Development of methods for studying the differentiation of human mesenchymal stem cells under cyclic compressive strain.

Human mesenchymal stem cells (hMSC) have numerous potential advantages over terminally differentiated cells and embryonic stem cells for use in tissue engineering applications. The aims of this study were to develop methods to test the hypothesis that hMSC could be differentiated using cyclic compressive strain alone. hMSC were successfully isolated, purified using D7-FIB antibody, cloned, and characterized. The cells were subsequently analyzed using fluorescence-activated cell sorting using a panel of antibodies and differentiation into multiple cell lineages. D7FIB-positive cells were then seeded into collagen-alginate scaffolds and subjected to 10% or 15% cyclic compressive strain for 4 out of 24 hours for up to 21 days in a bespoke servo-assisted displacement-controlled device. Cells were analyzed using adenosine triphosphate assay to determine cell number, live-dead cell assay, and quantitative real-time polymerase chain reaction at 7 and 21 days. Cloned D7-FIB-positive hMSCs showed evidence of differentiation to an osteogenic lineage under 10% cyclic compressive strain alone (core binding factor alpha 1 (CBFA-1) was significantly upregulated at 7 and 21 days by a factor of 18.3 and 32.2, respectively) and to an osteo-chondrogenic lineage under 15% cyclic compressive strain alone (increased expression of CBFA-1, Sox9, and aggrecan). A combination of a composite viscoelastic scaffold and controlled cyclic compressive strain may be useful for study of the differentiation of MSC.

Conservation value of clustered housing developments.

Traditionally, exurban lands in Colorado have been subdivided into a grid of parcels ranging from 2 to 16 ha. From an ecological perspective, this dispersed pattern of development effectively maximizes the individual influence of each home on the land. Clustered housing developments, designed to maximize open space, are assumed to benefit plant and wildlife communities of conservation interest. They have become a popular alternative for rural development despite the lack of empirical evidence demonstrating their conservation benefits. To better inform rural land-use planning, we evaluated clustered housing developments by comparing their spatial pattern with that of dispersed housing developments and by comparing their conservation value with that of both dispersed housing developments and undeveloped areas in Boulder County, Colorado. We used four indicators to assess conservation value: (1) densities of songbirds, (2) nest density and survival of ground-nesting birds, (3) presence of mammals, and (4) percent cover and proportion of native and non-native plant species. Clustered and dispersed housing developments did not differ on the majority of variables we examined. Both types of housing development had significantly higher densities of non-native and human-commensal species and significantly lower densities of native and human-sensitive species than undeveloped areas. More rigorous ecological guidelines and planning on a regional scale may help create clustered developments with higher conservation value.

Tissue engineering of cardiac valves: re-seeding of acellular porcine aortic valve matrices with human mesenchymal progenitor cells.

BACKGROUND AND AIM OF THE STUDY: Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study investigated the potential for re-seeding an acellular porcine heart valve matrix using human mesenchymal progenitor cells (MPC). METHODS: MPC were isolated from the bone marrow of patients undergoing hip replacement operations. Putative MPC were then cultured in several differentiation media in order to determine the multipotential differentiation capacity of the cells. The MPC were also characterized by FACS analysis. Cells at passage 8 were then seeded at between 1 x 10(4) and 1 x 10(5) cells/cm2 onto a decellularized porcine aortic valve matrix, and recellularization of the matrix was assessed. The phenotype of the re-seeded cells and re-seeded cell density was then determined by histology and immunohistochemistry. RESULTS: Putative MPC were successfully isolated and differentiated into cells of the adipogenic, neurogenic, and myogenic lineages. FACS analysis showed the cells to have a similar phenotype to those isolated by others (CD45-, CD13+, D7FIB+, CD105+, CD10+/-, LNGFR+/-, CD55+, BMP- and AP+/-). Cells seeded onto an acellular valve matrix penetrated the center of the tissue after four weeks to 2% of homograft cell density. Phenotypic analysis of the cells in the re-seeded matrix revealed the cells to have a similar phenotype to native valve interstitial cells (vimentin+, alpha-smooth muscle actin+, heavy chain myosin slow-, desmin-). However, re-seeded cells also expressed osteogenic markers (alkaline phosphatase, osteonectin, and osteopontin). CONCLUSION: This study has shown, for the first time, that human MPC have the capacity to infiltrate an acellular porcine valve matrix under static conditions in vitro. Future studies will comprise culture under pulsatile flow in a physiological heart valve bioreactor to maintain the desired cell phenotype and increase cell density.

Impacts of a Severe Drought on Grassland Birds in Western North Dakota.

We studied the effect of a severe drought on the population dynamics and community structure of grassland birds in western North Dakota. During the spring and summer of 1988 the northern Great Plains suffered one of the warmest, driest periods in its recorded history. We compared the changes in bird populations and nesting productivity over a 3-yr period before, during, and after the drought. Total grassland bird density declined 61% (P < .05) between June 1987 and June 1988. Densities of six of eight common species declined significantly during the drought. Populations of all but two species recovered in 1989 and total bird density in June 1989 did not differ significantly from June 1987. Species richness and species diversity both declined significantly during the drought and recovered to predrought levels in 1989. Species richness declined more on fair condition than on good condition range during the drought. Vesper Sparrow (Pooecetes gramineus) hatching success, number of young fledged per successful nest, and nesting success were significantly lower in 1988 than either 1987 or 1989. Clutch size did not differ among the three years. The decline in nesting success in 1988 was primarily due to nest abandonment during incubation. Nesting of Vesper Sparrows, Horned Larks (Eremophila alpestris), and Western Meadowlarks (Sturnella neglecta) ended abruptly in mid-June 1988 during a period of extremely hot weather. In 1987 and 1989, nesting continued into July. Despite substantial reductions in bird density and productivity during the drought, many species recovered to predrought levels 1 yr following the drought. This suggests that year-to-year fluctuations in densities of some of these species may not be tightly linked to short-term changes in local productivity. However, sequential years of low productivity may have more substantial effects on these short-lived species. Thus, if drought conditions in North American grasslands become more frequent, as some climate models predict, there could be related changes in the avifauna of the region.

Human Disturbance of an Avian Scavenging Guild.

In order to investigate the effects of human activities on relationships within foraging guilds, we examined inacanus dynamics of eagles, crows, and gulls scavenging on spawned salmon in the Pacific Northwest. We examined several hypotheses that postulate the asymmetric foraging relationships of the three guild members and that reveal the influence of competition and facilitation in these relationships. Spatial and temporal patterns of resource use by the three primary guild members varied with the presence and absence of human activity at experimental feeding stations. At control (undisturbed) stations, eagles preferred to feed >100 m from vegetative cover, whereas gulls fed <50 m from cover. At experimental (disturbed) stations, eagles rarely fed, and feeding activity by gulls increased at both near and far stations. Crows often fed on alternate food sources in fields adjacent to the river, especially when salmon carcasses were scarce, whereas eagles and gulls rarely did so. We also examined if and how the behavior of single guild members changes in the presence or absence of other guild members. In the absence of eagles, gulls and crows preferred stations far from cover, numbers of both increased at feeding stations, birds were distributed nearer to carcasses, and they fed more. We emphasize that guild theory lends important insights to our understanding of the effects of human disturbance on wildlife communities.