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PubMed2006

Multiphase microfluidics: from flow characteristics to chemical and materials synthesis.

We review transport characteristics of pressure-driven, multiphase flows through microchannel networks tens of nanometres to several hundred of micrometres wide with emphasis on conditions resulting in enhanced mixing and reduced axial dispersion. Dimensionless scaling parameters useful in characterizing multiphase flows are summarized along with experimental flow visualization techniques. Static and dynamic stability considerations are also included along with methods for stabilizing multiphase flows through surface modifications. Observed gas-liquid and immiscible liquid-liquid flows are summarized in terms of flow regime diagrams and the different flows are related to applications in chemistry and materials synthesis. Means to completely separate multiphase flows on the microscale and guidelines for design of scalable multiphase systems are also discussed.

Journal Article↗
PubMed2004

Shear-stress preconditioning and tissue-engineering-based paradigms for generating arterial substitutes.

In situ tissue engineering using shear-stress preconditioning and adhesive biomolecules is a new approach to autologous tissue engineering. In the present study, novel tissue-engineering grafts (TEGs) were preconditioned within an in vitro pulsatile flow circuit, with and without the addition of fibronectin (FN), to establish whether low-shear-stress conditions promoted endothelial cell (EC) retention and differentiation. TEGs ( n =24) were generated by the contraction and compaction of collagen(I) by porcine aortic smooth-muscle cells (SMCs) on to a compliant polyester graft scaffold. ECs were radiolabelled with [(111)In]indium tropolonate and seeded on to the luminal surface of the TEGs. Following organ culture in a bioreactor (7 days), TEGs were split into four groups ( n =six TEGs per group): Group A acted as controls with TEGs unmodified and seeded with radiolabelled ECs; Group B underwent luminal pre-coating with FN (75 microg/ml) prior to EC seeding; Group C underwent preconditioning within a pulsatile flow circuit at 10-20 microN (1-2 dyn)/cm(2) for 7 days prior to EC seeding, and Group D TEGs were preconditioned for 7 days at 1-2 dyn/cm(2), followed by luminal pre-coating with FN prior to EC seeding. The resistance to physiological shear stress of the seeded ECs was assessed using a gamma-radiation counter within a physiological flow circuit producing an arterial waveform with a mean shear stress of 93.2 microN (9.32 dyn)/cm(2). Environmental scanning electron microscopy (ESEM) was used to determine the distribution and degree of differentiation of the attached Ecs, and tissue-type-plasminogen-activator (tPA) assays provided a measure of function and viability. EC resistance to shear stress at 93.2 microN/cm(2) was significantly enhanced by a period of preconditioning (Group C) at 10-20 microN/cm(2), surface modification with FN (Group B), or both (Group D) when compared with control grafts (Group A). However, TEGs coated with FN whether preconditioned (Group D) or not (Group B) demonstrated the best results for EC retention. ESEM demonstrated near-confluent differentiated flattened ECs in both these cases. EC function was demonstrated by a steady increase in tPA production. Low-shear-stress preconditioning of TEGs enhances EC retention in vitro with an additional advantage demonstrated by pre-treatment with FN prior to endothelialization. These findings may be exploited in the development of tissue-engineered constructs to maintain a confluent endothelial lining.

Adaptation, Physiological↗
PubMed2000

A model to quantify encrustation on ureteric stents, urethral catheters and polymers intended for urological use.

OBJECTIVE: To validate an encrustation model and to quantify encrustation on currently used urological devices and polymers intended for urological use. Materials and methods An encrustation model was validated: (i) to measure the amount of calcium leaching from the glass model and from the polymer used; (ii) to determine whether the use of a single-source or pooled urine produced similar results; (iii) to determine in vitro encrustation; and (iv) to compare the results of in vivo implantation of the same materials into the bladders of rodents with the in vitro results. A test polymer (a ureteric stent, a urethral catheter or a biomaterial) and a control silicone polymer were housed separately but received human urine from the same reservoir and under the same conditions (pH 6.0 and 37 degrees C) for 5 days. The amount of calcium encrustation on each polymer was measured using atomic absorption spectroscopy. Each experiment was repeated at least four times and the results expressed as an encrustation index, defined as the ratio of encrustation of the test and reference polymers. RESULTS: The amount of calcium leaching from the glass model and polymers tested was insignificant. The use of a single-source or pooled urine gave the same results in the encrustation model. The in vitro results correlated with in vivo implantation of disks into the bladders of rats. Among the commonly used ureteric stents tested, the Cook C-Flex ureteric stents encrusted least. Hydrogel-coated ureteric stents encrusted more than uncoated stents. The Bard polytetrafluoroethylene short-term urethral catheter encrusted more than the Bard hydrogel-coated long-term catheter. A plasma-activated surface modification of a synthetic biomaterial with hyaluronic acid encrusted less than silicone, a long-term biomaterial widely regarded as the 'gold standard'. CONCLUSION: This validated encrustation model is the first to quantify encrustation on currently available ureteric stents and urethral catheters. A novel coating for a biomaterial was identified using the encrustation model, and which encrusted less than silicone.

