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Biomedical subjects

Karen J L Burg

Publications and source records attributed to Karen J L Burg.

8 recordsLinked to original sources

Grafting amine-terminated branched architectures from poly(L-lactide) film surfaces for improved cell attachment.

Poly(L-lactide) (PLL) has been used as a bioabsorbable material in the medical and pharmaceutical fields. The unmodified hydrophobic PLL surface generally has low cell affinity; thus, modification of PLL film surface properties is necessary to improve its use as a biomaterial. Our surface modification method involved the use of photografting and typical wet chemistry to create branched architectures containing amine functionalities on the periphery of the grafted layers. Amine (-NH2) groups were first introduced on the PLL film surface by photoinduced grafting of 4,4'-diaminobenzophenone and the grafted branched architectures were created by subsequent reactions with succinic acid and tris(2-aminoethyl) amine. The resulting film surface was analyzed using contact angle goniometry and X-ray photoelectron spectroscopy. MC3T3 fibroblasts were cultured on unmodified PLL film and PLL films grafted with the branched structures and the films were subsequently analyzed by optical microscopy. The contact angle goniometry results showed an initial decrease and subsequent plateau in the water contact angles for the PLL films with each successive generation of the branched architectures. The X-ray photoelectron spectroscopy data provided insight into the structure of the grafted layer and revealed an increase in the nitrogen content with each generation. Optical micrographs showed enhanced cell attachment and viability on the surface-modified PLL films.

Amines↗

Stem cells and adipose tissue engineering.

A large proportion of the plastic and reconstructive surgical procedures performed each year are to repair soft tissue defects that result from traumatic injury, tumor resection, and congenital defects. These defects typically result from the loss of a large volume of adipose tissue. To date, no ideal filler material which is successful in all cases has been developed. Additionally, the success of using autologous fat tissue grafts to repair soft tissue defects has been limited. Researchers are thus investigating strategies to engineer volumes of adipose tissue that may be used in these cases. A necessary component for engineering a viable tissue construct is an appropriate cell source. Attempts to engineer adipose tissue have involved the use of preadipocytes and adipocytes as the base cell source. Increased interest surrounding the research and development of stem cells as a source of cells for tissue engineering has, however, led to a new path of investigation for developing adipose tissue-engineering strategies. This manuscript serves as a review of the current state of adipose tissue-engineering methods and describes the shift toward tissue-engineering strategies using stem cells.

Adipocytes↗

Photopatterned polymer brushes promoting cell adhesion gradients.

The ability to spatially control cellular adhesion in a continuous manner on a biocompatible substrate is an important factor in designing new biomaterials for use in wound healing and tissue engineering applications. In this work, a novel method of engineering cell-adhesive RGD-ligand density gradients to control specific cell adhesion across a substrate is presented. Polymer brushes exhibiting spatially defined gradients in chain density are created and subsequently functionalized with RGD to create ligand density gradients capable of inducing cell adhesion on an otherwise weakly adhesive substrate. Cell studies indicate that these ligand-functionalized surfaces are noncytotoxic, with cellular adhesion increasing with RGD-ligand density across the gradient brush surface.

3T3 Cells↗

Biodegradable microparticles based on poly(D,L-lactide) as a protective transport system in ruminant digestion.

Despite its abundance in their diet, cattle are unable to directly digest cellulose. The bovine digestive tract overcomes this problem via the rumen, a portion of the stomach containing mixed anaerobic bacteria. These microbes, while breaking down foodstuffs, also perform undesirable processes such as biohydrogenation, in which unsaturated fatty acids become saturated, with deleterious cardiovascular effects. An approach to preventing this saturation entailing the use of polymeric microspheres to encapsulate feed supplements is proposed, with a single emulsion, solvent evaporation method used to formulate poly(D,L-lactide) microparticles for delivery of unsaturated fatty acids to ruminant abomasum.

Animals↗

Evaluation of polymer scaffolds to be used in a composite injectable system for intervertebral disc tissue engineering.

