Modulus Of Tissue Research Articles

Mechanical Properties of Inflamed Appendix Tissues

Background/Objectives: Histopathological examination enables visualization of morphological changes in cells and tissues. In recent years, there has been increasing interest in assessing the mechanical properties of tissues that cannot be determined by standard histopathological examinations. Mechanobiology is crucial in human physiology and holds promise for uncovering new diagnostic markers for disease processes such as carcinogenesis and inflammation. In this study, we concentrated on measuring the mechanical properties of appendix biopsy specimens to identify potential mechanomarkers of inflammation. Appendix tissues provided the opportunity to measure mechanical properties both with an atomic force microscope and a shear rheometer. Methods: The atomic force microscope AFM—NanoWizard 4 BioScience JPK/Bruker was used for the evaluation of the elastic modulus (i.e., Young’s modulus) of appendix tissues. Young’s modulus was derived from the Hertz-Sneddon model applied to force-indentation curves. The rheological properties of macroscopic samples were measured on a parallel-plate, strain-controlled shear rheometer Anton Paar MCR302. Results: The data collected suggest that elasticity, expressed as Young’s modulus and the storage modulus, could be considered a marker indicating appendix tissue inflammation. Young’s modulus of inflamed appendix tissues was found to be significantly lower than that of healthy ones, with an average reduction of 67%. Furthermore, it was observed that inflamed appendix tissues, in comparison to healthy ones, respond differently under varying axial and shear stresses, enabling their identification. Conclusions: Our findings suggest that the specific mechanical properties of inflamed vermiform appendices could serve as novel mechanomarkers for the early detection and monitoring of appendicitis.

Numerical Optimization of Functionally Graded Ti-HAP Material for Tibial Bone Fixation System.

Functionally graded materials (FGMs) are heterogeneous composites characterized by outstanding properties. They are built from two or more components with a gradient distribution of chemical composition along a given direction. A promising graded material for biomedical engineering as an implant could be a FGM made of titanium (Ti) and hydroxyapatite (HAP). It would allow us to counteract the difference between the stiffness modulus of pure titanium and bone tissue. Moreover, it can be a good solution to the problem of stress shielding for bone fixation plates made of conventional titanium or steel. The presented paper aims to perform micromechanical modeling and optimization of a functionally Ti-HAP graded plate, followed by numerical analysis of a fractured tibia stabilization system under specific boundary conditions. Finite element analysis was performed using ANSYS Workbench 2021 software. The models of the FGM plate and tibial fixation system were made using the Space Claim tool. The ANSYS software allowed the optimization of the model considered and the selection of the appropriate structural parameters of the FGM Ti-HAP material. In general, the results proved that the osteosynthesis plate built of graded Ti-HAP material resulted in lower bone stress compared to titanium and steel plates. The results obtained confirmed the validity of the design and the possibility to use functionally graded Ti-HAP bone fixation plates.

Geophysics-Inspired Nonlinear Stress-Strain Law for Biological Tissues and Its Applications in Compression Optical Coherence Elastography.

We propose a nonlinear stress-strain law to describe nonlinear elastic properties of biological tissues using an analogy with the derivation of nonlinear constitutive laws for cracked rocks. The derivation of such a constitutive equation has been stimulated by the recently developed experimental technique-quasistatic Compression Optical Coherence Elastography (C-OCE). C-OCE enables obtaining nonlinear stress-strain dependences relating the applied uniaxial compressive stress and the axial component of the resultant strain in the tissue. To adequately describe nonlinear stress-strain dependences obtained with C-OCE for various tissues, the central idea is that, by analogy with geophysics, nonlinear elastic response of tissues is mostly determined by the histologically confirmed presence of interstitial gaps/pores resembling cracks in rocks. For the latter, the nonlinear elastic response is mostly determined by elastic properties of narrow cracks that are highly compliant and can easily be closed by applied compressing stress. The smaller the aspect ratio of such a gap/crack, the smaller the stress required to close it. Upon reaching sufficiently high compressive stress, almost all such gaps become closed, so that with further increase in the compressive stress, the elastic response of the tissue becomes nearly linear and is determined by the Young's modulus of the host tissue. The form of such a nonlinear dependence is determined by the distribution of the cracks/gaps over closing pressures; for describing this process, an analogy with geophysics is also used. After presenting the derivation of the proposed nonlinear law, we demonstrate that it enables surprisingly good fitting of experimental stress-strain curves obtained with C-OCE for a broad range of various tissues. Unlike empirical fitting, each of the fitting parameters in the proposed law has a clear physical meaning. The linear and nonlinear elastic parameters extracted using this law have already demonstrated high diagnostic value, e.g., for differentiating various types of cancerous and noncancerous tissues.