Animals↗
PubMed2003

[In vitro culture of cells from respiratory mucosa on foils of collagen, poly-L-lactide (PLLA) and poly-3-hydroxy-butyrate (PHB)].

BACKGROUND: The replacement of respiratory mucosa after surgical resections at different locations in the ENT-field (trachea, nasal septum, sinus maxillaris) seems to be essential or at least of great benefit for patients. Presently a satisfying solution for this challenge does not exist. Therefore we have analysed the growth of cells from respiratory mucosa on different matrices. METHODS: Cell cultures are initiated mainly by growing out cells from tissue pieces but also by seeding of cells after enzymatic dissociation (sequential trypsinization) of mucosa. Cells were cultured with serum-containing and serum-free media on 3 different foils: made of collagen, of Poly-L-lactic acid (PLLA) and Poly-hydroxybutyric acid (PHB). Culturing time was on average about 4 - 5 weeks. RESULTS: On all tested materials fibroblasts and epithelial cells have grown in principle. PLLA was the material with best properties concerning the aim of this study whereas culturing of cells on PHB only happens after surface modification by amino-functionalisation with plasma treatment. Visualisation of cells on materials of collagen could not realised by light-microscopy due to the 3-dimensional structure and the optical properties of this material. Here the cultures were analysed by cell membrane staining and by REM. We could not find a differentiation of epithelial cells with beating cilia 6 weeks after starting the culture. However 20 days after starting the culture on PLLA (good conditions for observation with light-microscope) cells with beating cilia could be observed, in our opinion resulting from dissociated cells from tissue pieces and not from proliferated cells. CONCLUSIONS: In principle the analysed materials are useful for the culturing of cells from respiratory mucosa. However, for obtaining differentiated epithelial cells the culture conditions have to be modified.

Cell Adhesion↗
PubMed2004

[Synovial sarcoma of the hypopharynx - two case reports].

BACKGROUND: Synovial sarcoma is a rare soft tissue malignancy that most often occurs in the extremities of young adults. Approximately 3 % of these mesenchymal malignant tumors occur in the head and neck region. Up to now about 100 cases have been published. PATIENTS: We report on two cases of a synovial sarcoma of the hypopharynx. In the first case the patient died 3.5 years after beginning of therapy including all therapeutical efforts (surgery, chemotherapy, radiotherapy). The second patient with a synovial sarcoma of his left hypopharynx and a distant metastasis in his 8th thoracic vertebral body is actually treated by neoadjuvant chemotherapy and included into a clinical study to optimize therapy of advanced soft tissue sarcomas. Further surgical treatment depends on the success of this therapeutic approach. CONCLUSION: Despite multitherapeutic approaches over all 5-year survival rates for advanced synovial sarcomas of the head and neck have moderately improved in the recent decades and an ideal treatment has not yet surfaced. Modification of accepted treatment modalities is discussed and therapy options in controlled clinical studies are described.

Adult↗
PubMed2004

Apoptosis and survival of osteoblast-like cells are regulated by surface attachment.

