Biomedical subjects

Benjamin M Wu

Publications and source records attributed to Benjamin M Wu.

At least 19 recordsLinked to original sources

PubMed2007

Analysis of oxygen transport in a diffusion-limited model of engineered heart tissue.

Cardiac tissue engineering has made notable progress in recent years with the advent of an experimental model based on neonatal cardiomyocytes entrapped in collage gels and purified basement membrane extract, known as "engineered heart tissues" (EHTs). EHTs are a formidable display of tissue-level contractile function and cellular-level differentiation, although they suffer greatly from mass transport limitations due to the high density of metabolically active cells and the diffusion-limited nature of the hydrogel. In this report, a mathematical model was developed to predict oxygen levels inside a one-dimensional, diffusion-limited model of EHT. These predictions were then compared to values measured in corresponding experiments with a hypoxia-sensitive stain (pimonidazole). EHTs were cast between two plastic discs, which allowed for mass transfer with the culture medium to occur in only the radial direction. EHTs were cultured for up to 36 h in the presence of pimonidazole, after which time they were snap-frozen, histologically sectioned, and stained for bound pimonidazole. Quantitative image analysis was performed to measure the distance from the culture medium at which hypoxia first occurs under various conditions. As tested by variation of simple design parameters, the trends in oxygen profiles predicted by the model are in reasonable agreement with those obtained experimentally, although a number of ambiguities related to the specific model parameters led to a general overprediction of oxygen concentrations. Based on the sensitivity analysis in the present study, it is concluded that diffusion-reaction models may offer relatively precise predictions of oxygen concentrations in diffusion-limited tissue constructs.

Biological Transport↗

PubMed2006

Modulation of protein delivery from modular polymer scaffolds.

Growth factors are increasingly employed to promote tissue regeneration with various biomaterial scaffolds. In vitro release kinetics of protein growth factors from tissue engineering scaffolds are often investigated in aqueous environment, which is significantly different from in vivo environment. This study investigates the release of model proteins with net-positive (histone) and net-negative charge (bovine serum albumin, BSA) from various scaffolding surfaces and from encapsulated microspheres in the presence of ions, proteins, and cells. The release kinetics of proteins in media with varying concentrations of ions (NaCl) suggests stronger electrostatic interaction between the positively charged histone with the negatively charged substrates. While both proteins released slowly from hydrophobic PCL surfaces, plasma etching resulted in rapid release of BSA, but not histone. Interestingly, although negatively charged BSA released readily from negatively charged collagen (col), BSA released slowly from col-coated PCL scaffolds. Such electrostatic interaction effects were abolished in the presence of serum proteins and cells as evidenced by the rapid release of proteins from col-coated scaffolds. To achieve sustained release in the complex environment of serum proteins and cells, the model proteins were encapsulated into poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres, which were embedded within col-coated PCL scaffolds. Protein release from microspheres was modulated by changing the lactide-to-glycolide ratio of PLGA polymer. BSA adsorbed to col released faster than histone encapsulated in microspheres in the presence of serum and cells. Collectively, the data suggest that growth factor release is highly influenced by scaffold surface and the presence of ions, proteins, and cells in the media. Strategies to deliver multiple growth factors and studies which investigate their release should consider these important variables.

Animals↗

PubMed2006

Cell interaction with three-dimensional sharp-tip nanotopography.

Cells in their native microenvironment interact with three-dimensional (3D) nanofeatures. Despite many reports on the effects of substrate nanotopography on cells, the independent effect of 3D parameters has not been investigated. Recent advances in nanofabrication for precise control of nanostructure pattern, periodicity, shape, and height enabled this systematic study of cell interactions with 3D nanotopographies. Two distinct nanopatterns (posts and grates) with varying three-dimensionalities (50-600 nm in nanostructure height) were created, while maintaining the pattern periodicity (230 nm in pitch) and tip shape (needle- or blade-like sharp tips). Human foreskin fibroblasts exhibited significantly smaller cell size and lower proliferation on needle-like nanoposts, and enhanced elongation with alignment on blade-like nanogrates. These phenomena became more pronounced as the nanotopographical three-dimensionality (structural height) increased. The nanopost and nanograte architectures provided the distinct contact guidance for both filopodia extension and the formation of adhesion molecules complex, which was believed to lead to the unique cell behaviors observed.