Measurement of maize stalk shear moduli.

Maize is the most grown feed crop in the United States. Due to wind storms and other factors, 5% of maize falls over annually. The longitudinal shear modulus of maize stalk tissues is currently unreported and may have a significant influence on stalk failure. To better understand the causes of this phenomenon, maize stalk material properties need to be measured so that they can be used as material constants in computational models that provide detailed analysis of maize stalk failure. This study reports longitudinal shear modulus of maize stalk tissue through repeated torsion testing of dry and fully mature maize stalks. Measurements were focused on the two tissues found in maize stalks: the hard outer rind and the soft inner pith. Uncertainty analysis and comparison of multiple methodologies indicated that all measurements are subject to low error and bias. The results of this study will allow researchers to better understand maize stalk failure modes through computational modeling. This will allow researchers to prevent annual maize loss through later studies. This study also provides a methodology that could be used or adapted in the measurement of tissues from other plants such as sorghum, sugarcane, etc.

The rolling-disc milling texture-sorting of raw straw facilitates the reduction of steam explosion intensity

Lignocellulosic fermentable sugars have been attracting increasing attention due to the developments of synthetic biology, yet the heterogeneity of the raw material results in high pretreatment intensities and the generation of numerous fermentation inhibitors. In this study, the rolling-disc milling texture-sorting was established for raw material to mitigate inhibitors generation at its source, specifically targeting the variances in mechanical strength among corn straw tissues. Through the texture-sorting of raw corn straw, the low modulus of elasticity tissues in corn straw were crushed into soft-texture sorts and the high-strength tissues were retained as hard-texture sorts. The results showed that the steam explosion pretreatment intensity required for soft-texture sorts decreased by 77.34 %, from 1125.96 (180 °C, 5 min) to 255.10 (160 °C, 5 min), compared to the hard-texture sorts. The concentrations of inhibitors in the enzymatic hydrolysate of soft-texture sorts decreased by 55.10 %, from 8.13 g/L/(100 g/L total sugars) to 3.65 g/L/(100 g/L total sugars), indicating an improved suitability for enzymatic hydrolysate preparation. Therefore, the rolling-disc milling texture-sorting method was proposed to provide a new approach for the preparation of lignocellulosic fermentable sugars.

Corrosion and mechanical properties of bioresorbable composite based on Fe-Cu-hydroxyapatite powders

In this work, bimetallic Fe-Cu nanopowders with the addition of hydroxyapatite (HAp) were used for the fabrication of composite biodegradable material suitable for extrusion additive manufacturing processes. Varying the metal and organic binder constituents in the feedstock has led to high performance composite material. Increasing the organic binder content in the initial feedstock from 50 to 60 wt% contributes to the reduction of porosity of the obtained composites from 20.6 % to 8.9 %. The 45Fe-Cu-HAp samples were characterized by the highest mechanical properties in the tensile test: yield strength σ0.2 being 110 MPa and tensile strength σB being 150 MPa. The Young's modulus of all composites is close to that of the cortical bone tissue modulus (≈ 15 GPa). The microhardness of the composite has been shown to exceed that of pure iron more than in 2 times. Corrosion tests have demonstrated that the composite material with minimal content of the organic binder (50Fe-Cu-HAp) showed the highest corrosion rate, which makes it more attractive for its application in the fabrication of a biodegradable implant. From the findings, the presented composite material showed significant antibacterial activity (more than 99 % reduction) against S. aureus. The composites showed excellent biocompatibility against sensitive fibroblast cell line 3T3. The more than 90 % cell viability was observed after 24 hours incubated with sample.

Application of 3D printing to create an in vitro aneurysm rupture model.

