Quantifying Spatial Patterns of Tissue Stiffness Within the Embryonic Mouse Kidney.

Biophysical factors, including changes in mechanical stiffness, have been shown to influence the morphogenesis of developing organs. There is a lack of experimental techniques, however, that can probe the mechanical properties of embryonic tissues-especially those which are not mechanically or optically accessible, such as the visceral organs of the developing mouse embryo. Here, using the embryonic kidney as a model system, we describe a method to use microindentation to quantify tissue-level regional differences in the mechanical properties of an embryonic organ. This technique is generalizable and can be used to quantify patterns of tissue stiffness within other developing organ systems. Going forward, these data will enable new experimental studies of the role of biophysical cues during organogenesis.

Microhardness and Microstructure Analysis of the LPBF Additively Manufactured 18Ni300.

This research focuses on analysing the 18Ni300 maraging steel produced through laser powder bed fusion. Specifically, it aims to examine the phase components using X-ray diffraction, the microstructure through scanning electron microscopy, and the hardness of the different structures present in the manufactured material. The primary goal is to meticulously analyse the material and its microstructures. By doing so, a correlation between the hardness and each structure type, be it cellular or columnar, can be established. This will allow us to pinpoint any defects in the material before any surface chemical treatment is carried out and facilitate a thorough examination of its microstructure. A consistent pattern emerges across the samples through systematic measurement of microhardness distribution in various locations and detailed examination of the structure. The findings of the study reveal that the hardness of cellular and columnar structures exhibits a significant variation based on the location of the measurement about cell boundaries. The hardness value is notably higher in the combination of cellular and multiple layers, as the data indicate.

Citric Acid Cross-Linked Gelatin-Based Composites with Improved Microhardness.

The aim of this study is to investigate the influence of cross-linking and reinforcements in gelatin on the physico-mechanical properties of obtained composites. The gelatin-based composites cross-linked with citric acid (CA) were prepared: gelatin type B (GB) and ß-tricalcium phosphate (ß-TCP) and novel hybrid composite GB with ß-TCP and hydroxyapatite (HAp) particles, and their structure, thermal, and mechanical properties were compared with pure gelatin B samples. FTIR analysis revealed that no chemical interaction between the reinforcements and gelatin matrix was established during the processing of hybrid composites by the solution casting method, proving the particles had no influence on GB cross-linking. The morphological investigation of hybrid composites revealed that cross-linking with CA improved the dispersion of particles, which further led to an increase in mechanical performance. The microindentation test showed that the hardness value was increased by up to 449%, which shows the high potential of ß-TCP and HAp particle reinforcement combined with CA as a cross-linking agent. Furthermore, the reduced modulus of elasticity was increased by up to 288%. Results of the MTT assay on L929 cells have revealed that the hybrid composite GB-TCP-HA-CA was not cytotoxic. These results showed that GB cross-linked with CA and reinforced with different calcium phosphates presents a valuable novel material with potential applications in dentistry.

Effect of type 2 diabetes mellitus on the microstructural, compositional and mechanical properties of cartilages.

BACKGROUND: Osteoarthritis (OA) is a chronic and complicated degenerative disorder of joints, including several phenotypes. Type 2 diabetes mellitus (T2DM) is one of the major causes of OA. However, few studies on the mechanical behavior of diabetic cartilages have been conducted. METHODS: This study evaluated the microstructural, compositional, and mechanical properties of healthy and diabetic rat cartilages using scanning electronic microscopy, X-ray energy spectroscopy, histology staining, and microindentation tests. RESULTS: Our results indicated that the diabetic cartilages had a significantly higher elastic modulus and similar permeability (95%CI: 3.72-8.56 MPa and 3.16×10-6-1.83×10-5 mm4/N·s) compared to the healthy cartilages (95%CI: 0.741-3.58 MPa and 3.15×10-6-1.14×10-5 mm4/N·s). Their stress relaxation behaviors were similar regardless of the loading rate except for the stretching parameter under the fast loading. Furthermore, the stress relaxation behaviors of the diabetic cartilages were significantly affected by the loading rate, especially the equilibrium force ratio and time constant. These mechanical outcomes could be attributed to the increase of fibril diameters and calcium aggregation in the cartilage. CONCLUSIONS: This study deepens our understanding of how T2DM might facilitate OA in cartilages, which could contribute to the development of more scientific diagnosis and therapies for patients with diabetes.

Evaluation of gelatin bloom strength on gelatin methacryloyl hydrogel properties.

