Biomedical subjects

Stuart K Williams

Publications and source records attributed to Stuart K Williams.

15 recordsLinked to original sources

PubMed2006

Texture analysis of speckle in optical coherence tomography images of tissue phantoms.

Optical coherence tomography (OCT) is an imaging modality capable of acquiring cross-sectional images of tissue using back-reflected light. Conventional OCT images have a resolution of 10-15 microm, and are thus best suited for visualizing tissue layers and structures. OCT images of collagen (with and without endothelial cells) have no resolvable features and may appear to simply show an exponential decrease in intensity with depth. However, examination of these images reveals that they display a characteristic repetitive structure due to speckle. The purpose of this study is to evaluate the application of statistical and spectral texture analysis techniques for differentiating living and non-living tissue phantoms containing various sizes and distributions of scatterers based on speckle content in OCT images. Statistically significant differences between texture parameters and excellent classification rates were obtained when comparing various endothelial cell concentrations ranging from 0 cells/ml to 25 million cells/ml. Statistically significant results and excellent classification rates were also obtained using various sizes of microspheres with concentrations ranging from 0 microspheres/ml to 500 million microspheres/ml. This study has shown that texture analysis of OCT images may be capable of differentiating tissue phantoms containing various sizes and distributions of scatterers.

Algorithms↗

PubMed2006

Covalent modification of porous implants using extracellular matrix proteins to accelerate neovascularization.

Healing associated with many polymeric biomedical implants commonly involves the formation of an avascular fibrous capsule. The lack of either formation or persistence of blood vessels in formed fibrous capsules, as well as a lack of new blood vessels within porous polymeric implants, often results in poor performance of the implant. The current study evaluated the use of extracellular matrix protein modification of a commonly used biomedical implant material, expanded polytetrafluoroethylene (ePTFE), as a mechanism to increase the neovascularization both within these porous implants and in tissue that forms in the peri-implant area. Discs of ePTFE were covalently modified with different extracellular matrix proteins including collagen type IV, fibronectin, and laminin type I. Discs were implanted into the adipose tissue of adult rats, and following a 5-week implant phase, histologic analysis of peri-implant tissue angiogenesis and implant neovascularization was performed. Striking differences were observed in angiogenic and neovascularization responses to matrix-modified ePTFE when compared with control, untreated ePTFE. Fibronectin treatment resulted in an extensive inflammatory response but, relative to the degree of inflammation, limited evidence of tissue angiogenesis or polymer neovascularization. Collagen type IV treatment groups exhibited a significant increase in angiogenesis in the peri-implant tissue with minimal evidence of implant neovascularization. In contrast to all other implant modifications, laminin type 1-treated ePTFE samples stimulated an extensive peri-implant tissue angiogenic response and a coordinate neovascularization of the porous interstices of the biomaterial.

Adipose Tissue↗

PubMed2005

Gene expression in tissue associated with extracellular matrix modified ePTFE.

Previous studies have established that surface modification of ePTFE with extracellular matrix molecules promotes vascularization within and around the implanted material. To understand the molecular basis of this tissue response to modified ePTFE, we analyzed large-scale gene expression in nonmodified and extracellular matrix-modified ePTFE-associated healing. Using a microarray containing 15,000 unique mouse cDNAs and an ANOVA-based analysis, we identified 789 genes related to cell signaling, inflammation, matrix remodeling, and proliferation that were differentially expressed across time, between modifications, or both. Genes were clustered based upon similarity in gene expression, producing 7 unique temporal super-patterns of expression. The clustered data revealed 3 general expression patterns unique to tissue surrounding the nonmodified ePTFE, while 6 unique expression patterns were associated with extracellular matrix-modified ePTFE. The more diverse expression patterns associated with extracellular matrix-modified ePTFE suggests that the tissue surrounding the extracellular matrix-modified ePTFE is more dynamic in terms of transcriptional activity. Taken together, these clusters serve as a "genetic fingerprint" for tissue healing in response to a specific material or material modification. Use of these genetic profiles will aid in the pursuit of improved device biocompatibility and enhanced material function.

Animals↗

PubMed2005

Dual porosity expanded polytetrafluoroethylene for soft-tissue augmentation.