Currently available benchtop (in vitro) aneurysm models are inadequate for testing the efficacy of endovascular device treatments. Specifically, current models do not represent the mechanical instability of giant aneurysms (defined as aneurysms with 25 mm in height or width) and do not predictably rupture under simulated physiological conditions. Hence, in vitro aneurysm models with biomechanically relevant material properties and a predictable rupture timeframe are needed to accurately assess the efficacy of new medical device treatment options. Understanding the material properties of an aneurysm (e.g., shear and compression modulus) as it approaches rupture is a crucial step toward creating a pathologically relevant and sophisticated in vitro aneurysm rupture model. We investigated the change in material properties of a blood vessel, via enzymatic treatment, to simulate the degradation of an aneurysm wall and used this information to create a sophisticated aneurysm rupture model using the latest in additive manufacturing technologies (3D printing) with tissue-like materials. Mechanical properties (shear and compression modulus) of swine carotid vessels were evaluated before and after incubation with collagenase D enzyme (30 min at 37°C) to simulate the effect of biochemical activity on aneurysm wall approaching rupture compared to control vessels (untreated). Mechanical strength of a soft and flexible 3D-printed material (VCA-A30: 30 shore A hardness) was tested for comparison to these arterial vessels. This material was then used to create spherical shaped, giant-sized (25-mm diameter) aneurysm phantoms and were run under neurovascular pressures (120/80 ± 5 mmHg), beats per minute (BPM = 70) and flows representing the middle cerebral artery [MCA: 142.67 (±20.13) mL/min] using a blood analog [3.6 (±0.4) cP viscosity] with non-Newtonian shear-thinning properties. The shear modulus of swine carotid vessel before treatment was 12.2 (±2.7) KPa and compression modulus was 663.5 (±111.6) KPa. After enzymatic treatment by collagenase D, shear modulus of animal tissues reduced by 33% (p-value = .039) while compression modulus remained statistically unchanged (p-value = .615). Control group (untreated vessels) showed minimal reduction (13%, p-value = .226) in shear modulus and 78% increase (p-value = .034) in compression modulus. The shear modulus of the 3D-printed material was 228.59 (±24.82) KPa while its compression modulus was 668.90 (±13.16) KPa. This material was used to prototype a sophisticated in vitro giant aneurysm rupture model. When subjected to physiological pressures and flow rates, the untreated models consistently ruptured at ~12 min. These results indicate that aneurysm rupture can be recreated consistently in a benchtop in vitro model, utilizing the latest 3D-printed materials, connected to a physiologically relevant programmable pump. Further studies will investigate the optimization of various aneurysm dome thickness regions within the aneurysm, with tunable rupture times for comparison of aneurysm device deployment and benchtop controls based on the measurable effects of pressure and flow changes within the aneurysm models. These optimized in vitro rupture models could ultimately be used to test the efficacy of device treatment options and rupture risk by quantifying specific device rupture times and aneurysm rupture position.

MXene-Based Skin-Like Hydrogel Sensor and Machine Learning-Assisted Handwriting Recognition.

Conductive hydrogels are widely used in flexible sensors owing to their adjustable structure, good conductivity, and flexibility. The performance of excellent mechanical properties, high sensitivity, and elastic modulus compatible with human tissues is of great interest in the field of flexible sensors. In this paper, the functional groups of trisodium citrate dihydrate (SC) and MXene form multiple hydrogen bonds in the polymer network to prepare a hydrogel with mechanical properties (Young's modulus (23.5-92 kPa) of similar human tissue (0-100 kPa)), sensitivity (stretched GF is 4.41 and compressed S1 is 5.15 MPa-1), and durability (1000 cycles). The hydrogel is able to sensitively detect deformations caused by strain and stress and can be used in flexible sensors to detect human movement in real time such as fingers, wrists, and walking. In addition, the combination of matrix sensing and machine learning was successfully used for handwriting recognition with an accuracy of 0.9744. The combination of machine learning and flexible sensors shows great potential in areas such as healthcare, information security, and smart homes.

Fabrication of Apparatus Specialized for Measuring the Elasticity of Perioral Tissues.