Biocompatible Materials↗

PubMed2006

The osteoinductive properties of Nell-1 in a rat spinal fusion model.

BACKGROUND CONTEXT: Recombinant growth factors bone morphogenetic protein-2 (BMP-2) and BMP-7 are currently approved for human use but are associated with various adverse effects including ectopic bone formation and local inflammatory reaction. The development of alternative growth factors may help minimize the adverse effects of current osteoinductive therapeutics. Nell-1 (Nel-like molecule-1; Nel [a protein strongly expressed in neural tissue encoding epidermal growth factor like domain]) is a novel secretory molecule that appears to act more specifically on osteoblasts than the BMPs, which can act on multiple cell types. From a molecular point of view, Nell-1 is directly regulated by runt-related transcription factor 2 (Runx2/Cbfa1), a master regulatory gene controlling bone formation. Previous studies showed that Nell-1 accelerates osteogenic differentiation in vitro and calvarial bone formation in vivo. We hypothesize that Nell-1 may also effectively form bone in spinal fusion. PURPOSE: Our primary aim was to assess if direct adenoviral gene delivery with Nell-1 in a demineralized bone matrix (DBM) carrier can improve spinal fusion in a rat model. Because adenoviral vectors allow for sustained growth factor delivery, they were used for initial feasibility testing before protein studies. STUDY DESIGN/SETTING: Two groups of 20 athymic rats underwent posterolateral intertransverse process spinal fusion at L4-L5 with implanted DBM carrier containing either adenovirus coding for Nell-1 (AdNell-1) or control, Lac Z (AdLacZ). No cells were implanted. The 20 rats were sacrificed at 6 weeks for evaluation of spinal fusion. METHODS: All animals underwent Faxitron radiographs at 2, 4, and 6 weeks, manual spine palpation at 6 weeks, and high-resolution micro computerized tomography (microCT) at 6 weeks. Spinal fusion rate was assessed by: 1) 6-week Faxitron images; 2) manual palpation by three independent observers; 3) microCT; and 4) histology. New bone formation was assessed by hematoxylin-eosin and Masson trichrome staining on decalcified, coronally sectioned spine segments. RESULTS: All differences achieved statistical significance. After 6 weeks, direct application of adenoviral Nell-1 in a DBM carrier achieved significantly higher rates of spinal fusion over Lac Z controls: 60% Nell-1 versus 20% Lac Z by manual palpation and 70% Nell-1 versus 20% Lac Z by microCT and histology. Histological assessment of bone quality and maturity revealed more mature, higher quality bone in all the Nell-1 treated specimens relative to Lac Z at 6 weeks. CONCLUSIONS: Spinal fusion is more accurately assessed by microCT and histology than manual palpation. Direct application of adenoviral Nell-1 in a DBM carrier achieved significantly higher rates of spinal fusion over Lac Z controls at 6 weeks. Direct application of adenoviral Nell-1 in a DBM carrier also achieved significantly higher rates of spinal fusion over other reports in the literature using direct adenoviral BMP application. Direct application of adenoviral BMP in an allograft carrier achieved 8% fusion for BMP-2 and 16% fusion for BMP-7 at 8 weeks. These results indicate that Nell-1 may be a potent osteoinductive molecule. In addition, the regulation of Nell-1 by the master bone regulatory gene, Runx2 suggests that Nell-1 may exert its effects more specifically in osteoblastic cells than BMPs which affect multiple cell types. Overall, Nell-1 may fulfil a current need for an osteoinductive factor.

Adenoviridae↗

PubMed2006

Contractile function of the mechanically lengthened intestine.

