In vivo validation of highly customized cranial Ti-6AL-4V ELI prostheses fabricated through incremental forming and superplastic forming: an ovine model study.

Cranial reconstructions are essential for restoring both function and aesthetics in patients with craniofacial deformities or traumatic injuries. Titanium prostheses have gained popularity due to their biocompatibility, strength, and corrosion resistance. The use of Superplastic Forming (SPF) and Single Point Incremental Forming (SPIF) techniques to create titanium prostheses, specifically designed for cranial reconstructions was investigated in an ovine model through microtomographic and histomorphometric analyses. The results obtained from the explanted specimens revealed significant variations in bone volume, trabecular thickness, spacing, and number across different regions of interest (VOIs or ROIs). Those regions next to the center of the cranial defect exhibited the most immature bone, characterized by higher porosity, decreased trabecular thickness, and wider trabecular spacing. Dynamic histomorphometry demonstrated differences in the mineralizing surface to bone surface ratio (MS/BS) and mineral apposition rate (MAR) depending on the timing of fluorochrome administration. A layer of connective tissue separated the prosthesis and the bone tissue. Overall, the study provided validation for the use of cranial prostheses made using SPF and SPIF techniques, offering insights into the processes of bone formation and remodeling in the implanted ovine model.

Biomechanics of the Human Osteochondral Unit: A Systematic Review.

The damping system ensured by the osteochondral (OC) unit is essential to deploy the forces generated within load-bearing joints during locomotion, allowing furthermore low-friction sliding motion between bone segments. The OC unit is a multi-layer structure including articular cartilage, as well as subchondral and trabecular bone. The interplay between the OC tissues is essential in maintaining the joint functionality; altered loading patterns can trigger biological processes that could lead to degenerative joint diseases like osteoarthritis. Currently, no effective treatments are available to avoid degeneration beyond tissues' recovery capabilities. A thorough comprehension on the mechanical behaviour of the OC unit is essential to (i) soundly elucidate its overall response to intra-articular loads for developing diagnostic tools capable of detecting non-physiological strain levels, (ii) properly evaluate the efficacy of innovative treatments in restoring physiological strain levels, and (iii) optimize regenerative medicine approaches as potential and less-invasive alternatives to arthroplasty when irreversible damage has occurred. Therefore, the leading aim of this review was to provide an overview of the state-of-the-art-up to 2022-about the mechanical behaviour of the OC unit. A systematic search is performed, according to PRISMA standards, by focusing on studies that experimentally assess the human lower-limb joints' OC tissues. A multi-criteria decision-making method is proposed to quantitatively evaluate eligible studies, in order to highlight only the insights retrieved through sound and robust approaches. This review revealed that studies on human lower limbs are focusing on the knee and articular cartilage, while hip and trabecular bone studies are declining, and the ankle and subchondral bone are poorly investigated. Compression and indentation are the most common experimental techniques studying the mechanical behaviour of the OC tissues, with indentation also being able to provide information at the micro- and nanoscales. While a certain comparability among studies was highlighted, none of the identified testing protocols are currently recognised as standard for any of the OC tissues. The fibril-network-reinforced poro-viscoelastic constitutive model has become common for describing the response of the articular cartilage, while the models describing the mechanical behaviour of mineralised tissues are usually simpler (i.e., linear elastic, elasto-plastic). Most advanced studies have tested and modelled multiple tissues of the same OC unit but have done so individually rather than through integrated approaches. Therefore, efforts should be made in simultaneously evaluating the comprehensive response of the OC unit to intra-articular loads and the interplay between the OC tissues. In this regard, a multidisciplinary approach combining complementary techniques, e.g., full-field imaging, mechanical testing, and computational approaches, should be implemented and validated. Furthermore, the next challenge entails transferring this assessment to a non-invasive approach, allowing its application in vivo, in order to increase its diagnostic and prognostic potential.

Full-field strain distribution in hierarchical electrospun nanofibrous poly-L(lactic) acid/collagen scaffolds for tendon and ligament regeneration: A multiscale study.

