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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

PLGA nanometer surface features manipulate fibronectin interactions for improved vascular cell adhesion.

The largest cause of mortality in the Western world is atherosclerotic vascular disease. Many of these diseases require synthetic vascular grafts; however, their patency rate is only 30% in small (<6 mm) diameter vascular grafts after 5 years of implantation. In an effort to increase small diameter vascular graft success, researchers have been designing random nanostructured surface features which enhance vascular cell functions. However, for the present study, highly-controllable, nanostructured features on poly(lactic-co-glycolic acid) (PLGA) surfaces were formulated. To create ordered nanostructured roughness on PLGA surfaces, either 500, 200, or 100 nm polystyrene nanospheres were separately placed onto mica. These were then used as a template for creating an inverse poly(dimethylsiloxane) mold which was utilized to cast PLGA. Compared to all other PLGA films formulated, AFM results demonstrated greater initial fibronectin spreading on PLGA which possessed spherical 200 nm features. Compared to smooth PLGA, PLGA with 500 or 100 nm surface features, results further showed that PLGA with 200 nm spherical features promoted vascular cell (specifically, endothelial, and smooth muscle cell) adhesion. In this manner, the present study demonstrated a specific nanometer surface feature size that promoted fibronectin spreading and subsequent vascular cell adhesion; criteria critical to vascular graft success.

Animals↗

Increased osteoblast adhesion on nanograined hydroxyapatite and tricalcium phosphate containing calcium titanate.

Depending on the coating method utilized and subsequent heat treatments (such as through the use of plasma-spray deposition), inter-diffusion of atomic species across titanium (Ti) and hydroxyapatite (HA) coatings may result. These events may lead to structural and compositional changes that consequently cause unanticipated HA phase transformations which may clearly influence the performance of an orthopedic implant. Thus, the objective of the present in vitro study was to compare the cytocompatibility properties of chemistries that may form at the Ti:HA interface, specifically HA, tricalcium phosphate (TCP), HA doped with Ti, and those containing calcium titanate (CaTiO(3)). In doing so, results of this study showed that osteoblast (bone-forming cells) adhesion increased with greater CaTiO(3) substitutions in either HA or TCP. Specifically, osteoblast adhesion on HA and TCP composites with CaTiO(3) was almost 4.5 times higher than that over pure HA. Material characterization studies revealed that enhanced osteoblast adhesion on these compacts may be due to increasing shrinkage in the unit lattice parameters and decreasing grain size. Although all CaTiO(3) composites exhibited excellent osteoblast adhesion results, Ca(9)HPO(4)(PO(4))(5)OH phase transformation into TCP/CaTiO(3) increased osteoblast adhesion the most; because of these reasons, these materials should be further studied for orthopedic applications.

Bone Substitutes↗

Increased osteoblast adhesion on nanograined Ti modified with KRSR.

Peptide sequences such as lysine-arginine-serine-arginine (KRSR) selectively bind transmembrane proteoglycans (e.g. heparin sulfate) of osteoblasts (bone-forming cells) and are, therefore, actively being investigated for orthopedic applications. Further, nanophase materials (or materials with grain or particle sizes less than 100 nm) are promising new materials that promote new bone growth more than compared to conventional (that is, micron grain or particle size) materials. To combine the above two promising approaches for improving orthopedic implants, the objective of this in vitro study was to functionalize titanium (Ti) surfaces (both nanophase and conventional) with KRSR peptides and study their osteoblast cell adhesive properties. Materials were characterized by X-ray photoelectron spectroscopy, scanning electron microscopy, and atomic force microscopy. Results of this in vitro study provided evidence of increased osteoblast adhesion on nanophase compared to conventional Ti whether functionalized with KRSR or not. Results further showed that the immobilization of KRSR onto Ti (both nanophase and conventional) increased osteoblast adhesion compared to respective nonfunctionalized Ti and those functionalized with the negative control peptide KSRR. Most importantly, osteoblast adhesion on nonfunctionalized nanophase Ti increased compared to conventional Ti functionalized with KRSR. Further, select initial osteoblast adhesion was observed to occur at particle boundaries for any type of nanophase and conventional Ti formulated in this study. In summary, results provided evidence that not only should nonfunctionalized nanophase Ti be further studied for improved orthopedic applications but so should nanophase Ti functionalized with KRSR.

Biocompatible Materials↗

Nanobiomaterial applications in orthopedics.