BACKGROUND: A variety of nondegradable polymers have been evaluated for use as soft-tissue augmentation devices. This study compared a novel dual porosity expanded polytetrafluoroethylene with current, clinically used devices. METHODS: Studies were performed in a porcine model of soft-tissue healing with both histologic evaluations and determination of biomechanical strength of tissue incorporation. Five different samples of expanded polytetrafluoroethylene were used in this study. Control devices were clinically available soft-tissue augmentation devices manufactured by W. L. Gore and Associates (Newark, Del.). Atrium Medical Corporation (Hudson, NH) manufactured three test devices with modified porosities. A total of 12 animals were used with implant evaluations performed after 1, 3, 6, and 12 months. RESULTS: Significant differences in tissue incorporation were observed morphologically with the dual porosity material, including reduced inflammation and increased cellular and extracellular matrix incorporation of the material. Significant increases in both angiogenesis (new vessel formation in the peri-implant tissue) and neovascularization (blood vessel penetration into the interstices of the implants) were observed with the dual porosity expanded polytetrafluoroethylene material. CONCLUSIONS: This novel dual porosity expanded polytetrafluoroethylene is associated with reduced inflammation and more extensive tissue incorporation as compared with the currently available form. These results suggest a dual porosity expanded polytetrafluoroethylene may provide a superior material for soft-tissue augmentation.

Animals↗

PubMed2004

Laminin-5-enriched extracellular matrix accelerates angiogenesis and neovascularization in association with ePTFE.

The performance of biomedical implant devices is often limited by inappropriate tissue responses associated with synthetic materials used in device construction. Adverse healing responses, in particular the lack of an extensive vascular supply in the peri-implant tissue, are believed to lead to the ultimate failure of many of these medical devices. Accelerated formation of new blood vessels in the peri-implant tissue and within porous polymeric implants is hypothesized to improve the performance of such biomedical implant devices. The current study evaluated the use of cell-mediated, extracellular matrix modification of expanded polytetrafluoroethylene (ePTFE) to increase vessel growth in peri-implant tissue and within the pores of the implants. Discs of ePTFE were modified through cell-mediated matrix deposition using epithelial and endothelial cell lines with variable deposition of collagen types, fibronectin, and laminin types. Cell matrix-modified discs, Matrigel-coated discs, and nonmodified discs were implanted in both the adipose and subcutaneous tissues of the rat. Following a 5-week implant period, samples were removed and evaluated histologically and morphometrically for the presence of blood vessels in the peri-implant tissue and within the pores of the polymer as well as for the presence of activated macrophages and monocytes. A significantly increased presence of activated macrophages and monocytes was associated only with the samples modified with the matrix from a human microvessel endothelial cell line. Increased vessel density was identified in association with those ePTFE samples modified with either the 804-G, HaCaT, or II-4 cell matrices, all of which have extracellular matrices enriched in the protein laminin-5.

Adipose Tissue↗

PubMed2004

Rapid perfusion and network remodeling in a microvascular construct after implantation.

OBJECTIVE: We have previously demonstrated the ability to construct 3-dimensional microvascular beds in vitro via angiogenesis from isolated, intact, microvessel fragments that retain endothelial cells and perivascular cells. Our objective was to develop and characterize an experimental model of tissue vascularization, based on the implantation of this microvascular construct, which recapitulated angiogenesis, vessel differentiation, and network maturation. METHODS AND RESULTS: On implantation in a severe combined-immunodeficient mouse model, vessels in the microvascular constructs rapidly inosculated with the recipient host circulation. Ink perfusion of implants via the left ventricle of the host demonstrated that vessel inosculation begins within the first day after implantation. Evaluation of explanted constructs over the course of 28 days revealed the presence of a mature functional microvascular bed. Using a probe specific for the original microvessel source, 91.7%+/-11% and 88.6%+/-19% of the vessels by day 5 and day 28 after implantation, respectively, were derived from the original microvessel isolate. Similar results were obtained when human-derived microvessels were used to build the microvascular construct. CONCLUSIONS: With this model, we reproduce the important aspects of vascularization, angiogenesis, inosculation, and network remodeling. Furthermore, we demonstrate that the model accommodates human-derived vessel fragments, enabling the construction of human-mouse vascular chimeras.

Animals↗

PubMed2003

Accelerated endothelialization of interpositional 1-mm vascular grafts.