BACKGROUND: The application of longitudinal mechanical force induces lengthening of the small intestine. The purpose of this study is to evaluate the contractile function of the mechanically lengthened jejunum in a rodent model. MATERIALS AND METHODS: Three groups of rats including normal jejunum, isolated jejunal segment without mechanical lengthening, and isolated lengthened jejunal segment, were studied for contractile function. The isometric contractions of jejunal segments were investigated in organ baths for spontaneous activity and response to potassium chloride and cholinergic carbachol. RESULTS: The normal control group showed a basal spontaneous activity with an average frequency of 33 +/- 0.68 contractions per min (cpm). The basal spontaneous activity for the isolated group had an average frequency of 26 +/- 2.7 cpm and for the lengthened group had an average frequency of 24 +/- 5.7 cpm. Although the normal control group had a higher frequency of basal spontaneous activity as compared to either the isolated or lengthened groups, there was no statistically significant difference between the frequencies in the isolated and the lengthened groups. All three groups demonstrated a sustained increase in tension upon administration of either potassium chloride or carbachol. CONCLUSIONS: The response to pharmacological stimulation, as measured by total area under the tension curve and maximal change in basal tone, was larger in the normal group than in the isolated groups. The addition of longitudinal mechanical force to lengthen the isolated jejunal segment did not further alter this change.

Animals↗

PubMed2006

Tissue engineering for anterior cruciate ligament reconstruction: a review of current strategies.

The anterior cruciate ligament (ACL) is one the most commonly injured ligaments of the knee. Chronic ACL insufficiency can result in episodic instability, chondral and meniscal injury, and early osteoarthritis. The intra-articular environment of the ligament precludes normal healing and surgical replacement of the injured ligament is often mandated to restore stability. Current surgical strategies include the use of local autograft or allograft tissues for ligament reconstruction. These procedures have yielded superior long-term clinical results yet have the potential for serious associated morbidities. Existing limitations have prompted ongoing research designed to engineer a replacement ligament that will parallel the native ACL in both its biologic properties and mechanical durability. Ligament engineering necessitates the use of appropriate source cells and a growth matrix to support cell proliferation and collagen synthesis. The identification of appropriate growth modulators including both biochemical factors and mechanical stimuli are requisites for successful tissue growth. The characterization of the elements essential for successful graft development represents a significant challenge for investigators. This review examines the current literature regarding the potential and limitations of ligament engineering and describes the development of a novel 3-dimensional scaffold and bioreactor system at our institution.

Animals↗

PubMed2006

Distension enterogenesis: increasing the size and function of small intestine.

INTRODUCTION: The purpose of this study is to evaluate the feasibility of using saline infusion to lengthen small bowel while preserving intestinal enzymatic function. METHODS: Male Sprague-Dawley rats had a 3-cm jejunal segment taken out of continuity. A catheter was inserted in the proximal end, and the distal end was oversewn. Continuous infusion of saline into the isolated jejunal segment was started 2 weeks postoperatively. Segments were harvested 1 week later. Segment weights and lengths were measured preoperatively and at the time of harvest. Histology of harvested segments was performed. Alkaline phosphatase (ALP) and lactase assays were performed. Comparisons were made with normal jejunum from control animals. RESULTS: A 32% increase in length was achieved with saline distension of small intestine. The segment weight to length ratio was significantly increased by saline distension; however, the total protein-to-weight ratio was unchanged. Specific activities of ALP and lactase were not affected by saline distension. Because of the increased length and weight of the distended jejunal segments, total segment activities for both enzymes were significantly increased. CONCLUSIONS: Saline infusion appears to be a viable method for increasing small intestinal length without compromising enzymatic function. This phenomenon may provide a new method for the treatment of patients with short bowel syndrome in the future, and further study is warranted.

Animals↗

PubMed2006

Serum-free cultures of murine adrenal cortical cells.

