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At least 757 records · Page 42
All sourcesNASA NTRSDOE OSTI
NASA NTRS2012

Architecting the Human Space Flight Program with Systems Modeling Language (SysML)

The next generation of missions in NASA's Human Space Flight program focuses on the development and deployment of highly complex systems (e.g., Orion Multi-Purpose Crew Vehicle, Space Launch System, 21st Century Ground System) that will enable astronauts to venture beyond low Earth orbit and explore the moon, near-Earth asteroids, and beyond. Architecting these highly complex system-of-systems requires formal systems engineering techniques for managing the evolution of the technical features in the information exchange domain (e.g., data exchanges, communication networks, ground software) and also, formal correlation of the technical architecture to stakeholders' programmatic concerns (e.g., budget, schedule, risk) and design development (e.g., assumptions, constraints, trades, tracking of unknowns). This paper will describe how the authors have applied System Modeling Language (SysML) to implement model-based systems engineering for managing the description of the End-to-End Information System (EEIS) architecture and associated development activities and ultimately enables stakeholders to understand, reason, and answer questions about the EEIS under design for proposed lunar Exploration Missions 1 and 2 (EM-1 and EM-2).

scheduling↗
NASA NTRS2013

Unique Challenges Testing SDRs for Space

This paper describes the approach used by the Space Communication and Navigation (SCaN) Testbed team to qualify three Software Defined Radios (SDR) for operation in space and the characterization of the platform to enable upgrades on-orbit. The three SDRs represent a significant portion of the new technologies being studied on board the SCAN Testbed, which is operating on an external truss on the International Space Station (ISS). The SCaN Testbed provides experimenters an opportunity to develop and demonstrate experimental waveforms and applications for communication, networking, and navigation concepts and advance the understanding of developing and operating SDRs in space. Qualifying a Software Defined Radio for the space environment requires additional consideration versus a hardware radio. Tests that incorporate characterization of the platform to provide information necessary for future waveforms, which might exercise extended capabilities of the hardware, are needed. The development life cycle for the radio follows the software development life cycle, where changes can be incorporated at various stages of development and test. It also enables flexibility to be added with minor additional effort. Although this provides tremendous advantages, managing the complexity inherent in a software implementation requires a testing beyond the traditional hardware radio test plan. Due to schedule and resource limitations and parallel development activities, the subsystem testing of the SDRs at the vendor sites was primarily limited to typical fixed transceiver type of testing. NASA s Glenn Research Center (GRC) was responsible for the integration and testing of the SDRs into the SCaN Testbed system and conducting the investigation of the SDR to advance the technology to be accepted by missions. This paper will describe the unique tests that were conducted at both the subsystem and system level, including environmental testing, and present results. For example, test waveforms were developed to measure the gain of the transmit system across the tunable frequency band. These were used during thermal vacuum testing to enable characterization of the integrated system in the wide operational temperature range of space. Receive power indicators were used for Electromagnetic Interference tests (EMI) to understand the platform s susceptibility to external interferers independent of the waveform. Additional approaches and lessons learned during the SCaN Testbed subsystem and system level testing will be discussed that may help future SDR integrators

Chelmins, David↗
NASA NTRS2013

Unified Simulation and Analysis Framework for Deep Space Navigation Design

As the technology that enables advanced deep space autonomous navigation continues to develop and the requirements for such capability continues to grow, there is a clear need for a modular expandable simulation framework. This tool's purpose is to address multiple measurement and information sources in order to capture system capability. This is needed to analyze the capability of competing navigation systems as well as to develop system requirements, in order to determine its effect on the sizing of the integrated vehicle. The development for such a framework is built upon Model-Based Systems Engineering techniques to capture the architecture of the navigation system and possible state measurements and observations to feed into the simulation implementation structure. These models also allow a common environment for the capture of an increasingly complex operational architecture, involving multiple spacecraft, ground stations, and communication networks. In order to address these architectural developments, a framework of agent-based modules is implemented to capture the independent operations of individual spacecraft as well as the network interactions amongst spacecraft. This paper describes the development of this framework, and the modeling processes used to capture a deep space navigation system. Additionally, a sample implementation describing a concept of network-based navigation utilizing digitally transmitted data packets is described in detail. This developed package shows the capability of the modeling framework, including its modularity, analysis capabilities, and its unification back to the overall system requirements and definition.