On the human face, the lips are one of the most important anatomical elements, both morphologically and functionally. Morphologically, they have a significant impact on aesthetics, and abnormal lip morphology causes sociopsychological problems. Functionally, they play a crucial role in breathing, articulation, feeding, and swallowing. An apparatus that can accurately and easily measure the elastic modulus of perioral tissues in clinical tests was developed, and its measurement sensitivity was evaluated. The apparatus is basically a uniaxial compression apparatus consisting of a force sensor and a displacement sensor. The displacement sensor works by enhancing the restoring force due to the deformation of soft materials. Using the apparatus, the force and the displacement were measured for polyurethane elastomers with various levels of softness, which are a model material of human tissues. The stress measured by the developed apparatus increased in proportion to Young's modulus, and was measured by the compression apparatus at the whole region of Young's modulus, indicating that the relation can be used for calibration. Clinical tests using the developed apparatus revealed that Young's moduli for upper lip, left cheek, and right cheek were evaluated to be 45, 4.0, and 9.9 kPa, respectively. In this paper, the advantages of this apparatus and the interpretation of the data obtained are discussed from the perspective of orthodontics.

Biocompatible Co-organic Composite Thin Film Deposited by VHF Plasma-Enhanced Atomic Layer Deposition at a Low Temperature.

Although metal-organic thin films are required for many biorelated applications, traditional deposition methods have proven challenging in preparing these composite materials. Here, a Co-organic composite thin film was prepared by plasma-enhanced atomic layer deposition (PEALD) with cobaltocene (Co(Cp)2) on polydimethylsiloxane (PDMS), using two very high frequency (VHF) NH3 plasmas (60 and 100 MHz), for use as a tissue culture scaffold. VHF PEALD was employed to reduce the temperature and control the thickness and composition. In the result of the VHF PEALD process, the Young's modulus of the Co-organic composite thin film ranged from 82.0 ± 28.6 to 166.0 ± 15.2 MPa, which is similar to the Young's modulus of soft tissues. In addition, the deposited Co ion on the Co-organic composite thin film was released into the cell culture media under a nontoxic level for the biological environment. The proliferation of both L929, the mouse fibroblast cell line, and C2C12, the mouse myoblast cell line, increased to 164.9 ± 23.4% during 7 days of incubation. Here, this novel bioactive Co-organic composite thin film on an elastic PDMS substrate enhanced the proliferation of L929 and C2C12 cell lines, thereby expanding the application range of VHF PEALD in biological fields.

Association between Elastic Modulus of Foot Soft Tissues and Gait Characteristics in Young Individuals with Flatfoot.

Flatfoot is a common foot deformity, causing foot pain, osteoarthritis of the midfoot, and even knee and hip dysfunction. The elastic modulus of foot soft tissues and its association with gait biomechanics still remain unclear. For this study, we recruited 20 young individuals with flatfoot and 22 age-matched individuals with normal foot arches. The elastic modulus of foot soft tissues (posterior tibial tendon, flexor digitorum brevis, plantar fascia, heel fat pad) was obtained via ultrasound elastography. Gait data were acquired using an optical motion capture system. The association between elastic modulus and gait data was analyzed via correlation analysis. The elastic modulus of the plantar fascia (PF) in individuals with flatfoot was higher than that in individuals with normal foot arches. There was no significant difference in the elastic modulus of the posterior tibial tendon (PTT), the flexor digitorum brevis (FDB), or the heel fat pad (HFD), or the thickness of the PF, PTT, FDB, and HFD. Individuals with flatfoot showed greater motion of the hip and pelvis in the coronal plane, longer double-support phase time, and greater maximum hip adduction moment during walking. The elastic modulus of the PF in individuals with flatfoot was positively correlated with the maximum hip extension angle (r = 0.352, p = 0.033) and the maximum hip adduction moment (r = 0.429, p = 0.039). The plantar fascia is an important plantar structure in flatfoot. The alteration of the plantar fascia's elastic modulus is likely a significant contributing factor to gait abnormalities in people with flatfoot. More attention should be given to the plantar fascia in the young population with flatfoot.

Ultrasound Elastography Assessment of Knee Intra-Articular Adhesions at Varying Knee Angles.