Regeneration of injured tendons and ligaments (T/L) is a worldwide need. In this study electrospun hierarchical scaffolds made of a poly-L (lactic) acid/collagen blend were developed reproducing all the multiscale levels of aggregation of these tissues. Scanning electron microscopy, microCT and tensile mechanical tests were carried out, including a multiscale digital volume correlation analysis to measure the full-field strain distribution of electrospun structures. The principal strains (εp1 and εp3) described the pattern of strains caused by the nanofibers rearrangement, while the deviatoric strains (εD) revealed the related internal sliding of nanofibers and bundles. The results of this study confirmed the biomimicry of such electrospun hierarchical scaffolds, paving the way to further tissue engineering and clinical applications.

3D visualization of the human anterior cruciate ligament combining micro-CT and histological analysis.

PURPOSE: The study aimed to obtain a comprehensive 3D visualization of knee specimens, including the cruciate ligaments and corresponding femoral and tibial bone insertions using a non-destructive micro-CT method. METHODS: Knee specimens were fixed in anatomical positions and chemically dehydrated before being scanned using micro-CT with a voxel size of 17.5 µm. RGBA (red, green, blue, alpha) transfer functions were applied to virtually colorize each structure. Following micro-CT scanning, the samples were rehydrated, decalcified, and trimmed based on micro-CT 3D reconstructions as references. Histological evaluations were performed on the trimmed samples. Histological and micro-CT images were registered to morphologically and densitometrically assess the 4-layer insertion of the ACL into the bone. RESULTS: The output of the micro-CT images of the knee in extension and flexion allowed a clear differentiation of the morphologies of both soft and hard tissues, such as the ACL, femoral and tibial bones, and cartilage, and the subsequent creation of 3D composite models useful for accurately tracing the entire morphology of the ligament, including its fiber and bundle components, the trajectory between the femur and tibia, and the size, extension, and morphology of its insertions into the bones. CONCLUSION: The implementation of the non-destructive micro-CT method allowed complete visualization of all the different components of the knee specimens. This allowed correlative imaging by micro-CT and histology, accurate planning of histological sections, and virtual anatomical and microstructural analysis. The micro-CT approach provided an unprecedented 3D level of detail, offering a viable means to study ACL anatomy.

Histological and histomorphometric analysis of bone tissue using customized titanium meshes with or without resorbable membranes: A randomized clinical trial.

OBJECTIVES: To date, no clinical studies have investigated the effect of using resorbable collagen membrane in conjunction with customized titanium mesh to promote bone formation in guided bone regeneration. Therefore, a non-inferiority analysis (one-sided 95% CI approach) was designed to compare the augmented bone gained using meshes with and without collagen membranes, through histological and histomorphometric investigations. MATERIALS AND METHODS: Thirty patients undergoing bone augmentation procedures at both maxillary and mandible sites were randomly treated with customized titanium meshes alone (M-, n = 15) or covered with resorbable membrane (M+, n = 15), in both cases filled with autogenous bone and xenograft. After 6 months of healing, bone tissue biopsies were taken from the augmented region. The bone tissue (B.Ar), grafting material (G.Ar), and non-mineralized tissue (NMT.Ar) areas were quantified through histomorphometric analysis, as were the osteoid area (O.Ar) and its width. RESULTS: Collagen membrane did not appear to significantly influence the investigated parameters: B.Ar, G.Ar, NMT.Ar, and O.Ar were similar between Group M- (34.3%, 11.5%, 54.1%, 1.95 µm2 , respectively) and Group M+ (35.3%, 14.6%, 50.2%, and 1.75 µm2 , respectively). Considering the overall population, significantly higher rates of newly formed bone were obtained in mandibular sites, while non-mineralized and dense connective tissue rates were higher in the maxilla (p < .05). CONCLUSIONS: The application of collagen membrane over titanium mesh did not lead to significant results. Bone formation appeared significantly different in the maxilla compared with the mandible. Additional studies are required to further investigate the issues observed.