We tested the hypothesis that RGDS peptides regulate osteoblast survival in culture. Osteoblast-like MC3T3-E1 cells were allowed to attach to RGDS peptides that had been tethered to a silicone surface utilizing a previously described grafting technique. The RGDS-modified surface caused up-regulation of alpha(v)beta(3) integrin. We noted that there was an increase in expression of activated focal adhesion kinase and activated Akt. There was no change in the expression level of the anti-apoptotic protein Bcl-2, the pro-apoptotic protein Bad, or the inactivated form of Bad, pBad. Attachment to the RGDS-treated membrane completely abolished apoptosis induced by staurosporine, the Ca(2+).P(i) ion pair, and sodium nitroprusside. However, the surface modification did not interfere with apoptosis mediated by the free RGDS peptide or serum-free medium. When the activity of the phosphatidylinositol 3-kinase pathway was inhibited, RGDS-dependent resistance to apoptosis was eliminated. These results indicated that the binding of cells to RGDS abrogated apoptosis via the mitochondrial pathway and that the suppression of apoptosis was dependent on the activity of phosphatidylinositol 3-kinase.

3T3 Cells↗
PubMedDate not supplied

Ultrastructural abnormalities in cultured exostosis chondrocytes.

Hereditary multiple exostoses (HME) is an autosomal dominant disorder characterized by inappropriate chondrocyte proliferation and bone growth arising at the juxtaepiphyseal region of the long bones. HME is caused by mutations in the EXT 1 and EXT 2 genes, which have glycosyltransferase activity. These genes are responsible for synthesis of heparan sulfate (HS) chains, which are important signaling molecules in chondrocyte differentiation. HME chondrocytes in monolayer culture have been shown by transmission electron and deconvolution microscopy to contain enormous bundles of actin, cross-linked with muscle specific alpha-actinin. Here additional ultrastructural anomalies in HME chondrocytes are reported, including lobulated nuclei, shortened channels of rER, large numbers of cell processes and podosomes, nontypical junctions, elongated, bulbous-ended mitochondria, and reduced extracellular matrix. Microfilaments are present throughout the cytoplasm, compartmentalizing it, and isolating organelles. The excess microfilaments, attributed to increased cell adhesiveness, are likely to interfere with secretion and cytokinesis, and sterically hinder intracellular organelle differentiation. The observed surface modifications and cytoskeletal abnormalities are proposed to play a role in development of the mutant phenotype, via changes in cell adhesiveness and/or binding of signals to receptors, which results in loss of the unidirectionality of growth in the epiphyseal plate.

Cell Nucleus↗
PubMed1977

Distribution of anionic sites on the oviduct ciliary membrane.

Polycationic ferritin (PCF) was used as a visual probe for anionic sites on the oviduct ciliary membrane. The binding of PCF to ciliary membranes was dependent on the concentration of the probe in the incubation media. At low concentrations (0.08-0.16 mg/ml), PCF was bound exclusively to the tip of the cilium whereas at higher concentrations (0.32-0.64 mg/ml), ferritin was located at the tip and at the base around the transition region, with occasional scattered clumps on the remainder of the membrane. The base and tip binding was fount to be associated with special surface modifications of the membrane in these regions. At the tip, PCF was bound to a filamentous glycocalyx termed the ciliary crown. Base binding was associated with a system of five to six 140-A high ridges, each of which encircled the membrane of the transition region. The ridges were equally spaced (approxamately 245 A spacing) along the length of the transition region. Since pretreatment of oviduct with either neuraminidase or protease blocked the binding of the probe, the PCF-binding sites appear to be negatively charged glycoproteins or mucopolysaccharides.

Animals↗
PubMed2002

Porous poly(propylene fumarate) foam coating of orthotopic cortical bone grafts for improved osteoconduction.