Gelatin methacryloyl (GelMA) hydrogels are widely used for a variety of tissue engineering applications. The properties of gelatin can affect the mechanical properties of gelatin gels; however, the role of gelatin properties such as bloom strength on GelMA hydrogels has not yet been explored. Bloom strength is a food industry standard for describing the quality of gelatin, where higher bloom strength is associated with higher gelatin molecular weight. Here, we evaluate the role of bloom strength on GelMA hydrogel mechanical properties. We determined that both bloom strength of gelatin and weight percent of GelMA influenced both stiffness and viscoelastic ratio; however, only bloom strength affected diffusivity, permeability, and pore size. With this library of GelMA hydrogels of varying properties, we then encapsulated Swan71 trophoblast spheroids in these hydrogel variants to assess how bloom strength affects trophoblast spheroid morphology. Overall, we observed a decreasing trend of spheroid area and Feret diameter as bloom strength increased. In identifying clear relationships between bloom strength, hydrogel mechanical properties, and trophoblast spheroid morphology, we demonstrate that bloom strength should considered when designing tissue engineered constructs.

A Traumatic Impact Immediately Changes the Mechanical Properties of Articular Cartilage.

OBJECTIVE: To investigate whether and how a single traumatic impact changes the mechanical properties of talar articular cartilage. DESIGN: A marble was placed on the joint surface and a weight was dropped on both medial and lateral caprine talus to create a well-defined single focal impact. The mechanical properties of intact and impacted talar cartilage were measured with a micro-indenter. Elastic (storage) and viscous (loss) moduli were determined by oscillatory ramp and dynamic mechanical analysis protocols. RESULTS: We found significant differences between ankles and within the same ankle joint, with the medial talus having significantly higher storage- and loss moduli than the lateral talus. The storage- and loss moduli of intact articular cartilage increased with greater indentation depths. However, postimpact the storage- and loss moduli were significantly and consistently lower in all specimens indicating immediate posttraumatic damage. The deeper regions of talar cartilage were less affected by the impact than the more superficial regions. CONCLUSIONS: A single traumatic impact results in an immediate and significant decrease of storage- and loss moduli. Further research must focus on the development of non- or minimally invasive diagnostic tools to address the exact microdamage caused by the impact. We speculate that the traumatic impact damaged the collagen fibers that confine the water-binding proteoglycans and thereby decreasing the hydrostatic pressure of cartilage. As part of the treatment directly after a trauma, one could imagine a reduction or restriction of peak loads to prevent the progression of the cascade towards PTOA of the ankle joint.

The practicality of using bone impact microindentation in a population-based study of women: A Geelong-Osteoporosis Study.

Impact microindentation (IMI) is a minimally invasive technique that allows the assessment of bone material strength index (BMSi) in vivo, by measuring the depth of a micron-sized, spherical tip into cortical bone that is then indexed to the depth of the tip into a reference material. In this study, we aimed to assess the practicality of its application in 99 women aged 42-84 yr from the Geelong Osteoporosis Study. Impact microindentation was performed in the mid-shaft of the right tibia using the OsteoProbe. Immediately following measurement, each participant was requested to rate on a Visual Analogue Scale [0-10] the level of discomfort anticipated and experienced, any initial reluctance towards the measurement and whether they were willing to repeat the measurement. Of 99 potential participants who attended this assessment phase, 55 underwent IMI measurement. Reasons for non-measurement in 44 women were existing skin conditions (n = 8, 18.2 %) and excessive soft tissue around mid-tibial region (n = 32, 72.2 %). An additional four (9.1 %) participants did not provide any reasons for declining. For 55 participants who had underwent IMI, the expectation for pain when briefed about the procedure was low (2.28 ± 2.39), as was pain experienced during the measurement (0.72 ± 1.58). Participants were not reluctant to undergo the measurement (0.83 ± 1.67), and all indicated a willingness to repeat the measurement. Results of this study showed that the IMI technique is well tolerated and accepted by women participating in the Geelong Osteoporosis Study, suggesting that the technique shows promise in a research or clinical setting.

Natural, biphasic calcium phosphate from fish bones for enamel remineralization and dentin tubules occlusion.

