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Biomedical subjects

Thomas J Webster

Publications and source records attributed to Thomas J Webster.

At least 19 recordsLinked to original sources

Better osteoblast adhesion on nanoparticulate selenium- A promising orthopedic implant material.

Apart from problems such as poor osseointegration, stress shielding, and wear debris-associated bone cell death, a major concern of metallic orthopedic implants is that they slowly corrode under in vivo environments. It is possible that continuous tissue exposure to metallic corrosion products limits orthopedic implant efficacy; this is especially true for patients receiving implants due to bone cancer. To date, there is no metallic orthopedic implant available in the market that specifically deals with the prevention and/or recurring cancer that may happen in these patients. The objective of this study was to deal with these problems in an integrated way by introducing a new biomaterial to the orthopedic community with anticancer chemistry: selenium (Se). In this study, six types of Se compacts were tested for bone cell (osteoblast) adhesion under in vitro conditions. Two types of cylindrical compacts were made with conventional Se metal particles in the micron (6.539 +/- 1.364-microm diameter) and submicron (0.963 +/- 0.139-microm diameter) range. These two types of compacts were chemically etched with different concentrations of NaOH to create two additional types of Se particles in each category: conventional size particles with nanosurface roughness and nanometer particles (0.204- to 0.264-microm diameter). Results showed for the first time, enhanced osteoblast adhesion on particulate surfaces of the compacts made from conventional Se compared with reference nonparticulate wrought titanium sheets. More importantly, this study provided the first evidence that osteoblast density was further increased on the surfaces of the Se compacts with nanometer particles. These initial findings indicate that there may be a promising future for nanoparticulate Se as an anticancer biocompatible orthopedic material.

Biocompatible Materials↗

Increased osteoblast function on PLGA composites containing nanophase titania.

Nanotechnology creates materials that potentially outperform, at several boundaries, existing materials in terms of mechanical, electrical, catalytic, and optical properties. However, despite their promise to mimic the surface roughness cells experience in vivo, the use of nanophase materials in biological applications remains to date largely unexplored. The objective of the present in vitro study was, therefore, to determine whether when added to a polymer scaffold, nanophase compared to conventional ceramics enhance functions of osteoblasts (or bone-forming cells). Results from this study provided the first evidence that functions (specifically, adhesion, synthesis of alkaline phosphatase, and deposition of calcium-containing mineral) of osteoblasts increased on poly-lactic-co-glycolic acid (PLGA) scaffolds containing nanophase compared to conventional grain size titania with greater weight percentage (from 10-30 wt %). Because the chemistry, material phase, porosity (%), and pore size of the composites were similar, this study implies that the surface features created by adding nanophase compared to conventional titania was a key parameter that enhanced functions of osteoblasts. In this manner, the study adds another novel property of nanophase ceramics: their ability to promote osteoblast functions in vitro when added to a polymer scaffold. For this reason, nanophase ceramics (and nanomaterials in general) deserve further attention as orthopedic tissue engineering materials.

Biocompatible Materials↗

Mechanism(s) of increased vascular cell adhesion on nanostructured poly(lactic-co-glycolic acid) films.

Studies have shown that poly(lactic-co-glycolic acid) (PLGA) films with nanometer surface features promote vascular endothelial and smooth muscle cell adhesion. The objective of this in vitro research was to begin to understand the mechanisms behind this observed increase in vascular cell adhesion. Results provided evidence that nanostructured PLGA adsorbed significantly more vitronectin and fibronectin from serum compared to conventional (or those not possessing nanometer surface features) PLGA. When separately preadsorbing both vitronectin and fibronectin, increased vascular smooth muscle and endothelial cell density was observed on nanostructured (compared to conventional) PLGA. Additionally, blocking of cell-binding epitopes of fibronectin and vitronectin significantly decreased vascular cell adhesion on nanostructured (compared to conventional) PLGA. For this reason, results of the present in vitro study demonstrated that cell adhesive proteins adsorbed in different quantities and altered bioactivity on nanostructured compared to conventional PLGA topographies, which (at least in part) may account for the documented increased vascular cell adhesion on nanostructured PLGA. In this manner, this study continues to provide evidence for the promise of nanostructured PLGA in vascular tissue engineering applications.