Anzalone, Evan↗
NASA NTRS2014

Developing an Integration Infrastructure for Distributed Engine Control Technologies

Turbine engine control technology is poised to make the first revolutionary leap forward since the advent of full authority digital engine control in the mid-1980s. This change aims squarely at overcoming the physical constraints that have historically limited control system hardware on aero-engines to a federated architecture. Distributed control architecture allows complex analog interfaces existing between system elements and the control unit to be replaced by standardized digital interfaces. Embedded processing, enabled by high temperature electronics, provides for digitization of signals at the source and network communications resulting in a modular system at the hardware level. While this scheme simplifies the physical integration of the system, its complexity appears in other ways. In fact, integration now becomes a shared responsibility among suppliers and system integrators. While these are the most obvious changes, there are additional concerns about performance, reliability, and failure modes due to distributed architecture that warrant detailed study. This paper describes the development of a new facility intended to address the many challenges of the underlying technologies of distributed control. The facility is capable of performing both simulation and hardware studies ranging from component to system level complexity. Its modular and hierarchical structure allows the user to focus their interaction on specific areas of interest.

distributed control↗
NASA NTRS2016

An Ultra-Wideband Millimeter-Wave Phased Array

Wideband millimeter-wave arrays are of increasing importance due to their growing use in high data rate systems, including 5G communication networks. In this paper, we present a new class of ultra-wideband millimeter wave arrays that operate from nearly 20 GHz to 90 GHz. The array is based on tightly coupled dipoles. Feeding designs and fabrication challenges are presented, and a method for suppressing feed resonances is provided.

Tightly Coupled Dipole Array↗
NASA NTRS2016

Systems and Methods for Collaboratively Controlling at Least One Aircraft

An unmanned vehicle management system includes an unmanned aircraft system (UAS) control station controlling one or more unmanned vehicles (UV), a collaborative routing system, and a communication network connecting the UAS and the collaborative routing system. The collaborative routing system being configured to receive flight parameters from an operator of the UAS control station and, based on the received flight parameters, automatically present the UAS control station with flight plan options to enable the operator to operate the UV in a defined airspace.

Estkowski, Regina I.↗
NASA NTRS2016

Real-time Identification and Control of Satellite Signal Impairments Solution and Application of the Stratonovich Equation Part 1. Theoretical Development

As satellite communications systems become both more complex and reliant with respect to their operating environment, it has become imperative to be able to identify, during real-time operation, the onset of one or more impairments to the quality of overall communications system integrity. One of the most important aspects to monitor of a satellite link operating within the Earth's atmosphere is the signal fading due to the occurrence of rain and/or phase scintillations. This, of course, must be done in the presence of the associated measurement uncertainty or potentially faulty measurement equipment such as in the Advanced Communication Technology Satellite (ACTS) experiment. In the present work, an approach originally suggested in 1991, and apparently still considered iconoclastic, will be significantly developed and applied to the satellite communications link on which the deleterious composite signal fade is the result of one or many component fade mechanisms. Through the measurement (with the attendant uncertainty or 'error' in the measurement) of such a composite fading satellite signal, it is desired to extract the level of each of the individual fading mechanisms so they can be appropriately mitigated before they impact the overall performance of the communications network. Rather than employing simple-minded deterministic filtering to the real-time fading, the present approach is built around all the models and/or descriptions used to describe the individual fade components, including their dynamic evolution. The latter is usually given by a first-order Langevin equation. This circumstance allows the description of the associated temporal transition probability densities of each of the component processes. By using this description, along with the real-time measurements of the composite fade (along with the measurement errors), one can obtain statistical estimates of the levels of each of the component fading mechanisms as well as their predicted values into the future. This is all accomplished by the use of the well-known Stratonovich integro-differential equation that results from the model of the measured signal fade that is also tailored to adaptively adjust the values of the parameters used in the statistical models of the individual fade mechanisms. Three examples of increasing complexity are addressed and solved for the iterative determination of fade component levels from the measured composite signal fade in the presence of measurement error and, in the last case, with uncertainty in the model parameters.

