Ultrasound attenuation of cortical bone correlates with biomechanical, microstructural, and compositional properties.

BACKGROUND: We investigated the relationship of two commonly used quantitative ultrasound (QUS) parameters, speed of sound (SoS) and attenuation coefficient (α), with water and macromolecular contents of bovine cortical bone strips as measured with ultrashort echo time (UTE) magnetic resonance imaging (MRI). METHODS: SoS and α were measured in 36 bovine cortical bone strips utilizing a single-element transducer with nominal 5 MHz center frequency based on the time of flight principles after accommodating for reflection losses. Specimens were then scanned using UTE MRI to measure total, bound, and pore water proton density (TWPD, BWPD, and PWPD) as well as macromolecular proton fraction and macromolecular transverse relaxation time (T2-MM). Specimens were also scanned using microcomputed tomography (µCT) at 9-µm isometric voxel size to measure bone mineral density (BMD), porosity, and pore size. The elastic modulus (E) of each specimen was measured using a 4-point bending test. RESULTS: α demonstrated significant positive Spearman correlations with E (R = 0.69) and BMD (R = 0.44) while showing significant negative correlations with porosity (R = -0.41), T2-MM (R = -0.47), TWPD (R = -0.68), BWPD (R = -0.67), and PWPD (R = -0.45). CONCLUSIONS: The negative correlation between α and T2-MM is likely indicating the relationship between QUS and collagen matrix organization. The higher correlations of α with BWPD than with PWPD may indicate that water organized in finer structure (bound to matrix) provides lower acoustic impedance than water in larger pores, which is yet to be investigated thoroughly. RELEVANCE STATEMENT: This study highlights the importance of future investigations exploring the relationship between QUS measures and all major components of the bone, including the collagenous matrix and water. Investigating the full potential of QUS and its validation facilitates a more affordable and accessible tool for bone health monitoring in clinics. KEY POINTS: • Ultrasound attenuation demonstrated significant positive correlations with bone mechanics and mineral density. • Ultrasound attenuation demonstrated significant negative correlations with porosity and bone water contents. • This study highlights the importance of future investigations exploring the relationship between QUS measures and all major components of the bone.

Correlations between elastic modulus and ultrashort echo time (UTE) adiabatic T1ρ relaxation time (UTE-Adiab-T1ρ) in Achilles tendons and entheses.

Patients with psoriatic arthritis commonly have abnormalities of their entheses, which are the connections between tendons and bone. There are shortcomings with the use of conventional magnetic resonance imaging (MRI) sequences for the evaluation of entheses and tendons, whereas ultrashort echo time (UTE) sequences are superior for the detection of high signals, and can also be used for non-invasive quantitative assessments of these structures. The combination of UTE-MRI with an adiabatic-T1ρ preparation (UTE-Adiab-T1ρ) allows for reliable assessment of entheses and tendons with decreased susceptibility to detrimental magic angle effects. This study aimed to investigate the relationship between quantitative UTE-MRI measures and the biomechanical properties of Achilles tendons and entheses. In total, 28 tendon-enthesis sections were harvested from 11 fresh-frozen human cadaveric foot-ankle specimens (52 ± years old). Tendon-enthesis sections were scanned using the UTE-Adiab-T1ρ and UTE-T1 sequences on a clinical 3 T scanner. MRI-based measures and indentation tests were performed on the enthesis, transitional, and tensile tendon zones of the specimens. Hayes' elastic modulus showed significant inverse correlations (Spearman's) with UTE-Adiab-T1ρ in all zones (R= - 0.46, - 0.54, and - 0.61 in enthesis, transition, and tensile tendon zones, respectively). Oliver-Pharr's elastic modulus showed significant inverse correlations with UTE-Adiab-T1ρ in transition (R= - 0.52) and tensile tendon zone (R=- 0.60). UTE-T1 did not show significant correlations with the elastic modulus. UTE-MRI and elastic modulus were significantly lower in the tensile tendon compared with the enthesis regions This study highlights the potential of the UTE-Adiab-T1ρ technique for the non-invasive evaluation of tendons and enthuses.

Determining the Relationship between Mechanical Properties and Quantitative Magnetic Resonance Imaging of Joint Soft Tissues Using Patient-Specific Templates.

