Strain-dependent elastography of cancer cells reveals heterogeneity and stiffening due to attachment.

Because cells vary in thickness and in biomechanical properties, the use of a constant force trigger during atomic force microscopy (AFM) stiffness mapping produces a varied nominal strain that can obfuscate the comparison of local material properties. In this study, we measured the biomechanical spatial heterogeneity of ovarian and breast cancer cells by using an indentation-dependent pointwise Hertzian method. Force curves and surface topography were used together to determine cell stiffness as a function of nominal strain. By recording stiffness at a particular strain, it may be possible to improve comparison of the material properties of cells and produce higher contrast representations of cell mechanical properties. Defining a linear region of elasticity that corresponds to a modest nominal strain, we were able to clearly distinguish the mechanics of the perinuclear region of cells. We observed that, relative to the lamelopodial stiffness, the perinuclear region was softer for metastatic cancer cells than their nonmetastatic counterparts. Moreover, contrast in the strain-dependent elastography in comparison to conventional force mapping with Hertzian model analysis revealed a significant stiffening phenomenon in the thin lamellipodial region in which the modulus scales inversely and exponentially with cell thickness. The observed exponential stiffening is not affected by relaxation of cytoskeletal tension, but finite element modeling indicates it is affected by substrate adhesion. The novel cell mapping technique explores cancer cell mechanical nonlinearity that results from regional heterogeneity, which could help explain how metastatic cancer cells can show soft phenotypes while simultaneously increasing force generation and invasiveness.

Pathologic mechanobiological interactions between red blood cells and endothelial cells directly induce vasculopathy in iron deficiency anemia.

The correlation between cardiovascular disease and iron deficiency anemia (IDA) is well documented but poorly understood. Using a multi-disciplinary approach, we explore the hypothesis that the biophysical alterations of red blood cells (RBCs) in IDA, such as variable degrees of microcytosis and decreased deformability may directly induce endothelial dysfunction via mechanobiological mechanisms. Using a combination of atomic force microscopy and microfluidics, we observed that subpopulations of IDA RBCs (idRBCs) are significantly stiffer and smaller than both healthy RBCs and the remaining idRBC population. Furthermore, computational simulations demonstrated that the smaller and stiffer idRBC subpopulations marginate toward the vessel wall causing aberrant shear stresses. This leads to increased vascular inflammation as confirmed with perfusion of idRBCs into our "endothelialized" microfluidic systems. Overall, our multifaceted approach demonstrates that the altered biophysical properties of idRBCs directly lead to vasculopathy, suggesting that the IDA and cardiovascular disease association extends beyond correlation and into causation.

Nanofiber-Based Delivery of Bioactive Lipids Promotes Pro-regenerative Inflammation and Enhances Muscle Fiber Growth After Volumetric Muscle Loss.

Volumetric muscle loss (VML) injuries after extremity trauma results in an important clinical challenge often associated with impaired healing, significant fibrosis, and long-term pain and functional deficits. While acute muscle injuries typically display a remarkable capacity for regeneration, critically sized VML defects present a dysregulated immune microenvironment which overwhelms innate repair mechanisms leading to chronic inflammation and pro-fibrotic signaling. In this series of studies, we developed an immunomodulatory biomaterial therapy to locally modulate the sphingosine-1-phosphate (S1P) signaling axis and resolve the persistent pro-inflammatory injury niche plaguing a critically sized VML defect. Multiparameter pseudo-temporal 2D projections of single cell cytometry data revealed subtle distinctions in the altered dynamics of specific immune subpopulations infiltrating the defect that were critical to muscle regeneration. We show that S1P receptor modulation via nanofiber delivery of Fingolimod (FTY720) was characterized by increased numbers of pro-regenerative immune subsets and coincided with an enriched pool of muscle stem cells (MuSCs) within the injured tissue. This FTY720-induced priming of the local injury milieu resulted in increased myofiber diameter and alignment across the defect space followed by enhanced revascularization and reinnervation of the injured muscle. These findings indicate that localized modulation of S1P receptor signaling via nanofiber scaffolds, which resemble the native extracellular matrix ablated upon injury, provides great potential as an immunotherapy for bolstering endogenous mechanisms of regeneration following VML injury.

Modulating local S1P receptor signaling as a regenerative immunotherapy after volumetric muscle loss injury.