We aimed to verify the feasibility of using shear wave elastography (SWE) to quantify knee scars and the elastic modulus of scar tissues. Overall, 16 participants underwent SWE assessments and range-of-motion measurement and completed the Knee Injury and Osteoarthritis Outcome Score. The inter-rater reliability for SWE in the suprapatellar bursa, below the patellar tendon, and in the medial and lateral trochlear groove remained within 0.861-0.907. The SWE values in the four regions increased with increasing knee angle, and significant differences were observed between the values for below the patellar tendon and the suprapatellar bursa at knee flexion angles of 60° and 90°. The SWE values of the medial and lateral trochlear groove at 30°, 60°, and 90° knee flexion were higher on the affected side. A negative correlation was observed between the SWE values for the lateral trochlear groove at 0°, 30°, and 60° and those for below the patellar tendon at 0° and the suprapatellar bursa at 30° with both active and passive knee extension. The suprapatellar bursa value at 60° exhibited a positive correlation with both knee flexion and passive knee flexion, whereas that of the suprapatellar bursa at 90° exhibited a positive correlation with both the range of motion and passive range of motion. SWE is a replicable and effective method for detecting scar strength in the knee joint.

Mechanical and structural properties of articular cartilage and subchondral bone in human osteoarthritic knees.

Knee osteoarthritis (OA), characterized by multiple joint tissue degenerations, remains a significant clinical challenge. Recent evidence suggests that crosstalk within the osteochondral unit may drive OA progression. Although structural-biomechanical properties of bone and cartilage have been studied, potential interaction within the osteochondral unit in the context of OA has yet to be investigated. We performed comprehensive structural and biomechanical quantification of the cartilage, subchondral bone plate (SBP), and subchondral trabecular bone (STB) using 101 osteochondral cores collected from tibial plateaus of 12 control human cadavers (CT, 5 male/7 female) and 19 patients undergoing total knee replacement (OA, 6 male/13 female). For each sample, we quantified SBP microstructure, plate-and-rod morphological properties of the STB using individual trabecula segmentation, and morphological and compositional properties of the articular cartilage. We also performed indentation testing on each compartment of the osteochondral unit to extract the respective structural-mechanical properties. Cartilage thickness was lower in moderate and severe OA regions, while Osteoarthritis Research Society International score was higher only in severe OA regions. GAG content did not change in any OA region. Aggregate and shear moduli were lower only in severe OA regions, while permeability was lower only in moderate OA regions. In the SBP, thickness and tissue mineral density were higher in moderate and severe OA regions. Tissue modulus of STB was lower in moderate OA regions despite a thicker and more mineralized SBP; this deterioration was not observed in severe OA regions. Regression analysis revealed strong correlations between cartilage and STB properties in CT; these correlations were also found in moderate OA regions but were not observed in severe OA regions. In summary, our findings comprehensively characterize the human OA osteochondral unit. Importantly, uncoupling cartilage and subchondral bone structural-mechanical properties may be a hallmark of OA.

Tuning the rheological properties of chitosan/alginate hydrogels for tissue engineering application

Biomaterials that mimic biological tissues are in great demand in tissue engineering. Hydrogels are rising as a conducive candidate in tissue engineering on account of their water-holding capacity, mechanical strength, and elasticity. Nevertheless, the development of hydrogels that mimic biological tissues with the desired stability and mechanical strength, remains a notable barrier. In this investigation, chitosan/alginate hydrogels were prepared by tuning the chitosan concentration from 0.5 % to 2.5 %w/v with a fixed 1 %w/v alginate concentration using 0.1 % glutaraldehyde as a gelling agent. The hydrogel system forms through covalent bonding between chitosan and alginate, mediated by aldehyde groups of the glutaraldehyde. The mechanical strength of the hydrogel was elucidated by probing its viscoelastic behavior through rheological analysis. The rheological investigations revealed that the prepared chitosan/alginate hydrogels are profoundly stable with shear-thinning characteristics. The increase in chitosan concentration enhanced the stability and viscoelastic attributes of the hydrogels. The yield stress values ranging from 0.93 kPa to 14.24 kPa indicate the possibility of using these hydrogels in bioadhesives and soft and bone tissue engineering. The complex modulus of the prepared chitosan/alginate hydrogels resembles the shear modulus of liver tissues, osteogenic cells, and articular cartilage, and hence, these hydrogels pose as suitable candidates in liver, bone, and cartilage tissue engineering. The chitosan/alginate hydrogels possess outstanding self-healing ability, along with biocompatibility, stability, and mechanical strength, rendering them highly advantageous for applications in biomedical tissue engineering and bioadhesives.