Nanomechanical Mapping of Three Dimensionally Printed Poly-ε-Caprolactone Single Microfibers at the Cell Scale for Bone Tissue Engineering Applications.

Poly-ε-caprolactone (PCL) has been widely used in additive manufacturing for the construction of scaffolds for bone tissue engineering. However, its use is limited by its lack of bioactivity and inability to induce cell adhesion, hence limiting bone tissue regeneration. Biomimicry is strongly influenced by the dynamics of cell-substrate interaction. Thus, characterizing scaffolds at the cell scale could help to better understand the relationship between surface mechanics and biological response. We conducted atomic force microscopy-based nanoindentation on 3D-printed PCL fibers of ~300 µm thickness and mapped the near-surface Young's modulus at loading forces below 50 nN. In this non-disruptive regime, force mapping did not show clear patterns in the spatial distribution of moduli or a relationship with the topographic asperities within a given region. Remarkably, we found that the average modulus increased linearly with the logarithm of the strain rate. Finally, a dependence of the moduli on the history of nanoindentation was demonstrated on locations of repeated nanoindentations, likely due to creep phenomena capable of hindering viscoelasticity. Our findings can contribute to the rational design of scaffolds for bone regeneration that are capable of inducing cell adhesion and proliferation. The methodologies described are potentially applicable to various tissue-engineered biopolymers.

Relations between Structure/Composition and Mechanics in Osteoarthritic Regenerated Articular Tissue: A Machine Learning Approach.

In the context of a large animal model of early osteoarthritis (OA) treated by orthobiologics, the purpose of this study was to reveal relations between articular tissues structure/composition and cartilage viscoelasticity. Twenty-four sheep, with induced knee OA, were treated by mesenchymal stem cells in various preparations-adipose-derived mesenchymal stem cells (ADSCs), stromal vascular fraction (SVF), and amniotic endothelial cells (AECs)-and euthanized at 3 or 6 months to evaluate the (i) biochemistry of synovial fluid; (ii) histology, immunohistochemistry, and histomorphometry of articular cartilage; and (iii) viscoelasticity of articular cartilage. After performing an initial analysis to evaluate the correlation and multicollinearity between the investigated variables, this study used machine learning (ML) models-Variable Selection Using Random Forests (VSURF) and Extreme Gradient Boosting (XGB)-to classify variables according to their importance and employ them for interpretation and prediction. The experimental setup revealed a potential relation between cartilage elastic modulus and cartilage thickness (CT), synovial fluid interleukin 6 (IL6), and prostaglandin E2 (PGE2), and between cartilage relaxation time and CT and PGE2. SVF treatment was the only limit on the deleterious OA effect on cartilage viscoelastic properties. This work provides indications to future studies aiming to highlight these and other relationships and focusing on advanced regeneration targets.

Medial Meniscal Posterior Horn Suturing Influences Tibial Internal-External Rotation in ACL-Reconstructed Knees.

Background: The role of meniscal lesions and repair in combination with anterior cruciate ligament (ACL) injury and reconstruction has not been extensively investigated in vivo and under weightbearing conditions. Purpose: The purposes of this study were to (1) compare the in vivo knee kinematics between patients with ACL tear and those with combined ACL and medial meniscal tears and (2) investigate kinematic differences between isolated ACL reconstruction and ACL reconstruction plus medial meniscal repair (MR). It was hypothesized that concomitant posterior horn medial meniscal tear and ACL deficiency would affect knee internal-external rotation and anterior-posterior translation but MR would restore these parameters. Study Design: Controlled laboratory study. Methods: Nineteen patients who underwent ACL reconstruction were included: 10 had intact menisci (IM group) and 9 had a medial meniscal injury that was repaired during ACL reconstruction using an all-inside technique (MR group). Preoperatively and 18 months postoperatively, active knee kinematics under weightbearing conditions was evaluated during a single-leg squat using a dynamic biplane x-ray imaging system. The general linear model was used to investigate the differences between group (IM vs MR) and time (preoperative vs follow-up) and their interactions. Results: Tibial internal rotation was higher in the MR group than the IM group both before and after surgery (P = .007). Knee valgus rotation was higher in the MR group preoperatively (P < .001), while no differences were found postoperatively because of an increase of valgus rotation in the IM group, which was significant in the descendant phase (P < .001). Preoperatively, the IM group showed a more medial tibial translation compared with the MR group in the descendant phase (P = .006). Conclusion: When performing a single-leg squat, patients with ACL-deficient knees and a medial meniscal tear demonstrated a more valgus rotation, tibial internal rotation, and lateral tibial translation versus those with intact menisci. After ACL reconstruction and MR, these patients demonstrated significantly higher tibial internal rotation when compared with patients who underwent isolated ACL reconstruction. Clinical Relevance: Surgeons should be aware that MR does not fully restore knee kinematics in vivo and under weightbearing conditions in the context of ACL reconstruction.