To determine whether the mechanical properties of joint soft tissues such as cartilage can be calculated from quantitative magnetic resonance imaging (MRI) data, we investigated whether the mechanical properties of articular cartilage and meniscus scheduled to be resected during arthroplasty are correlated with the T2 relaxation time on quantitative MRI at the same location. Six patients who had undergone knee arthroplasty and seven who had undergone hip arthroplasty were examined. For the knee joint, the articular cartilage and lateral meniscus of the distal lateral condyle of the femur and proximal lateral tibia were examined, while for the hip joint, the articular cartilage above the femoral head was studied. We investigated the relationship between T2 relaxation time by quantitative MRI and stiffness using a hand-made compression tester at 235 locations. The patient-individualized template technique was used to align the two measurement sites. The results showed a negative correlation (from -0.30 to -0.35) in the less severely damaged articular cartilage and meniscus. This indicates that tissue mechanical properties can be calculated from T2 relaxation time, suggesting that quantitative MRI is useful in determining when to start loading after interventional surgery on cartilage tissue and in managing the health of elderly patients.

Ultrashort Echo Time (UTE) MRI porosity index (PI) and suppression ratio (SR) correlate with the cortical bone microstructural and mechanical properties: Ex vivo study.

Ultrashort echo time (UTE) MRI can image and consequently enable quantitative assessment of cortical bone. UTE-MRI-based evaluation of bone is largely underutilized due to the high cost and time demands of MRI in general. The signal ratio in dual-echo UTE imaging, known as porosity index (PI), as well as the signal ratio between UTE and inversion recovery UTE (IR-UTE) imaging, known as the suppression ratio (SR), are two rapid UTE-based bone evaluation techniques (∼ 5 mins scan time each), which can potentially reduce the time demand and cost in future clinical studies. This study aimed to investigate the correlations of PI and SR measures with cortical bone microstructural and mechanical properties. Cortical bone strips (n = 135) from tibial and femoral midshafts of 37 donors (61 ± 24 years old) were scanned using a dual-echo 3D Cones UTE sequence and a 3D Cones IR-UTE sequence for PI and SR calculations, respectively. Average bone mineral density, porosity, and pore size were measured using microcomputed tomography (µCT). Bone mechanical properties were measured using 4-point bending tests. The µCT measures showed significant correlations with PI (moderate to strong, R = 0.68-0.71) and SR (moderate, R = 0.58-0.68). Young's modulus, yield stress, and ultimate stress demonstrated significant moderate correlations with PI and SR (R = 0.52-0.62) while significant strong correlations with µCT measures (R > 0.7). PI and SR can potentially serve as fast and noninvasive (non-ionizing radiation) biomarkers for evaluating cortical bone in various bone diseases.

Pneumatospinning Biomimetic Scaffolds for Meniscus Tissue Engineering.

Nanofibrous scaffolds fabricated via electrospinning have been proposed for meniscus tissue regeneration. However, the electrospinning process is slow, and can only generate scaffolds of limited thickness with densely packed fibers, which limits cell distribution within the scaffold. In this study, we explored whether pneumatospinning could produce thicker collagen type I fibrous scaffolds with higher porosity, that can support cell infiltration and neo-fibrocartilage tissue formation for meniscus tissue engineering. We pneumatospun scaffolds with solutions of collagen type I with thicknesses of approximately 1 mm in 2 h. Scanning electron microscopy revealed a mix of fiber sizes with diameters ranging from 1 to 30 µm. The collagen scaffold porosity was approximately 48% with pores ranging from 7.4 to 100.7 µm. The elastic modulus of glutaraldehyde crosslinked collagen scaffolds was approximately 45 MPa, when dry, which reduced after hydration to 0.1 MPa. Mesenchymal stem cells obtained from the infrapatellar fat pad were seeded in the scaffold with high viability (>70%). Scaffolds seeded with adipose-derived stem cells and cultured for 3 weeks exhibited a fibrocartilage meniscus-like phenotype (expressing COL1A1, COL2A1 and COMP). Ex vivo implantation in healthy bovine and arthritic human meniscal explants resulted in the development of fibrocartilage-like neotissues that integrated with the host tissue with deposition of glycosaminoglycans and collagens type I and II. Our proof-of-concept study indicates that pneumatospinning is a promising approach to produce thicker biomimetic scaffolds more efficiently that electrospinning, and with a porosity that supports cell growth and neo-tissue formation using a clinically relevant cell source.

