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Duke, P. J.

Publications and source records attributed to Duke, P. J..

Human Bone-Forming Chondrocytes Cultured in the Hydrodynamic Focusing Bioreactor Retain Matrix Proteins: Similarities to Spaceflight Results

Fracture healing, crucial to a successful Mars mission, involves formation of a cartilaginous fracture callus which differentiates, mineralizes, ossifies and remodels via the endochondral process. Studies of spaceflown and tailsuspended rats found that, without loading, fracture callus formation and cartilage differentiation within the callus were minimal. We found delayed differentiation of chondrocytes within the rat growth plate on Cosmos 1887, 2044, and Spacelab 3. In the current study, differentiation of human bone-forming chondrocytes cultured in the hydrodynamic focusing bioreactor (HFB) was assessed. Human costochondral chondrocytes in suspension were aggregated overnight, then cultured in the HFB for 25 days. Collagen Type II, aggrecan and unsulfated chondroitin were found extracellularly and chondroitin sulfates 4 and 6 within the cell. Lack of secretion was also found in pancreatic cells of spaceflown rats, and in our SL3 studies. The HFB can be used to study cartilage differentiation in simulated microgravity.

Duke, P. J.

[Dentists on Mars]

The oral health of astronauts, last studied in the mid- 1970s on Skylab, has not been a priority area of study for NASA due to the short length of shuttle stays. But with longer stays on the International Space Station, and planning for a trip to Mars, investigations into how spaceflight affects oral health are needed. The objective of this symposium is to introduce the dental research community to changes occurring in humans who go into space, and how these changes might impact oral health. Dr. Millie Hughes-Fulford, an astronaut-scientist, will review what happens to humans who go into space, and the difficulties of living, and carrying out experiments in space. She will then discuss her research on cell cultures of osteoblasts in space, and in hypergravity Dr. Gerald Sonnenfeld will review immunological changes that occur in spaceflight, and relate the observed decreases in various types of immunological responses to possible effects on oral immunological factors. Dr. Marian Lewis will discuss the effects of spaceflight on gene expression using results from her spaceflight experiments on various cell types. Dr. Jack van Loon, fiom the Dutch Experiment Support Center, will review what is known about bone loss in humans and rats and metatarsal cultures which go into space, and review ground based models (head down bedrest, and tail suspension) that simulate the unloading of spaceflight. Attendees will gain a knowledge of spaceflight research, and information on getting their own experiments in space. The symposium is supported by the NASA Office of Bioastronautics

Duke, P. J.

Galacturonomannan and Golgi-derived membrane linked to growth and shaping of biogenic calcite

The coccolithophores are valuable models for the design and synthesis of composite materials, because the cellular machinery controlling the nucleation, growth, and patterning of their calcitic scales (coccoliths) can be examined genetically. The coccoliths are formed within the Golgi complex and are the major CaCO(3) component in limestone sediments-particularly those of the Cretaceous period. In this study, we describe mutants lacking a sulfated galacturonomannan and show that this polysaccharide in conjunction with the Golgi-derived membrane is directly linked to the growth and shaping of coccolith calcite but not to the initial orientated nucleation of the mineral phase.

NASA Discipline Cell Biology

The Spacelab 3 simulation: basis for a model of growth plate response in microgravity in the rat

Data from Spacelab 3 (SL3) suggested that spaceflight significantly reduces the activity of the rat tibial growth plate. Animal processing after SL3 began twelve hours post-landing, so data reflect post-flight re-adaptation in addition to spaceflight effects. To determine if a twelve-hour period of weight bearing after seven days of unloading could affect the physes of spaceflown rats, the present study assessed the growth plate response to unloading with or without a reloading period. Rats were subjected to hind-limb suspension for seven days and then euthanized, with or without twelve hours of reloading. Activity of the growth plate was assessed by morphometric analysis. Rats suspended without reloading had reserve zone (RZ) height greater than controls, and shorter hypertrophy/calcification zone (HCZ) with fewer cells. The greater RZ was associated with a larger cell area, indicating a possible mitotic delay or secretion defect. Twelve hours of reloading decreased RZ height and cell number, and restored the number of cells in HCZ to control values, but the number of cells in the proliferative zone and height in HCZ were reduced. These results suggest the rebound response to preserve/restore skeletal function after a period of unloading involves an acceleration of growth associated with a decreased cell cycle time in PZ. Changes during the reloading period in this simulation support our hypothesis that the effects of spaceflight on SL3 growth plates were altered by changes that occurred post-landing. The similarities in response to unloading by suspension or during spaceflight are used to propose a model of growth plate response during spaceflight.