Countermeasure↗
NASA NTRS2017

Osteogenic Transcription Regulated by Exaggerated Stretch Loading via Convergent Wnt Signaling

Cell and animal studies conducted onboard the International Space Station and formerly the Shuttle flights have provided data illuminating the deleterious biological response of bone to mechanical unloading. Down regulation of proliferative mechanisms within stem cell populations of the osteogenic niche is a suggested mechanism for loss of bone mass. However the intercellular communicative cues from osteoblasts and osteocytes in managing stem cell proliferation and osteogenic differentiation are largely unknown. In this investigation, MLO-Y4 osteocyte-like and MC3T3-E1 osteoblast-like cells, are co-culture under dynamic tensile conditions and evaluated for phenotypic expression of biochemical signaling proteins influential in driving stem cell differentiation. MLO-Y4 and MC3T3-E1 were co-cultured on polyethersulfone membrane with a 0.45m porosity to permit soluble factor transfer and direct cell-cell gap junction signaling. Cyclic tensile stimulation was applied for 48 h at a frequency of 0.1Hz and strain of 0.1. Total Live cell counts indicate mechanical activation of MC3T3-E1s inhibits proliferation while MLO-Y4s increase in number. However, the percent of live MLO-Y4s within the population is low (46.3 total count, *p0.05, n4) suggesting a potential apoptotic signaling cascade. Immunofluorescence demonstrated that stimulation of co-cultures elicits increased gap junction communication. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggest that an osteocyte-osteoblast signaling feedback mechanism may regulate mechanotransduction of an apoptotic cascade within osteocytes and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.

mechanical stimulation↗
NASA NTRS2016

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture.

Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesion/attachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.

mechanical loading↗
NASA NTRS2016

Experimenting Galileo on Board the International Space Station

The SCaN Testbed is an advanced integrated communications system and laboratory facility installed on the International Space Station (ISS) in 2012. The testbed incorporates a set of new generation of Software Defined Radio (SDR) technologies intended to allow researchers to develop, test, and demonstrate new communications, networking, and navigation capabilities in the actual environment of space. Qascom, in cooperation with ESA and NASA, is designing a Software Defined Radio GalileoGPS Receiver capable to provide accurate positioning and timing to be installed on the ISS SCaN Testbed. The GalileoGPS waveform will be operated in the JPL SDR that is constituted by several hardware components that can be used for experimentations in L-Band and S-Band. The JPL SDR includes an L-Band Dorne Margolin antenna mounted onto a choke ring. The antenna is connected to a radio front end capable to provide one bit samples for the three GNSS frequencies (L1, L2 and L5) at 38 MHz, exploiting the subharmonic sampling. The baseband processing is then performed by an ATMEL AT697 processor (100 MIPS) and two Virtex 2 FPGAs. The JPL SDR supports the STRS (Space Telecommunications Radio System) that provides common waveform software interfaces, methods of instantiation, operation, and testing among different compliant hardware and software products. The standard foresees the development of applications that are modular, portable, reconfigurable, and reusable. The developed waveform uses the STRS infrastructure-provided application program interfaces (APIs) and services to load, verify, execute, change parameters, terminate, or unload an application. The project is divided in three main phases. 1)Design and Development of the GalileoGPS waveform for the SCaN Testbed starting from Qascom existing GNSS SDR receiver. The baseline design is limited to the implementation of the single frequency Galileo and GPS L1E1 receiver even if as part of the activity it will be to assess the feasibility of a dual frequency implementation (L1E1+L5E5a) in the same SDR platform.2)Qualification and test the GalileoGPS waveform using ground systems available at the NASA Glenn Research Center. Experimenters can have access to two SCaN Testbed ground based systems for development and verification: the Experimenter Development System (EDS) that is intended to provide initial opportunity for software testing and basic functional validation and the Ground Integration Unit (GIU) that is a high fidelity version of the SCaN Testbed flight system and is therefore used for more controlled final development testing and verification testing.3)Perform in-orbit validation and experimentation: The experimentation phase will consists on the collection of raw measurements (pseudorange, Carrier phase, CN0) in space, assessment on the quality of the measurements and the receiver performances in terms of signal acquisition, tracking, etc. Finally computation of positioning in space (Position, Velocity and time) and assessment of its performance.(Complete abstract in attached document).

