Human vascular endothelial cells (ECs) are exposed to various levels of hemodynamic forces, cycli... more Human vascular endothelial cells (ECs) are exposed to various levels of hemodynamic forces, cyclic strain, and shear stress in vivo. Here, we examined the in vitro effects of the various levels (0-6%, 7-16%, and 17-25%) of strain at 60, 30, and 15 cycles per minute (cpm) on human monocyte adherence to endothelial cells and extracellular matrix protein preabsorbed surfaces. Monocyte adhesion to endothelial cells under cyclic strain significantly increased. At both 30 and 60 cpm, ECs under strains of 7-16% and 17-25% showed >52% and >117% higher monocyte adhesion than endothelial cells under static condition when monocytes were added for 0.5 h. This increase in monocyte adhesion to ECs under cyclic strain remained significantly higher even after 24 h of incubation. Human monocyte adhesion to extracellular matrix protein preabsorbed surfaces differed depending on the specific extracellular matrix protein. Monocytes adhered to collagen type I and fibronectin preabsorbed surfaces &...
ABSTRACT This chapter presents our efforts to develop a better mechanistic understanding of how b... more ABSTRACT This chapter presents our efforts to develop a better mechanistic understanding of how biomaterial interactions with blood components lead to alteration of the basic pathophysiologic mechanisms, in particular, inflammation and the foreign body response, which increase the probability of bacterial interactions, colonization, biofilm formation, and infection. In particular, we present perspectives on mechanisms of S. epidermidis biofilm formation, the role of surface chemistry on biofilm formation, the role of bacterial slime production in device infections, the apoptosis of adherent polymorphonuclear leukocytes in the acute inflammatory response, neutrophil mobility and phagocytosis of bacteria on biomaterials, generation of reactive oxygen and nitrogen species by biomaterial-adherent neutrophils, and quorum sensing in S. epidermidis biofilm formation. Our work has focused on infection mechanisms of cardiovascular prostheses and devices where blood hemodynamics and shear stress play important roles in inflammatory cell interactions with biomaterial surfaces. However, we believe that much of our results also are applicable to static implant situations found in orthopedic, cosmetic (plastic), and other surgical areas.
A novel biofabrication modality, electrophoretic compaction with macromolecular alignment, was ut... more A novel biofabrication modality, electrophoretic compaction with macromolecular alignment, was utilized to make collagen threads that mimic the native tendon's structure and mechanical properties. A device with kinematic electrodes was designed to fabricate collagen threads in continuous length. For the first time, a 3D-biotextile was woven purely from collagen. Mechanical properties and load-displacement behavior of the biotextile mimicked those of the native tendon while presenting a porosity of 80%. The open pore network facilitated cell seeding across the continuum of the bioscaffold. Mesenchymal stem cells (MSCs) seeded in the woven scaffold underwent tenogenic differentiation in the absence of growth factors and synthesized a matrix that was positive for tenomodulin, COMP and type I collagen. Up-regulation of tenomodulin, a tendon specific marker, was 11.6 ± 3.5 fold, COMP was up-regulated 16.7 ± 5.5 fold, and Col I was up-regulated 6.9 ± 2.7 fold greater on ELAC threads when compared to randomly oriented collagen gels. These results demonstrate that a bioscaffold woven by using collagen threads with densely compacted and anisotropically aligned substrate texture stimulates tenogenesis topographically, rendering the electrochemically aligned collagen as a promising candidate for functional repair of tendons and ligaments.
Despite relatively sparse data regarding their outcomes in the setting of infection, biologic gra... more Despite relatively sparse data regarding their outcomes in the setting of infection, biologic grafts have gained rapid acceptance by the surgical community for complex hernia repair. These materials are heterogeneous in their procurement and processing techniques, which may ultimately have an impact in their ability to withstand infection. The objective of this study is to evaluate the impact of varying levels of contamination on biologic graft performance in a chronic ventral hernia animal model. Four commonly applied biologic grafts were used in the repair of a chronic ventral hernia rat model (n = 218). Each material was repaired in the setting of 1 of 4 surgical wound classifications (clean, clean contaminated, contaminated, dirty infected) with Staphylococcus aureus as our inoculum agent. After a 30-day survival, repairs underwent quantitative cultures, histological, and biomechanical testing. Marked differences were observed in biologic graft bacterial burden, biomechanical an...