Knee Joint Contact Forces during High-Risk Dynamic Tasks: 90° Change of Direction and Deceleration Movements.

Pivoting sports expose athletes to a high risk of knee injuries, mainly due to mechanical overloading of the joint which shatters overall tissue integrity. The present study explored the magnitude of tibiofemoral contact forces (TFCF) in high-risk dynamic tasks. A novel musculoskeletal model with modifiable frontal plane knee alignment was developed to estimate the total, medial, and lateral TFCF developed during vigorous activities. Thirty-one competitive soccer players performing deceleration and 90° sidestepping tasks were assessed via 3D motion analysis by using a marker-based optoelectronic system and TFCF were assessed via OpenSim software. Statistical parametric mapping was used to investigate the effect of frontal plane alignment, compartment laterality, and varus-valgus genu on TFCF. Further, in consideration of specific risk factors, sex influence was also assessed. A strong correlation (R = 0.71 ÷ 0.98, p < 0.001) was found between modification of compartmental forces and changes in frontal plane alignment. Medial and lateral TFCF were similar throughout most of the tasks with the exception of the initial phase, where the lateral compartment had to withstand to higher loadings (1.5 ÷ 3 BW higher, p = 0.010). Significant sex differences emerged in the late phase of the deceleration task. A comprehensive view of factors influencing the mediolateral distribution of TFCF would benefit knee injury prevention and rehabilitation in sport activities.

A Comprehensive Framework to Evaluate the Effects of Anterior Cruciate Ligament Injury and Reconstruction on Graft and Cartilage Status through the Analysis of MRI T2 Relaxation Time and Knee Laxity: A Pilot Study.

BACKGROUND: Anterior cruciate ligament (ACL) tear represents a common orthopedic traumatic issue that often leads to an early development of osteoarthritis. To improve the diagnostic and prognostic techniques involved in the assessment of the joint after the trauma and during the healing process, the present work proposes a multi-parametric approach that aims to investigate the relationship between joint function and soft tissue status before and after ACL reconstruction. METHODS: Thirteen consecutive patients who underwent ACL reconstruction were preliminarily enrolled in this study. Joint laxity assessment as well as magnetic resonance imaging with T2 mapping were performed in the pre-operative stage, at four and 18 months after surgery to acquire objective information to correlate knee function and soft tissue condition. RESULTS: Correlations were found between graft and cartilage T2 signal, suggesting an interplay between these tissues within the knee joint. Moreover, graft maturation resulted in being connected to joint laxity, as underlined by the correlation between the graft T2 signal and the temporal evolution of knee function. CONCLUSIONS: This preliminary study represents a step forward in assessing the effects of ACL graft maturation on knee biomechanics, and vice versa. The presented integrated framework underlines the possibility to quantitatively assess the impact of ACL reconstruction on trauma recovery and cartilage homeostasis. Moreover, the reported findings-despite the preliminary nature of the clinical impacts-evidence the possibility of monitoring the surgery outcomes using a multi-parametric prognostic investigation tool.

Determination of the Spatial Anisotropy of the Surface MicroStructures of Different Implant Materials: An Atomic Force Microscopy Study.