GPS↗
NASA NTRS2016

Experimenting Galileo on Board the International Space Station

The SCaN Testbed is an advanced integrated communications system and laboratory facility installed on the International Space Station (ISS) in 2012. The testbed incorporates a set of new generation of Software Defined Radio (SDR) technologies intended to allow researchers to develop, test, and demonstrate new communications, networking, and navigation capabilities in the actual environment of space. Qascom, in cooperation with ESA and NASA, is designing a Software Defined Radio GalileoGPS Receiver capable to provide accurate positioning and timing to be installed on the ISS SCaN Testbed. The GalileoGPS waveform will be operated in the JPL SDR that is constituted by several hardware components that can be used for experimentations in L-Band and S-Band. The JPL SDR includes an L-Band Dorne Margolin antenna mounted onto a choke ring. The antenna is connected to a radio front end capable to provide one bit samples for the three GNSS frequencies (L1, L2 and L5) at 38 MHz, exploiting the subharmonic sampling. The baseband processing is then performed by an ATMEL AT697 processor (100 MIPS) and two Virtex 2 FPGAs. The JPL SDR supports the STRS (Space Telecommunications Radio System) that provides common waveform software interfaces, methods of instantiation, operation, and testing among different compliant hardware and software products. The standard foresees the development of applications that are modular, portable, reconfigurable, and reusable. The developed waveform uses the STRS infrastructure-provided application program interfaces (APIs) and services to load, verify, execute, change parameters, terminate, or unload an application. The project is divided in three main phases. 1)Design and Development of the GalileoGPS waveform for the SCaN Testbed starting from Qascom existing GNSS SDR receiver. The baseline design is limited to the implementation of the single frequency Galileo and GPS L1E1 receiver even if as part of the activity it will be to assess the feasibility of a dual frequency implementation (L1E1+L5E5a) in the same SDR platform.2)Qualification and test the GalileoGPS waveform using ground systems available at the NASA Glenn Research Center. Experimenters can have access to two SCaN Testbed ground based systems for development and verification: the Experimenter Development System (EDS) that is intended to provide initial opportunity for software testing and basic functional validation and the Ground Integration Unit (GIU) that is a high fidelity version of the SCaN Testbed flight system and is therefore used for more controlled final development testing and verification testing.3)Perform in-orbit validation and experimentation: The experimentation phase will consists on the collection of raw measurements (pseudorange, Carrier phase, CN0) in space, assessment on the quality of the measurements and the receiver performances in terms of signal acquisition, tracking, etc. Finally computation of positioning in space (Position, Velocity and time) and assessment of its performance.(Complete abstract in attached document).

Galileo↗
NASA NTRS2017

Recent Technology Advances in Distributed Engine Control

This presentation provides an overview of the work performed at NASA Glenn Research Center in distributed engine control technology. This is control system hardware technology that overcomes engine system constraints by modularizing control hardware and integrating the components over communication networks.

Distributed Control↗
NASA NTRS2017

Considerations for an Integrated UAS CNS Architecture

The National Aeronautics and Space Administration (NASA) Glenn Research Center (GRC) is investigating revolutionary and advanced universal, reliable, always available, cyber secure and affordable Communication, Navigation, Surveillance (CNS) options for all altitudes of UAS operations. In Spring 2015, NASA issued a Call for Proposals under NASA Research Announcements (NRA) NNH15ZEA001N, Amendment 7 Subtopic 2.4. Boeing was selected to conduct a study with the objective to determine the most promising candidate technologies for Unmanned Air Systems (UAS) air-to-air and air-to-ground data exchange and analyze their suitability in a post-NextGen NAS environment. The overall objectives are to develop UAS CNS requirements and then develop architectures that satisfy the requirements for UAS in both controlled and uncontrolled air space. This contract is funded under NASAs Aeronautics Research Mission Directorates (ARMD) Aviation Operations and Safety Program (AOSP) Safe Autonomous Systems Operations (SASO) project and proposes technologies for the Unmanned Air Systems Traffic Management (UTM) service.There is a need for accommodating large-scale populations of Unmanned Air Systems (UAS) in the national air space. Scale obviously impacts capacity planning for Communication, Navitation, and Surveillance (CNS) technologies. For example, can wireless communications data links provide the necessary capacity for accommodating millions of small UASs (sUAS) nationwide? Does the communications network provide sufficient Internet Protocol (IP) address space to allow air traffic control to securely address both UAS teams as a whole as well as individual UAS within each team? Can navigation and surveillance approaches assure safe route planning and safe separation of vehicles even in crowded skies?Our objective is to identify revolutionary and advanced CNS alternatives supporting UASs operating at all altitudes and in all airspace while accurately navigating in the absence of navigational aids. These CNS alternatives must be reliable, redundant, always available, cyber-secure, and affordable for all types of vehicles including small UAS to large transport category aircraft. The approach will identify CNS technology candidates that can meet the needs of the range of UAS missions to specific air traffic management applications where they will be most beneficial and cost effective.