Advances in experimental medicine and biology, 2011
Macrophages undergo fusion with other macrophages to form the hallmark multinucleated giant cells... more Macrophages undergo fusion with other macrophages to form the hallmark multinucleated giant cells of chronic inflammation. However, neither the existence of distinct morphological types of giant cells, the signaling pathways that induce their formation, the molecular mechanism(s) of macrophage fusion, nor the significance of macrophage multinucleation at chronic inflammatory sites are well understood. Our efforts have been focused on these unknowns, particularly as they relate to the foreign body-type giant cells that form on implanted biomaterials and biomedical devices. We have pursued the discoveries of human macrophage fusion factors (interleukin-4, interleukin-13, α-tocopherol) with emphasis on foreign body giant cells, and identified adhesion receptors and signaling intermediates, as well as an adhesion protein substrate (vitronectin) that supports macrophage fusion. Studies on the molecular mechanism of macrophage fusion have revealed it to be a mannose receptor-mediated phag...
This paper summarizes our recent e †orts to better understand the e †ects of antioxidants, the e ... more This paper summarizes our recent e †orts to better understand the e †ects of antioxidants, the e †ects of strain-state, mechanistic studies of soft segment cleavage by reactive oxygen radicals, and the e †ects of di †erent soft segment chemistries on the biostability/biodegradation of polyether polyurethanes (PEUUs). In vivo cage implant system studies and in vitro cobalt ion/ hydrogen peroxide studies have been carried out on PEUUs and the polymers have been analysed by attenuated total reÑectance and Fourier transform infrared (ATR-FTIR) spectroscopy, and scanning electron microscopic (SEM) characterization of the PEUU surfaces. The natural antioxidant, vitamin E, has been shown to inhibit biodegradation and enhance biostability of PEUUs. Studies of the e †ect of stress state on PEUU biodegradation demonstrate that stress can inhibit biodegradation. While polyether soft segments may be cleaved by the presence of reactive oxygen radicals, the presence of oxygen has a profound e †ect in accelerating biodegradation. The biodegradation of polyurethanes may be inhibited by substituting di †erent chemistries such as polydimethylsiloxanes, polycarbonates, and hydrocarbon soft segments for the polyether soft segments. To safely utilize polyurethanes in long-term biomedical devices, the biodegradation mechanisms of polyurethane elastomers must be fully understood and subsequently prevented. 1998 SCI.
The cyclic deformation of two polyurethane elastomers that differed in soft segment content and m... more The cyclic deformation of two polyurethane elastomers that differed in soft segment content and molecular weight was investigated. The microphase-separated morphology of the polyurethane with higher soft segment content consisted of hard segment domains dispersed in a soft segment matrix. In the polyurethane with lower soft segment content, the hard segment domains appeared to be partially cocontinuous. Following an initial 'conditioning' cycle, both polyurethanes exhibited reversible elastomeric behavior. Structural changes that occurred during conditioning were investigated using atomic force microscopy and Fourier transform infrared dichroism. The results provided the basis of a structural model for the deformation behavior. Yielding and reorganization of hard domains resulted in a highly oriented microfibrous morphology. Subsequent unloading and reloading were associated with reversible relaxation and reformation of the microfibrous entities. The elastic behavior of the conditioned polyurethanes was satisfactorily described by classical rubber theory with inextensibility. The structural model proposed here extended previous efforts to describe the deformation processes of polyurethanes during cyclic loading.
Empiric amphotericin B therapy was compared to central venous catheter Githdrawal in a prospectiv... more Empiric amphotericin B therapy was compared to central venous catheter Githdrawal in a prospective randomized trial. Of 32 febrile, neutropenic patients with indwelling Broviac catheters and without documented infection, 14 had persistent fever while receiving broad spectrum antibacterial drugs. Six patients were randomized to catheter removal and eight patients received amphotericin B empirically. None of six patients responded to catheter removal and six of eight defervesced after receiving antifungal therapy (p less than 0.01). Of the six patients in whom catheters were removed, two later became afebrile while on subsequent therapy with amphotericin B. Culture and histologic evaluation of the removed catheters failed to implicate the prosthesis as an infectious source. Central venous catheters in a persistently febrile neutropenic host need not be removed, unless local difficulties or bacteremia with skin commensal organisms occur. Amphotericin B can be infused through a central venous catheter in febrile, neutropenic patients unresponsive to empiric antibacterial drugs, with many patients becoming afebrile as a result of this therapy.
Journal of Materials Science: Materials in Medicine, 2006
The purpose of this communication is to present the author&am... more The purpose of this communication is to present the author's perspectives on the future of biomedical materials that were presented at the Larry L. Hench Retirement Symposium held at Imperial College, London, in late September 2005. The author has taken a broad view of the future of biomedical materials and has presented key ideas, concepts, and perspectives necessary for the future research and development of biomedical polymers and their future role as an enabling technology for the continuing progress of tissue engineering, regenerative medicine, prostheses, and medical devices. This communication, based on the oral presentation, is meant to be provocative and generate discussion. In addition, it is targeted for students and young scientists who will play an ever-increasing role in the future of biomedical materials.