Absorbable Implants↗

Non-uniform flow behavior in a parallel plate flow chamber : alters endothelial cell responses.

Arterial flow characteristics determine vessel health by modulating vascular endothelial cells. One system used to study these interactions is the parallel plate flow chamber. The present in vitro study quantified the uniformity of fluid flow across a parallel plate flow chamber and characterized plate-location dependent endothelial cell gene expression. More specifically, shear stress varied by as much as 11% across the chamber area, which caused non-uniform ecNOS (p < 0.05) and COX-2 (p < 0.05) mRNA expression across the plate area. Results herein suggest that chamber variations may result during construction or assembly, which ultimately affect flow-sensitive cell responses (including mRNA expression). Therefore, these limitations should be considered when reporting endothelial cell responses to fluid flow using parallel plate flow chambers.

Animals↗

Increased osteoblast functions on theta + delta nanofiber alumina.

Nanophase materials, or materials with grain sizes less than 100 nm in at least one direction, are promising materials for various implant applications since our tissues are composed of nanometer components (i.e., proteins and/or inorganics). Specifically, bone is comprised of nanostructured hydroxyapatite and collagen fibers which continuously provide an extracellular matrix surface to bone-forming cells (osteoblasts) with a high degree of nanometer roughness. Despite this fact, materials currently utilized for orthopedic implants, whether metallic or ceramic, have constituent grain sizes in the non-biologically inspired micron regime. For this reason, the objective of the present in vitro study was to determine osteoblast functions on one classification of nanomaterials for orthopedic applications: nanofiber alumina. Various crystalline forms of nanofiber alumina were tested in this study. To obtained different crystalline structured nanofiber alumina, boehmite nanofiber alumina was sintered at either 400 degrees C, 600 degrees C, 800 degrees C, 1000 degrees C, or 1200 degrees C for 2 h in air. X-ray diffraction results provided evidence that boehmite nanofiber alumina remained boehmite when sintered at 400 degrees C but changed crystalline phases to gamma, gamma + delta, theta + delta, and alpha when sintered at 600 degrees C, 800 degrees C, 1000 degrees C, and 1200 degrees C, respectively. Moreover, compared to any other alumina formulation tested in this study, osteoblast functions (as measured by alkaline phosphatase activity and calcium deposition) were the greatest on theta + delta crystalline phase nanofiber alumina after 14 days of culture. Boehmite had the next greatest amount of calcium deposition by osteoblasts followed by gamma + delta. Gamma crystalline phase then followed and was greater than alpha crystalline phase nanofiber alumina which promoted osteoblast functions the least of all the compacts with the exception of borosilicate glass (reference substrate). For this reason, this study suggests that theta+delta nanofiber alumina should be further investigated in orthopedic applications.

Aluminum Oxide↗

Enhanced osteoblast adhesion on hydrothermally treated hydroxyapatite/titania/poly(lactide-co-glycolide) sol-gel titanium coatings.

Sol-gel processing was used to coat titanium substrates with hydroxyapatite (HA), TiO2, and poly(DL-lactic-glycolic acid). Coating surface characteristics were analyzed with XRD, EDS, AFM, SEM, and water contact angle measurements which indicated that the coatings had a high degree of crystallinity and good resistance to cracking. Coatings were also evaluated by cytocompatibility testing with osteoblast-like cells (or bone-forming cells). The cytocompatibility of the HA composite coatings prepared in the present in vitro study was compared to that of a traditional plasma-sprayed HA coating. Results showed that osteoblast-like cell adhesion was promoted on the novel HA sol-gel coating compared to the traditional plasma-sprayed HA coating. In addition, hydrothermal treatment of the sol-gel coating improved osteoblast-like cell adhesion. Since osteoblast adhesion is a necessary prerequisite for subsequent formation of bone, these results provided evidence that hydrothermally sol-gel processed HA may improve bonding of titanium implants to juxtaposed bone and, thus, warrants further investigation.

Cell Adhesion↗

Three-dimensional, nano-structured PLGA scaffolds for bladder tissue replacement applications.