A porous biodegradable scaffold coating for perforated and demineralized cortical bone allografts could maintain immediate structural recovery and subsequently allow normal healing and remodeling by promoting bony ingrowth and avoiding accelerated graft resorption. This new type of osteoconductive surface modification should improve allograft incorporation by promoting new bone growth throughout the biodegradable scaffold, hence encasing the graft with the recipient's own bone. We investigated the feasibility of augmenting orthotopically transplanted cortical bone grafts with osteoconductive biodegradable polymeric scaffold coatings. Five types of bone grafts were prepared: type I, untreated fresh-frozen cortical bone grafts (negative control); type II, perforated and partially demineralized cortical bone grafts without additional coating (positive control); type III, perforated and partially demineralized cortical bone coated with a low-porosity poly(propylene fumarate) (PPF) foam; type IV, perforated and partially demineralized cortical bone coated with a medium-porosity PPF foam; and type V, perforated and partially demineralized cortical bone coated with a high-porosity PPF foam. Grafts were implanted into the rat tibial diaphysis. Fixation was achieved with an intramedullary threaded K-wire. Two sets of animals were operated on. Animals were killed in groups of eight with one set being killed 12 weeks, and the other 16 weeks, postoperatively. Radiographic, histologic, and histomorphometric analyses of grafts showed that the amount of new bone forming around the foam-coated grafts was significantly higher than that in the type I control group (uncoated) or that in type II group (perforated and partially demineralized cortical bone grafts). Although all foam formulations appeared initially equally osteoconductive, histologic evaluation of medium-porosity PPF foam-based coatings appeared to result in a sustained response 16 weeks postoperatively. Significant resorption was present in perforated and partially demineralized cortical bone graft allografts, with some accompanying new bone formation occurring primarily within the laser holes. Therefore, PPF foam-coated cortical bone grafts appeared to be better protected from excessive bone resorption, as frequently seen with invasion of fibrovascular tissue. Biomechanical analysis of the PPF foam-coated grafts corroborated findings of the morphometric analysis in that the failure strength at the allograft-host bone junction sites of all PPF-coated cortical bone grafts was higher than in the uncoated controls.

Animals↗
PubMed2006

Co-culture of bone marrow fibroblasts and endothelial cells on modified polycaprolactone substrates for enhanced potentials in bone tissue engineering.

The creation of a vascularized bed makes the survival of seeded cells on 3-dimensional scaffolds much more likely. However, relying purely on random capillary ingrowth into the porous scaffolds from the host may compromise vascularization of a scaffold. One solution is to transplant cells capable of differentiating into new blood vessels into the scaffolds to accelerate the creation of a vascularized scaffold. Because endothelial cells are the key cells involved in blood vessel formation, the present study was designed to investigate the development of a biomaterial surface that supports endothelial cell attachment and proliferation. The subsequent effects of the material surface modifications on the differentiation and proliferation of human bone marrow-derived fibroblasts (HBMFs) when grown in co-culture with a human bone marrow endothelial cell line (HBMEC-60) were studied. Endothelialization studies showed that the gelatin-coated and hydroxyapatite-coated substrates were superior for HBMEC-60 attachment and proliferation to hydrolyzed-only or untreated polycaprolactone substrates. Co-culture studies showed that the presence of the HBMEC-60 specifically enhanced HBMF cell proliferation and differentiation and that this effect was not observed with co-culture with skin fibroblasts. It is concluded that the co-culture of endothelial cells with HBMFs could be a promising culture system for bone tissue- engineering applications.

Bone Marrow Cells↗
PubMed1976

Ultrastructural studies of surface features of human normal and tumor cells in tissue culture by scanning and transmission electron microscopy.

Human tumors of a variety of histopathologic types have been established in tissue culture. The surface features of these cell lines were investigated by scanning electron microscopy (SEM) with the use of new techniques for specimen preparation. Tumor cells demonstrated striking degrees of surface activity with numerous microvilli, filopodia, blebs, and ruffles. Intercellular contacts were also prominent in cultures of most solid tumors observed by SEM. At low cell density, normal human fibroblasts exhibited some surface features such as microvilli and blebs, but at higher cell density they lacked extensive surface modifications. By transmission electron microscopy (TEM), the cytoskeleton of normal fibroblasts was shown to be well organized, with parallel orientation of microfilaments, filaments, and microtubules. These structures were also in tumor cells, but they lacked the degree of organization of fibroblasts. Desmosomes were readily demonstrated in normal fibroblasts and carcinoma cells in culture but not in sarcomas, melanomas, or tumors of neural origin. These studies have provided the first correlative SEM and TEM analyses of solid human tumor cells of diverse pathologic types in vitro.

Carcinoma↗
PubMed2000

Effect of low-friction ion-treated femoral heads on polyethylene wear rates.