PURPOSE: The feasibility of culturing murine adrenal cortical cells before transplantation was investigated in this study. METHOD: Primary murine adrenal cortical cells were maintained in either fetal bovine and horse sera-containing media or serum-free media. Real-time polymerase chain reaction was used to quantify the levels of adrenal cortical gene expression in the cultured cells. RESULTS: The use of sera-containing media led to the growth of many cells in the culture, but the expression of Sf-1, Dax-1, and Cyp11b1 in such cultures declined rapidly. In contrast, there was no significant cell growth in the serum-free culture medium. Culturing murine adrenal cortical cells in the serum-free medium resulted in higher levels of Sf-1, Dax-1, and Cyp11b1 gene expression. In the serum-free medium, adrenal cortical cells also responded to adrenocorticotropic hormone by increasing the expression of Cyp11b1 and suppressing the expression of Dax-1 in a dose-dependent manner. The addition of basic fibroblast growth factor to the serum-free medium maintained the expression of Sf-1, Dax-1, and Cyp11b1 for 4 weeks. CONCLUSION: Adrenal cortical cells isolated from adult mice were successfully maintained in a serum-free culture medium with basic fibroblast growth factor. This culture system may be suitable for further manipulation of adrenal cortical cells in vitro before transplantation.

Adrenal Cortex↗

PubMed2006

The behavior of human mesenchymal stem cells in 3D fibrin clots: dependence on fibrinogen concentration and clot structure.

Natural biopolymers such as human fibrin are appealing to tissue engineers, because fibrin is biocompatible, bioresorbable, and essential in normal wound healing. There have been numerous studies to date to develop a fibrin-based injectable cell delivery system, albeit with varying success. We propose that the outcome of fibrin cell delivery can, in part, be attributed to the relative concentrations of fibrinogen and thrombin solutions (i.e., formulations) and the structure of the final 3D fibrin clot. Formulation-dependent proliferation of human mesenchymal stem cells (hMSCs) within 3D fibrin clots was investigated in vitro. Our results indicate that hMSCs are viable in all fibrin sealant formulations investigated, and proliferation rates vary with fibrin formulations. Furthermore, the fibrinogen solution, not thrombin, was found to have a more dominant role on hMSC proliferation, with dilute fibrinogen solutions promoting greater hMSC proliferation. Confocal and electron microscopy reveal formulation dependence on 3D fibrin clot structure, with dilute fibrinogen solutions yielding more open, homogeneous microstructures. This study suggests that the concentrations of fibrinogen and thrombin solutions must be carefully considered for cell delivery because they affect 3D fibrin clot structure and cell proliferation.

Blood Coagulation↗

PubMed2006

Human mesenchymal stem cell proliferation and osteogenic differentiation in fibrin gels in vitro.

This study analyzed human mesenchymal stem cell (hMSC) behavior in a fibrin sealant. hMSC morphology, proliferation, and osteogenic differentiation were analyzed after up to 28 days of incubation in eight different formulations of fibrin gels (Tisseel) prepared with various concentrations of fibrinogen complex (FC) and thrombin. Cell morphology and distribution within the gels were observed by fluorescence microscopy after cell staining with calcein dye. Cell proliferation was assessed by measuring the fluorescence intensity of the cell suspension stained with calcein dye after dissolution of the gels. A standard alkaline phosphatase (ALP) assay, von Kossa staining, and real-time reverse transcriptase-polymerase chain reaction (RT-PCR) were used to analyze hMSC osteogenic differentiation. Cell behavior varied depending on the gel formulation. Proliferation was higher in the formulations containing a low FC concentration, but ALP activity was higher in the formulations containing a high FC concentration. Variations in thrombin concentration had a lesser effect. Small nodules of mineralization were observed at days 21 and 28 in a formulation containing a high FC concentration, in addition to a marked increase in bone sialoprotein (BSP) gene expression level as well as a lower increase in ALP and osteopontin (OPN) levels. However, there was no significant increase in osteocalcin (OCN) expression, a late marker of osteogenic differentiation, up to day 28. In conclusion, this study demonstrated that hMSC morphology, proliferation, and osteogenic differentiation in fibrin gels depended on the FC/thrombin ratio. hMSCs appeared to undergo osteogenic differentiation when seeded in Tisseel fibrin sealant containing a high FC concentration, but they did not fully differentiate into mature osteoblasts.

