Non-destructive, continuous monitoring of biochemical, mechanical, and structural maturation in engineered tissue.

Regulatory guidelines for tissue engineered products require stringent characterization during production and necessitate the development of novel, non-destructive methods to quantify key functional parameters for clinical translation. Traditional assessments of engineered tissues are destructive, expensive, and time consuming. Here, we introduce a non-destructive, inexpensive, and rapid sampling and analysis system that can continuously monitor the mechanical, biochemical, and structural properties of a single sample over extended periods of time. The label-free system combines the imaging modalities of fluorescent lifetime imaging and ultrasound backscatter microscopy through a fiber-based interface for sterile monitoring of tissue quality. We tested the multimodal system using tissue engineered articular cartilage as an experimental model. We identified strong correlations between optical and destructive testing. Combining FLIm and UBM results, we created a novel statistical model of tissue homogeneity that can be applied to tissue engineered constructs prior to implantation. Continuous monitoring of engineered tissues with this non-destructive system has the potential for in-process monitoring of tissue engineered products, reducing costs and improving quality controls in research, manufacturing, and clinical applications.

Multimodal Label-Free Imaging for Detecting Maturation of Engineered Osteogenic Grafts.

There is a critical need to develop noninvasive, nondestructive methods for assessing the quality of engineered constructs prior to implantation. Currently, the composition and maturity of engineered tissues are assessed using destructive, costly, and time-consuming biochemical and mechanical analyses. The goal of this study was to use noninvasive, multimodal imaging to monitor osteogenic differentiation and matrix deposition by human mesenchymal stem/stromal cells (MSCs) during in vitro culture. MSCs were encapsulated in alginate hydrogels and cultured in osteogenic conditions for 4 weeks. Samples were evaluated using fluorescence lifetime imaging (FLIm) and ultrasound backscatter microscopy (UBM) prior to traditional biochemical and mechanical testing. Using linear regression analysis, we identified strong correlations between imaging parameters (e.g., fluorescence lifetime and acoustic attenuation coefficient) and destructive mechanical and biochemical tests to assess the maturation of osteogenically induced constructs. These data demonstrate the promise of nondestructive label-free imaging techniques to noninvasively ascertain the progression and maturity of tissue engineered bone grafts.

Impact of Shoulder Abduction Loading on Brain-Machine Interface in Predicting Hand Opening and Closing in Individuals With Chronic Stroke.

BACKGROUND: Many individuals with moderate and severe stroke are unable to use their paretic hand. Currently, the effect of conventional therapy on regaining meaningful hand function in this population is limited. Efforts have been made to use brain-machine interfaces (BMIs) to control hand function. To date, almost all BMI classification algorithms are designed for detecting hand movements with a resting arm. However, many functional movements require simultaneous movements of the arm and hand. Arm movement will possibly affect the detection of intended hand movements, specifically for individuals with chronic stroke who have muscle synergies. The most prevalent upper-extremity synergy-flexor synergy-is expressed as an abnormal coupling between shoulder abductors and elbow/wrist/finger flexors. OBJECTIVE: We hypothesized that because of flexor synergy, shoulder abductor activity would affect the detection of the hand-opening (a movement inhibited by flexion synergy) but not the hand-closing task (a movement facilitated by the flexion synergy). METHODS: We evaluated the accuracy of a BMI classification algorithm in detecting hand-opening versus closing after reaching a target with 2 different shoulder-abduction loads in 6 individuals with stroke. RESULTS: We found a decreased accuracy in detecting hand opening when an individual with stroke intends to open the hand while activating shoulder abductors. However, such decreased accuracy with increased shoulder loading was not shown while detecting a hand-closing task. CONCLUSIONS: This study supports the idea that one should consider the effect of shoulder abduction activity when designing BMI classification algorithms for the purpose of restoring hand function in individuals with moderate to severe stroke.

Characterization of an improved polyimide-etalon all-optical transducer for high-resolution ultrasound imaging.

