3D printed needleless injector based on thermocavitation: analysis of impact and penetration depth in skin phantoms in a repetitive regime.

A global issue that requires attention is the duality between the shortage of needles for regular vaccination campaigns and the exponential increase in syringe and needle waste from such campaigns, which has been exacerbated by the COVID-19 pandemic. In response to this problem, this study presents a 3D printed needleless injector based on thermocavitation. The work focused on investigating the interaction of the resulting liquid jets with skin phantoms at different concentrations (1-2%), emphasizing their impact and penetration depth in a repetitive regime. The injector was designed and fabricated from a semi-transparent polymer using a high-resolution 3D printer, allowing the ejection of liquid jets with velocities up to ~ 73 m/s. The impact of these jets on skin phantoms was evaluated using a high-speed camera. After 6 consecutive liquid jets (1% concentration), a maximum penetration depth of ~ 2.5 mm was achieved, delivering approximately 4.7 µL. For the highest concentration (2.0%) and the same number of shots, the penetration depth was reduced to ~ 0.6 mm with a delivered volume of ~ 0.7 µL. An important finding of this study is that the liquid jet with the highest pressure does not cause the maximum penetration depth, but is the result of a series of successive shots. In addition, the velocity and shape of the ejected jet are determined by the amount of solution and the meniscus formed inside the injector. These findings advance the development of precise and efficient thermocavitation-based injectors with broad potential applications in medical and pharmaceutical fields.

Bubble dynamics and speed of jets for needle-free injections produced by thermocavitation.

Significance: The number of injections administered has increased dramatically worldwide due to vaccination campaigns following the COVID-19 pandemic, creating a problem of disposing of syringes and needles. Accidental needle sticks occur among medical and cleaning staff, exposing them to highly contagious diseases, such as hepatitis and human immunodeficiency virus. In addition, needle phobia may prevent adequate treatment. To overcome these problems, we propose a needle-free injector based on thermocavitation. Aim: Experimentally study the dynamics of vapor bubbles produced by thermocavitation inside a fully buried 3D fused silica chamber and the resulting high-speed jets emerging through a small nozzle made at the top of it. The injected volume can range from ∼0.1 to 2 µL per shot. We also demonstrate that these jets have the ability to penetrate agar skin phantoms and ex-vivo porcine skin. Approach: Through the use of a high-speed camera, the dynamics of liquid jets ejected from a microfluidic device were studied. Thermocavitation bubbles are generated by a continuous wave laser (1064 nm). The 3D chamber was fabricated by ultra-short pulse laser-assisted chemical etching. Penetration tests are conducted using agar gels (1%, 1.25%, 1.5%, 1.75%, and 2% concentrations) and porcine tissue as a model for human skin. Result: High-speed camera video analysis showed that the average maximum bubble wall speed is about 10 to 25 m/s for almost any combination of pump laser parameters; however, a clever design of the chamber and nozzle enables one to obtain jets with an average speed of ∼70 m/s. The expelled volume per shot (0.1 to 2 µl) can be controlled by the pump laser intensity. Our injector can deliver up to 20 shots before chamber refill. Penetration of jets into agar of different concentrations and ex-vivo porcine skin is demonstrated. Conclusions: The needle-free injectors based on thermocavitation may hold promise for commercial development, due to their cost and compactness.

Adaptive Feature Extraction for Blood Vessel Segmentation and Contrast Recalculation in Laser Speckle Contrast Imaging.

Microvasculature analysis in biomedical images is essential in the medical area to evaluate diseases by extracting properties of blood vessels, such as relative blood flow or morphological measurements such as diameter. Given the advantages of Laser Speckle Contrast Imaging (LSCI), several studies have aimed to reduce inherent noise to distinguish between tissue and blood vessels at higher depths. These studies have shown that computing Contrast Images (CIs) with Analysis Windows (AWs) larger than standard sizes obtains better statistical estimators. The main issue is that larger samples combine pixels of microvasculature with tissue regions, reducing the spatial resolution of the CI. This work proposes using adaptive AWs of variable size and shape to calculate the features required to train a segmentation model that discriminates between blood vessels and tissue in LSCI. The obtained results show that it is possible to improve segmentation rates of blood vessels up to 45% in high depths (≈900 µm) by extracting features adaptively. The main contribution of this work is the experimentation with LSCI images under different depths and exposure times through adaptive processing methods, furthering the understanding the performance of the different approaches under these conditions. Results also suggest that it is possible to train a segmentation model to discriminate between pixels belonging to blood vessels and those belonging to tissue. Therefore, an adaptive feature extraction method may improve the quality of the features and thus increase the classification rates of blood vessels in LSCI.

Intrinsic fluorescence and mechanical testing of articular cartilage in human patients with osteoarthritis.

