Identifying a site for maximum delivery of oxygen to transplanted cells

For in vivo cell implantation techniques to be successful, the energy and metabolic substrate requirement of the cells being grown must be met. Certain cells with high-energy requirements (e.g., hepatocytes, pancreatic island cells) experience a high degree of cell death after implantation due to a limited supply of oxygen. We proposed that the pleural cavity might be an oxygen-rich environment and hence an excellent site for cell implantation. To test the hypothesis that the delivery of oxygen to the pleural cavity is directly proportional to the inspired oxygen concentration we measured the pO(2) of saline instilled in the pleural cavity as compared to that of the peritoneal cavity. We postulated that the physiologic basis for any difference was the result of direct diffusion of oxygen into the pleural space across the alveoli. The study was conducted on sheep (n = 6), after induction of general anesthesia, in two phases, control and experimental. Saline was instilled into the peritoneal and pleural cavities via catheters, after equilibration at given FiO(2), the pO(2) of the paline aspirated from the two cavities was compared. In the experimental group, animals were sacrificed (no circulation) and ventilated. The same sequence of steps as in the control phase were repeated. In the control group, the pO(2) of saline aspirated from the pleural cavity approached the arterial pO(2) at all FiO(2) levels. The pO(2) of the peritoneal saline aspirate fell over time. In the experimental phase (no circulation), the pO(2) of the pleural cavity saline rose to \u3e400 mm Hg. We conclude that this is a result of direct diffusion and is a potential source of unlimited oxygen supply not dependent on vascular supply

In Vitro Analog Of Human Bone Marrow From 3D Scaffolds With Biomimetic Inverted Colloidal Crystal Geometry

In vitro replicas of bone marrow can potentially provide a continuous source of blood cells for transplantation and serve as a laboratory model to examine human immune system dysfunctions and drug toxicology. Here we report the development of an in vitro artificial bone marrow based on a 3D scaffold with inverted colloidal crystal (ICC) geometry mimicking the structural topology of actual bone marrow matrix. To facilitate adhesion of cells, scaffolds were coated with a layer of transparent nanocomposite. After seeding with hematopoietic stem cells (HSCs), ICC scaffolds were capable of supporting expansion of CD34+ HSCs with B-lymphocyte differentiation. Three-dimensional organization was shown to be critical for production of B cells and antigen-specific antibodies. Functionality of bone marrow constructs was confirmed by implantation of matrices containing human CD34+ cells onto the backs of severe combined immunodeficiency (SCID) mice with subsequent generation of human immune cells. © 2008 Elsevier Ltd

A composite tissue-engineered trachea using sheep nasal chondrocyte and epithelial cells

This study evaluates the feasibility of producing a composite engineered tracheal equivalent composed of cylindrical cartilaginous structures with lumens lined with nasal epithelial cells. Chondrocytes and epithelial cells isolated from sheep nasal septum were cultured in Ham\u27s F12 media. After 2 wk, chondrocyte suspensions were seeded onto a matrix of polyglycolic acid. Cell-polymer constructs were wrapped around silicon tubes and cultured in vitro for 1 wk, followed by implanting into subcutaneous pockets on the backs of nude mice. After 6 wk, epithelial cells were suspended in a hydrogel and injected into the embedded cartilaginous cylinders following removal of the silicon tube. Implants were harvested 4 wk later and analyzed. The morphology of implants resembles that of native sheep trachea. HandE staining shows the presence of mature cartilage and formation of a pseudo-stratified columnar epithelium, with a distinct interface between tissue-engineered cartilage and epithelium. Safranin-O staining shows that tissue-engineered cartilage is organized into lobules with round, angular lacunae, each containing a single chondrocyte. Proteoglycan and hydroxyproline contents are similar to native cartilage. This study demonstrates the feasibility of recreating the cartilage and epithelial portion of the trachea using tissue harvested in a single procedure. This has the potential to facilitate an autologous repair of segmental tracheal defects

Testosterone injection stimulates net protein synthesis but not tissue amino acid transport

Testosterone administration (T) increases lean body mass and muscle protein synthesis. We investigated the effects of short-term T on leg muscle protein kinetics and transport of selected amino acids by use of a model based on arteriovenous sampling and muscle biopsy. Fractional synthesis (FSR) and breakdown (FBR) rates of skeletal muscle protein were also directly calculated. Seven healthy men were studied before and 5 days after intramuscular injection of 200 mg of testosterone enanthate. Protein synthesis increased twofold after injection (P less than 0.05), whereas protein breakdown was unchanged. FSR and FBR calculations were in accordance, because FSR increased twofold (P less than 0.05) without a concomitant change in FBR. Net balance between synthesis and breakdown became more positive with both methodologies (P less than 0.05) and was not different from zero. T injection increased arteriovenous essential and nonessential nitrogen balance across the leg (P less than 0.05) in the fasted state, without increasing amino acid transport. Thus T administration leads to an increased net protein synthesis and reutilization of intracellular amino acids in skeletal muscle