Biomedical subjects

W Mark Saltzman

Publications and source records attributed to W Mark Saltzman.

28 records · Page 2Linked to original sources

PubMed2003

Gene expression and mucosal immune responses after vaginal DNA immunization in mice using a controlled delivery matrix.

IgA antibodies in the vaginal tract are essential as a first defense line against microorganisms that enter the body via mucosal surfaces. Several studies have shown that direct application of DNA to the vaginal mucosal surface can induce secretion of IgA molecules specific to the expressed protein. The major challenge of formulating effective vaccines is to overcome the barriers to DNA administration caused by the estrus cycle and physical environment of the vaginal tract. In this study, we investigated whether controlled delivery of DNA to the vaginal surface would induce long-term IgA antibody production by applying controlled delivery matrices to the vaginal tract. The controlled DNA delivery matrices were composed of poly(ethylene-co-vinyl acetate) (EVAc) and loaded with a model plasmid encoding sperm-specific lactate dehydrogenase C(4) (LDH-C(4)). These EVAc matrices provided a controlled and sustained DNA release to the vaginal mucosal surface. The DNA released from the EVAc disks was functionally active and capable of transfecting vaginal tissues. When inserted into the vaginal tract of mice, the DNA-loaded EVAc matrices triggered the immune system and induced specific IgA to LDH-C(4) in the vaginal secretions. These results demonstrate that the EVAc disks are efficient and convenient vehicles for delivering DNA to the vaginal tract and providing long-term local immunity.

Administration, Intravaginal↗

PubMed2003

Diffusion of nerve growth factor in rat striatum as determined by multiphoton microscopy.

Neurotrophins such as nerve growth factor (NGF) may be useful for treating diseases in the central nervous system; our ability to harness the potential therapeutic benefit of NGF is directly related to our understanding of the fate of exogenously supplied factors in brain tissue. We utilized multiphoton microscopy to quantify the dynamic behavior of NGF in coronal, 400- micro m thick, fresh rat brain tissue slices. We administered a solution containing bioactive rhodamine nerve growth factor conjugate via pressure injection and monitored the dispersion in the striatal region of the coronal slices. Multiphoton microscopy facilitated repeated imaging deep ( approximately 200 micro m) into tissue slices with minimal photodamage of tissue and photobleaching of label. The pressure injection paradigm approximated diffusion from a point source, and we therefore used the corresponding solution to the diffusion equation to estimate an apparent diffusion coefficient in brain tissue (D(b)(34 degrees C)) of 2.75 +/- 0.24 x 10(-7) cm(2)/s (average +/- SE). In contrast, we determined a corresponding free diffusion coefficient in buffered solution (D(f)(34 degrees C)) of 12.6 +/- 0.9 x 10(-7) cm(2)/s using multiphoton fluorescence photobleaching recovery. The tortuosity, defined as the square root of the ratio of D(f) to D(b), was 2.14 and moderate in magnitude.

Algorithms↗

PubMed2003

Biomimetic design in microparticulate vaccines.

Current efforts to improve the effectiveness of microparticle vaccines include incorporating biomimetic features into the particles. Many pathogens use surface molecules to target specific cell types in the gut for host invasion. This observation has inspired efforts to chemically conjugate cell-type targeting ligands to the surfaces of microparticles in order to increase the efficiency of uptake, and therefore the effectiveness, of orally administered microparticles. Bio-mimicry is not limited to the exterior surface of the microparticles. Anti-idiotypic antibodies, cytokines or other biological modifiers can be encapsulated for delivery to sites of interest as vaccines or other therapeutics. Direct mucosal delivery of microparticle vaccines or immunomodulatory agents may profoundly enhance mucosal and systemic immune responses compared to other delivery routes.

Animals↗

PubMed2002

Topographical control of human neutrophil motility on micropatterned materials with various surface chemistry.