In total, approximately 400 million people worldwide suffer from urinary bladder cancer (Nat Biotechnol 17 (1999) 149). When radical cysectomy is required as treatment, a replacement material is clearly necessitated. For this purpose, three-dimensional poly(lactic-co-glycolic acid) (PLGA) scaffolds were constructed using solvent casting and salt leaching processes. These scaffolds were manipulated to possess nano-dimensional surface features by soaking in sodium hydroxide at select concentrations and for various periods of time. Human bladder smooth muscle cells were then seeded onto these nano-dimensional scaffolds; adhesion and longer-term cell growth experiments were performed for either 4 h, or 1, 3, and 5 days, respectively. Additionally, collagen and elastin production was quantified following each experiment. In all cases, control cells were placed in an incubator and subjected to normal atmospheric pressure, while experimental cells were placed in a pressure chamber and subjected to a sustained pressure of 10 cm H(2)O. Results of this study provided evidence that porous, nano-dimensional polymeric scaffolds enhanced cell adhesion and growth, while also promoting increased elastin and collagen production. Moreover, in general, exposure to pressure did not alter cellular adhesion, growth, or extracellular matrix protein production, which suggests that the scaffolds and their resident cells will fair well in the complex mechanical environment of the bladder wall. In combination, these results provide evidence that the nano-dimensional PLGA scaffolds created in this research are promising as the next generation of bladder wall replacement materials.

Biocompatible Materials↗

Accelerated chondrocyte functions on NaOH-treated PLGA scaffolds.

Compared to conventional poly(lactic-co-glycolic acid) (PLGA), previous studies have shown that NaOH-treated PLGA two-dimensional substrates enhanced functions of osteoblasts (bone-forming cells), vascular and bladder smooth muscle cells, and chondrocytes (cartilage-synthesizing cells). In this same spirit, the purpose of this in vitro study was to fabricate three-dimensional NaOH-treated PLGA scaffolds and determine their efficacy toward articular cartilage applications. To improve functions of chondrocytes including their adhesion, growth, differentiation, and extracellular matrix synthesis, PLGA scaffolds were modified via chemical etching techniques using 1N NaOH for 10 min. Results demonstrated that NaOH-treated PLGA three-dimensional scaffolds enhanced chondrocyte functions compared to non-treated scaffolds. Specifically, chondrocyte numbers, total intracellular protein content, and the amount of extracellular matrix components (such as glycosaminoglycans and collagens) were significantly greater on NaOH-treated than on non-treated PLGA scaffolds. Underlying material properties that may have enhanced chondrocyte functions include a more hydrophilic surface (due to hydrolytic degradation of PLGA by NaOH), increased surface area, altered porosity (both percent and diameter of individual pores), and a greater degree of nanometer roughness. For these reasons, this study adds a novel tissue-engineering scaffold to the cartilage biomaterial community: NaOH-treated PLGA. Clearly, such modifications to PLGA may ultimately enhance the efficacy of tissue-engineering scaffolds for articular cartilage repair.

Biocompatible Materials↗

Analytically derived material properties of multilaminated extracellular matrix devices using the ball-burst test.

Xenogeneic extracellular matrices (ECMs) have been shown to be effective as naturally occurring scaffolds for soft-tissue repair. As acellular tissue substitutes at the time of surgical implantation, ECMs are subjected to the mechanical forces and micro-environmental conditions representative of the anatomical location in which they are placed. Ideally such natural scaffolds would possess mechanical properties that allow for normal tissue function in and around the implant site. The ball-burst test was used to simulate biaxial forces and to determine the strength of the ECM scaffold under a relevant physiological loading condition. The ball-burst test, in itself, does not quantify intrinsic mechanical properties and therefore a methodology was developed to determine the maximum stress resultant tangent modulus (MSRTM) or the maximum stress tangent modulus (MSTM), stress to failure (sigma(f)), failure stress resultant (N(f)), ball-burst pressure (P), and maximum elongation (lambda(max)) from the raw ball-burst data obtained at a constant-rate of transverse. The analytical methodology was compared to finite element simulations and showed good correlation with the analytical solution presented. The proposed approximations were used to compute biaxial failure properties for a variety of multilaminate ECM devices with varying number of layers, disinfection and sterilization, and organ origin.

Animals↗

Helical rosette nanotubes: a biomimetic coating for orthopedics?