Correlation between the elastic modulus of anterior cruciate ligament (ACL) and quantitative ultrashort echo time (UTE) magnetic resonance imaging.

Conventional magnetic resonance imaging (MRI) often acquires no signal in anterior cruciate ligament (ACL) due to the short apparent transverse relaxation time of ACL. Ultrashort echo time (UTE) MRI is capable of imaging ACL with high signal which enables quantitative ACL assessment. This study aimed to investigate the correlations of the mechanical and microstructural properties of human ACL specimens with quantitative three-dimensional UTE Cones (3D-UTE-Cones) MRI measures. ACL specimens were harvested from cadaveric knee joints of 13 (50.9 ± 21.1 years old, 11 males and 2 female) donors. Specimens were scanned using a series of quantitative 3D-UTE-Cones T2 * (UTE-T2 *), T1 (UTE-T1 ), Adiabatic T1ρ (UTE-Adiab-T1ρ ), and magnetization transfer (UTE-MT) sequences in a wrist coil on a clinical 3T scanner. ACL elastic modulus was measured using a uniaxial tensile mechanical test. Histomorphometry analysis was performed to measure the average fascicle specific surface, fascicle size, and number of cells per unit area. Spearman's rank correlations of UTE-MRI biomarkers with mechanical and histomorphometry measures were investigated. The elastic modulus of ACL showed significant moderate correlations with UTE-Adiab-T1ρ (R = -0.59, p = 0.01), macromolecular fraction from MT modeling (R = 0.54, p = 0.01), magnetization transfer ratio (R = 0.53, p = 0.01), UTE-T2* (R = -0.53, p = 0.01), and average fascicle specific surface (R = 0.54, p = 0.01). UTE-MRI showed nonsignificant correlations with histomorphometry measures. UTE-MRI biomarkers may be useful noninvasive tools for the ACL mechanical assessment.

Intraoperative knee kinematics measured by computer-assisted navigation and intraoperative ligament balance have the potential to predict postoperative knee kinematics.

This study was designed to analyze the effects of type of activity and cruciate ligament resection on knee kinematics and ligament balance after total knee arthroplasty (TKA), and to determine if intraoperative passive kinematics are associated with active kinematics. Fresh-frozen human cadaveric knees were examined. The knees were mounted on a quadriceps-driven simulator. Cruciate-retaining (CR-TKA) and posterior-substituting (PS-TKA) TKA was performed using a contemporary knee system. Active flexion (closed-kinetic chain [CKC] and open-kinetic-chain [OKC]) and passive flexion were analyzed by recording the knee kinematics using a specifically developed application of an imageless navigation system. An electronic ligament balancer was used to measure the tibiofemoral gap under constant distraction pressure. The femur rotated externally relative to the tibia during passive and active CKC flexion. The femur translated anteriorly from 10° to 50° of flexion after TKA. Beyond 50° of flexion, the femur translated posteriorly in all surgical conditions. The femoral location during active CKC flexion was posterior relative to that during active OKC. Femoral rotation and translation during passive knee flexion correlated significantly with that during active knee flexion. Posterior tilt of the electronic ligament balancer was greater with CR-TKA than with PS-TKA and correlated significantly with the anteroposterior position of the femur. Statement of Clinical Significance: Intraoperative knee kinematics measured by computer-assisted navigation and intraoperative ligament balance have the potential to predict postoperative knee kinematics.

A Useful Combination of Quantitative Ultrashort Echo Time MR Imaging and a Probing Device for Biomechanical Evaluation of Articular Cartilage.