Aseptic loosening of orthopedic implants is thought to be caused primarily by osteoclast differen... more Aseptic loosening of orthopedic implants is thought to be caused primarily by osteoclast differentiation induced by bone resorptive cytokines produced in response to phagocytosis of implant-derived wear particles. This study examined whether adherent endotoxin on the wear particles is responsible for inducing osteoclast differentiation as well as production of interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor a (TNF-alpha). Removal of adherent endotoxin almost completely inhibited the responses to titanium (Ti) particles by both murine marrow cells and human peripheral blood monocytes. In vivo experiments showed that endotoxin removal reduced particle-induced osteolysis by 50-70%. Addition of lipopolysaccharide (LPS) to the "endotoxin-free" particles restored their ability to induce cytokine production and osteoclast differentiation in vitro. Moreover, marrow cells from mice that are hyporesponsive to endotoxin because of mutation of Toll-like receptor 4 induced significantly less cytokine production and osteoclast differentiation in response to Ti particles with adherent endotoxin than did marrow cells from normoresponsive mice. This mutation also resulted in significantly less particle-induced osteolysis in vivo. Taken together, these results show that adherent endotoxin is involved in many of the biological responses induced by orthopedic wear particles and should stimulate development of new approaches designed to reduce the activity of adherent endotoxin in patients with orthopedic implants.
Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2014
Coating of various synthetic, absorbable, and biologic meshes with mesenchymal stem cells (MSCs) ... more Coating of various synthetic, absorbable, and biologic meshes with mesenchymal stem cells (MSCs) and fibroblasts was analyzed qualitatively and quantitatively. Five hernia meshes-light weight monofilament polypropylene (Soft Mesh), polyester (Parietex-TET), polylactide composite (TIGR), heavy weight monofilament polypropylene (Marlex), and porcine dermal collagen (Strattice)-were coated with three cell lines: human dermal fibroblasts (HFs), rat kidney fibroblasts (NRKs), and rat MSCs. Cell densities were determined at different time points. Samples also underwent histology and transmission electron microscopic (TEM) analyses. It required HFs 3 weeks to cover the entire mesh, while only 2 weeks for NRKs and MSCs to do so. MSCs had no preference for any of the meshes and produced the highest cell densities on Parietex and TIGR. Substrate-preference accounted for the significantly lower fibroblast densities on TIGR than Parietex. Fibroblasts failed to coat Marlex. Strattice, which had the least surface area, generated comparable cell densities to Parietex. Both histology and TEM confirmed cell coating of mesh surface. Various prosthetics can be coated by certain cell strains. Both mesh composition and cell preference dramatically influence the coating process. This methodology provides foundation for novel avenues of modulation of host response to various modern synthetic and biologic meshes.
The kinetics of attachment of NIL B and SV-NIL cells to glass, siliconized glass, and surfaces co... more The kinetics of attachment of NIL B and SV-NIL cells to glass, siliconized glass, and surfaces coated with random copolypeptides have been studied. It was found that in the absence of serum proteins, neither the rate nor the extent of attachment of cells is affected by the nature of the surface. In the presence of bovine serum albumin, the total uptake and rate of attachment of both NIL B and SV-NIL cells to the neutral, hydrophobic, and negatively charged copolymers is decreased compared with attachment to the same surfaces in the absence of protein. In contrast, the attachment of NIL B and SV-NIL cells to the positively charged (lysyl) copolymers was not decreased in the presence of protein. It was shown that the effect of protein resulted from its adsorption to the surface rather than to the cells. It was also concluded that both the NIL €3 and SV-NIL cells consist of a single cell population with respect to adhesiveness, and that both cell lines reach a kinetic equilibrium with the surfaces. This work represents one of the first studies to vary copolypeptide compositions systematically from negatively to neutral to positively charged surfaces and to examine these substrates without any mediating effects from various serum proteins. The results of this study support the concept that while cells bind to an adsorbed layer of protein rather than directly to the surface, the underlying surface can modify the attachment process by its effect on the protein adsorbed.