Polyethylene wear is a major contributor to osteolysis and subsequent aseptic loosening of prosthetic components in total hip arthroplasty. Use of ion implantation as a surface modification to the metallic bearing component of orthopaedic implants may be an effective means of reducing wear debris at the bearing interface. In July 1991, low friction ion treated femoral heads were introduced. This study evaluates the effect of the low friction ion treated femoral head on polyethylene wear. Fifty-five total hip arthroplasties (53 patients) with low friction ion treated femoral heads followed up a minimum of 3 years were matched with 55 total hip arthroplasties (47 patients) without low friction ion treated femoral heads for the same postoperative period. Socket wear was evaluated radiographically. Case matching and strict inclusion criteria were used to control for known factors influencing polyethylene wear. These criteria included: (1) cases matched for gender and age within 2 years; (2) diagnosis limited to osteoarthritis or avascular necrosis of the femoral head only; (3) femoral head diameter limited to 26 or 28 mm only; (4) hydroxyapatite coated femoral stem of the same design and a metal backed socket of the one of two designs with the same polyethylene insert; and (5) minimum followup of 3 years. The linear wear rate of polyethylene was 0.161 +/- 0.095 mm per year in the group without the low friction ion treated heads and 0.116 +/- 0.101 mm per year in the low friction ion treated group. The volumetric wear rates were 74.5 +/- 44.3 mm3 per year for the group without the low friction ion treated heads and 57.8 +/- 51.1 mm3 per year for the low friction ion treated group. Assuming the sensitivity of these measurements can detect these small differences in wear accurately, these results suggest low friction ion treated prosthetic heads are useful in reducing polyethylene wear at 3-year minimum followup.

Adult↗
PubMed1997

Biodistribution and computed tomography blood-pool imaging properties of polyethylene glycol-coated iopromide-carrying liposomes.

RATIONALE AND OBJECTIVES: Surface-modified contrast-carrying liposomes potentially are useful as computed tomography (CT) blood-pool agents. The biodistribution and CT-imaging behavior of conventional as well as polyethylene glycol (PEG)-coated iopromide-carrying liposomes were tested. Two different types of PEG-ylated lipids were used to demonstrate possible differences. METHODS: Iopromide-containing liposomes were prepared by a continuous high-pressure extrusion method and subsequently PEG-ylated by simple mixing with either DSPE-PEG2000 or CHHS-PEG2000. The resulting liposomes were investigated in rats (biodistribution) and rabbits (imaging). RESULTS: Surface modification with CHHS-PEG consistently resulted in less effective stabilization of liposomes in the blood than with DSPE-PEG. In the biodistribution study, no significant differences in blood concentration could be found 1 hour after injection between the different formulations at a dose of 250 mg total iodine/kg body weight (approximately 500 mg lipid/kg). At this dose, the unmodified as well as the DSPE-PEG liposomes displayed prolonged blood circulation with CT density differences above 70 Hounsfield units (aorta) for up to 20 minutes (n = 1). CONCLUSIONS: DSPE-PEG-coated and unmodified liposomes proved to be useful for CT blood-pool imaging displaying favorable imaging properties. Future studies will have to demonstrate whether PEG-ylation offers diagnostic or toxicologic advantages over conventional vesicles in this indication.

Animals↗
PubMed1991

Intraocular lenses.

In reviewing the 1990 literature on intraocular lenses, it is evident that the increased popularity of phacoemulsification in North America has stimulated interest in lens designs that can be inserted through small incisions, and various methods for folding soft lenses. Critical evaluation of various multifocal lenses has now resulted in greater understanding of their advantages and drawbacks, and it is becoming apparent that there are guidelines of suitable candidates for such lenses. Use of multifocals has stimulated interest in contrast sensitivity testing, and new more accurate lens power formulas. Heparin surface modification of polymethylmethacrylate lenses now seems to be accepted as having value in reducing pigment deposits after surgery. Clinical trials of lenses for patients suffering from age-related macular degeneration hold some promise for this large group of unhappy patients. However, the use of anterior chamber lenses to correct high myopia in phakic eyes is now on hold because of reports of progressive endothelial cell loss in some eyes. More reports are available on the technique and results of suturing posterior chamber lenses to the sclera where there is absent or insufficient capsule to support a conventional sulcus fixated posterior chamber lens. Intraocular lenses remain alive and well in 1990 with increased usage worldwide.

Astigmatism↗
PubMed1999

Transfusion practice for patients with sickle cell disease.