In this study, we combined quantitative ultrashort echo time (UTE) magnetic resonance (MR) imaging and an investigation by a probing device with tri-axial force sensor to seek correlations with mechanical properties of human patellar cartilage for in situ evaluation of biomechanical properties. Cartilage blocks (15 × 20 × 5 mm3) were dissected from the patella of six donors; 5 mm square regions of interest from the cartilage blocks were imaged using UTE-MR imaging sequences (T2* and magnetization transfer ratio (MTR)), and mechanical properties were measured using a micro indentation device. Then, the vertical reaction force on the cartilage surface was measured while push-probing forward 3 mm with the probing device at a 30° tilt to the horizontal plane. The results showed a positive correlation between stiffness/elastic modulus and each predictor variable (UTE-T2* (r = 0.240 and 0.255, respectively, UTE-MTR (r = 0.378 and 0.379, respectively), and probing device force (r = 0.426 and 0.423, respectively). Furthermore, multiple linear regression analysis showed the combination of the three predictors had stronger correlation (adjusted r2 = 0.314 (stiffness), 0.323 (elastic), respectively). Our results demonstrate the potential for these non- and less- invasive methods for in situ evaluation of the mechanical properties of cartilage tissue.

Nanofiber Scaffolds by Electrospinning for Rotator Cuff Tissue Engineering.

Rotator cuff tears continue to be at risk of retear or failure to heal after surgical repair, despite the use of various surgical techniques, which stimulate development of novel scaffolding strategies. They should be able to address the known causes of failure after the conventional rotator cuff repair: (1) failure to reproduce the normal tendon healing process, (2) resultant failure to reproduce four zones of the enthesis, and (3) failure to attain sufficient mechanical strength after repair. Nanofiber scaffolds are suited for this application because they can be engineered to mimic the ultrastructure and properties of the native rotator cuff tendon. Among various methods for tissue-engineered nanofibers, electrospinning has recently been highlighted in the rotator cuff field. Electrospinning can create fibrous and porous structures that resemble natural tendon's extracellular matrix. Other advantages include the ability to create relatively large surface-to-volume ratios, the ability to control fiber size from the micro to the nano scale, and the flexibility of material choices. In this review, we will discuss the anatomical and mechanical features of the rotator cuff tendon, their potential impacts on improper healing after repair, and the current knowledge of the use of electrospinning for rotator cuff tissue engineering.

The Resistance Force of the Anterior Cruciate Ligament during Pull Probing Is Related to the Mechanical Property.

There are various methods for reconstructing the anterior cruciate ligament (ACL) from other muscles or tendons. Initial tension of the reconstructed ACL is one of the key elements affecting postoperative outcomes. However, tension cannot be measured after graft fixation. The only intraoperative assessment is pull probing, which is performed by pulling joint soft tissues with the arthroscopic probe and can be measured quantitatively. Therefore, its value might be used as an alternative value for the mechanical property of the ACL. Using a probing device one author developed to measure the resistance force of soft tissues quantitatively while probing, we measured the resistance force of dissected ACLs and used tensile testing to investigate the correlation between the resistance force and the mechanical property of the ligaments. According to the results, when a certain amount of tension (strain; 16.6%) was applied, its mechanical properties were moderately correlated (r = 0.56 [p = 0.045]) with the probing force. Therefore, the tension of the reconstructed ACL after fixation under real ACL reconstruction surgery can be derived from the value of the probing device.

Development of an Ex Vivo Murine Osteochondral Repair Model.

OBJECTIVE: Mouse models are commonly used in research applications due to the relatively low cost, highly characterized strains, as well as the availability of many genetically modified phenotypes. In this study, we characterized an ex vivo murine osteochondral repair model using human infrapatellar fat pad (IPFP) progenitor cells. DESIGN: Femurs from euthanized mice were removed and clamped in a custom multidirectional vise to create cylindrical osteochondral defects 0.5 mm in diameter and 0.5 mm deep in both condyles. The IPFP contains progenitors that are a promising cell source for the repair of osteochondral defects. For proof of concept, human IPFP-derived progenitor cells, from osteoarthritic (OA) patients, cultured as pellets, were implanted into the defects and cultured in serum-free medium with TGFß3 for 3 weeks and then processed for histology and immunostaining. RESULTS: The custom multidirectional vise enabled reproducible creation of osteochondral defects in murine femoral condyles. Implantation of IPFP-derived progenitor cells led to development of cartilaginous tissue with Safranin O staining and deposition of collagen type II in the extracellular matrix. CONCLUSIONS: We showed feasibility in creating ex vivo osteochondral defects and demonstrated the regenerative potential of OA human IPFP-derived progenitors in mouse femurs. The murine model can be used to study the effects of aging and OA on tissue regeneration and to explore molecular mechanisms of cartilage repair using genetically modified mice.