Many biomaterials' surfaces exhibit directional properties, i.e., possess spatial anisotropy on a range of spatial scales spanning from the domain of the naked eye to the sub-micrometer level. Spatial anisotropy of surface can influence the mechanical, physicochemical, and morphological characteristics of the biomaterial, thus affecting its functional behavior in relation, for example, to the host tissue response in regenerative processes, or to the efficacy of spatially organized surface patterns in avoiding bacterial attachment. Despite the importance of the availability of quantitative data, a comprehensive characterization of anisotropic topographies is generally a hard task due to the proliferation of parameters and inherent formal complications. This fact has led so far to excessive simplification that has often prevented researchers from having comparable results. In an attempt to overcome these issues, in this work a systematic and multiscale approach to spatial anisotropy is adopted, based on the determination of only two statistical parameters of surface, namely the texture aspect ratio Str and the roughness exponent H, extracted from atomic force microscopy images of the surface. The validity on this approach is tested on four commercially available implant materials, namely titanium alloy, polyethylene, polyetheretherketone and polyurethane, characterized by textured surfaces obtained after different machining. It is found that the "two parameters" approach is effective in describing the anisotropy changes on surfaces with complex morphology, providing a simple quantitative route for characterization and design of natural and artificial textured surfaces at spatial scales relevant to a wide range of bio-oriented applications.

Dynamic Radiostereometry Evaluation of 2 Different Anterior Cruciate Ligament Reconstruction Techniques During a Single-Leg Squat.

BACKGROUND: Lateral extra-articular tenodesis in the context of anterior cruciate ligament (ACL) reconstruction (ACLR) is performed to better control anterolateral knee instability in patients with high-grade preoperative pivot shift. However, some authors believe these procedures may cause lateral compartment overconstraint, affecting knee motion in daily life. PURPOSE/HYPOTHESIS: The primary aim of the present study was to identify kinematic differences during the execution of an activity under weightbearing conditions between knees having undergone ACLR using anatomic single-bundle (SB) versus single-bundle plus lateral plasty (SBLP) techniques. The secondary aim was to compare the postoperative kinematic data with those from the same knees before ACLR and from the healthy contralateral knees in order to investigate if ACLR was able to restore physiologic knee biomechanics during squat execution. The hypotheses were that (1) the SBLP technique would allow a better restoration of internal-external (IE) knee rotation than would SB and (2) regardless of the technique, ACLR would not fully restore physiologic knee biomechanics. STUDY DESIGN: Randomized controlled trial; Level of evidence, 2. METHODS: In total, 32 patients (42 knees) were included in the study. Patients were asked to perform a single-leg squat before surgery (ACL-injured group, n = 32; healthy contralateral group, n = 10) and at minimum 18-month follow-up after ACLR (SB group, n = 9; SBLP group, n = 18). Knee motion was determined using a validated model-based tracking process that matched patient-specific magnetic resonance imaging bone models to dynamic biplane radiographic images under the principles of roentgen stereophotogrammetric analysis. Data processing was performed using specific software. The authors compared IE and varus-valgus rotations and anterior-posterior and medial-lateral translations among the groups. RESULTS: The mean follow-up period was 21.7 ± 4.5 months. No kinematic differences were found between the SB and SBLP groups (P > .05). A more medial tibial position (P < .05) of the ACL-injured group was reported during the entire motor task and persisted after ACLR in both the SB and the SBLP groups. Differences in IE and varus-valgus rotations were found between the ACL-injured and healthy groups. CONCLUSION: There were no relevant kinematic differences between SBLP and anatomic SB ACLR during the execution of a single-leg squat. Regardless of the surgical technique, ACLR failed in restoring knee biomechanics. REGISTRATION: NCT02323386 (ClinicalTrials.gov identifier).

Extra-Corporeal Membrane Oxygenation Cadaver Donors: What about Tissues Used as Allografts?