Regeneration of skeletal muscle after volumetric injury is thought to be impaired by a dysregulated immune microenvironment that hinders endogenous repair mechanisms. Such defects result in fatty infiltration, tissue scarring, chronic inflammation, and debilitating functional deficits. Here, we evaluated the key cellular processes driving dysregulation in the injury niche through localized modulation of sphingosine-1-phosphate (S1P) receptor signaling. We employ dimensionality reduction and pseudotime analysis on single cell cytometry data to reveal heterogeneous immune cell subsets infiltrating preclinical muscle defects due to S1P receptor inhibition. We show that global knockout of S1P receptor 3 (S1PR3) is marked by an increase of muscle stem cells within injured tissue, a reduction in classically activated relative to alternatively activated macrophages, and increased bridging of regenerating myofibers across the defect. We found that local S1PR3 antagonism via nanofiber delivery of VPC01091 replicated key features of pseudotime immune cell recruitment dynamics and enhanced regeneration characteristic of global S1PR3 knockout. Our results indicate that local S1P receptor modulation may provide an effective immunotherapy for promoting a proreparative environment leading to improved regeneration following muscle injury.

Reflection sensitivity of 1.3 µm quantum dot lasers epitaxially grown on silicon.

We present measurements of relative intensity noise versus various levels of optical feedback for 1.3 µm quantum dot lasers epitaxially grown on silicon for the first time. A systematic comparison is made with heterogeneously integrated 1.55 µm quantum well lasers on silicon. Our results indicate up to 20 dB reduced sensitivity of the quantum dot lasers on silicon compared to the quantum wells.

Electrically pumped continuous wave quantum dot lasers epitaxially grown on patterned, on-axis (001) Si.

High performance III-V lasers at datacom and telecom wavelengths on on-axis (001) Si are needed for scalable datacenter interconnect technologies. We demonstrate electrically injected quantum dot lasers grown on on-axis (001) Si patterned with {111} v-grooves lying in the [110] direction. No additional Ge buffers or substrate miscut was used. The active region consists of five InAs/InGaAs dot-in-a-well layers. We achieve continuous wave lasing with thresholds as low as 36 mA and operation up to 80°C.

Electrically pumped continuous-wave 1.3 µm quantum-dot lasers epitaxially grown on on-axis (001) GaP/Si.

We demonstrate the first electrically pumped continuous-wave (CW) III-V semiconductor lasers epitaxially grown on on-axis (001) silicon substrates without offcut or germanium layers, using InAs/GaAs quantum dots as the active region and an intermediate GaP buffer between the silicon and device layers. Broad-area lasers with uncoated facets achieve room-temperature lasing with threshold current densities around 860 A/cm2 and 110 mW of single-facet output power for the same device. Ridge lasers designed for low threshold operations show maximum lasing temperatures up to 90°C and thresholds down to 30 mA.

1.3-µm InAs quantum-dot micro-disk lasers on V-groove patterned and unpatterned (001) silicon.

We report comparison of lasing dynamics in InAs quantum dot (QD) micro-disk lasers (MDLs) monolithically grown on V-groove patterned and planar Si (001) substrates. TEM characterizations reveal abrupt interfaces and reduced threading dislocations in the QD active regions when using the GaAs-on-V-grooved-Si template. The improved crystalline quality translates into lower threshold power in the optically pumped continuous-wave MDLs. Concurrent evaluations were also made with devices fabricated simultaneously on lattice-matched GaAs substrates. Lasing behaviors from 10 K up to room temperature have been studied systematically. The analyses spotlight insights into the optimal epitaxial scheme to achieve low-threshold lasing in telecommunication wavelengths on exact Si (001) substrates.

Optically pumped 1.3 µm room-temperature InAs quantum-dot micro-disk lasers directly grown on (001) silicon.

Direct integration of high-performance laser diodes on silicon will dramatically transform the world of photonics, expediting the progress toward low-cost and compact photonic integrated circuits (PICs) on the mainstream silicon platform. Here, we report, to the best of our knowledge, the first 1.3 µm room-temperature continuous-wave InAs quantum-dot micro-disk lasers epitaxially grown on industrial-compatible Si (001) substrates without offcut. The lasing threshold is as low as hundreds of microwatts, similar to the thresholds of identical lasers grown on a GaAs substrate. The heteroepitaxial structure employed here does not require the use of an absorptive germanium buffer and/or dislocation filter layers, both of which impede the efficient coupling of light from the laser active regions to silicon waveguides. This allows for full compatibility with the extensive silicon-on-insulator (SOI) technology. The large-area virtual GaAs (on Si) substrates can be directly adopted in various mature in-plane laser configurations, both optically and electrically. Thus, this demonstration represents a major advancement toward the commercial success of fully integrated silicon photonics.