Adult porcine nucleus pulposus cells were seeded onto gelatin, demineralized bone matrix (DBM), and polylactide scaffolds and cultured in vitro. Cellular behavior in response to the scaffolds was analyzed using biochemical assays, histology, and real-time quantitative reverse transcriptase-polymerase chain reaction. Scanning electron microscopy showed pronounced differences in surface texture of the scaffolds. Nucleus pulposus cells attached and assumed an elongated fibroblast-like morphology on the gelatin and DBM scaffolds. The cells cultured on the gelatin and DBM were metabolically active and expressed types I and II collagen and aggrecan. Detached cellular aggregates with a rounded morphology were noted in the culture tubes containing the polylactide scaffolds. Both surface chemistry and texture likely had a role in causing differences in cellular behavior in response to scaffold material. Promising results were observed using the gelatin and demineralized bone scaffolds, but the behavior of cells cultured on these scaffolds will need to be characterized further. This initial research will be used to direct future work involved in developing this composite injectable tissue engineering system.

Animals↗

Characterization of cellular carriers for use in injectable tissue-engineering composites.

Injectable composite tissue-engineering scaffolds are systems that incorporate individual cell carriers within a gel delivery matrix. This study assessed low-temperature casting as a possible method to produce synthetic cell-carrier beads. Porous poly-L-lactide beads were manufactured by low-temperature casting. Two porogens, either glucose or sodium chloride, were incorporated into the beads and subsequently leached. Beads were seeded with primary culture aortic smooth muscle rat cells and were evaluated over a 13-day period using a series of chemical, biochemical, and histological assays. Results indicate that low-temperature casting is a viable technique to produce injectable beads on the order of 1.5-2.0 mm. The manufactured beads supported smooth muscle cell attachment and proliferation; where the beads formed with sodium chloride allowed enhanced proliferation. Differences in physical qualities, namely buoyancy and topography, were dependent on porogen selection and may provide a mechanism for bead and composite customization.

Animals↗

Design and evaluation of a novel flow chamber for measuring cell adhesion to absorbable polymer films.

There is great interest in improving cellular attachment to synthetic materials, particularly for developing small diameter tissue-engineered vascular grafts. However, limited research has been conducted to evaluate the adhesion characteristics of different cell types to absorbable substrates. Tissue engineered vessels typically fail as a result of delamination of the endothelial cell layer when exposed to fluid or blood flow. The focus of this research was to design and evaluate a flow chamber, using fibroblasts, smooth muscle cells, and endothelial cells, to probe the bounds of the system. A flow chamber was designed and fabricated to compare the relative adhesion characteristics of cells to absorbable polymer films. A preliminary investigation of mouse fibroblast (3T3M) adhesion to semicrystalline poly-L-lactide (PLL) films was conducted to determine general operating specifications. Cell coverage on films was evaluated using a live-dead assay and image analysis; following exposure to flow, tests were similarly conducted. Based on these results, additional studies were conducted to compare the adhesion of rat aortic smooth muscle cells (SMC) and endothelial cells (EC) on PLL films.

3T3 Cells↗

Evaluation of smooth muscle cell response using two types of porous polylactide scaffolds with differing pore topography.

The goal of tissue engineering is to create bioartificial tissues for the replacement of failed or nonfunctional tissue. Porous tissue-engineered scaffolds may be created through a solvent-casting/porogen-leaching technique. Almost exclusively, sodium chloride (NaCl) is the porogen of choice. Previous studies have demonstrated the importance of porosity and pore size in cell adhesion and tissue development, yet the impact of porogen morphology and the chemical effect of porogen residual has not been fully explored. Poly-L-lactide (PLLA) scaffolds were manufactured by a solvent-casting, particulate-leaching method with either glucose or NaCl porogen in an effort to vary pore characteristics and, subsequently, cell adhesion and tissue development. Porogen influence on scaffold morphology and topography was compared via histological techniques and qualitative surface characteristics. Using an in vitro model, scaffolds were seeded with rat aortic smooth muscle cells (SMCs) and evaluated over a 28-day period. Cell attachment and proliferation were subsequently evaluated. Results indicate that initial SMC attachment is higher for scaffolds manufactured with NaCl rather than glucose. The proliferation of SMCs was higher for scaffolds manufactured with glucose and, by day 28, scaffolds manufactured with glucose supported a higher cell population than those processed using NaCl porogen.

Animals↗