Biocompatible Materials↗

PubMed2006

Analysis of cell growth in three-dimensional scaffolds.

The in vitro growth of pre-osteoblasts in multi-layer, three-dimensional scaffolds was determined from experimental measurements and was compared to a mathematical model. Immediately following cell seeding, the initial cell density was uniform throughout the scaffold. After 10 days, the cell density increased from 2.1 x 10(5) cells/cm(3) to 1.3 x 10(7) cells/cm(3) at the fluid-scaffold interface. The increase in cell density was largely confined to the outermost 200 microm from the fluid-scaffold interface. The cell density profile was in good agreement with a mathematical model that simulated the cell growth based on the local oxygen tension. The improved understanding derived from this mathematical model may be useful in the design of three-dimensional scaffolds that can support more uniform growth of cells.

3T3 Cells↗

PubMed2005

Modulation of gene expression in neonatal rat cardiomyocytes by surface modification of polylactide-co-glycolide substrates.

Myocardial tissue engineering presents a potential treatment option for heart disease. Cardiomyocytes isolated at various stages of development retain the ability to form contractile networks in vitro, which suggests that it should be possible to reconstitute viable myocardium given the appropriate architecture, stimuli, and cardiomyogenic cell source. This study investigates the effects of modifying substrate surface energy (by plasma etching) and protein coating (by fibronectin adsorption) on neonatal rat ventricular myocyte (NRVM) function. Primary NRVMs were cultured for 96 h on modified and control films of a common degradable polymer, polylactide-co-glycolide. Cultures were analyzed for cell spreading, protein content, and mRNA expression of atrial natriuretic factor and beta-myosin heavy chain. The results demonstrate that NRVMs cultured on etched films significantly increased in spreading, myofibril development, protein content, and gene expression of atrial natriuretic factor and beta-myosin heavy chain compared with unetched films, and that this surface energy effect is overwhelmed by the addition of fibronectin. Conclusions from this study are that surface energy and protein adsorption influence the gene expression of adherent NRVMs, and may be important for modulating the function of engineered myocardium.

Animals↗

PubMed2005

Gelatin-embedded cell-polymer constructs for histological cryosectioning.

Many tissue-engineering strategies involve the delivery of cells via porous polymer scaffolds. Obtaining histological sections of the emerging tissue is often necessary to analyze numerous characteristics of the microscopic environment. However, difficulties arise upon applying standard histological techniques to cell-seeded polymer scaffolds. This report describes a simple and reliable method for cryosectioning cell-polymer constructs embedded in gelatin. Solvent-soluble (PLGA) and insoluble (PGA) scaffolds were cultured in vitro with preosteoblasts, followed by histological processing with paraffin, OCT, or gelatin. Although paraffin-embedded PGA scaffolds withstood standard sectioning and rinsing steps, paraffin-embedded PLGA scaffolds were partially dissolved during the clearing step. OCT-embedded scaffolds produced sections that did not adhere well to slides, and most of the sample was lost during rinsing steps. In contrast, gelatin-embedded scaffolds exhibited adequate structural integrity during cryosectioning, adhered well to the slides, retained the actual polymer morphology, and exhibited compatibility with common stains.

Animals↗

PubMed2005

In vitro response of MC3T3-E1 pre-osteoblasts within three-dimensional apatite-coated PLGA scaffolds.