Assessment of mechanical properties of articular cartilage with quantitative three-dimensional ultrashort echo time (UTE) cones magnetic resonance imaging.

Conventional magnetic resonance imaging (MRI) is not capable of detecting signal from the deep cartilage due to its short transverse relaxation time (T2). Moreover, several quantitative MRI techniques are significantly influenced by the magic angle effect. The combinations of ultrashort echo time (UTE) MRI with magnetization transfer (UTE-MT) and Adiabatic T1ρ (UTE-AdiabT1ρ) imaging allow magic angle-insensitive assessments of all regions of articular cartilage. The purpose of this study was to investigate the correlations between quantitative three-dimensional UTE MRI biomarkers and mechanical properties of human tibiofemoral cartilage specimens. In total, 40 human tibiofemoral cartilage specimens were harvested from three male and four female donors (64 ± 18 years old). Cartilage samples were scanned using a series of quantitative 3D UTE Cones T2* (UTE-T2*), T1 (UTE-T1), UTE-AdiabT1ρ, and UTE-MT sequences in a standard knee coil on a clinical 3T scanner. UTE-MT data were acquired with a series of MT powers and frequency offsets to calculate magnetization transfer ratio (MTR), as well as macromolecular fraction (MMF) and macromolecular T2 (T2mm) through modeling. Cartilage stiffness and Hayes elastic modulus were measured using indentation tests. Correlations of 3D UTE Cones MRI measurements in the superficial layer, deep layer, and global regions of interest (ROIs) with mechanical properties were investigated. Cartilage mechanical properties demonstrated highest correlations with UTE measures of the superficial layer of cartilage. AdiabT1ρ, MTR, and MMF in superficial layer ROIs showed significant correlations with Hayes elastic modulus (p < 0.05, R = -0.54, 0.49, and 0.66, respectively). These UTE measures in global ROIs showed significant, though slightly lower, correlations with Hayes elastic modulus (p < 0.05, R = -0.37, 0.52, and 0.60, respectively). Correlations between other UTE MRI measurements (T2*, T1, and T2mm) and mechanical properties were non-significant. The 3D UTE-AdiabT1ρ and UTE-MT sequences were highlighted as promising surrogates for non-invasive assessment of cartilage mechanical properties. MMF from UTE-MT modeling showed the highest correlations with cartilage mechanics.

Sagittal orientation of coracoclavicular ligament reconstruction affects the stability of surgical repair.

BACKGROUND: The variation in the anatomic relationship between the coracoid and the clavicle affects the biomechanical stability of coracoclavicular ligament reconstruction (CCLR). METHODS: Three-dimensional computed tomography reconstruction of 85 patients was analyzed. Anatomic landmarks were used to derive the coracoclavicular sagittal reconstruction angle (sRA). The lateral concave angle, which indicated the shape of the distal clavicle, and the offsets between the clavicle and coracoid were also measured. To investigate the biomechanical effects of the sRA on CCLR, 7 computed tomography scans with different sRAs were 3D printed. Two reconstructions, a single trans-coracoclavicular tunnel and a looped reconstruction technique, were performed sequentially. Models were cyclically loaded at 70 N in the anterior, posterior, and superior directions. RESULTS: The mean sRA was 68° ± 9.3° (range, 47°-85°). The superoinferior offset between the clavicle and the coracoid and the lateral concave angle positively correlated with the sRA (r = 0.359 and 0.837, respectively; P ≤ .001), whereas the anteroposterior offset had a negative correlation (r = -0.925; P < .001). The sRA had a negative correlation with the anterior displacement of the clavicle (rho = -0.96; P < .001) and a positive correlation with the posterior displacement for both surgical techniques (rho = 1.0; P < .001). CONCLUSION: The anatomic orientation of the native coracoclavicular ligaments is highly variable in the sagittal plane. Low sagittal angles can reduce anterior stability, whereas high sagittal angles can reduce posterior stability of CCLR.