Advancements in nanobiotechnology are revolutionizing our capability to understand biological intricacies and resolve biological and medical problems by developing subtle biomimetic techniques. Nanocomposites and nanostructured materials are believed to play a pivotal role in orthopedic research since bone itself is a typical example of a nanocomposite. This article reviews current strategies using nanobiomaterials to improve current orthopedic materials and examines their applications in bone tissue engineering. Preliminary investigations support the potential of nanobiomaterials in orthopedic applications; however, significant advancements are necessary to achieve clinical use. Overall, current trends in nanobiotechnology foreshadow a bright future through the use of nanobiomaterials in the orthopedic domain.

Biocompatible Materials↗

Nanostructured biomaterials for tissue engineering bone.

Advances in several critical research fields (processing, catalytic, optical, actuation, electrical, mechanical, etc.) have started to benefit from nanotechnology. Nano-technology can be broadly defined as the use of materials and systems whose structures and components exhibit novel and significantly changed properties when control is gained at the atomic, molecular, and supramolecular levels. Specifically, such advances have been found for materials when particulate size is decreased to below 100 nm. However, to date, relatively few advantages have been described for biological applications (specifically, those involving bone tissue engineering). This chapter elucidates several promising examples of how nanophase materials can be used to improve orthopedic implant applications. These include mechanical advantages as well as altered cell functions, leading to increased bone tissue regeneration on a wide range of nanophase materials including ceramics, polymers, metals, and composites thereof. Such advances were previously unimaginable with conventional materials possessing large micron-sized particulates.

Animals↗

Increased osteoblast functions among nanophase titania/poly(lactide-co-glycolide) composites of the highest nanometer surface roughness.

Currently, the scientific challenges for bone tissue engineering lie in the development of suitable scaffold materials that can improve bone cell adhesion, proliferation, and differentiation. The design of nanophase titania/poly(lactide-co-glycolide) (PLGA) composites offers an exciting approach to combine the advantages of a degradable polymer with nanosize ceramic particles to optimize the physical and biological properties necessary for bone regeneration. Moreover, because of the presence of nanosized ceramics, such composites can be formulated to match the surface roughness of bone. For these reasons, the objective of the present in vitro study was to investigate osteoblast (bone-forming cell) adhesion and long-term functions on nanophase titania/PLGA composites that mimic the surface roughness of bone. Various sonication powers were applied in this study to manipulate titania dispersions in PLGA and consequently control their surface roughness. Most importantly, results correlated better osteoblast adhesion and long-term functions (such as collagen, alkaline phosphatase activity, and calcium-containing mineral deposition) among nanophase titania/PLGA composites that had surface roughness values closer to natural bone. In this manner, this present study demonstrated that the nanophase titania/PLGA composites sonicated to have nanometer surface roughness values can improve osteoblast functions necessary for enhanced bone tissue engineering applications.

Alkaline Phosphatase↗

Increased osteoblast adhesion on nanograined hydroxyapatite and partially stabilized zirconia composites.

To improve the mechanical properties of hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)) for orthopedic applications, numerous investigators have proposed combining HA with high strength materials, specifically zirconia. Despite the fact that, compared to pure HA, it is now well-established that zirconia and HA composites have improved mechanical properties, the cytocompatibility properties of this composite remain largely uninvestigated. For these reasons, the objective of the present in vitro study was to synthesize HA and partially stabilized zirconia composites for osteoblast (bone-forming cell) adhesion assays. Various sintering temperatures and amounts of zirconia in HA composites were used in order to ascertain their influence on osteoblast adhesion. Results demonstrated increased interactions between HA and partially stabilized zirconia, when either higher sintering temperatures (between 900 and 1,300 degrees C for 1 h) or higher zirconia contents (between 10 and 40 wt %) were used during material synthesis. More importantly, greater osteoblast adhesion was measured on HA-zirconia composites sintered either at lower temperatures (specifically, 900 degrees C) or with lower amounts of zirconia added to HA composites (specifically, 10 wt %). Results further indicated that when sintered at lower temperatures the composites possessed smaller nanometer grain sizes with increased surface roughness and a more stable HA phase. For these reasons, this study suggests that to optimize osteoblast adhesion on HA and partially stabilized zirconia composites for orthopedic applications, low sintering temperatures and low amounts of zirconia should be used. This suggests that a delicate balance must be reached between increasing mechanical properties of HA without decreasing osteoblast cytocompatibility properties through zirconia addition.

Animals↗

Increased osteoblast and decreased Staphylococcus epidermidis functions on nanophase ZnO and TiO2.

