Viscoelasticity in 3D Cell Culture and Regenerative Medicine: Measurement Techniques and Biological Relevance.

The field of mechanobiology is gaining prominence due to recent findings that show cells sense and respond to the mechanical properties of their environment through a process called mechanotransduction. The mechanical properties of cells, cell organelles, and the extracellular matrix are understood to be viscoelastic. Various technologies have been researched and developed for measuring the viscoelasticity of biological materials, which may provide insight into both the cellular mechanisms and the biological functions of mechanotransduction. Here, we explain the concept of viscoelasticity and introduce the major techniques that have been used to measure the viscoelasticity of various soft materials in different length- and timescale frames. The topology of the material undergoing testing, the geometry of the probe, the magnitude of the exerted stress, and the resulting deformation should be carefully considered to choose a proper technique for each application. Lastly, we discuss several applications of viscoelasticity in 3D cell culture and tissue models for regenerative medicine, including organoids, organ-on-a-chip systems, engineered tissue constructs, and tunable viscoelastic hydrogels for 3D bioprinting and cell-based therapies.

Gelatin methacryloyl and Laponite bioink for 3D bioprinted organotypic tumor modeling.

Three-dimensional (3D)in vitrotumor models that can capture the pathophysiology of human tumors are essential for cancer biology and drug development. However, simulating the tumor microenvironment is still challenging because it consists of a heterogeneous mixture of various cellular components and biological factors. In this regard, current extracellular matrix (ECM)-mimicking hydrogels used in tumor tissue engineering lack physical interactions that can keep biological factors released by encapsulated cells within the hydrogel and improve paracrine interactions. Here, we developed a nanoengineered ion-covalent cross-linkable bioink to construct 3D bioprinted organotypic tumor models. The bioink was designed to implement the tumor ECM by creating an interpenetrating network composed of gelatin methacryloyl (GelMA), a light cross-linkable polymer, and synthetic nanosilicate (Laponite) that exhibits a unique ionic charge to improve retention of biological factors released by the encapsulated cells and assist in paracrine signals. The physical properties related to printability were evaluated to analyze the effect of Laponite hydrogel on bioink. Low GelMA (5%) with high Laponite (2.5%-3.5%) composite hydrogels and high GelMA (10%) with low Laponite (1.0%-2.0%) composite hydrogels showed acceptable mechanical properties for 3D printing. However, a low GelMA composite hydrogel with a high Laponite content could not provide acceptable cell viability. Fluorescent cell labeling studies showed that as the proportion of Laponite increased, the cells became more aggregated to form larger 3D tumor structures. Reverse transcription-polymerase chain reaction (RT-qPCR) and western blot experiments showed that an increase in the Laponite ratio induces upregulation of growth factor and tissue remodeling-related genes and proteins in tumor cells. In contrast, cell cycle and proliferation-related genes were downregulated. On the other hand, concerning fibroblasts, the increase in the Laponite ratio indicated an overall upregulation of the mesenchymal phenotype-related genes and proteins. Our study may provide a rationale for using Laponite-based hydrogels in 3D cancer modeling.

Skin-interfaced electronics: A promising and intelligent paradigm for personalized healthcare.

Skin-interfaced electronics (skintronics) have received considerable attention due to their thinness, skin-like mechanical softness, excellent conformability, and multifunctional integration. Current advancements in skintronics have enabled health monitoring and digital medicine. Particularly, skintronics offer a personalized platform for early-stage disease diagnosis and treatment. In this comprehensive review, we discuss (1) the state-of-the-art skintronic devices, (2) material selections and platform considerations of future skintronics toward intelligent healthcare, (3) device fabrication and system integrations of skintronics, (4) an overview of the skintronic platform for personalized healthcare applications, including biosensing as well as wound healing, sleep monitoring, the assessment of SARS-CoV-2, and the augmented reality-/virtual reality-enhanced human-machine interfaces, and (5) current challenges and future opportunities of skintronics and their potentials in clinical translation and commercialization. The field of skintronics will not only minimize physical and physiological mismatches with the skin but also shift the paradigm in intelligent and personalized healthcare and offer unprecedented promise to revolutionize conventional medical practices.

Fabrication and Characterization of a Miniaturized 15-MHz Side-Looking Phased-Array Transducer Catheter.