Vertical and Rotational Stiffness of Coracoclavicular Ligament Reconstruction: A Biomechanical Study of 3 Different Techniques.

PURPOSE: To compare the biomechanical stability of 3 different coracoclavicular reconstruction techniques under rotational and vertical loading using a cadaveric model. METHODS: In total, 12 cadaveric shoulders were used for testing. The native state was first tested then followed by 3 different reconstruction configurations using suture tapes and cortical buttons: coracoid loop (CL), single-bundle (SB), and double-bundle (DB). Superior displacement was measured by cycling an inferiorly directed force of 70 N to the scapula. The rotational stiffness of the scapula was determined by cycling the scapula in rotational displacement control between 15° of internal and external rotation. The rotational stiffness of the clavicle was determined by rotating the clavicle around its long axis 20° anteriorly and 30° posteriorly in rotational displacement control. All measurements were captured over 10 cycles at a rate of 200 Hz. RESULTS: Both the CL and SB techniques demonstrated significantly less internal scapular rotation stiffness. (intact: 19.70 ± 9.07 cNm/deg, CL: 3.70 ± 2.63 cNm/deg, SB:4.30 ± 2.66 cNm/deg, P <.001) External scapular rotation stiffness was significantly decreased in all techniques (intact: 17.70 ± 4.43 cNm/deg, CL: 3.30 ± 1.37 cNm/deg, SB: 4.50 ± 1.56 cNm/deg, DB: 4.67 ± 1.99 cNm/deg, P < .001). The CL and SB reconstructions were significantly less stiff with regards to posterior rotation of the clavicle (intact: 5.60 ± 1.80 cNm/deg, CL: 2.90 ± 1.10 cNm/deg, SB: 1.40 ± 0.65 cNm/deg, P < .001). Anterior rotation stiffness of the clavicle was significantly lower in all of the reconstructions (intact: 6.95 ± 1.90 cNm/deg, CL: 3.08 ± 0.84 cNm/deg, SB: 3.64 ± 0.93 cNm/deg, DB: 4.48 ± 1.21 cNm/deg, P < .001). CONCLUSIONS: None of the described techniques provided equivalent rotational stability in all planes compared with the native state. DB reconstruction presented stiffness characteristics closest to the native state under cyclic loading during internal scapular and posterior clavicular rotation. CLINICAL RELEVANCE: Additional procedures such as tendon grafting or acromioclavicular ligament reconstruction may be required to control rotational stability.

Cartilage tissue engineering combining microspheroid building blocks and microneedle arrays.

Purpose: Scaffold-free cartilage tissue engineering circumvents issues with scaffold seeding, potential toxicity response, and impaired host integration. However, precisely controlling and maintaining a scaffold-free construct shape have been challenging. We explored the feasibility of microneedle arrays to print tissue using cellular microspheroids as building blocks.Materials and Methods: Human embryonic-derived mesenchymal stem cells or infrapatellar fat pad mesenchymal stem cells were used to create microspheroids of 500 µm in diameter, which were assembled on microneedle arrays in a predefined arrangement using a robotic system under computer vision. Microspheroids on microneedles were cultured to permit fusion into a tissue construct. Infrapatellar fat pad mesenchymal stem cell constructs were either implanted into chondral defects created in human osteoarthritic cartilage explants or maintained on the microneedle array for 3 weeks. Embryonic-derived mesenchymal stem cell constructs were designed to be press-fit into 3 mm subchondral defects in New Zealand White rabbits and maintained for up to 8 weeks to assess retention, early tissue repair, and more mature cartilage regeneration.Results: Microspheroids of both cell types fused together in culture to form neotissues of predefined shape and size. Infrapatellar fat pad mesenchymal stem cell neotissues expressed high levels of chondrogenic genes and integrated with the surrounding osteoarthritic host cartilage. Embryonic-derived mesenchymal stem cell constructs generated chondrogenic neotissue in vivo as early as 2 weeks and more mature tissue by 8 weeks with increased glycosaminoglycan deposition.Conclusions: We constructed defined scaffold-free shapes by bioprinting and fusing microspheroids. Proof of concept was shown in the repair of ex vivo osteoarthritic human cartilage and in vivo rabbit osteochondral (OC) defects.