manned

Studies of chondrogenesis in rotating systems

A great deal of energy has been exerted over the years researching methods for regenerating and repairing bone and cartilage. Several techniques, especially bone implants and grafts, show great promise for providing a remedy for many skeletal disorders and chondrodystrophies. The bioreactor (rotating-wall vessel, RWV) is a cell culture system that creates a nurturing environment conducive to cell aggregation. Chondrocyte cultures have been studied as implants for repair and replacement of damaged and missing bone and cartilage since 1965 [Chesterman and Smith, J Bone Joint Surg 50B:184-197, 1965]. The ability to use large, tissue-like cartilage aggregates grown in the RWV would be of great clinical significance in treating skeletal disorders. In addition, the RWV may provide a superior method for studying chondrogenesis and chondrogenic mutations. Because the RWV is also reported to simulate many of the conditions of microgravity it is a very useful ground-based tool for studying how cell systems will react to microgravity.

Non-NASA Center

Cartilage formation in the CELLS 'double bubble' hardware

The CELLS experiment scheduled to be flown on the first International Microgravity Laboratory is designed to study the effect of microgravity on the cartilage formation, by measuring parameters of growth in a differentiating cartilage cell culture. This paper investigates the conditions for this experiment by studying cartilage differentiation in the 'bubble exchange' hardware with the 'double bubble' design in which the bubbles are joined by a flange which also overlays the gasket. Four types of double bubbles (or double gas permeable membranes) were tested: injection-molded bubbles 0.01- and 0.005-in. thick, and compression molded bubbles 0.015- and 0.01-in. thick. It was found that double bubble membranes of 0.005- and 0.010-in. thickness supported cartilage differentiation, while the 0.015-in. bubbles did not. It was also found that nodule count, used in this study as a parameter, is not the best measure of the amount of cartilage differentiation.

Duke, P. J.

Experiment K-6-06. Morphometric and EM analyses of tibial epiphyseal plates from Cosmos 1887 rats

Light and electron microscopy studies were carried out on decalcified tibial epiphyseal plates of rats flown aboard Cosmos 1887 (12.5d flight plus 53.5h recovery). Analysis of variance showed that the proliferative zone of flight animals was significantly higher than that of synchronous controls, while the hypertrophic/calcification zone was significantly reduced. Flight animals had more cells than synchronous controls in the proliferative zone, and less in the hypertrophic/calcification region. The total number of cells, however, was significantly higher in flight animals. No differences were found for perimeter or shape factor of growth plates, but area was significantly lower in flight animals in comparison to synchronous controls. Collagen fibrils in flight animals were shorter and wider than in synchronous controls. The time required for a cell to cycle through the growth plate is 2 to 3 days, so most of the cells and matrix present were formed after the animals had returned to 1 g, and probably represent stages of recovery from microgravity exposure, which in itself is an interesting question.

Duke, P. J.

Histomorphometric and electron microscopic analyses of tibial epiphyseal plates from Cosmos 1887 rats

Previous studies have shown that the changes seen in the bones of growing rats exposed to microgravity are due in part to changes that occur in the growth plate during spaceflight. In this study, growth plates of rats flown aboard Cosmos 1887 (12.5-day flight plus 53.5-h recovery at 1 g) were analyzed using light and electron microscopy and computerized planimetry. The proliferative zone of flight animals was found to be significantly (P less than or equal to 0.01) larger than that of controls, while the reserve and hypertrophic/calcification zones were significantly reduced. Flight animals also had more cells per column in the proliferative zone than did controls and less in the hypertrophic/calcification region. The total number of cells, however, was significantly greater in flight animals. No difference was found in perimeter or in shape factor, but area was significantly less in flight animals. Electron microscopy showed that collagen fibrils in flight animals were wider than in controls. Since the time required for a cell to cycle through the growth plate is 2-3 days at 1 g, the results reported here represent both the effects of exposure to microgravity and the initial stages of recovery from that exposure.

Non-NASA Center

Growth and differentiation of mammalian embryonic tissues exposed to hypergravity in vivo and in vitro

In about 10 years or so, men and women from Earth will be long-term inhabitants of a space station aboard which plants and animals will be growing and developing in gravities other than that of Earth. The effect of gravitational changes on development was examined. It is indicated that differentiation is speeded up under excess G and slowed in low or null G. The effects of exposure to excess gravity on fusion of the embryonic mouse secondary palate were studied. During fusion, the palatal shelves first adhere by means of glycoproteins appearing along the medial epithelial edge (MEE). The contacting epithelia then reorganize and undergo programmed cell death, allowing the underlying mesenchymes to come in contact. The process of cell death occurs in vitro at about the same rate that it occurs in vivo.

Duke, P. J.