Helical rosette nanotubes (HRN) are obtained through an entropically driven self-assembly process of low-molecular-weight synthetic modules under physiological conditions. Counter-intuitively, these materials undergo extensive self-assembly under the effect of temperature, resulting in networks of very long nanotubes. We have previously shown, using an in vitro model, that titanium (Ti) coated with HRN containing a lysine side chain (HRN-K1) displayed enhanced osteoblast (OB) adhesion when compared to uncoated Ti (p < 0.01). Because it has been widely known that proteins play a critical role in OB adhesion on nanophase materials, here we examine OB adhesion on heated (+T) and unheated (-T) HRN-K1-coated Ti under serum (+S, presence of proteins) and serum-free (-S, absence of proteins) conditions. The results demonstrated that (a) while proteins enhanced OB adhesion on +T HRN-K1-coated Ti, they had no effect on -T HRN-K1-coated Ti, suggesting an active role played by the rosette nanotubes in promoting OB adhesion, and (b) under -S conditions, +T HRN-K1 induced the same level of OB adhesion as uncoated Ti under +S conditions, suggesting that +T HRN-K1 acts as a protein substitute. Finally, transmission electron microscopy and atomic force microscopy studies of +T and -T HRN-K1-coated Ti revealed a significant change in surface coverage, density and hierarchical organization of the nanotubes upon heating, which was correlated with their ability to promote cell adhesion.

Adsorption↗

Nanometer surface roughness increases select osteoblast adhesion on carbon nanofiber compacts.

Carbon nanofibers have exceptional theoretical mechanical properties (such as low weight-to-strength ratios) that, along with possessing nanoscale fiber dimensions similar to crystalline hydroxyapatite found in bone, suggest strong possibilities for use as an orthopedic/dental implant material. To determine, for the first time, cytocompatibility properties pertinent for bone prosthetic applications, osteoblast (bone-forming cells), fibroblast (cells contributing to callus formation and fibrous encapsulation events that result in implant loosening), chondrocyte (cartilage-forming cells), and smooth muscle cell (for comparison purposes) adhesion were determined on carbon nanofibers in the present in vitro study. Results provided evidence that, compared to conventional carbon fibers, nanometer dimension carbon fibers promoted select osteoblast adhesion. Moreover, adhesion of other cells was not influenced by carbon fiber dimensions. In fact, smooth muscle cell, fibroblast, and chondrocyte adhesion decreased with an increase in either carbon nanofiber surface energy or simultaneous change in carbon nanofiber chemistry. To determine properties that selectively enhanced osteoblast adhesion, similar cell adhesion assays were performed on polymer (specifically, poly-lactic-co-glycolic; PLGA) casts of carbon fiber compacts previously tested. Compared to PLGA casts of conventional carbon fibers, results provided the first evidence of enhanced select osteoblast adhesion on PLGA casts of nanophase carbon fibers. The summation of these results demonstrate that due to a high degree of nanometer surface roughness, carbon fibers with nanometer dimensions may be optimal materials to selectively increase osteoblast adhesion necessary for successful orthopedic/dental implant applications.

Alloys↗

Decreased fibroblast cell density on chemically degraded poly-lactic-co-glycolic acid, polyurethane, and polycaprolactone.

Select prolonged functions of fibroblasts leading to extensive fibrous tissue encapsulation can be detrimental to numerous implant applications, including materials designed for the bladder, vasculature, and bone. Specifically, overextended functions of fibroblasts at the tissue-implant interface for orthopedic applications lead to callus formation, fibrous encapsulation events, and ultimately soft (not desirable hard-bony) tissue juxtaposition. Such events result in insufficient regeneration of bone and compromise the overall success of the implant. The objective of the present in vitro study was to determine, for the first time, fibroblast densities on NaOH-treated poly-lactic-co-glycolic acid co-polymers (PLGA), HNO(3)-treated polyurethane (PU), and NaOH-treated polycaprolactone (PCL). Previous studies have demonstrated increased bladder, vascular, and bone cell densities on chemically treated compared to unaltered PLGA, PU, and PCL films. Results of this study provided evidence of decreased fibroblast numbers on chemically treated PLGA, PU, and PCL after time periods of up to 5 days. Examination of these substrates revealed that all chemically modified polymers possessed a high degree of nanometer surface roughness compared to their respective unaltered polymers. In contrast, other material properties (such as chemistry and wettability) were different when comparing chemically treated PLGA, PU, and PCL films. Since fibroblasts are anchorage-dependent cells whose adhesion is a critical prerequisite to the prolonged, extensive formation of a fibrous-tissue containing extracellular matrix, the present in vitro results of decreased fibroblast densities on chemically degraded PLGA, PU, and PCL suggest that these materials may be suitable materials for numerous tissue-engineering applications and, thus, deserve further investigation.