Poly(etherurethane) elastomers are useful materials in medical devices because of their mechanica... more Poly(etherurethane) elastomers are useful materials in medical devices because of their mechanical properties and biocompatibility. However, it is necessary to stabilize these elastomers against the oxidation of their ether soft segments. Synthetic antioxidants such as Santowhite and Irganox are often satisfactory; however, particularly for biomedical applications, it was of interest to test the natural antioxidant vitamin E in poly(etherurethane urea) (PEUU) elastomers in vivo. The alpha-tocopherol form of vitamin E was added to PEUU at 5% by weight. Biaxially strained PEUU specimens with and without vitamin E were tested in vivo in the cage implant system. The influence of vitamin E on PEUU biostability was analyzed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy and scanning electron microscopic (SEM) characterization of the PEUU surface. ATR-FTIR results showed that vitamin E prevented chemical degradation of the PEUU surface up to 5 weeks implantation, and at 10 weeks 82% of the ether remained. In contrast, without an antioxidant, only 18% of the ether remained after 10 weeks. No surface pitting or cracking was observed by SEM on PEUU with vitamin E; PEUU without antioxidant ruptured owing to extensive pitting and cracking. It was concluded that the antioxidant properties of vitamin E prevented oxidation of strained PEUU elastomers in vivo. The influence of vitamin E on PEUU biocompatibility was characterized by exudate leukocyte counts, density of leukocytes adherent to the PEUU, and morphology of adherent leukocytes. These results indicated decreased leukocyte counts in the exudate and less active adherent cells on the PEUU with vitamin E compared to PEUU without antioxidant. A proposed cell-polymer feedback system demonstrates how vitamin E improves both biostability and biocompatibility of PEUU elastomers in vivo.
Photochemical immobilization technology was utilized to covalently couple polymers to silicone ru... more Photochemical immobilization technology was utilized to covalently couple polymers to silicone rubber either at multiple points along a polymer backbone or at the endpoint of an amphiphilic chain. The coating variants then were tested in vitro and in vivo for improvement of desired responses compared to uncoated silicone rubber. All coating variants suppressed the adsorption of fibrinogen and immunoglobulin G, and most also inhibited fibroblast growth by 90-99%. None of the coating variants inhibited monocyte or neutrophil adhesion in vitro. However, the surfaces that supported the highest levels of monocyte adhesion also elicited the lowest secretion of pro-inflammatory cytokines. None of the materials elicited a strong inflammatory response or significantly (p< 0.05) reduced the thickness of the fibrous capsule when implanted subcutaneously in rats. Overall, the most passivating coating variant was an endpoint immobilized polypeptide that reduced protein adsorption, inhibited fibroblast growth by 90%, elicited low cytokine secretion from monocytes, and reduced fibrous encapsulation by 33%. In general, although some coating variants modified the adsorption of proteins and the behavior of leukocytes or fibroblasts in vitro, none abolished the development of a fibrous capsule in vivo.
The adhesion of bacteria on a biomaterial surface is believed to be the first step in the develop... more The adhesion of bacteria on a biomaterial surface is believed to be the first step in the development of biomaterial-related infection. The goal of this study was to investigate the mechanisms that permit adherent bacteria to persist on the surface of an implanted cardiovascular biomaterial. We hypothesized that circulating leukocytes are unable to adhere to the biomaterial surface under physiologic shear stress conditions, and this prevents them from interacting with adherent bacteria. To address this hypothesis, we investigated the adhesion profiles of Staphylococcus epidermidis and polymorphonuclear leukocytes (PMN), incubated under controlled shear stress conditions with the test biomaterial. We found that bacteria could adhere on the biomaterial surface, even when their concentration in the test medium was as low as 10(3) cfu/mL. At this concentration, the bacteria did not induce significant complement activation. PMN adhesion on the biomaterial surface was sensitive to shear stress and minimal at shear stress > 10 dynes/cm2. Low concentrations of bacteria could induce a significant increase in the expression of PMN adhesion molecules CD11b and CD11c. We conclude that the presence of bacteria induces PMN activation but does not increase PMN adhesion on biomaterial surfaces under physiologic shear stress conditions. This could be a major mechanism that protects adherent bacteria from PMN antibacterial activity.
It is generally accepted that biodegradation of poly(etheruethane urea) (PEUU) involves oxidation... more It is generally accepted that biodegradation of poly(etheruethane urea) (PEUU) involves oxidation of the polyether segments on the surface where leukocytes are adhered. The influence of dissolved oxygen, which is known to control oxidation of polymers in more traditional environments, was explored in this study. Specimens treated in vitro with hydrogen peroxide-cobalt chloride for 12 days exhibited a brittle, degraded surface layer about 10 microm thick. Attenuated total reflectance-Fourier transform infrared spectroscopy of the surface revealed that the ether absorbance at 1110 cm(-1) gradually decreased with in vitro treatment time to 30% of its initial value after 12 days. In contrast, 6 days in vitro followed by 6 days in air produced a decrease to 12% of the initial volume. Therefore, removing a specimen from the in vitro solution after 6 days and exposing it to air for the remainder of the 12 days actually resulted in more oxidation than leaving it in the in vitro solution for the entire 12 days. These results suggest that PEUU degrades by an autooxidation mechanism sustained by oxygen. By successfully modeling the depth of the surface degraded layer with a diffusion-reaction model, it was demonstrated that PEUU biodegradation is controlled by diffusion of oxygen into the polymer.