Several studies demonstrated the efficacy of post-mortem extracorporeal membrane oxygenation (ECMO) on donors in preserving organ function addressing organ transplantation. Nevertheless, no common and shared evidence was reached about the possibility of using ECMO donors in tissue harvesting. Therefore, this work aimed first to review the current scientific literature about ECMO donors, and then to focus on the use of ECMO tissues as allografts, mainly addressing musculoskeletal tissues, which are of the most interest for reconstruction. A search was conducted on the current scientific literature, focusing on the keywords "ECMO" and "Donor". Several online databases were used, including PubMed, Scopus, and Web of Science. From the preliminary search, 478 articles were obtained, out of which 173 specifically reported the use of ECMO for donation and transplantation purposes. Literature reported extensive analyses of ECMO organs-overall from the abdomen-both in pre- and post-transplantation studies. On the other hand, ECMO tissues were explanted only in a very limited number of cases; moreover, no information was referred about their status and use. A revision of the current scientific literature highlighted the lack of information concerning ECMO tissues and the necessity to perform preclinical, ex vivo studies to compare allografts from ECMO donors, with respect to standard donors, and, thus, to verify whether they can be harvested and implanted safely and with efficacy.

Anisotropy and inhomogeneity of permeability and fibrous network response in the pars intermedia of the human lateral meniscus.

Within the human tibiofemoral joint, meniscus plays a key role due to its peculiar time-dependent mechanical characteristics, inhomogeneous structure and compositional features. To better understand the pathophysiological mechanisms underlying this essential component, it is mandatory to analyze in depth the relationship between its structure and the function it performs in the joint. Accordingly, the aim of this study was to evaluate the behavior of both solid and fluid phases of human meniscus in response to compressive loads, by integrating mechanical assessment and histological analysis. Cubic specimens were harvested from seven knee lateral menisci, specifically from anterior horn, pars intermedia and posterior horn; unconfined compressive tests were then performed according to three main loading directions (i.e., radial, circumferential and vertical). Fibril modulus, matrix modulus and hydraulic permeability of the tissue were thence estimated through a fibril-network-reinforced biphasic model. Tissue porosity and collagen fibers arrangement were assessed through histology for each region and related to the loading directions adopted during mechanical tests. Regional and strain-dependent constitutive parameters were finally proposed for the human lateral meniscus, suggesting an isotropic behavior of both the horns, and a transversely isotropic response of the pars intermedia. Furthermore, the histological findings supported the evidences highlighted by the compressive tests. Indeed, this study provided novel insights concerning the functional behavior of human menisci by integrating mechanical and histological characterizations and thus highlighting the key role of this component in knee contact mechanics and presenting fundamental information that can be used in the development of tissue-engineered substitutes. STATEMENT OF SIGNIFICANCE: This work presents an integration to the approaches currently used to model the mechanical behavior of the meniscal tissue. This study assessed in detail the regional and directional contributions of both the meniscal solid and fluid phases during compressive response, providing also complementary histological evidence. Within this updated perspective, both knee computational modeling and meniscal tissue engineering can be improved to have an effective impact on the clinical practice.

Composite Scaffolds for Bone Tissue Regeneration Based on PCL and Mg-Containing Bioactive Glasses.

Polycaprolactone (PCL) is widely used in additive manufacturing for the construction of scaffolds for tissue engineering because of its good bioresorbability, biocompatibility, and processability. Nevertheless, its use is limited by its inadequate mechanical support, slow degradation rate and the lack of bioactivity and ability to induce cell adhesion and, thus, bone tissue regeneration. In this study, we fabricated 3D PCL scaffolds reinforced with a novel Mg-doped bioactive glass (Mg-BG) characterized by good mechanical properties and biological reactivity. An optimization of the printing parameters and scaffold fabrication was performed; furthermore, an extensive microtopography characterization by scanning electron microscopy and atomic force microscopy was carried out. Nano-indentation tests accounted for the mechanical properties of the scaffolds, whereas SBF tests and cytotoxicity tests using human bone-marrow-derived mesenchymal stem cells (BM-MSCs) were performed to evaluate the bioactivity and in vitro viability. Our results showed that a 50/50 wt% of the polymer-to-glass ratio provides scaffolds with a dense and homogeneous distribution of Mg-BG particles at the surface and roughness twice that of pure PCL scaffolds. Compared to pure PCL (hardness H = 35 ± 2 MPa and Young's elastic modulus E = 0.80 ± 0.05 GPa), the 50/50 wt% formulation showed H = 52 ± 11 MPa and E = 2.0 ± 0.2 GPa, hence, it was close to those of trabecular bone. The high level of biocompatibility, bioactivity, and cell adhesion encourages the use of the composite PCL/Mg-BG scaffolds in promoting cell viability and supporting mechanical loading in the host trabecular bone.