All-optical transduction of ultrasound provides high-frequency (>20 MHz) operation in the absence of electrical noise and distortion that hinders small-scale piezoelectric probes. Although fabrication of an all-optical 2-D array suitable for in vivo imaging remains incomplete, a thin-film structure integrating a polyimide film with a Fabry-Perot (etalon) receiver has been shown to be a viable candidate. We present here incremental improvements in the performance of a polyimide-etalon transducer and demonstrate imaging with an array configuration alternative to our previous study. We first show that a gain of more than 30% in output pressure is achieved when increasing the thickness of a bare polyimide film from 3 to 15 µm. This motivated the choice of polyimide as the etalon medium--a configuration made possible by utilizing a dielectric mirror that transmits wavelengths used for generation of ultrasound (ultraviolet) and reflects those for detection (near infrared). The increased reflectivity of the dielectric mirror resulted in a 2-fold decrease in noise-equivalent pressure to 3.3 kPa over a bandwidth of 47.5 MHz (0.48 Pa/√Hz). The transmit/receive center frequency increased from 37 to 49 MHz with a -6-dB bandwidth of 126%, and a maximum pressure of 213 kPa was produced using a 43 µm UV spot. A 2 x 2 mm synthetic array of 957 transmitters centered on a 1 x 1 mm synthetic array of four receivers was used to image two wire targets. Offline reconstruction indicated lateral resolutions of 70 and 114 µm at depths of 2.4 and 5.8 mm, respectively, with an average axial resolution of 35 µm. Finally, we explore the challenges of imaging in this configuration, which provides the best opportunity for real-time performance pending further development.

A polyimide-etalon thin film structure for all-optical high-frequency ultrasound transduction.

In this work, we have designed, fabricated, and tested an all-optical ultrasound transducer by integrating a photoabsorptive polyimide thin film into a Fabry-Pérot (etalon) high-frequency receiver. A 5-ns UV pulse was used for thermoelastic ultrasound generation in the polyimide film, and the transmission had a maximum amplitude of 4.3 MPa centered at 27 MHz with a fractional bandwidth of 107%. The device attained a noise-equivalent pressure of 1.3 Pa/√Hz in receive-only mode. When used in pulse-echo mode, the -6-dB upper cutoff frequency of the transmit/receive response reached 47 MHz. Basic imaging capabilities were also investigated by scanning the near-infrared probe beam across the device to create a 2 × 2 mm synthetic aperture. The imaging of targets placed at depths of 1.8 and 5.2 mm yielded estimates of 71 and 145 µm, respectively, for the lateral resolution and 35 and 38 µm, respectively, for the axial resolution. Finally, a design concept for a forward-viewing intravascular imager is presented that entails the coupling of light to a rotating, linear array of optical fibers on top of which are deposited polyimide-etalon transducers. Such a design would allow for a flexible and compact high-resolution imager well-suited for intravascular applications, such as guidance of treatment in the case of chronic total occlusion.

Usage of the ACT Robot in a Brain Machine Interface for Hand Opening and Closing in Stroke Survivors.

At six months after brain injury, about 65% of stroke survivors have been shown to be unable to incorporate the affected hand into activities of daily living (ADL). Using a reliable Brain-Machine-Interface (BMI) together with Neural Electronic Stimulation (NES) is a possible solution for the restoration of hand function in severely impaired hemiparetic stroke survivors. However, discoordination, i.e. the abnormal coupling between adjacent joints, causes an expected reduction in the performance of BMI algorithms. In this study, we test whether the active support of an ACT(3D) robot can increase the performance of two brain-machine-interface (BMI) algorithms in separating the subject's intention to open or close the impaired hand during reach. Improvement in recognition rate was obtained in 4 chronic hemiparetic stroke subjects when support from the robot was available. Further analysis on one subject suggests that such an improvement is related to quantitative changes in cortical activity. This result suggests that the ACT(3D) robot can be used to train severely impaired stroke subjects to use a BMI-controlled NES device.