BACKGROUND: There is an increased need for alternative, synthetic, small-diameter vascular grafts due to a growing segment of the population who suffer from ischemic heart disease and lack suitable autologous vein grafts for use in coronary artery bypass grafting (CABG). We hypothesized that a cell-mediated extracellular matrix (ECM) modification of ePTFE would stimulate increased vascularization within the graft and thus promote lumenal endothelialization in a 1-mm rat abdomenal aortic implant model. METHODS AND RESULTS: Expanded polytetrafluoroethylene (ePTFE) vascular grafts (1 mm i.d.) were modified on the ablumenal surface with ECM deposited by the HaCaT or II-4 cell lines and implanted intrapositionally into the descending aorta of rats. Five weeks after implantation, all samples were patent and examination of the grafts demonstrated that the ECM modified samples exhibited extensive ablumenal vascularization and tissue incorporation compared to nonmodified samples. Also, ECM modified grafts had a cellular lining, while the nonmodified grafts were void of a cellular lining except for a limited pannus ingrowth. CONCLUSION: HaCaT and II-4 cell ECM modifications of ePTFE increase new blood vessel growth in association with the graft, and the II-4 cell modification results in formation of an endothelial monlayer on the lumenal surface of the graft.

Animals↗

PubMed2002

A comparative evaluation of the tissue responses associated with polymeric implants in the rat and mouse.

End product application is an important consideration when evaluating a material in an in vivo setting (Didisheim, Cardiovasc Pathol 1993;2:1S-2S). Small animal models allow high through-put evaluation of biocompatability. Previous preclinical evaluations have often used a rat subcutaneous model for the characterization of material-tissue interaction. Recent advances in genetic manipulation have provided mouse models with selective expression of a wide range of critical proteins. The rat model does not have many of the resources (i.e., knockouts, SCID, nude) that are present in mouse strains. The availability of these mice provides a resource to delineate the mechanisms regulating the healing associated with implants. However, before the mouse models can be used, they must be validated with respect to their ability to accurately assess tissue responses to materials. In this study the tissue responses after the implantation of expanded polytetrafluoroethylene (ePTFE) were compared between rat and mouse. Discs of ePTFE (30-microm internodal distance) were implanted in subcutaneous and epididymal fat tissue of rats (Sprague-Dawley) and mice (129-SVJ). After 5 weeks the samples were removed and evaluated for vascular density, inflammation, and fibrous encapsulation. No difference in the vessel density was observed within the peri-implant subcutaneous and adipose tissue or within the porous material. However, a significant difference was found in the number of activated macrophages and giant cells between these two species. Implants in the rat exhibited greater numbers of activated inflammatory cells in the peri-implant tissue. The data indicate that the mouse and rat provide a comparable model for evaluating angiogenesis and neovascularization associated with synthetic porous implants.

Adipose Tissue↗

PubMed2002

Characterization of angiogenesis and inflammation surrounding ePTFE implanted on the epicardium.

The response of epicardial tissue to the implantation of expanded polytetrafluoroethylene (ePTFE) was evaluated and compared with identical material implanted within subcutaneous and adipose tissues. These two tissue environments were selected for comparison with epicardial implants because they represent tissue often involved in device implantation. Discs of ePTFE (6 mm) were implanted into three different tissue sites in Sprague-Dawley rats. At 5 weeks, polymers and surrounding tissues were harvested and processed for light microscopy. General histology and histochemistry data indicated all polymers to be well incorporated with new tissue. Subcutaneous implants were covered by a dense fibrous capsule (55-70 microm). Epicardial and adipose implants had no fibrous capsule and a significantly greater number of microvessels (arterioles, capillaries, and venules) within the surrounding tissues compared with subcutaneous implants. An increased level of inflammation was also observed around epicardial implants compared with the other implants. Additionally, the new vasculature surrounding epicardially implanted ePTFE revealed an altered microvessel density and vessel type distribution compared with normal (control) epicardium. These results suggest that epicardial tissue responds to implanted ePTFE with a robust inflammatory response that may support the formation of a new microvasculature that is uniquely different from the native epicardial microvasculature.

Adipose Tissue↗

PubMed2002

Angiogenesis and neovascularization associated with extracellular matrix-modified porous implants.