OBJECTIVES: A calcium phosphate extracted from fish bones (CaP-N) was evaluated for enamel remineralization and dentinal tubules occlusion. METHODS: CaP-N was characterized by assessing morphology by SEM, crystallinity by PXRD, and composition by ICP-OES. CaP-N morphology, crystallinity, ion release, and pH changes over time in neutral and acidic solutions were studied. CaP-N was then tested to assess remineralization and dentinal tubules occlusion on demineralized human enamel and dentin specimens (n = 6). Synthetic calcium phosphate in form of stoichiometric hydroxyapatite nanoparticles (CaP-S) and tap water were positive and negative controls, respectively. After treatment (brush every 12 h for 5d and storage in Dulbecco's modified PBS), specimens' morphology and surface composition were assessed (by SEM-EDS), while the viscoelastic behavior was evaluated with microindentation and DMA. RESULTS: CaP-N consisted of rounded microparticles (200 nm - 1 µm) composed of 33 wt% hydroxyapatite and 67 wt% ß-tricalcium phosphate. In acidic solution, CaP-N released calcium and phosphate ions thanks to the preferential ß-tricalcium phosphate phase dissolution. Enamel remineralization was induced by CaP-N comparably to CaP-S, while CaP-N exhibited a superior dentinal tubule occlusion than CaP-S, forming mineral plugs and depositing new nanoparticles onto demineralized collagen. This behavior was attributed to its bigger particle size and increased solubility. DMA depth profiling and SEM showed an excellent interaction between the newly formed mineralized structures and the pristine tissue, particularly at the exposed collagen fibrils. SIGNIFICANCE: CaP-N demonstrated very good remineralizing and occlusive activity in vitro, comparable to CaP-S, thus could be a promising circular economy alternative therapeutic agent for dentistry.

Characterization of the mechanical properties of the mouse Achilles tendon enthesis by microindentation. Effects of unloading and subsequent reloading.

The fibrocartilaginous tendon enthesis, i.e. the site where a tendon is attached to bone through a fibrocartilaginous tissue, is considered as a functionally graded interface. However, at local scale, a very limited number of studies have characterized micromechanical properties of this transitional tissue. The first goal of this work was to characterize the micromechanical properties of the mineralized part of the healthy Achilles tendon enthesis (ATE) through microindentation testing and to assess the degree of mineralization and of carbonation of mineral crystals by Raman spectroscopy. Since little is known about enthesis biological plasticity, our second objective was to examine the effects of unloading and reloading, using a mouse hindlimb-unloading model, on both the micromechanical properties and the mineral phase of the ATE. Elastic modulus, hardness, degree of mineralization, and degree of carbonation were assessed after 14 days of hindlimb suspension and again after a subsequent 6 days of reloading. The elastic modulus gradually increased along the mineralized part of the ATE from the tidemark to the subchondral bone, with the same trend being found for hardness. Whereas the degree of carbonation did not differ according to zone of measurement, the degree of mineralization increased by >70 % from tidemark to subchondral bone. Thus, the gradient in micromechanical properties is in part explained by a mineralization gradient. A 14-day unloading period did not appear to affect the gradient of micromechanical properties of the ATE, nor the degree of mineralization or carbonation. However, contrary to a short period of unloading, early return to normal mechanical load reduced the micromechanical properties gradient, regardless of carbonate-to-phosphate ratios, likely due to the more homogeneous degree of mineralization. These findings provide valuable data not only for tissue bioengineering, but also for musculoskeletal clinical studies and microgravity studies focusing on long-term space travel by astronauts.

Effects of surface profile on porcine dural mechanical properties.

BACKGROUND: Cerebrospinal fluid leakage through the spinal meninges is difficult to diagnose and treat. Moreover, its underlying mechanism remains unknown. Considering that the dura mater is structurally the strongest and outermost membrane among the three-layered meninges, we hypothesized that a dural mechanical tear would trigger spontaneous cerebrospinal fluid leakage, especially when a traumatic loading event is involved. Thus, accurate biomechanical properties of the dura mater are indispensable for improving computational models, which aid in predicting blunt impact injuries and creating artificial substitutes for transplantation and surgical training. METHOD: We characterized the surface profile of the spinal dura and its mechanical properties (Young's moduli) with a distinction of its inherent anatomical sites (i.e., the cervical and lumbar regions as well as the dorsal and ventral sides of the spinal cord). FINDINGS: Although the obtained Young's moduli exhibited no considerable difference between the aforementioned anatomical sites, our results suggested that the wrinkles structurally formed along the longitudinal direction would relieve stress concentration on the dural surface under in vivo and supraphysiological conditions, enabling mechanical protection of the dural tissue from a blunt impact force that was externally applied to the spine. INTERPRETATION: This study provides fundamental data that can be used for accurately predicting cerebrospinal fluid leakage due to blunt impact trauma.