How preconditioning and pretensioning of grafts used in ACLigaments surgical reconstruction are influenced by their mechanical time-dependent characteristics: Can we optimize their initial loading state?

PURPOSE: Consensus about a pre-implant preparation protocol adaptable to any graft used in Anterior Cruciate Ligament reconstruction is still lacking. In fact, there is not agreement on reliable metrics that consider both specific graft dimensional characteristics, such as its diameter, and the inherent properties of its constitutive material, i.e. ligaments or tendons. Aim of the present study was to investigate and propose the applied engineering stress as a possible metrics. METHODS: Preconditioning and pretensioning protocol involved groups of grafts with different section (10 or 32 mm2) and materials (i.e. human patellar and hamstring tendons, and synthetic grafts). A 140 N load was applied to the grafts and maintained for 100 s. Initial stress and following stress-relaxation (a mechanical characteristic that can be related to knee laxity) were specifically analysed. FINDINGS: Initial stress, ranging between 4 and 12 MPa, was affected primarily by the graft cross-section area and secondarily by the choice of the graft material. In terms of loss of the initial stress, stress-relaxation behaviour varied instead on a narrower range, namely 13-17%. INTERPRETATION: Engineering stress can be identified as the correct metrics to optimize the initial state of each graft to avoid excessive stiffness, laxity or fatigue rupture phenomena.

Knee position at the moment of bone bruise could reflect the late phase of non-contact anterior cruciate ligament injury rather than the mechanisms leading to ligament failure.

PURPOSE: The aim of the present study was to trace knee position at the time of bone bruise (BB) and investigate how much this position departed from the knee biomechanics of an in vivo flexion-extension. METHODS: From an original cohort of 62 patients, seven (11%) presented bicompartmental edemas and were included in the study. 3D models of bones and BB were obtained from MRI. Matching bone edemas, a reconstruction of the knee at the moment of BB was obtained. For the same patients, knee kinematics of a squat was calculated using dynamic Roentgen sterephotogrammetric analysis (RSA). Data describing knee position at the moment of BB were compared to kinematics of the same knee extrapolated from RSA system. RESULTS: Knee positions at the moment of BB was significantly different from the kinematics of the squat. In particular, all the patients' positions were out of squat range for both anterior and proximal tibial translation, varus-valgus rotation (five in valgus and two in varus), tibial internal-external rotation (all but one, five externally and one internally). A direct comparison at same flexion angle between knee at the moment of BB (average 46.1° ± 3.8°) and knee during squat confirmed that tibia in the former was significantly more anterior (p < 0.0001), more externally rotated (6.1 ± 3.7°, p = 0.04), and valgus (4.1 ± 2.4°, p = 0.03). CONCLUSION: Knee position at the moment of Bone bruise position was out of physiological in-vivo knee range of motion and could reflect a locked anterior subluxation occurring in the late phase of ACL injury rather than the mechanism leading to ligament failure. LEVEL OF EVIDENCE: Level IV.

Evaluation of cartilage biomechanics and knee joint microenvironment after different cell-based treatments in a sheep model of early osteoarthritis.