Therapies directed toward stimulation of angiogenesis seek to accelerate the development of new blood vessels in tissues rendered dysfunctional because of an insufficient microvascular supply. The goal of the current study was the stimulation of an angiogenic response around and within porous biomedical implants, such as vascular grafts, constructed from a base polymer composed of expanded polytetrafluoroethylene (ePTFE). Similar to many biomaterials, ePTFE does not elicit a significant angiogenic response and the porous interstices of this material remain avascular after implantation. Studies were performed to evaluate the ability of a tumorigenic cell line, the 804-G rat kidney cell to secrete an angiogenic extracellular matrix on and within the porous structures of ePTFE. A rat model was used to evaluate and compare implant-associated healing responses between nonmodified materials and extracellular matrix-modified ePTFE. Results demonstrated that, in contrast to untreated ePTFE, the matrix-modified ePTFE stimulated both angiogenesis in implant-associated tissue and neovascularization of the pores within the ePTFE interstices. Deposition of an insoluble matrix stimulates an angiogenic response and has a potential application for the improvement of medical device function.

Adipose Tissue↗

PubMed2002

Abdominal aortic healing associated with a thin-walled Dacron-covered endovascular graft in a canine model.

PURPOSE: To characterize the healing response associated with an experimental endovascular graft (EVG) by examining the lumen of the prosthesis and the native vessel wall responses after implantation in a canine model. METHODS: An endovascular graft (EVG) constructed of hookless self-expanding nitinol springs covered by an ultrathin Dacron fabric was placed in the abdominal aorta of 8 dogs following femoral artery catheterization. After 12 weeks, specimens were subjected to histological and immunocytochemical testing to quantitatively and qualitatively analyze the cellular makeup of the luminal and abluminal tissues. RESULTS: Gross examination revealed a glistening, thrombus-free luminal surface on all prostheses, which was confirmed by scanning electron microscopy. The EVGs were well incorporated into the aortic wall, leaving a concentric, smooth flow surface. Immunocytochemistry verified the presence of von Willebrand factor-positive endothelial cells on the luminal surface and alpha-smooth muscle cell actin-positive smooth muscle and/or fibroblast cells in the subendothelial space. A sparse inflammatory response and a paucity of proliferating nuclear cell antigen-positive cells were noted within the neomedial lining adjacent to the graft material. In addition, a rich vaso vasorum-like plexus of microvessels was evident within the neomedial tissue. The native vessel wall upon which the EVG impinged was largely unaffected. CONCLUSIONS; These data demonstrate that placement of an experimental EVG in the canine abdominal aorta elicits a healing response that is distinct from interpositional vascular grafts. Results from this study have particular relevance to the proximal and distal anastomotic neck regions.

Actins↗

PubMed2002

Erythropoietin stimulates vasculogenesis in neonatal rat mesenteric microvascular endothelial cells.

Human breast milk is a rich source of growth factors, including erythropoietin (Epo), the endogenous hormonal stimulant of erythropoiesis. Recombinant human Epo (rhEpo) has been shown to stimulate 1) angiogenesis, the process of new blood vessel growth from preexisting vessels; 2) vasculogenesis, tubule formation from single-cell suspensions; and 3) endothelial cell proliferation in immortalized endothelial cells and vessel explants. We hypothesized that Epo would induce mitogenesis and stimulate vasculogenesis in primary cultures of microvascular endothelial cells (MVECs) from neonatal rat mesentery. Isolation, purification, characterization, and culture of MVECs were performed. Cell proliferative effects of rhEpo were studied by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in cultured MVECs. Vasculogenic effects of rhEpo were examined on cultured MVECs plated on either hormone-rich Matrigel substratum or the extracellular matrix protein, type I collagen. Our findings show that MVECs are isolated and purified, and that rhEpo stimulates MVEC proliferation, with maximal proliferation seen with a concentration of 50 IU/mL rhEpo. Tubule formation assays reveal that an rhEpo concentration of 50 IU/mL produces maximal tubule formation after 12 h on both Matrigel and the simple substratum, type I collagen. Our study is the first to examine the effects of rhEpo on the endothelium of the neonatal gastrointestinal tract. These data suggest that Epo may have a trophic effect on the vasculature of the gastrointestinal tract early in development. Furthermore, as Epo has been measured in breast milk, and its receptor has been shown to exist on the mucosa and gastrointestinal vasculature, Epo may be an endogenous stimulant of vessel growth during neonatal gastrointestinal development.