Templin, Fred L.↗
NASA NTRS2017

Time-Coordination Strategies and Control Laws for Multi-Agent Unmanned Systems

Time-critical coordination tools for unmanned systems can be employed to enforce the type of temporal constraints required in terminal control areas, ensure minimum distance requirements among vehicles are satisfied, and successfully perform coordinated missions. In comparison with previous literature, this paper presents an ampler spectrum of coordination and temporal specifications for unmanned systems, and proposes a general control law that can enforce this range of constraints. The constraint classification presented con- siders the nature of the desired arrival window and the permissible coordination errors to define six different types of time-coordination strategies. The resulting decentralized coordination control law allows the vehicles to negotiate their speeds along their paths in response to information exchanged over the communication network. This control law organizes the different members in the fleet hierarchically per their behavior and informational needs as reference agent, leaders, and followers. Examples and simulation results for all the coordination strategies presented demonstrate the applicability and efficacy of the coordination control law for multiple unmanned systems.

Puig-Navarro, Javier↗
NASA NTRS2017

Considerations for an Integrated UAS CNS Architecture

The National Aeronautics and Space Administration (NASA) Glenn Research Center (GRC) is investigating revolutionary and advanced universal, reliable, always available, cyber secure and affordable Communication, Navigation, Surveillance (CNS) options for all altitudes of UAS operations. In Spring 2015, NASA issued a Call for Proposals under NASA Research Announcements (NRA) NNH15ZEA001N, Amendment 7 Subtopic 2.4. Boeing was selected to conduct a study with the objective to determine the most promising candidate technologies for Unmanned Air Systems (UAS) air-to-air and air-to-ground data exchange and analyze their suitability in a post-NextGen NAS environment. The overall objectives are to develop UAS CNS requirements and then develop architectures that satisfy the requirements for UAS in both controlled and uncontrolled air space. This contract is funded under NASAs Aeronautics Research Mission Directorates (ARMD) Aviation Operations and Safety Program (AOSP) Safe Autonomous Systems Operations (SASO) project and proposes technologies for the Unmanned Air Systems Traffic Management (UTM) service.There is a need for accommodating large-scale populations of Unmanned Air Systems (UAS) in the national air space. Scale obviously impacts capacity planning for Communication, Navigation, and Surveillance (CNS) technologies. For example, can wireless communications data links provide the necessary capacity for accommodating millions of small UASs (sUAS) nationwide? Does the communications network provide sufficient Internet Protocol (IP) address space to allow air traffic control to securely address both UAS teams as a whole as well as individual UAS within each team? Can navigation and surveillance approaches assure safe route planning and safe separation of vehicles even in crowded skies?Our objective is to identify revolutionary and advanced CNS alternatives supporting UASs operating at all altitudes and in all airspace while accurately navigating in the absence of navigational aids. These CNS alternatives must be reliable, redundant, always available, cyber-secure, and affordable for all types of vehicles including small UAS to large transport category aircraft. The approach will identify CNS technology candidates that can meet the needs of the range of UAS missions to specific air traffic management applications where they will be most beneficial and cost effective.

Templin, Fred L.↗
NASA NTRS2018

An Overview of Science Challenges Pertaining to our Understanding of Extreme Geomagnetically Induced Currents

Vulnerability of man-made infrastructure to Earth-directed space weather events is a serious concern for today's technology-dependent society. Space weather-driven geomagnetically induced currents (GICs) can disrupt operation of extended electrically conducting technological systems. The threat of adverse impacts on critical technological infrastructure, like power grids, oil and gas pipelines, and communication networks, has sparked renewed interest in extreme space weather. Because extreme space weather events have low occurrence rate but potentially high impact, this presents a major challenge for our understanding of extreme GIC activity. In this chapter, we discuss some of the key science challenges pertaining to our understanding of extreme events. In addition, we present an overview of GICs including highlights of severe impacts over the last 80 years and recent U.S. Federal actions relevant to this community.

Ngwira, Chigomezyo M.↗
NASA NTRS2016

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture

Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesionattachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.

Juran, C. M.↗