The degeneration of articular cartilage is the main cause of osteoarthritis (OA), a common cause of disability among elderly patients. The aim of this study is to understand the correlation between intrinsic fluorescence of articular cartilage and its biomechanical properties in patients with osteoarthritis. Cylindrical samples of articular cartilage 6 mm in diameter were extracted via biopsy punch from the femoral condyles of 6 patients with advanced OA undergoing knee replacement surgery. The mechanical stiffness and fluorescence of each cartilage plug were measured by indentation test and spectrofluorometry. Maps of fluorescence intensity, at excitation/emission wavelengths of 240-520/290-530 nm, were used to identify wavelengths of interest. The mechanical stiffness and fluorescence intensity were correlated using a Spearman analysis. The excitation/emission maps demonstrated three fluorescence peaks at excitation/emission wavelength pairs 330/390, 350/430 and 370/460 nm. The best correlation between the fluorescence intensity and stiffness of cartilage was obtained for the 330 nm excitation band [R=0.82, p=0.04]. The intrinsic fluorescence of articular cartilage may have application in optically assessing the state of cartilage in patients with osteoarthritis.

Changes in endogenous UV fluorescence and biomechanical stiffness of bovine articular cartilage after collagenase digestion are strongly correlated.

A significant source of morbidity in the elderly population of the United States is osteoarthritis (OA), a disease caused by the breakdown and loss of articular cartilage. The exact causes of OA remain unknown, though biomechanical forces and biochemical alterations are important factors. There exists an unmet need for an imaging tool to identify early lesions of OA via metabolic, chemical or structural changes. Our work aims to characterize changes in the intensity of UV fluorescent bands associated with known structural proteins of cartilage. We employed an OA model in which bovine osteochondral plugs were digested in collagenase of varying concentrations. UV fluorescence before and after proteolytic digestion was measured using a spectrofluorimeter. The elastic modulus (EM) of each sample was measured using an indentation apparatus. Hydroxypyridinoline crosslink (330/390 nm) fluorescence intensity after digestion correlated with cartilage EM (R = 0.922, p = 0.026), as did tryptophan (290/350 nm) fluorescence intensity after digestion and EM (R = 0.949, p = 0.014) and tyrosine (290/310 nm) fluorescence intensity after digestion and EM (R = 0.946, p = 0.015). Loss of endogenous UV fluorescence correlated with cartilage degradation in an in-vitro model of OA, and may serve as a sensitive optical biomarker for the state of cartilage.

Evaluation of cell and matrix mechanics using fluorescence excitation spectroscopy: Feasibility study in collagen gels containing fibroblasts.

BACKGROUND AND OBJECTIVE: Collagen gels containing cells are commonly used in tissue engineering, wound healing, and cancer research to investigate the interplay between cells and the extracellular matrix (ECM), as changes in the density and stiffness of the microenvironment are known to play a role in many diseases or pathological conditions. In these gels, the stiffness is regularly determined using destructive methods, such as indentation and tensile tests. Certain molecules native to cells and the ECM display fluorescence upon irradiation with ultraviolet light. The objective of the present study was to investigate the feasibility of using the endogenous, or innate, fluorescence of collagen gels containing fibroblasts as an optical marker to evaluate changes in the mechanical properties of the ECM. MATERIALS AND METHODS: Human foreskin fibroblasts cells at concentrations of 50,000 and 100,000 cells/ml were cultured in three-dimensional gels of type I collagen for 16 days. Fibroblast cells remodeled the ECM, contracting and increasing the stiffness of the gel. During this remodeling process, changes in mechanical properties and fluorescence were measured with an indentation test and a spectrofluorometer, respectively. Force and displacement measurements from the indentation test were used to calculate the elastic modulus of the gel. Maps of fluorescence intensity, at excitation/emission of 240-520/290-530 nm, were used to identify the wavelengths of interest. RESULTS: Fluorescence excitation/emission maps exhibited two distinct excitation/emission bands whose intensities increased as the fibroblasts remodeled and increased the stiffness of the ECM: The 290/340 nm band ascribed to tryptophan and the 330/390 nm band ascribed to cross-links of collagen (pepsin-digestible cross-links). A Spearman correlation analysis, between the elastic modulus of the gel containing fibroblasts and the fluorescence of cross-links of collagen, resulted in R = 0.95 (P < 0.05) and R = 0.77 (P = 0.12) for 50,000 and 100,000 cells/ml, respectively. CONCLUSIONS: The endogenous fluorescence intensity ascribed to pepsin-digestible cross-links of collagen may serve as an optical marker to evaluate changes in the mechanical properties of the ECM; this is relevant to collagenous tissues for which pathological states are related to mechanical alterations, such as keratoconus in cornea and osteoarthritis in articular cartilage.