Animals↗

PubMedDate not supplied

Three-dimensional bioassembly tool for generating viable tissue-engineered constructs.

The primary emphasis of tissue engineering is the design and fabrication of constructs for the replacement of nonfunctional tissue. Because tissue represents a highly organized interplay of cells and extracellular matrix, the fabrication of replacement tissue should mimic this spatial organization. This report details studies evaluating the use of a three-dimensional, direct-write cell deposition system to construct spatially organized viable structures. A direct-write bioassembly system was designed and fabricated to permit layer-by-layer placement of cells and extracellular matrix on a variety of material substrates. Human fibroblasts suspended in polyoxyethylene/polyoxypropylene were coextruded through a positive displacement pen delivery onto a polystyrene slide. After deposition, approximately 60% of the fibroblasts remained viable. Bovine aortic endothelial cells (BAECs) suspended in soluble collagen type I were coextruded via microdispense pen delivery onto the hydrophilic side of flat sheets of polyethylene terephthalate. After deposition with a 25-gauge tip, approximately 86% of the BAECs were viable. When maintained in culture for up to 35 days, the constructs remained viable and maintained their original spatial organization. These results indicate the potential for utilizing a direct-write, three-dimensional bioassembly tool to create viable, patterned tissue-engineered constructs.

Animals↗

PubMedDate not supplied

Stimulated endothelial cell adhesion and angiogenesis with laminin-5 modification of expanded polytetrafluoroethylene.

Biomedical implants often exhibit poor clinical performance due to the formation of a periimplant avascular fibrous capsule. Surface modification of synthetic materials has been evaluated to accelerate the formation of functional microcirculation in association with implants. The current study used a flow-mediated protein deposition system to modify expanded polytetrafluoroethylene (ePTFE) with a laminin-5-rich conditioned growth medium and with medium from which laminin-5 had been selectively removed. An in vitro model of endothelial cell adherence determined that laminin-5 modification resulted in significantly increased adhesion of human microvessel endothelial cells to ePTFE. In vivo studies evaluating the periimplant vascular response to laminin-5-treated samples indicated that absorption of laminin-5-rich conditioned medium supported accelerated neovascularization of ePTFE implants. A flow system designed to treat porous implant materials facilitates laminin-5 modification of commercially available ePTFE, resulting in increased endothelial cell adhesion in vitro and increased vascularization in vivo.

Adipose Tissue↗

PubMedDate not supplied

Cardiac patch constructed from human fibroblasts attenuates reduction in cardiac function after acute infarct.

The current experiments used a scaffold-based, three-dimensional, human dermal fibroblast culture (3DFC) as a cardiac patch to stimulate revascularization and preserve left ventricular (LV) function of the infarcted LV in severe combined immunodeficient (SCID) mice. The 3DFC contains viable cells that secrete angiogenic growth factors and has been previously shown to stimulate angiogenesis. The hypothesis tested was that a 3DFC cardiac patch would attenuate a reduction in LV function of infarcted hearts. Five groups of mice were studied, including normal SCID mice (n = 13), normal SCID mice with 3DFC (n = 6), infarcted SCID mice (n = 6), infarcted mice with nonviable 3DFC (n = 6), and infarcted SCID mice with 3DFC (n = 6). An occlusion of a branch of the left anterior descending (LAD) coronary artery was performed by thermal ligation, and 3DFC was sized to the damaged area and implanted onto the epicardium at the site of tissue injury. Fourteen days postsurgery, LV mechanics were characterized with the Millar conductance catheter system (CCS). The data demonstrated that 3DFC-treated infarcted myocardium had significantly higher ejection fractions (EFs) compared with infarct-only mice (58.9 +/- 10.8 versus 31.0 +/- 5.8%, respectively; p < 0.05). Preload recruitable stroke work (PRSW) parameters were significantly higher in 3DFC-treated mice compared with infarct-only mice (64.6 +/- 11.9 versus 36.8 +/- 6.4 mmHg, respectively; p < 0.05). These results show that the 3DFC as a cardiac patch functioned to attenuate further loss of LV function accompanying acute myocardial infarct and that this may be related in part to myocardial revascularization.

Animals↗