Many engineers and surgeons trace implant failure to poor osseointegration (or the bonding of an orthopedic implant to juxtaposed bone) and/or bacteria infection. By using novel nanotopographies, researchers have shown that nanostructured ceramics, carbon fibers, polymers, metals, and composites enhance osteoblast adhesion and calcium/phosphate mineral deposition. However, the function of bacteria on materials with nanostructured surfaces remains largely uninvestigated. This is despite the fact that during normal surgical insertion of an orthopedic implant, bacteria from the patient's own skin and/or mucosa enters the wound site. These bacteria (namely, Staphylococcus epidermidis) irreversibly adhere to an implant surface while various physiological stresses induce alterations in the bacterial growth rate leading to biofilm formation. Because of their integral role in determining the success of orthopedic implants, the objective of this in vitro study was to examine the functions of (i) S. epidermidis and (ii) osteoblasts (or bone-forming cells) on ZnO and titania (TiO(2)), which possess nanostructured compared to microstructured surface features. ZnO is a well-known antimicrobial agent and TiO(2) readily forms on titanium once implanted. Results of this study provided the first evidence of decreased S. epidermidis adhesion on ZnO and TiO(2) with nanostructured when compared with microstructured surface features. Moreover, compared with microphase formulations, results of this study showed increased osteoblast adhesion, alkaline phosphatase activity, and calcium mineral deposition on nanophase ZnO and TiO(2). In this manner, this study suggests that nanophase ZnO and TiO(2) may reduce S. epidermidis adhesion and increase osteoblast functions necessary to promote the efficacy of orthopedic implants.

Anti-Bacterial Agents↗

Using hydroxyapatite nanoparticles and decreased crystallinity to promote osteoblast adhesion similar to functionalizing with RGD.

Better materials are needed to promote bone growth. For this reason, the present study created nanometer crystalline hydroxyapatite (HA) and amorphous calcium phosphate compacts functionalized with the arginine-glycine-aspartic acid (RGD) peptide sequence. Crystalline HA and amorphous calcium phosphate nanoparticles were synthesized by a wet chemical process followed by a hydrothermal treatment for 2 h at 200 degrees C and 70 degrees C, respectively. Resulting particles were then pressed into compacts. For the preparation of conventional HA particles (or those with micron diameters), the aforementioned pressed compacts were sintered at 1,100 degrees C for 2 h. Peptide functionalization was conducted by means of a three step reaction procedure: silanization with 3-aminopropyltriethoxysilane (APTES), cross-linking with N-succinimidyl-3-maleimido propionate (SMP), and finally peptide immobilization. The three step reaction procedure was characterized by a novel 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) fluorescence technique. For all materials, results showed that the immobilization of the cell adhesive RGD sequence increased osteoblast (bone-forming cell) adhesion compared to those non-functionalized and those functionalized with the noncell adhesive control peptide (RGE) after 4 h. However, surprisingly, results also showed that the adhesion of osteoblasts on non-functionalized amorphous nanoparticulate calcium phosphate was similar to conventional HA functionalized with RGD. Osteoblast adhesion on nanocrystalline HA (unfunctionalized and functionalized with RGD) was below that of the respective functionalized amorphous calcium phosphate but above that of the respective functionalized conventional HA. In this manner, results of this study suggest that decreasing the particulate size into the nanometer regime and reducing crystallinity of calcium phosphate based materials may promote osteoblast adhesion to the same degree as the well-established techniques of functionalizing conventional HA with RGD.

Calcium Phosphates↗

Select bladder smooth muscle cell functions were enhanced on three-dimensional, nano-structured poly(ether urethane) scaffolds.

Bladder wall resection is often required as a treatment for invasive bladder cancer. When this happens, a suitable replacement material is needed. The present study, therefore, created three-dimensional, porous, nano-structured poly(ether urethane) (PU) matrices for use as bladder tissue-engineering scaffolds. Select cytocompatibility experiments (specifically adhesion and long-term growth studies) were performed on these scaffolds using human bladder smooth muscle cells (BdSMCs). In addition, the amount of total collagen and elastin present in each cell-seeded scaffold was determined since the production of these extracellular matrix (ECM) proteins is essential for the health and survival of cells and for the functionality of the replaced organ. Finally, to better understand how these scaffolds and resident cells would perform in the complex mechanical environment of the bladder wall, scaffolds and cells were subjected to 10 cmH2O pressure using a computer-controlled pressure chamber. Results provided evidence that compared to conventionally used, micro-dimensional PU scaffolds, the novel, nanodimensional scaffolds created in this research increased cell adhesion, growth, and ECM protein production. Additionally, scaffolds and resident cells were not affected by exposure to 10 cmH2O pressure (compared to controls maintained under atmospheric conditions). These results are promising and provide evidence that the nano-dimensional PU scaffolds created in this research are suitable bladder replacement materials that may outperform materials currently used for such purposes.

Animals↗

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↗