Absorbable Implants↗

Osteoblast response to hydroxyapatite doped with divalent and trivalent cations.

The present in vitro study doped hydroxyapatite (HA) with various metal cations (Mg(2+), Zn(2+), La(3+), Y(3+), In(3+), and Bi(3+)) in an attempt to enhance properties of HA pertinent to orthopedic and dental applications. X-ray diffraction material characterization indicated that the metal cations may have substituted for calcium in the HA crystal structure and that all of the doped HA formulations were single-phase and crystalline. Scanning electron microscopy analysis revealed a variety of grain sizes, depending on the dopant utilized. Energy-dispersive spectroscopy confirmed that the dopants added during synthesis were present and that all of the HA formulations synthesized were within the defined range of HA phase in the CaO-P(2)O(5)-H(2)O system. Lastly, Bi-doped HA had a slower dissolution rate than either undoped HA or HA doped with other cations when exposed to simulated physiological conditions for 21 days. In terms of cell function, results provided the first evidence that osteoblasts, bone-forming cells, adhered and differentiated (as measured by alkaline phosphatase synthesis) in response to HA doped with trivalent cations (specifically, La(3+), Y(3+), In(3+), Bi(3+)) at earlier time points than either HA doped with divalent cations (Mg(2+), Zn(2+)) or undoped HA. Of the dopants examined, Bi(3+) most enhanced osteoblast long-term calcium-containing mineral deposition. For these reasons, this study revealed for the first time the potential benefits of doping HA with Bi(3+) according to criteria critical for bone prosthetic clinical success.

Alkaline Phosphatase↗

Biaxial strength of multilaminated extracellular matrix scaffolds.

Xenogeneic extracellular matrix (ECM) can be harvested and configured to function as a bioscaffold for tissue and organ reconstruction. The mechanical properties of the ECM vary depending upon the tissue from which it is harvested. Likewise, the manufacturing steps required to develop ECMs into medical grade devices will affect the surface morphology and the mechanical properties of the bioscaffold; important properties for constructive tissue remodeling. The present study compared the ball-burst strength of five different ECM scaffolds before and after treatment with peracetic acid (PAA): porcine small intestinal submucosa (SIS), porcine urinary bladder submucosa (UBS), porcine urinary bladder matrix (UBM), a composite of UBS + UBM, and canine stomach submucosa (SS). This study also compared the mechanical properties of 2- and 4-layer ECM scaffolds. Results showed 2-layer SS devices had the highest ball-burst value of all 2-layer ECM devices. Moreover, all 4-layer ECM devices had similar ball-burst strength except for 4-layer UBM devices which was the weakest. PAA-treatment decreased the ball-burst strength of SS and increased the ball-burst strength of UBS 2-layer devices. This study showed the material properties of the ECM scaffolds could be engineered to mimic those of native soft tissues (i.e. vascular, musculotendinous, etc) by varying the number of layers and modifying the disinfection/sterilization treatments used for manufacturing.

Absorbable Implants↗

Increased viable osteoblast density in the presence of nanophase compared to conventional alumina and titania particles.

In the present in vitro study, osteoblast (bone-forming cells) viability and cell density were investigated when cultured in the presence of nanophase compared to conventional (i.e. micron) alumina and titania particles at various concentrations (from 10,000 to 100 microg/ml of cell culture media) for up to 6h. Results confirmed previous studies of the detrimental influences of all ceramic particulates on osteoblast viability and cell densities. For the first time, however, results provided evidence of increased apoptotic cell death when cultured in the presence of conventional compared to nanophase alumina and titania particles. Moreover, since material characterization studies revealed that the only difference between respective ceramic particles was nanometer- and conventional-dimensions (specifically, phase and chemical properties were similar between respective nanophase and conventional alumina as well as titania particles), these results indicated that osteoblast viability and densities were influenced solely by particle size. Such nanometer particulate wear debris may result from friction between articulating components of orthopedic implants composed of novel nanophase ceramic materials. Results of a less detrimental effect of nanometer--as compared to conventional-dimensioned particles on the functions of osteoblasts provide additional evidence that nanophase ceramics may become the next generation of bone prosthetic materials with increased efficacy and, thus, deserve further testing.