Correlations of cortical bone microstructural and mechanical properties with water proton fractions obtained from ultrashort echo time (UTE) MRI tricomponent T2* model.

Mechanical and microstructural evaluations of cortical bone using ultrashort echo time magnetic resonance imaging (UTE-MRI) have been performed increasingly in recent years. UTE-MRI acquires considerable signal from cortical bone and enables quantitative bone evaluations. Fitting bone apparent transverse magnetization (T2*) decay using a bicomponent model has been regularly performed to estimate bound water (BW) and pore water (PW) in the quantification of bone matrix and porosity, respectively. Human cortical bone possesses a considerable amount of fat, which appears as MRI T2* signal oscillation and can subsequently lead to BW overestimation when using a bicomponent model. Tricomponent T2* fitting model has been developed to improve BW and PW estimations by accounting for fat contribution in the MRI signal. This study aimed to investigate the correlations of microstructural and mechanical properties of human cortical bone with water pool fractions obtained from a tricomponent T2* model. 135 cortical bone strips (~4 × 2 × 40 mm3 ) from tibial and femoral midshafts of 37 donors (61 ± 24 years old) were scanned using ten sets of dual-echo 3D-UTE-Cones sequences (TE = 0.032-24.0 ms) on a 3 T MRI scanner for T2* fitting analyses. Average bone porosity and pore size were measured using microcomputed tomography (µCT) at 9 µm voxel size. Bone mechanical properties were measured using 4-point bending tests. Using a tricomponent model, bound water fraction (FracBW ) showed significant strong (R = 0.70, P < 0.01) and moderate (R = 0.58-0.62, P < 0.01) correlations with porosity and mechanical properties, respectively. Correlations of bone microstructural and mechanical properties with water pool fractions were higher for tricomponent model results compared with the bicomponent model. The tricomponent T2* fitting model is suggested as a useful technique for cortical bone evaluation where the MRI contribution of bone fat is accounted for.

Assessing cortical bone mechanical properties using collagen proton fraction from ultrashort echo time magnetization transfer (UTE-MT) MRI modeling.

Cortical bone shows as a signal void when using conventional clinical magnetic resonance imaging (MRI). Ultrashort echo time MRI (UTE-MRI) can acquire high signal from cortical bone, thus enabling quantitative assessments. Magnetization transfer (MT) imaging combined with UTE-MRI can indirectly assess protons in the organic matrix of bone. This study aimed to examine UTE-MT MRI techniques to estimate the mechanical properties of cortical bone. A total of 156 rectangular human cortical bone strips were harvested from the tibial and femoral midshafts of 43 donors (62 ±â€¯22 years old, 62 specimens from females, 94 specimens from males). Bone specimens were scanned using UTE-MT sequences on a clinical 3 T MRI scanner and on a micro-computed tomography (µCT) scanner. A series of MT pulse saturation powers (400°, 600°, 800°) and frequency offsets (2, 5, 10, 20, 50 kHz) was used to measure the macromolecular fraction (MMF) utilizing a two-pool MT model. Failure mechanical properties of the bone specimens were measured using 4-point bending tests. MMF from MRI results showed significant strong correlations with cortical bone porosity (R = -0.72, P < 0.01) and bone mineral density (BMD) (R = +0.71, P < 0.01). MMF demonstrated significant moderate correlations with Young modulus, yield stress, and ultimate stress (R = 0.60-0.61, P < 0.01). These results suggest that the two-pool UTE-MT model focusing on the organic matrix of bone can potentially serve as a novel tool to detect the variations of bone mechanical properties and intracortical porosity.

Evaluation of normal cadaveric Achilles tendon and enthesis with ultrashort echo time (UTE) magnetic resonance imaging and indentation testing.