Human vascular endothelial cells (ECs) are exposed to various levels of hemodynamic forces, cycli... more Human vascular endothelial cells (ECs) are exposed to various levels of hemodynamic forces, cyclic strain, and shear stress in vivo. Here, we examined the in vitro effects of the various levels (0-6%, 7-16%, and 17-25%) of strain at 60, 30, and 15 cycles per minute (cpm) on human monocyte adherence to endothelial cells and extracellular matrix protein preabsorbed surfaces. Monocyte adhesion to endothelial cells under cyclic strain significantly increased. At both 30 and 60 cpm, ECs under strains of 7-16% and 17-25% showed >52% and >117% higher monocyte adhesion than endothelial cells under static condition when monocytes were added for 0.5 h. This increase in monocyte adhesion to ECs under cyclic strain remained significantly higher even after 24 h of incubation. Human monocyte adhesion to extracellular matrix protein preabsorbed surfaces differed depending on the specific extracellular matrix protein. Monocytes adhered to collagen type I and fibronectin preabsorbed surfaces &...
ABSTRACT This chapter presents our efforts to develop a better mechanistic understanding of how b... more ABSTRACT This chapter presents our efforts to develop a better mechanistic understanding of how biomaterial interactions with blood components lead to alteration of the basic pathophysiologic mechanisms, in particular, inflammation and the foreign body response, which increase the probability of bacterial interactions, colonization, biofilm formation, and infection. In particular, we present perspectives on mechanisms of S. epidermidis biofilm formation, the role of surface chemistry on biofilm formation, the role of bacterial slime production in device infections, the apoptosis of adherent polymorphonuclear leukocytes in the acute inflammatory response, neutrophil mobility and phagocytosis of bacteria on biomaterials, generation of reactive oxygen and nitrogen species by biomaterial-adherent neutrophils, and quorum sensing in S. epidermidis biofilm formation. Our work has focused on infection mechanisms of cardiovascular prostheses and devices where blood hemodynamics and shear stress play important roles in inflammatory cell interactions with biomaterial surfaces. However, we believe that much of our results also are applicable to static implant situations found in orthopedic, cosmetic (plastic), and other surgical areas.
A novel biofabrication modality, electrophoretic compaction with macromolecular alignment, was ut... more A novel biofabrication modality, electrophoretic compaction with macromolecular alignment, was utilized to make collagen threads that mimic the native tendon's structure and mechanical properties. A device with kinematic electrodes was designed to fabricate collagen threads in continuous length. For the first time, a 3D-biotextile was woven purely from collagen. Mechanical properties and load-displacement behavior of the biotextile mimicked those of the native tendon while presenting a porosity of 80%. The open pore network facilitated cell seeding across the continuum of the bioscaffold. Mesenchymal stem cells (MSCs) seeded in the woven scaffold underwent tenogenic differentiation in the absence of growth factors and synthesized a matrix that was positive for tenomodulin, COMP and type I collagen. Up-regulation of tenomodulin, a tendon specific marker, was 11.6 ± 3.5 fold, COMP was up-regulated 16.7 ± 5.5 fold, and Col I was up-regulated 6.9 ± 2.7 fold greater on ELAC threads when compared to randomly oriented collagen gels. These results demonstrate that a bioscaffold woven by using collagen threads with densely compacted and anisotropically aligned substrate texture stimulates tenogenesis topographically, rendering the electrochemically aligned collagen as a promising candidate for functional repair of tendons and ligaments.
Despite relatively sparse data regarding their outcomes in the setting of infection, biologic gra... more Despite relatively sparse data regarding their outcomes in the setting of infection, biologic grafts have gained rapid acceptance by the surgical community for complex hernia repair. These materials are heterogeneous in their procurement and processing techniques, which may ultimately have an impact in their ability to withstand infection. The objective of this study is to evaluate the impact of varying levels of contamination on biologic graft performance in a chronic ventral hernia animal model. Four commonly applied biologic grafts were used in the repair of a chronic ventral hernia rat model (n = 218). Each material was repaired in the setting of 1 of 4 surgical wound classifications (clean, clean contaminated, contaminated, dirty infected) with Staphylococcus aureus as our inoculum agent. After a 30-day survival, repairs underwent quantitative cultures, histological, and biomechanical testing. Marked differences were observed in biologic graft bacterial burden, biomechanical an...