Biomimetic apatites have been reported to promote osteogenic activities in numerous in vivo and in vitro models, but the precise mechanism by which the apatite microenvironment promotes such activities is not well understood. Such mechanistic studies require reproducible model systems that are relevant to tissue engineering practices. Although two-dimensional (2D) apatite-coated polystyrene culture dishes provide practicality and reproducibility, they do not simulate the effects of the three-dimensional (3D) microenvironment and degrading polymeric substrates. A simple 3D model system to address these relevant effects, and its utilization in the investigation of apatite-promoted osteoblastic differentiation in vitro is reported in this paper. Apatite coating was achieved by sequentially immersing poly(lactide-co-glycolide) (PLGA) scaffolds into different simulated body fluids (SBF). SEM, EDX, FTIR, TEM electron diffraction confirmed the apatite coating to comprise of calcium-deficient carbonated hydroxyapatite crystals. While both apatite-coated and non-coated PLGA scaffolds supported MC3T3-E1 attachment, spreading, and proliferation, significant differences in osteoblastic differentiation were observed. Relative to non-coated controls, quantitative real-time PCR revealed significant apatite-associated suppression of alkaline phosphatase (ALP), early upregulation of osteopontin (OPN) at 3 days, and upregulation of osteocalcin (OCN) and bone sialoprotein (BSP) at 4 weeks. In summary, apatite-promoted osteoblastic differentiation can be observed in a 3D model system that is relevant to tissue engineering.

Animals↗

PubMed2005

The effect of biomimetic apatite structure on osteoblast viability, proliferation, and gene expression.

The conventional biomimetic apatite coating process can be accelerated by immersing substrates into concentrated simulated body fluid (5 x SBF) at 37 degrees C to form an initial coating of apatite precursor spheres, and transform the precursors into plate-like apatite structures. Depending on processing parameters, different apatite structures can be created over the same substrate. The purpose of this study is to investigate the effects of the different apatite microenvironment on cell spreading, viability, proliferation, and gene expression. MC3T3-E1 preosteoblasts were cultured on five surfaces: conventional apatite (CA), precursor apatite spheres (PreA), large plate-like apatites (LgA), small plate-like apatites (SmA), and tissue culture grade polystyrene (TCPS). PreA induced significantly higher cell death during the first two weeks. TCPS supported more uniform spreading (1 day) and higher proliferation (2 weeks) than CA, LgA, and SmA. Apatites restricted spreading and promoted the extension of cellular projections along the textured surfaces under confocal microscopy observation. By 3 weeks, LgA induced highest expression of mature osteogenic markers osteocalcin (OCN) and bone sialoprotein (BSP) in both regular and osteogenic culture media based on quantitative real-time RT-PCR. The results of this study suggest differential cell responses to subtle changes in apatite microenvironment.

3T3 Cells↗

PubMed2005

Scaffold fabrication by indirect three-dimensional printing.

Three-dimensional printing (3DP) has been employed to fabricate porous scaffolds by inkjet printing liquid binder droplets onto particulate matter. Direct 3DP, where the final scaffold materials are utilized during the actual 3DP process, imposes several limitations on the final scaffold structure. This study describes an indirect 3DP protocol, where molds are printed and the final materials are cast into the mold cavity to overcome the limitations of the direct technique. To evaluate the resolution available in this technique, scaffolds with villi features (500 microm diameter, 1 mm height) were produced by solvent casting into plaster molds, followed by particulate leaching. Scanning electron microscope (SEM) showed highly open, well interconnected, uniform pore architecture ( approximately 100-150 microm). The ability of these scaffolds to support intestinal epithelial cell (IEC6) culture was investigated in vitro. IEC6 cells attached to scaffolds uniformly in vitro and grew preferentially in the villi region. To exploit the freeform nature of this technique with large pore size, anatomically shaped zygoma scaffolds with 300-500 microm interconnected pores were produced and characterized. Indirect 3DP provides an alternative method to complement other direct solid freeform fabrication methods.

Biocompatible Materials↗

PubMed2005

An evaluation of the effects of handpiece speed, abrasive characteristics, and polishing load on the flexural strength of polished ceramics.