This paper describes the development of a miniaturized 15-MHz side-looking phased-array transducer catheter. The array features a 2-2 linear composite with 64 piezoelectric elements mechanically diced into a piece of PMN-30%PT single crystal and separated by non-conductive epoxy kerfs at a 50-µm pitch, yielding a total active aperture of 3.2 mm in the azimuth direction and 1.8 mm in the elevation direction, with an elevation natural focal depth of 8.1 mm. The array includes non-conductive epoxy backing and two front matching layers. A custom flexible circuit connects the array piezoelectric elements to a bundle of 64 individual 48-AWG micro-coaxial cables enclosed within a 1.5-m long 10F catheter. Performance characterization was evaluated via finite element analysis simulations and afterwards compared against obtained measurement results, which showed an average center frequency of 17.7 MHz, an average bandwidth of 52.2% at -6 dB, and crosstalk less than -30 dB. Imaging of a tungsten fine-wire phantom resulted in axial and lateral spatial resolutions of approximately 90 µm and 420 ìm, respectively. The imaging capability was further evaluated with colorectal tissue-mimicking phantoms, demonstrating the potential suitability of the proposed phased-array transducer for the intraoperative assessment of surgical margins during minimally invasive colorectal surgery procedures.

Fabrication and Characterization of a 20-MHz Microlinear Phased-Array Transducer for Intervention Guidance.

This paper describes the design and fabrication of a miniature ultrasonic phased-array transducer used for intervention guidance. Currently, ultrasound probes are often placed at the body surface of the patients, leading to several drawbacks including the limitation of penetration and image quality. In order to improve the reliability of the guiding process, we propose a miniature phased-array transducer that can be placed adjacent to the intervention device during the interventional procedure. In this paper, we report the work that has been carried out on the development of this miniature phased-array transducer. It comprised 48 elements housed in a 3-mm-diameter needle. A specially designed flexible circuit was used for accommodating the transducer array in the long, thin needle housing. The center frequency and the fractional bandwidth were approximately 20 MHz and 42%, respectively, with an average crosstalk lower than -30 dB. The axial and azimuth resolutions were approximately 80 and [Formula: see text], respectively. The imaging capability of the transducer was further evaluated by acquiring the B-mode images of a needle in a cow liver. The performance of the proposed phased-array transducer demonstrates the feasibility of such an approach for interventional guidance.

High-Frequency Ultrasound Array Designed for Ultrasound-Guided Breast Biopsy.

This paper describes the development of a miniaturized high-frequency linear array that can be integrated within a core biopsy needle to improve tissue sampling accuracy during breast cancer biopsy procedures. The 64-element linear array has an element width of [Formula: see text], kerf width of [Formula: see text], element length of 1 mm, and element thickness of [Formula: see text]. The 2-2 array composite was fabricated using deep reactive ion etching of lead magnesium niobate-lead titanate (PMN-PT) single crystal material. The array composite fabrication process as well as a novel high-density electrical interconnect solution are presented and discussed. Array performance measurements show that the array had a center frequency and fractional bandwidth ([Formula: see text]) of 59.1 MHz and 29.4%, respectively. Insertion loss and adjacent element crosstalk at the center frequency were -41.0 and [Formula: see text], respectively. A B-mode image of a tungsten wire target phantom was captured using a synthetic aperture imaging system and the imaging test results demonstrate axial and lateral resolutions of 33.2 and [Formula: see text], respectively.

High-frequency ultrasound imaging for breast cancer biopsy guidance.

Image-guided core needle biopsy is the current gold standard for breast cancer diagnosis. Microcalcifications, an important radiographic finding on mammography suggestive of early breast cancer such as ductal carcinoma in situ, are usually biopsied under stereotactic guidance. This procedure, however, is uncomfortable for patients and requires the use of ionizing radiation. It would be preferable to biopsy microcalcifications under ultrasound guidance since it is a faster procedure, more comfortable for the patient, and requires no radiation. However, microcalcifications cannot reliably be detected with the current standard ultrasound imaging systems. This study is motivated by the clinical need for real-time high-resolution ultrasound imaging of microcalcifications, so that biopsies can be accurately performed under ultrasound guidance. We have investigated how high-frequency ultrasound imaging can enable visualization of microstructures in ex vivo breast tissue biopsy samples. We generated B-mode images of breast tissue and applied the Nakagami filtering technique to help refine image output so that microcalcifications could be better assessed during ultrasound-guided core biopsies. We describe the preliminary clinical results of high-frequency ultrasound imaging of ex vivo breast biopsy tissue with microcalcifications and without Nakagami filtering and the correlation of these images with the pathology examination by hematoxylin and eosin stain and whole slide digital scanning.