Advances in experimental medicine and biology, 2011
Macrophages undergo fusion with other macrophages to form the hallmark multinucleated giant cells... more Macrophages undergo fusion with other macrophages to form the hallmark multinucleated giant cells of chronic inflammation. However, neither the existence of distinct morphological types of giant cells, the signaling pathways that induce their formation, the molecular mechanism(s) of macrophage fusion, nor the significance of macrophage multinucleation at chronic inflammatory sites are well understood. Our efforts have been focused on these unknowns, particularly as they relate to the foreign body-type giant cells that form on implanted biomaterials and biomedical devices. We have pursued the discoveries of human macrophage fusion factors (interleukin-4, interleukin-13, α-tocopherol) with emphasis on foreign body giant cells, and identified adhesion receptors and signaling intermediates, as well as an adhesion protein substrate (vitronectin) that supports macrophage fusion. Studies on the molecular mechanism of macrophage fusion have revealed it to be a mannose receptor-mediated phag...
This paper summarizes our recent e †orts to better understand the e †ects of antioxidants, the e ... more This paper summarizes our recent e †orts to better understand the e †ects of antioxidants, the e †ects of strain-state, mechanistic studies of soft segment cleavage by reactive oxygen radicals, and the e †ects of di †erent soft segment chemistries on the biostability/biodegradation of polyether polyurethanes (PEUUs). In vivo cage implant system studies and in vitro cobalt ion/ hydrogen peroxide studies have been carried out on PEUUs and the polymers have been analysed by attenuated total reÑectance and Fourier transform infrared (ATR-FTIR) spectroscopy, and scanning electron microscopic (SEM) characterization of the PEUU surfaces. The natural antioxidant, vitamin E, has been shown to inhibit biodegradation and enhance biostability of PEUUs. Studies of the e †ect of stress state on PEUU biodegradation demonstrate that stress can inhibit biodegradation. While polyether soft segments may be cleaved by the presence of reactive oxygen radicals, the presence of oxygen has a profound e †ect in accelerating biodegradation. The biodegradation of polyurethanes may be inhibited by substituting di †erent chemistries such as polydimethylsiloxanes, polycarbonates, and hydrocarbon soft segments for the polyether soft segments. To safely utilize polyurethanes in long-term biomedical devices, the biodegradation mechanisms of polyurethane elastomers must be fully understood and subsequently prevented. 1998 SCI.
The cyclic deformation of two polyurethane elastomers that differed in soft segment content and m... more The cyclic deformation of two polyurethane elastomers that differed in soft segment content and molecular weight was investigated. The microphase-separated morphology of the polyurethane with higher soft segment content consisted of hard segment domains dispersed in a soft segment matrix. In the polyurethane with lower soft segment content, the hard segment domains appeared to be partially cocontinuous. Following an initial 'conditioning' cycle, both polyurethanes exhibited reversible elastomeric behavior. Structural changes that occurred during conditioning were investigated using atomic force microscopy and Fourier transform infrared dichroism. The results provided the basis of a structural model for the deformation behavior. Yielding and reorganization of hard domains resulted in a highly oriented microfibrous morphology. Subsequent unloading and reloading were associated with reversible relaxation and reformation of the microfibrous entities. The elastic behavior of the conditioned polyurethanes was satisfactorily described by classical rubber theory with inextensibility. The structural model proposed here extended previous efforts to describe the deformation processes of polyurethanes during cyclic loading.
Empiric amphotericin B therapy was compared to central venous catheter Githdrawal in a prospectiv... more Empiric amphotericin B therapy was compared to central venous catheter Githdrawal in a prospective randomized trial. Of 32 febrile, neutropenic patients with indwelling Broviac catheters and without documented infection, 14 had persistent fever while receiving broad spectrum antibacterial drugs. Six patients were randomized to catheter removal and eight patients received amphotericin B empirically. None of six patients responded to catheter removal and six of eight defervesced after receiving antifungal therapy (p less than 0.01). Of the six patients in whom catheters were removed, two later became afebrile while on subsequent therapy with amphotericin B. Culture and histologic evaluation of the removed catheters failed to implicate the prosthesis as an infectious source. Central venous catheters in a persistently febrile neutropenic host need not be removed, unless local difficulties or bacteremia with skin commensal organisms occur. Amphotericin B can be infused through a central venous catheter in febrile, neutropenic patients unresponsive to empiric antibacterial drugs, with many patients becoming afebrile as a result of this therapy.
Journal of Materials Science: Materials in Medicine, 2006
The purpose of this communication is to present the author&am... more The purpose of this communication is to present the author's perspectives on the future of biomedical materials that were presented at the Larry L. Hench Retirement Symposium held at Imperial College, London, in late September 2005. The author has taken a broad view of the future of biomedical materials and has presented key ideas, concepts, and perspectives necessary for the future research and development of biomedical polymers and their future role as an enabling technology for the continuing progress of tissue engineering, regenerative medicine, prostheses, and medical devices. This communication, based on the oral presentation, is meant to be provocative and generate discussion. In addition, it is targeted for students and young scientists who will play an ever-increasing role in the future of biomedical materials.