PURPOSE: Aiming to prevent cartilage damage during early osteoarthritis (OA), the therapeutic challenge is to restore and maintain the physiological and functional properties of such a tissue with minimally invasive therapeutic strategies. METHODS: Accordingly, an in vivo model of early OA in sheep was here treated through three different cell therapies (culture expanded ADSCs, SVF, and culture expanded AECs) thus to preserve the joint surface from the progression of the pathology. Three months after the treatment injections, their performance was assessed through mechanical automated mapping (Young's modulus and cartilage thickness), gross evaluation of articular surfaces, and biochemical analysis of the synovial fluid. RESULTS: No severe degeneration was observed after three months from OA induction. Cartilage mechanical properties were crucial to identify early degeneration. All the treatments improved the macroscopic cartilage surface aspect and reduced pro-inflammatory cytokines in the synovial fluid. Among the three treatments, SVF highlighted the best performance while ADSCs the worst. CONCLUSION: Despite that the evaluated experimental time is an early follow-up and, thus, longer trial is mandatory to properly assess treatments effectiveness, the proposed multidisciplinary approach allowed to obtain preliminary, but also crucial, results concerning the reduction in OA signs on cartilage properties, in osteophyte development and in all the inflammatory markers.

Cartilage mechanical tests: Evolution of current standards for cartilage repair and tissue engineering. A literature review.

BACKGROUND: Repair procedures and tissue engineering are solutions available in the clinical practice for the treatment of damaged articular cartilage. Regulatory bodies defined the requirements that any products, intended to regenerate cartilage, should have to be applied. In order to verify these requirements, the Food and Drug Administration (FDA, USA) and the International Standard Organization (ISO) indicated some Standard tests, which allow evaluating, in a reproducible way, the performances of scaffolds/treatments for cartilage tissue regeneration. METHODS: A review of the literature about cartilage mechanical characterization found 394 studies, from 1970 to date. They were classified by material (simulated/animal/human cartilage) and method (theoretical/applied; static/dynamic; standard/non-standard study), and analyzed by nation and year of publication. FINDINGS: While Standard methods for cartilage mechanical characterization still refer to studies developed in the eighties, expertise and interest on cartilage mechanics research are evolving continuously and internationally, with studies both in vitro - on human and animal tissues - and in silico, dealing with tissue function and modelling, using static and dynamic loading conditions. INTERPRETATION: there is a consensus on the importance of mechanical characterization that should be considered to evaluate cartilage treatments. Still, relative Standards need to be updated to describe advanced constructs and procedures for cartilage regeneration in a more exhaustive way. The use of the more complex, fibre-reinforced biphasic model, instead of the standard simple biphasic model, to describe cartilage response to loading, and the standardisation of dynamic tests can represent a first step in this direction.

Design of a novel procedure for the optimization of the mechanical performances of 3D printed scaffolds for bone tissue engineering combining CAD, Taguchi method and FEA.

In order to increase manufacturing and experimental efficiency, a certain degree of control over design performances before realization phase is recommended. In this context, this paper presents an integrated procedure to design 3D scaffolds for bone tissue engineering. The procedure required a combination of Computer Aided Design (CAD), Finite Element Analysis (FEA), and Design methodologies Of Experiments (DOE), firstly to understand the influence of the design parameters, and then to control them. Based on inputs from the literature and limitations imposed by the chosen manufacturing process (Precision Extrusion Deposition), 36 scaffold architectures have been drawn. The porosity of each scaffold has been calculated with CAD. Thereafter, a generic scaffold material was considered and its variable parameters were combined with the geometrical ones according to the Taguchi method, i.e. a DOE method. The compressive response of those principal combinations was simulated by FEA, and the influence of each design parameter on the scaffold compressive behaviour was clarified. Finally, a regression model was obtained correlating the scaffold's mechanical performances to its geometrical and material parameters. This model has been applied to a novel composite material made of polycaprolactone and innovative bioactive glass. By setting specific porosity (50%) and stiffness (0.05 GPa) suitable for trabecular bone substitutes, the model selected 4 of the 36 initial scaffold architectures. Only these 4 more promising geometries will be realized and physically tested for advanced indications on compressive strength and biocompatibility.