Entheses are regions where tendons and ligaments attach to bone, and are the primary target in seronegative and other diseases of the musculoskeletal (MSK) system. MRI has been widely used for visualizing features of inflammatory and degenerative MSK disease; however, normal tendons and entheses have short transverse relaxation times (T2 ), and show little or no signal with conventional clinical MRI pulse sequences, making it difficult to investigate their MR properties. In this study we examined the normal MR morphology of the cadaveric Achilles tendon and enthesis at 3 T using novel three-dimensional ultrashort echo time (3D UTE) Cones sequences, and at 11.7 T using conventional MRI sequences. We also studied the MR properties of the Achilles tendon and enthesis including T2 *, T1 , and magnetization transfer ratio (MTR). In addition, MT modeling of macromolecular proton fractions was investigated using 3D UTE Cones sequences at 3 T. Indentation testing was performed to investigate the mechanical properties of the tendons and entheses, and this was followed by histological examination. In total five specimens (<50 years) were investigated. On average, tendons and entheses respectively had T2 * values of 0.93 ± 0.48 ms and 2.77 ± 0.79 ms, T1 values of 644 ± 22 ms and 780 ± 55 ms, MTRs of 0.373 ± 0.03 and 0.244 ± 0.009 with an MT power of 1000° and frequency offset of 2 kHz, and macromolecular proton fractions of 18.0 ± 2.2% and 13.9 ± 1.9%. Compared with the tendon, the enthesis generally had a longer T2 *, a longer T1 , a lower MTR, and a lower macromolecular proton fraction as well as both a higher Young's modulus and stiffness. Results from this study are likely to provide a useful baseline for identifying deviations from the normal in seronegative arthritis and other disease of the entheses.

Detecting stress injury (fatigue fracture) in fibular cortical bone using quantitative ultrashort echo time-magnetization transfer (UTE-MT): An ex vivo study.

Bone stress injury (BSI) incidents have been increasing amongst athletes in recent years as a result of more intense sporting activities. Cortical bone in the tibia and fibula is one of the most common BSI sites. Nowadays, clinical magnetic resonance imaging (MRI) is the recommended technique for BSI diagnosis at an early stage. However, clinical MRI focuses on edema observations in surrounding soft tissues, rather than the injured components of the bone. Specifically, both normal and injured bone are invisible in conventional clinical MRI. In contrast, ultrashort echo time (UTE)-MRI is able to detect the rapidly decaying signal from the bone. This study aimed to employ UTE-MRI for fatigue fracture detection in fibula cortical bone through an ex vivo investigation. Fourteen human fibular samples (47 ± 20 years old, four women) were subjected to cyclic loading on a four-point bending setup. The loading was displacement controlled to induce -5000 ± 1500 µ-strain at 4 Hz. Loading was stopped when bone stiffness was reduced by 20%. Fibula samples were imaged twice, using UTE-MRI and micro-computed tomography (µCT), first pre-loading and second post-loading. After loading, the macromolecular fraction (MMF) from UTE-MT modeling demonstrated a significant decrease (12% ± 20%, P = 0.02) on average. Single-component T2 * also decreased significantly by BSI (12% ± 11%, P = 0.01) on average. MMF reduction is hypothesized to be a result of collagenous matrix rupture and water increase. However, faster T2 * decay might be a result of water shifts towards newly developed microcracks with higher susceptibility. Despite this good sensitivity level of the UTE-MRI technique, the µCT-based porosity at a voxel size of 9 µm was not affected by loading. UTE-MRI shows promise as a new quantitative technique to detect BSI.

Control of Retinal Ganglion Cell Positioning and Neurite Growth: Combining 3D Printing with Radial Electrospun Scaffolds.

Retinal ganglion cells (RGCs) are responsible for the transfer of signals from the retina to the brain. As part of the central nervous system, RGCs are unable to regenerate following injury, and implanted cells have limited capacity to orient and integrate in vivo. During development, secreted guidance molecules along with signals from extracellular matrix and the vasculature guide cell positioning, for example, around the fovea, and axon outgrowth; however, these changes are temporally regulated and are not the same in the adult. Here, we combine electrospun cell transplantation scaffolds capable of RGC neurite guidance with thermal inkjet 3D cell printing techniques capable of precise positioning of RGCs on the scaffold surface. Optimal printing parameters are developed for viability, electrophysiological function and, neurite pathfinding. Different media, commonly used to promote RGC survival and growth, were tested under varying conditions. When printed in growth media containing both brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF), RGCs maintained survival and normal electrophysiological function, and displayed radial axon outgrowth when printed onto electrospun scaffolds. These results demonstrate that 3D printing technology may be combined with complex electrospun surfaces in the design of future retinal models or therapies.