The practice of transfusion for patients with sickle cell disease is changing. Diminished childhood mortality is resulting in greater patient longevity and a higher prevalence of chronic systemic complications. Many of these chronic complications, as well as several acute ones, are treatable with erythrocyte transfusion. Proactive or preventive uses for transfusion are being considered for selected patients. Immunologic phenomena, hemosiderosis, and risk for transmission of infectious agents are the three most noticeable risks associated with repetitive transfusion. Evolving work with alternative oxygen carriers and surface modification of erythrocytes is also promising and relevant for patients with sickle cell disease.

Adult↗
PubMed2005

The microenvironment around total hip replacement prostheses.

The metal stem of the totally replaced hip carries load and resists fatigue, but it is electrochemically corroded. Metallic atoms act as haptens, induce type 1 T-helper cells/Th1-type immune responses and enhance periprosthetic osteolysis. Stiff metal implants, which do not have the same elasticity as the surrounding bone, cause stress shielding. Cyclic loading and lack of ligamentous support lead to mechanical and ischemia reperfusion injury and particle formation from bone, polymethylmethacrylate, and porous implant surfaces, which accelerate third-body polyethylene wear. Surgical injury and micromotion induce the formation of a fibrous capsule interface. Type-B lining cells produce lubricin and surface-active phospholipids to promote solid-to-solid lubrication but may loosen the implant from bone. The pumping action of the cyclically loaded joint and synovial fluid pressure waves dissect the implant-host interface and transports polyethylene particles and pro-inflammatory mediators to the interface. Hyaluronan induces formation of a synovial lining like layer. Because of its localization close to bone, foreign body inflammation at the interface stimulates osteoclastogenesis and peri-implant bone loss. Metal-on-metal and ceramic-on-ceramic pairs might minimize third body wear, but can lead to high-impact load of the acetabulum. Diamond coating of a metal-on-polyethylene couple might solve both of these problems. The basic biomaterial solutions allow good mechanical performance and relatively long life in-service, but surface modifications (porous coating, hydroxyapatite, diamond, bioglass, and others) may facilitate performance of the implant and improve the biomaterial and body interfaces.

Arthroplasty, Replacement, Hip↗
PubMed2005

An investigational study on the healing process of anterior spinal arthrodesis using a bioactive ceramic spacer and the change in load-sharing of spinal instrumentation.

STUDY DESIGN: Ceramic anterior lumbar interbody arthrodesis was performed using an in vivo sheep model. Observations of fusion status and the load-sharing of spinal instrumentation were studied at sequential intervals for 1 year after surgery. OBJECTIVES: To elucidate the healing process of spinal arthrodesis performed with a bioactive ceramic spacer and the change in load-sharing of anterior spinal instrumentation. SUMMARY OF BACKGROUND DATA: With the improved development of spinal instrumentation, anterior spinal arthrodesis has become a standard spinal reconstruction technique; however, the mechanistic basis underlying the healing process is not well documented. Moreover, it remains unclear how load-distribution through the fusion mass and spinal instrumentation change throughout the healing process. METHODS: Using 24 sheep, a two-level anterior lumbar interbody fusion (L2-L3, L4-L5) was performed using a smooth surface and a porous surface-modified bioactive ceramic, with each segment instrumented using a one-rod anterior spinal instrumentation system. Four animals each were killed at 2, 4, 8, 12, 24, and 52 weeks after surgery. Postmortem analysis included quantification of anterior rod strain under multidirectional flexibility testing and radiographic and histologic analyses of the arthrodesed segments. RESULTS: From 0 to 8 weeks after surgery, the bending strain of the rod gradually decreased despite no obvious bone formation. From 8 to 24 weeks after surgery, the rod strain markedly decreased with the development of bridging trabeculated bone formation between vertebral bodies. After 24 weeks after surgery, minimal changes were observed in rod strain; however, the fusion mass volumetrically increased with corresponding facet joint atrophy. The porous surface-modification of ceramic did not influence the histologic healing process, despite the improvement of interface osseous union rate. CONCLUSIONS: In anterior spinal arthrodesis, spinal instrumentation is mainly exposed to bending stress, with decreased load-sharing with corresponding development of the spinal fusion. Continuous bone remodeling of the anterior fusion mass results in concurrent decreases in spinal instrumentation and posterior spinal element load-transmission. The principal healing mechanism of ceramic anterior interbody spinal fusion is not an osseous union between the ceramic and vertebral body, but bridging bone formation around the ceramic, which directly connects the vertebral bodies above and below the disc.

Animals↗