Controlling cell responses to an implantable material is essential to tissue engineering. Because the surface is in direct contact with cells, both chemical and topographical properties of a material surface can play a crucial role. In this study, parallel ridges/grooves were micropatterned on glass surfaces using photosensitive polyimide to create transparent substrates. The migratory behavior of live human neutrophils on the patterned surfaces was observed using a light microscope with transmitted light source. The width (2 microm) and length (400 microm) of the ridges were kept constant. The height (5 or 3 microm) and the repeat spacing (6-14 microm) of the ridges were systematically changed to investigate the effect of microgeometry on neutrophil migration. In addition, the effect of surface chemistry on neutrophil migration was studied by deposition of a thin layer of "inert", biocompatible metal such as Au-Pd alloy and titanium on patterned substrates. More than 95% of neutrophils moved in the direction of the long axis of ridges/grooves regardless of the topographical geometry and chemistry, consistent with a phenomenon termed "contact guidance". Therefore, cell migration was characterized using a one-dimensional persistent random walk. The rate of cell movement was strongly dependent on the topographical microgeometry of the ridges. The random motility coefficient mu, 9.8 x 10(-9) cm2/s, was the greatest at a ridge height of 5 microm and spacing of 10 microm, about 10 times faster than on smooth glass surface. The Au-Pd coating did not change neutrophil migratory behavior on patterned surfaces, whereas titanium decreased cell motility substantially. The results of this study suggest that optimization of both surface chemistry and topography may be important when designing biomaterials for tissue engineering. In addition, parallel ridges/grooves can be used to control the direction and rate of cell migration on the surface.

Biocompatible Materials↗

PubMed2002

Building drug delivery into tissue engineering.

The creation of efficient methods for manufacturing biotechnology drugs--many of which influence fundamental but complex cell behaviours, such as proliferation, migration and differentiation--is creating new opportunities for tissue repair. Many agents are potent and multifunctional; that is, they produce different effects within different tissues. Therefore, control of tissue concentration and spatial localization of delivery is essential for safety and effectiveness. Synthetic systems that can control agent delivery are particularly promising as materials for enhancing tissue regeneration. This review discusses the state of the art in controlled-release and microfluidic drug delivery technologies, and outlines their potential applications for tissue engineering.

Drug Delivery Systems↗

PubMed2002

Pharmacokinetics of the carmustine implant.

Controlled release delivery of carmustine from biodegradable polymer wafers was approved as an adjunct to surgical resection in the treatment of recurrent glioblastoma multiforme after it was shown in clinical trials to be well tolerated and effective. Given the localised nature of the drug in the brain tissue, no direct pharmacokinetic measurements have been made in humans after implantation of a carmustine wafer. However, drug distribution and clearance have been extensively studied in both rodent and non-human primate brains at various times after implantation. In addition, studies to characterise the degradation of the polymer matrix, the release kinetics of carmustine and the metabolic fate of the drug and polymer degradation products have been conducted both in vitro and in vivo. GLIADEL wafers have been shown to release carmustine in vivo over a period of approximately 5 days; when in continuous contact with interstitial fluid, wafers should degrade completely over a period of 6 to 8 weeks. Metabolic elimination studies of the polymer degradation products have demonstrated that sebacic acid monomers are excreted from the body in the form of expired CO(2), whereas 1,3-bis-(p-carboxyphenoxy)propane monomers are excreted primarily through the urine. Carmustine degradation products are also excreted primarily through the urine. Pharmacokinetic studies in animals and associated modelling have demonstrated the capability of this modality to produce high dose-delivery (millimolar concentrations) within millimetres of the polymer implant, with a limited penetration distance of carmustine from the site of delivery. The limited spread of drug is presumably due to the high transcapillary permeability of this lipophilic molecule. However, the presence of significant convective flows due to postsurgical oedema may augment the diffusive transport of drug in the hours immediately after wafer implantation, leading to a larger short-term spread of drug. Additionally, in non-human primates, the presence of significant doses in more distant regions of the brain (centimetres away from the implant) has been shown to persist over the course of a week. The drug in this region was presumed to be transported from the implant site by either cerebral blood flow or cerebrospinal fluid flow, suggesting that although drug is able to penetrate the blood-brain barrier at the site of delivery, it may re-enter within the confines of the brain tissue.

Animals↗

PubMedDate not supplied

Improving the expansion and neuronal differentiation of mesenchymal stem cells through culture surface modification.