Aseptic loosening of orthopedic implants is thought to be caused primarily by osteoclast differen... more Aseptic loosening of orthopedic implants is thought to be caused primarily by osteoclast differentiation induced by bone resorptive cytokines produced in response to phagocytosis of implant-derived wear particles. This study examined whether adherent endotoxin on the wear particles is responsible for inducing osteoclast differentiation as well as production of interleukin-1beta (IL-1beta), IL-6, and tumor necrosis factor a (TNF-alpha). Removal of adherent endotoxin almost completely inhibited the responses to titanium (Ti) particles by both murine marrow cells and human peripheral blood monocytes. In vivo experiments showed that endotoxin removal reduced particle-induced osteolysis by 50-70%. Addition of lipopolysaccharide (LPS) to the "endotoxin-free" particles restored their ability to induce cytokine production and osteoclast differentiation in vitro. Moreover, marrow cells from mice that are hyporesponsive to endotoxin because of mutation of Toll-like receptor 4 induced significantly less cytokine production and osteoclast differentiation in response to Ti particles with adherent endotoxin than did marrow cells from normoresponsive mice. This mutation also resulted in significantly less particle-induced osteolysis in vivo. Taken together, these results show that adherent endotoxin is involved in many of the biological responses induced by orthopedic wear particles and should stimulate development of new approaches designed to reduce the activity of adherent endotoxin in patients with orthopedic implants.
Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2014
Coating of various synthetic, absorbable, and biologic meshes with mesenchymal stem cells (MSCs) ... more Coating of various synthetic, absorbable, and biologic meshes with mesenchymal stem cells (MSCs) and fibroblasts was analyzed qualitatively and quantitatively. Five hernia meshes-light weight monofilament polypropylene (Soft Mesh), polyester (Parietex-TET), polylactide composite (TIGR), heavy weight monofilament polypropylene (Marlex), and porcine dermal collagen (Strattice)-were coated with three cell lines: human dermal fibroblasts (HFs), rat kidney fibroblasts (NRKs), and rat MSCs. Cell densities were determined at different time points. Samples also underwent histology and transmission electron microscopic (TEM) analyses. It required HFs 3 weeks to cover the entire mesh, while only 2 weeks for NRKs and MSCs to do so. MSCs had no preference for any of the meshes and produced the highest cell densities on Parietex and TIGR. Substrate-preference accounted for the significantly lower fibroblast densities on TIGR than Parietex. Fibroblasts failed to coat Marlex. Strattice, which had the least surface area, generated comparable cell densities to Parietex. Both histology and TEM confirmed cell coating of mesh surface. Various prosthetics can be coated by certain cell strains. Both mesh composition and cell preference dramatically influence the coating process. This methodology provides foundation for novel avenues of modulation of host response to various modern synthetic and biologic meshes.
The kinetics of attachment of NIL B and SV-NIL cells to glass, siliconized glass, and surfaces co... more The kinetics of attachment of NIL B and SV-NIL cells to glass, siliconized glass, and surfaces coated with random copolypeptides have been studied. It was found that in the absence of serum proteins, neither the rate nor the extent of attachment of cells is affected by the nature of the surface. In the presence of bovine serum albumin, the total uptake and rate of attachment of both NIL B and SV-NIL cells to the neutral, hydrophobic, and negatively charged copolymers is decreased compared with attachment to the same surfaces in the absence of protein. In contrast, the attachment of NIL B and SV-NIL cells to the positively charged (lysyl) copolymers was not decreased in the presence of protein. It was shown that the effect of protein resulted from its adsorption to the surface rather than to the cells. It was also concluded that both the NIL €3 and SV-NIL cells consist of a single cell population with respect to adhesiveness, and that both cell lines reach a kinetic equilibrium with the surfaces. This work represents one of the first studies to vary copolypeptide compositions systematically from negatively to neutral to positively charged surfaces and to examine these substrates without any mediating effects from various serum proteins. The results of this study support the concept that while cells bind to an adsorbed layer of protein rather than directly to the surface, the underlying surface can modify the attachment process by its effect on the protein adsorbed.