Experimental extraction of Young's modulus of gastric tissue with development of spherical, cylindrical, and crowned rollers contact theories

Nanotechnology has been considered with the aim of recognizing the structural and mechanical properties as well as improving the treatment and diagnostic process in the field of medicine. The process of nanomanipulation by examining healthy and cancerous tissues in nanoscale is one of the processes used in this field. Therefore, in this article, considering the importance of recognizing the properties of cancerous and healthy tissues in improving the treatment and diagnosis process, one of the most common types of cancer has been studied. Young modulus has been used as a parameter in the diagnosis of cancerous tissue and its value has been calculated for gastric cancerous tissue. To achieve this goal, atomic force microscopy (AFM) was used during the manipulation process. This tool with the ability to study cancerous tissues in different environments and with the least amount of damage to the target tissue, is one of the effective tools in the field of nanomanipulation. The parameter studied in this study is the geometry of gastric cancer tissue. Therefore, the simulations have been performed by considering contact models with spherical, cylindrical and crowned rollers geometries. The force-indentation depth diagram for gastric tissue is plotted experimentally and compared with theoretical results. According to the experimental work done after reviewing the recorded topographic images, the approximate range of the Young's modulus value for gastric tissue has been calculated according to different geometries. Since the geometry of the crowned rollers is closer to the geometry of the gastric tissue, it has a higher accuracy and the values of the Young's modulus have been calculated according to this geometry in the range of 316–310 KPa.

In vivo mechanical response of thigh soft tissues under compression: A two-layer model allows an improved representation of the local tissue kinematics

Biomechanical parameters have the potential to be used as physical markers for prevention and diagnosis. Finite Element Analysis (FEA) is a widely used tool to evaluate these parameters in vivo. However, the development of clinically relevant FEA requires personalisation of the geometry, boundary conditions, and constitutive parameters. This contribution focuses on the characterisation of mechanical properties in vivo which remains a significant challenge for the community. The aim of this retrospective study is to evaluate the sensitivity of the computed elastic parameters (shear modulus of fat and muscle tissues) derived by inverse analysis as a function of the geometrical modelling assumption (homogenised monolayer vs bilayer) and the formulation of the cost function. The methodology presented here proposes to extract the experimental force-displacement response for each tissue layer (muscle and fat) and construct the associated Finite Element Model for each volunteer, based on data previously collected in our group (N = 7 volunteers) as reported in (Fougeron et al., 2020). The sensitivity analysis indicates that the choice of the cost function has minimal impact on the topology of the response surface in the parametric space. Each surface displays a valley of parameters that minimises the cost function. The constitutive properties of the thigh (reported as median ± interquartile range) were determined to be (μ=198±322Pa,α=37) for the monolayer and (μmuscle=1675±1127Pa,αmuscle=22±14,μfat=537±1131Pa,αfat=32±7) for the bilayer. A comparison of the homogenised monolayer and bilayer models showed that adding a layer reduces the error on the local force displacement curves, increasing the accuracy of the local kinematics of soft tissues during indentation. This allows for an increased understanding of load transmission in soft tissue. The comparison of the two models in terms of strains indicates that the modelling choice significantly influences the localization of maximal compressive strains. These results support the idea that the biomechanical community should conduct further work to develop reliable methodologies for estimating in vivo strain in soft tissue.

Treadmill exercise promotes bone tissue recovery in rats subjected to high + Gz loads.

High + Gz loads, the gravitational forces experienced by the body in hypergravity environments, can lead to bone loss in pilots and astronauts, posing significant health risks. To explore the effect of treadmill exercise on bone tissue recovery, a study was conducted on 72 male Wistar rats. These rats were subjected to four weeks of varying levels of periodic high + Gz loads (1G, 8G, 20G) experiments, and were subsequently divided into the treadmill group and the control group. The treadmill group underwent a continuous two-week treadmill experiment, while the control group rested during this period. The mechanical properties, microstructure, and molecular markers of their tibial bone tissue were measured using three-point bending, micro-CT, and PCR. The results showed that treadmill exercise improved the elastic modulus, ultimate deflection, and ultimate load of rat bone tissue. It also increased the number, density, and volume fraction of bone trabeculae, and decreased their separation. Moreover, treadmill exercise enhanced osteogenesis and inhibited osteoclastogenesis. This study demonstrates that treadmill exercise can promote the recovery of bone tissue in rats subjected to high + Gz loads, providing a potential countermeasure for bone loss in pilots and astronauts.