Poly-D-lysine, poly-L-lysine, collagen, laminin, fibronectin, and Matrigel were compared with standard tissue grade polystyrene for their impact on the expansion and neuronal differentiation of mesenchymal stem cells (MSCs). Among these substrates, adsorption of Matrigel at 5 microg/cm2 did not enhance cell proliferation but gave rise to the highest percentage of MSC-derived neuron-like cells with the best morphological differentiation. Matrigel at a higher coating density of 50 microg/cm2 not only further enhanced the differentiation but also significantly improved cell expansion. In contrast, poly-D-lysine did not effectively support the growth of MSCs. Hence the expansion and neuronal differentiation of MSCs both depend on surface properties of the culture substrate. These results could lead to a culture process with improved yield of MSC-derived neuron-like cells and to novel biomaterials for tissue engineering.

Cell Culture Techniques↗

PubMedDate not supplied

In vitro cytotoxicity and in vivo distribution after direct delivery of PEG-camptothecin conjugates to the rat brain.

Low water solubility and rapid elimination from the brain inhibits local delivery via implants and other delivery systems of most therapeutic drugs to the brain. We have conjugated the chemotherapy drug, camptothecin (CPT), to poly(ethylene glycol) (PEG) of molecular weight 3400 using previously established protocols. These new conjugates are very water-soluble and hydrolyze at a pH-dependent rate to release the active parent drug. We have studied the uptake of these conjugates by cells in vitro and quantified their cytotoxicity toward gliosarcoma cells. These conjugates were loaded into biodegradable polymeric controlled-release implants, and their release characteristics were studied in vitro. We implanted similar polymeric disks into rat brains and used a novel sectioning scheme to determine the concentration profile of CPT in comparison to conjugated CPT in the brain after 1, 7, 14, and 28 days. We have found that PEGylation greatly increases the maximum achievable drug concentration and greatly enhances the distribution properties of CPT, compared to corelease of CPT with PEG. Although only one percent of CPT in the conjugate system was found in the hydrolyzed, active form, drug concentrations were still significantly above cytotoxic levels over a greater distance for the conjugate system. On the basis of these results, we believe that PEGylation shows great promise toward increasing drug distribution after direct, local delivery in the brain for enhanced efficacy in drug treatment.

Animals↗

PubMedDate not supplied

Cellular fate of a modular DNA delivery system mediated by silica nanoparticles.

Development of efficient molecular medicines, including gene therapeutics, RNA therapeutics, and DNA vaccines, depends on efficient means of transfer of DNA or RNA into the cell. Potential problems, including toxicity and immunogenicity, surrounding viral methods of DNA delivery have necessitated the use of nonviral, synthetic carriers. To better design synthetic carriers, or transfection reagents, the modular design of viruses has inspired a modular approach to DNA and RNA delivery. Each modular component can be designed to circumvent each of the many barriers. The modular approach will allow modification of individual components for a specific application. By utilizing a dense silica nanoparticle to form a ternary complex, transfection efficiency of a DNA-transfection reagent complex was increased by a factor of approximately 10 by concentrating the DNA at the surface of cells. Surface modification of the silica nanoparticles allowed determination of the cellular uptake mechanism with only minor alteration of transfection efficiency. Nanoparticles are internalized by an endosome-lysosomal route followed by perinuclear accumulation. The modification mechanism confirms that surface modification of the modular system can allow specific moieties to be incorporated into the modular system without significant alteration of the transfection efficiency. By showing that the modular system based upon concentration of DNA at the level of the cell can be used to increase transfection efficiency, we have shown that further modification of the system may better target DNA delivery and overcome other barriers of DNA expression.

Animals↗

PubMedDate not supplied

Cost-reducing protein production and delivery for sexually transmitted disease prevention.

Table 2 summarizes our two approaches: vaginal delivery of antibodies to immunize passively and edible microspheres for active immunization. These approaches have different roles in disease prevention in communities and each may be most useful for different kinds of diseases. The passive approach could easily be extended to lots of different sexually transmitted diseases by just changing the specificity of the antibody incorporated into the device. The active immunization approach requires more specific development steps for each particular type of disease. Detailed analysis of the cost-reduction potential of these approaches is not yet available, but estimates of the cost to manufacture these products can be made, based on the initial laboratory studies (Table 4). The results to date suggest that both products are feasible and that they could be manufactured as cheaply as other products (such as condoms and oral contraceptives) that people currently use for sexual health. All products, once made available, could have a significant impact on the reduction of healthcare costs.

Administration, Intravaginal↗