatibility of metallic plates used in reconstructive maxillofacial surgery, analyzing the effect of titanium, nickel-chromium-molybdenum and stainless steel plates on some functional features of human erythrocytes and platelets, being rheological properties and platelet responsiveness pivotal for the perfusion of the tissues surroding the metallic implants. Our data underline that the tested plates are not responsible for significant alterations of the erythrocyte rheological features and of platelet reactivity.

In vitro effects of calcium phosphate biomaterials on fibroblastic cell behavior
Gregoire, M., I. Orly, et al. (1987), Biol Cell 59(3): 255-60.
Abstract: The effect of synthetic granular hydroxyapatite (HAP) on cultured fibroblastic cells (L929, human bone and gingiva cells) was studied. Phagocytosis of HAP particles and resulting morphological cell changes were demonstrated by microscopic examinations. Cell counts and [3H]thymidine uptake indicated significant increases in cell proliferation and DNA synthesis. These results could account for some of the alterations of the fibroblast behavior induced by changes in intracellular levels of calcium ions released from the material.

In vitro effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 cells
Qi, L. F., Z. R. Xu, et al. (2005), World J Gastroenterol 11(33): 5136-41.
Abstract: AIM: To investigate the effects of chitosan nanoparticles on proliferation of human gastric carcinoma cell line MGC803 in vitro and the possible mechanisms involved. METHODS: Chitosan nanoparticles were characterized by particle size, zeta potential, and morphology. After treatment with various concentrations of chitosan nanoparticles (25, 50, 75, 100 microg/mL) at various time intervals, cell proliferation, ultrastructural changes, DNA fragmentation, mitochondrial membrane potential (MMP), cell cycle phase distribution and apoptotic peaks of MGC803 cells were analyzed by MTT assay, electron microscopy, DNA agarose gel electrophoresis, and flow cytometry. RESULTS: Chitosan nanoparticles exhibited a small particle size as 65 nm and a high surface charge as 52 mV. Chitosan nanoparticles markedly inhibited cell proliferation of MGC803 cells with an IC50 value of 5.3 microg/mL 48 h after treatment. After treatment with chitosan nanoparticles, the typical necrotic cell morphology was observed by electron microscopy, a typical DNA degradation associated with necrosis was determined by DNA agarose electrophoresis. Flow cytometry showed the loss of MMP and occurrence of apoptosis in chitosan nanoparticles-treated cells. CONCLUSION: Chitosan nanoparticles effectively inhibit the proliferation of human gastric carcinoma cell line MGC803 in vitro through multiple mechanisms, and may be a beneficial agent against human carcinoma.

In vitro efficacy of glutaraldehyde-crosslinked chitosan microspheres against the fish-pathogenic ciliate Philasterides dicentrarchi
Parama, A., A. Luzardo, et al. (2005), Dis Aquat Organ 64(2): 151-8.
Abstract: Philasterides dicentrarchi is a protozoan ciliate which causes significant economic losses in fish aquaculture. This study investigated the effects of chitosan microspheres cross linked with glutaraldehyde and containing beta-cyclodextrin (betaCD) on the survival of this parasite in 7 d cultures. When used alone in assays, neither chitosan nor betaCD showed any activity, whereas free glutaraldehyde was strongly toxic to the parasite. Microspheres were likewise strongly toxic, at total glutaraldehyde concentrations much lower than with free glutaraldehyde: near-100% ciliate death was obtained (1) with 50 microg ml(-1) of microspheres prepared with 5% glutaraldehyde and no betaCD, or (2) with 10 microg ml(-1) of microspheres prepared with 0.15% glutaraldehyde and 0.1% betaCD. This suggests that the main active component is glutaraldehyde, but that the presence of small amounts of betaCD enhances efficacy. This high efficacy, together with the low toxicity to fish and rapid biodegradability of the individual components, suggest that these microspheres may be an attractive alternative to the formaldehyde baths traditionally used for the control of this parasite.

In vitro enzymatic biodegradation of amino acid based poly(ester amide)s biomaterials
Tsitlanadze, G., T. Kviria, et al. (2004), J Mater Sci Mater Med 15(2): 185-90.
Abstract: A systematic in vitro biodegradation study of regular poly(ester amide)s (PEAs) composed of naturally occurring hydrophobic alpha-amino acids, fatty diols and dicarboxylic acids was carried out in the presence of hydrolases like trypsin, alpha-chymotrypsin, and lipase. An automatic potentiometric titration method was used to examine the biodegradation property of the PEAs. Spontaneous immobilization (absorption) of the enzymes onto the PEAs films surfaces was observed. The surface immobilized enzyme not only accelerated the erosion of the PEAs but also was able to catalyze the hydrolysis of both low-molecular-weight (ATEE) and high-molecular-weight (protein) external substrates. It was found that the enzyme surface absorption process is reversible by nature. A kinetic method for a quantitative determination of the enzyme desorbed from the film surface was developed. The enzymes could also be impregnated into the PEAs to make them "self-destructive" at a target rate. A comparison of the PEAs' in vitro biodegradation data with polylactide (PDLLA) showed that PEAs exhibited a far more tendency toward enzyme catalyzed biodegradation than PDLLA.

In vitro evaluation of biocompatibility of beta-tricalcium phosphate-reinforced high-density polyethylene; an orthopedic composite
Homaeigohar, S., M. A. Shokrgozar, et al. (2005), J Biomed Mater Res A 75(1): 14-22.
Abstract: Beta-tricalcium phosphate-reinforced high-density polyethylene (beta-TCP/HDPE) is a new biomaterial which was made as a copy of bone composition with the aim of replacement of bony tissues. The composite samples were prepared using medical grade TCP powder and granular polyethylene. The raw materials were first compounded and the resulting composite preforms were compression molded into desired shape. The biocompatibility of composite samples with different volume fractions of TCP (20, 30, and 40 vol %) was assessed by proliferation, alkaline phosphatase (ALP), and cell adhesion assays using G-292 osteoblast cells. Cell-material interaction on the surface of the composites was observed by scanning electron microscopy (SEM). The effect of beta-TCP/HDPE on the behavior of G-292 cells was compared with those of a composite and a negative control samples. Results showed the composite samples had a higher proliferation rate of G-292 cells in the presence of composite samples as compared to the composite control sample after 3, 7, and 14 days of incubation period. ALP production after incubation in the presence of composite samples was seen to peak on the day 7. The number of adhered cells on the composite samples was higher than the numbers adhered on composite and negative control samples after the above incubation periods. Morphology investigation of adhered cells by SEM indicated a normal morphology and also many of the cells were in the process of cell division. The above results indicate that beta-TCP/HDPE samples are biocompatible, nontoxic, and in some cases show an increase in the proliferation rate of the cells, ALP production, and cell adhesion as compared to the control counterparts.


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