High Quality Tissue Formation Method

One of the central objectives of this project was the development and characterization of a 3D mineralized tissue model system in which the effects of mechanical load (e.g., compression loading, tension, vibration, etc.) on the cellular responses of osteoblasts and osteoclasts could be investigated. After introducing mineralization agents to the culture, the constructs take on a bone-like appearance and have a more rigid structure suitable for being tested. Testing of the mineralized constructs confirmed the presence of calcium through a crystalline matrix histochemical stain. The central core is void of necrotic material, instead filled by a crystalline matrix with embedded nucleated cells. Remarkably, the nucleated cells do not express osteoblast markers, indicating differentiation to the in vivo cell type known as the osteocyte. In addition, as is characteristic to native periosteum, osteoclast precursor cells were imaged and proven to naturally arrange as an outer layer of the mineralized bone tissue construct. Development of this model will provide a unique venue for testing proposed countermeasures to space flight-induced bone loss. It will also allow a mechanistic approach in the modulation of cell signaling at the cellular level within the bone matrix. The Development And Characterization Of A Three-Dimensional Tissue Culture Model Of Bone is a technology readiness level (TRL) 6 (system/subsystem prototype demonstrated in a relevant environment). The innovation is now available for your company to license. Please note that NASA does not manufacture products itself for commercial sale.

Current scaffold designs and materials do not provide all of the appropriate cues necessary to mimic in-vivo conditions for tissue engineering and stem cell engineering applications. It has been hypothesized that many biomaterials, such as bone, muscle, brain and heart tissue exhibit piezoelectric and ferroelectric properties. Typical cell seeding environments incorporate biochemical cues and more recently mechanical stimuli, however, electrical cues have just recently been incorporated in standard in-vitro examinations. In order to develop their potential further, novel scaffolds are required to provide adequate cues in the in-vitro environment to direct stem cells to differentiate down controlled pathways or develop novel tissue constructs. This invention is for a scaffold that provides for such cues by mimicking the native biological environment, including biochemical, topographical, mechanical and electrical cues.

In vitro three-dimensional (3D) human broncho-epithelial (HBE) tissue-like assemblies (3D HBE TLAs or TLAs) were engineered in modeled microgravity using rotating wall vessel technology (pictured above) to mimic the characteristics of in vivo tissue. The TLAs were bioengineered onto collagen-coated cyclodextran beads using primary human mesenchymal bronchial-tracheal cells (HBTC) as the foundation matrix and an adult human broncho-epithelial immortalized cell line (BEAS-2B) as the overlying component. The resulting TLAs share significant characteristics with in vivo human respiratory epithelium including polarization, tight junctions, desmosomes, and microvilli. The presence of tissue-like differentiation markers including villi, keratins, and specific lung epithelium markers, as well as the production of tissue mucin, further confirm these TLAs have differentiated into tissues functionally like in vivo tissues. TLAs mimic aspects of the human respiratory epithelium and provide a unique capability to study the interactions of respiratory viruses and their primary target tissue independent of the host's immune system. The innovation "Methods For Growing Tissue-Like 3D Assemblies Of Human Broncho-Epithelial Cells" is at Technology Readiness Level (TRL) 6 (which means system/subsystem prototype demonstration in a relevant environment) and the related patent is now available to license for development into a commercial product. Please note that NASA does not manufacture products itself for commercial sale.

JSC's technology provides hazard-free, microgravity-compatible hardware for DNA/RNA isolation. It also allows PCR analysis to be used outside the lab in environments where pipetting is difficult and/or where hazardous chemicals must be confined to an enclosed container, such as military settings and remote clinical operations. This self-contained device for isolating DNA/RNA, proteins, and cells is a component system that includes syringes and pistons, membranes of different capacities, reagents, four-way valves, and small pumps. The pre-filled reagents are the same as those used in conventional PCR laboratory isolation analysis. The DNA and RNA isolation kits are novel and process small sample amounts using a self-enclosed and pipette-free technique. Multiple kits can be stacked to allow several samples to be processed simultaneously. The system can be used in conjunction with existing analysis modules, such as commercially available DNA instruments. The process can be fully automated and programmed and can potentially be applied to other biological processes. The JSC innovation will permit the extension of laboratory isolation protocols to many applications. This NASA Technology is available for your company to license and develop into a commercial product. NASA does not manufacture products for commercial sale.

Once genes for a desired material type, delivery mode, control method and affinity have been chosen, assembling the genetic components and creating the cell lines can be done with well-established synthetic biology techniques. A 3D microdeposition system is used to make a 3D array of these cells in a precise, microstructure pattern and shape. The engineered cells are suspended in a printable 'ink'. The 3D microdeposition system deposits minute droplets of the cells onto a substrates surface in a designed print pattern. Additional printer passes thicken the material. The cell array is fed nutrients and reagents to activate the engineered genes within the cells to create and deposit the desired molecules. These molecules form the designed new material. If desired, the cells may be removed by flushing. The end product is thus a 3D composite microstructure comprising the novel material. This innovation provides a fast, controlled production of natural, synthetic, and novel biomaterials with minimum resource overhead and reduced pre- and post-processing requirements.