Aluminum Oxide↗

Increased osteoblast adhesion on nanophase metals: Ti, Ti6Al4V, and CoCrMo.

Previous studies have demonstrated increased functions of osteoblasts (bone-forming cells) on nanophase compared to conventional ceramics (specifically, alumina, titania, and hydroxyapatite), polymers (such as poly lactic-glycolic acid and polyurethane), carbon nanofibers/nanotubes, and composites thereof. Nanophase materials are unique materials that simulate dimensions of constituent components of bone since they possess particle or grain sizes less than 100 nm. However, to date, interactions of osteoblasts on nanophase compared to conventional metals remain to be elucidated. For this reason, the objective of the present in vitro study was to synthesize, characterize, and evaluate osteoblast adhesion on nanophase metals (specifically, Ti, Ti6Al4V, and CoCrMo alloys). Such metals in conventional form are widely used in orthopedic applications. Results of this study provided the first evidence of increased osteoblast adhesion on nanophase compared to conventional metals. Interestingly, osteoblast adhesion occurred preferentially at surface particle boundaries for both nanophase and conventional metals. Since more particle boundaries are present on the surface of nanophase compared to conventional metals, this may be an explanation for the measured increased osteoblast adhesion. Lastly, material characterization studies revealed that nanometal surfaces possessed similar chemistry and only altered in degree of nanometer surface roughness when compared to their respective conventional counterparts. Because osteoblast adhesion is a necessary prerequisite for subsequent functions (such as deposition of calcium-containing mineral), the present study suggests that nanophase metals should be further considered for orthopedic implant applications.

Alloys↗

Endothelial and vascular smooth muscle cell function on poly(lactic-co-glycolic acid) with nano-structured surface features.

Biomaterials that successfully integrate into surrounding tissue should match not only the tissue's mechanical properties, but also its topography. The cellular response to a biomaterial may be enhanced in synthetic polymer formulations by mimicking the surface roughness created by the associated nano-structured extra-cellular matrix components of natural tissue. As a first step towards this endeavor, the goal of the present in vitro study was to use these design parameters to develop a synthetic, nano-structured, polymeric biomaterial that promotes cell adhesion and growth for vascular applications. In a novel manner, poly(lactic-co-glycolic acid) (PLGA) (50/50wt% mix) was synthesized to possess a range (from micron to nanometer) of surface features. Reduction of surface features was accomplished by treating conventional PLGA with various concentrations of NaOH for select periods of time. Results from cell experiments indicated that, compared to conventional PLGA, NaOH treated PLGA enhanced vascular smooth muscle cell adhesion and proliferation. However, PLGA prepared by soaking in NaOH decreased endothelial cell adhesion and proliferation compared to conventional PLGA. After further investigation, this finding was determined to be a result of chemical (and not topographical) changes during polymer synthesis. Surface chemistry effects were removed while retaining nano-structured topography by using polymer/elastomer casting methods. Results demonstrated that endothelial and smooth muscle cell densities increased on nano-structured cast PLGA. For these reasons, the present in vitro study provided the first evidence that nano-structured surface features can significantly improve vascular cell densities; such design criteria can be used in the synthesis of the next-generation of more successful tissue-engineered vascular grafts.

Animals↗

Nanobiotechnology: implications for the future of nanotechnology in orthopedic applications.

Nanotechnology involves the use of materials with components, such as fibers, grains and particles, that have dimensions of less than 100 nm. While numerous advantages of nanomaterials have been elucidated for catalytic, processing, mechanical, electrical, and optical applications, few have been described for orthopedic applications. Better orthopedic biomaterials are needed since the average lifetime of a bone biomaterial is less than 15 years. This review discusses recent studies that have been conducted to determine the efficacy of nanophase materials as bone implants. In doing so, it is suggested that nanophase materials can be synthesized to possess similar nanometer dimensions to components of bone tissue to promote new bone formation, compared with conventional orthopedic implant materials.

Biotechnology↗