STATEMENT OF PROBLEM: Many studies on the strengthening effects of grinding and polishing, as well as heat treatment on ceramics, are not well standardized or use commercially available industrial polishing systems. The reported effectiveness of these strengthening mechanisms on ceramics may not be applicable to clinical dentistry. PURPOSE: The purpose of this study was to evaluate the effects of controlled polishing on the flexural strength of dental ceramics by using a custom-made machine that applied standardized loads and speeds that coincided with the mean loads and speeds used by experienced prosthodontists. MATERIAL AND METHODS: A total of 140 aluminous dental ceramic bar-shaped specimens (Vitadur Alpha Enamel) measuring 1.5 x 2.0 x 25 mm were fabricated and divided into 12 groups (for most groups, n=10). Specimens were untreated, polished with different polishing systems, polished at different speeds, ground and autoglazed, polished and autoglazed, autoglazed and polished, polished with loose (paste) and bonded abrasives, or overglazed. Simulated clinical polishing was performed on the ceramic specimens by using a customized polishing apparatus that allowed independent control over the relevant polishing parameters (abrasive hardness, applied load, linear speed, rotational velocity, and wheel stiffness). Flexural strength (MPa) was measured with a 4-point bending test, and subjective surface roughness was assessed with scanning electron microscopy. Autoglazing was performed at various stages of the polishing sequence to determine the effects of polishing on surface stresses. Mean values, standard deviations, independent-sample t tests, 1-way and 2-way analyses of variance, Dunnett t tests and Kruskal-Wallis tests were applied to the data (alpha=.05). RESULTS: Under a clinical load of 0.6 N for a coarse polishing wheel, 1.0 N for a medium polishing wheel, and 1.3 N for a fine polishing wheel, a linear speed of 499 mm/min, and a rotational velocity of 10,000 rpm, the use of clinical polishing instruments did not affect the flexural strength of the aluminous ceramics studied (P=.274). At higher rotational velocity (20,000 rpm), specimens polished with the diamond polishing system produced statistically weaker specimens compared with those that had been polished at 10,000 rpm (P=.019). Autoglazing treatment of the diamond-polished specimens did not reverse the strength degradation (P=.125). Conversely, diamond polishing of the autoglazed specimens resulted in significant flexural strength reduction (P=.029). Fine-diamond-bonded abrasive significantly reduced flexural strength (P=.025). CONCLUSIONS: Simulated clinical polishing at 10,000 rpm did not appear to substantially strengthen or weaken the ceramic specimens. Polishing at 20,000 rpm reduced flexural strength of the ceramic bars.

Aluminum Oxide↗

PubMed2005

Osteogenic differentiation is inhibited and angiogenic expression is enhanced in MC3T3-E1 cells cultured on three-dimensional scaffolds.

Osteogenic differentiation of osteoprogenitor cells in three-dimensional (3D) in vitro culture remains poorly understood. Using quantitative real-time RT-PCR techniques, we examined mRNA expression of alkaline phosphatase, osteocalcin, and vascular endothelial growth factor (VEGF) in murine preosteoblastic MC3T3-E1 cells cultured for 48 h and 14 days on conventional two-dimensional (2D) poly(l-lactide-co-glycolide) (PLGA) films and 3D PLGA scaffolds. Differences in VEGF secretion and function between 2D and 3D culture systems were examined using Western blots and an in vitro Matrigel-based angiogenesis assay. Expression of both alkaline phosphatase and osteocalcin in cells cultured on 3D scaffolds was significantly downregulated relative to 2D controls in 48 h and 14 day cultures. In contrast, elevated levels of VEGF expression in 3D culture were noted at every time point in short- and long-term culture. VEGF protein secretion in 3D cultures was triple the amount of secretion observed in 2D controls. Conditioned medium from 3D cultures induced an enhanced level of angiogenic activity, as evidenced by increases in branch points observed in in vitro angiogenesis assays. These results collectively indicate that MC3T3-E1 cells commit to osteogenic differentiation at a slower rate when cultured on 3D PLGA scaffolds and that VEGF is preferentially expressed by these cells when they are cultured in three dimensions.

Alkaline Phosphatase↗