Poly(etherurethane) elastomers are useful materials in medical devices because of their mechanica... more Poly(etherurethane) elastomers are useful materials in medical devices because of their mechanical properties and biocompatibility. However, it is necessary to stabilize these elastomers against the oxidation of their ether soft segments. Synthetic antioxidants such as Santowhite and Irganox are often satisfactory; however, particularly for biomedical applications, it was of interest to test the natural antioxidant vitamin E in poly(etherurethane urea) (PEUU) elastomers in vivo. The alpha-tocopherol form of vitamin E was added to PEUU at 5% by weight. Biaxially strained PEUU specimens with and without vitamin E were tested in vivo in the cage implant system. The influence of vitamin E on PEUU biostability was analyzed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy and scanning electron microscopic (SEM) characterization of the PEUU surface. ATR-FTIR results showed that vitamin E prevented chemical degradation of the PEUU surface up to 5 weeks implantation, and at 10 weeks 82% of the ether remained. In contrast, without an antioxidant, only 18% of the ether remained after 10 weeks. No surface pitting or cracking was observed by SEM on PEUU with vitamin E; PEUU without antioxidant ruptured owing to extensive pitting and cracking. It was concluded that the antioxidant properties of vitamin E prevented oxidation of strained PEUU elastomers in vivo. The influence of vitamin E on PEUU biocompatibility was characterized by exudate leukocyte counts, density of leukocytes adherent to the PEUU, and morphology of adherent leukocytes. These results indicated decreased leukocyte counts in the exudate and less active adherent cells on the PEUU with vitamin E compared to PEUU without antioxidant. A proposed cell-polymer feedback system demonstrates how vitamin E improves both biostability and biocompatibility of PEUU elastomers in vivo.
Photochemical immobilization technology was utilized to covalently couple polymers to silicone ru... more Photochemical immobilization technology was utilized to covalently couple polymers to silicone rubber either at multiple points along a polymer backbone or at the endpoint of an amphiphilic chain. The coating variants then were tested in vitro and in vivo for improvement of desired responses compared to uncoated silicone rubber. All coating variants suppressed the adsorption of fibrinogen and immunoglobulin G, and most also inhibited fibroblast growth by 90-99%. None of the coating variants inhibited monocyte or neutrophil adhesion in vitro. However, the surfaces that supported the highest levels of monocyte adhesion also elicited the lowest secretion of pro-inflammatory cytokines. None of the materials elicited a strong inflammatory response or significantly (p< 0.05) reduced the thickness of the fibrous capsule when implanted subcutaneously in rats. Overall, the most passivating coating variant was an endpoint immobilized polypeptide that reduced protein adsorption, inhibited fibroblast growth by 90%, elicited low cytokine secretion from monocytes, and reduced fibrous encapsulation by 33%. In general, although some coating variants modified the adsorption of proteins and the behavior of leukocytes or fibroblasts in vitro, none abolished the development of a fibrous capsule in vivo.
The adhesion of bacteria on a biomaterial surface is believed to be the first step in the develop... more The adhesion of bacteria on a biomaterial surface is believed to be the first step in the development of biomaterial-related infection. The goal of this study was to investigate the mechanisms that permit adherent bacteria to persist on the surface of an implanted cardiovascular biomaterial. We hypothesized that circulating leukocytes are unable to adhere to the biomaterial surface under physiologic shear stress conditions, and this prevents them from interacting with adherent bacteria. To address this hypothesis, we investigated the adhesion profiles of Staphylococcus epidermidis and polymorphonuclear leukocytes (PMN), incubated under controlled shear stress conditions with the test biomaterial. We found that bacteria could adhere on the biomaterial surface, even when their concentration in the test medium was as low as 10(3) cfu/mL. At this concentration, the bacteria did not induce significant complement activation. PMN adhesion on the biomaterial surface was sensitive to shear stress and minimal at shear stress > 10 dynes/cm2. Low concentrations of bacteria could induce a significant increase in the expression of PMN adhesion molecules CD11b and CD11c. We conclude that the presence of bacteria induces PMN activation but does not increase PMN adhesion on biomaterial surfaces under physiologic shear stress conditions. This could be a major mechanism that protects adherent bacteria from PMN antibacterial activity.
It is generally accepted that biodegradation of poly(etheruethane urea) (PEUU) involves oxidation... more It is generally accepted that biodegradation of poly(etheruethane urea) (PEUU) involves oxidation of the polyether segments on the surface where leukocytes are adhered. The influence of dissolved oxygen, which is known to control oxidation of polymers in more traditional environments, was explored in this study. Specimens treated in vitro with hydrogen peroxide-cobalt chloride for 12 days exhibited a brittle, degraded surface layer about 10 microm thick. Attenuated total reflectance-Fourier transform infrared spectroscopy of the surface revealed that the ether absorbance at 1110 cm(-1) gradually decreased with in vitro treatment time to 30% of its initial value after 12 days. In contrast, 6 days in vitro followed by 6 days in air produced a decrease to 12% of the initial volume. Therefore, removing a specimen from the in vitro solution after 6 days and exposing it to air for the remainder of the 12 days actually resulted in more oxidation than leaving it in the in vitro solution for the entire 12 days. These results suggest that PEUU degrades by an autooxidation mechanism sustained by oxygen. By successfully modeling the depth of the surface degraded layer with a diffusion-reaction model, it was demonstrated that PEUU biodegradation is controlled by diffusion of oxygen into the polymer.
Uploads
Papers by James Anderson