Compression cycling of 3D-printed meniscal tissues in vitro using a custom bioreactor

An estimated 750,000 arthroscopic knee operations are performed in the United States each year, and many are due to a torn meniscus. Transplantation with donor tissue is the gold standard of care in cases where the meniscus cannot be repaired. However, there is a limited supply of transplantable tissue, which may not be the ideal size or shape for the recipient. 3D printing and tissue engineering have been used to produce replacement tissue of specified shape and size, but none offer the compressive modulus or durability of adult-derived tissue. While biomechanical loading of engineered tissues is known to increase mechanical strength, no current paradigms provide sufficient strength. Instead, a combinatorial approach addressing both physiological form and function has emerged as a promising strategy. In this work, anisotropic menisci were bioprinted using ink composed of collagen types I & II, chondroitin sulfate, and mesenchymal stem cells. After printing, a custom bioreactor was used to apply cyclic compression within an incubator throughout the culture period. Compression cycled prints containing cells maintained viability for 3 weeks, while the mechanical strength of cellularized prints increased after 1 week. However, print dimensions and mass of cellular prints decreased over time independent of compression, while glycosaminoglycans were lost from the prints into the culture media. The expression of eight genes were significantly altered due to compression cycling. This work demonstrated that bioprinted menisci containing live cells can be successfully compressed over long time periods in culture without cell death, and despite changing print dimensions, cells under compression contributed to meniscal strengthening whereas acellular prints consistently weaken. By optimizing structure, culture conditions, and compression paradigms, the strength of bioprinted menisci may approach that of native tissue, and this combinatorial approach may reduce or eliminate the need for cadaveric tissues for allograft transplants.

An Apparatus for Measuring Combined Shear-Tensile Loading in Fibrous Tissues Ex Vivo.

Soft tissues such as tendon and ligament undergo a combination of shear and tensile loading in vivo due to their boundary conditions at muscle and/or bone. Current experimental protocols are limited to pure tensile loading, biaxial loading, or simple shear, and thus may not fully characterize the mechanics of these tissues under physiological loading scenarios. Our objective was to create an experimental protocol to determine the shear modulus of fibrous tissues at different tensile loads. We assembled a four-actuator experimental system that facilitated shear deformation to be superimposed on a tissue subjected to an axial preload. We measured shear modulus in axially loaded electrospun nanofiber scaffolds with either randomly oriented or aligned fibers. We found that shear modulus in the nanofiber phantoms was shear-strain stiffening and dependent on both the axial load (p < 0.001) and fiber alignment (p < 0.001) of the scaffold. The proposed system can enhance our understanding of microstructure and functional mechanics in soft tissues, while also providing a platform to investigate the behavior of electrospun scaffolds for tissue regeneration. Our experimental protocol for determining loaded shear modulus would be further useful as a method to gauge tissue mechanics under loading conditions that are more representative of physiological loads applied to tendon and ligament.

Fabrication and Characterization of Quad-Component Bioinspired Hydrogels to Model Elevated Fibrin Levels in Central Nervous Tissue Scaffolds.

Multicomponent interpenetrating polymer network (mIPN) hydrogels are promising tissue-engineering scaffolds that could closely resemble key characteristics of native tissues. The mechanical and biochemical properties of mIPNs can be finely controlled to mimic key features of target cellular microenvironments, regulating cell-matrix interactions. In this work, we fabricated hydrogels made of collagen type I (Col I), fibrin, hyaluronic acid (HA), and poly (ethylene glycol) diacrylate (PEGDA) using a network-by-network fabrication approach. With these mIPNs, we aimed to develop a biomaterial platform that supports the in vitro culture of human astrocytes and potentially serves to assess the effects of the abnormal deposition of fibrin in cortex tissue and simulate key aspects in the progression of neuroinflammation typically found in human pathologies such as Alzheimer's disease (AD), Parkinson's disease (PD), and tissue trauma. Our resulting hydrogels closely resembled the complex modulus of AD human brain cortex tissue (~7.35 kPa), promoting cell spreading while allowing for the modulation of fibrin and hyaluronic acid levels. The individual networks and their microarchitecture were evaluated using confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Human astrocytes were encapsulated in mIPNs, and negligible cytotoxicity was observed 24 h after the cell encapsulation.