Classical target coronal alignment in high tibial osteotomy demonstrates validity in terms of knee kinematics and kinetics in a computer model.

PURPOSE: The purpose of this study was to determine the ideal coronal alignment under dynamic conditions after open-wedge high tibial osteotomy (OWHTO). It was hypothesised that, although the classical target alignment was based on experimental evidence, it would demonstrate biomechanical validity. METHODS: Musculoskeletal computer models were analysed with various degrees of coronal correction in OWHTO during gait and squat, specifically with the mechanical axis passing through points at 40%, 50%, 60%, 62.5%, 70%, and 80% of the tibial plateau from the medial edge, defined as the weight-bearing line percentage (WBL%). The peak load on the lateral tibiofemoral (TF) joint, the medial collateral ligament (MCL), and anterior cruciate ligament (ACL) tensions, and knee kinematics with or without increased posterior tibial slope (PTS) were evaluated. RESULTS: The classical alignment with WBL62.5% achieved sufficient load on the lateral TF joint and maintained normal knee kinematics after OWHTO. However, over-correction with WBL80% caused an excessive lateral load and non-physiological kinematics. Increased WBL% resulted in increased MCL tension due to lateral femoral movement against the tibia. With WBL80%, abnormal contact between the medial femoral condyle and the medial intercondylar eminence of the tibia occurred at knee extension. The screw-home movement around knee extension and the TF rotational angle during flexion were reduced as WBL% increased. Increased PTS was associated with increased ACL tension and decreased TF rotation angle because of ligamentous imbalance. CONCLUSIONS: The classical target alignment demonstrated validity in OWHTO, and over-correction should be avoided as it negatively impacts clinical outcome. LEVEL OF EVIDENCE: IV.

Neofunction of ACVR1 in fibrodysplasia ossificans progressiva.

Fibrodysplasia ossificans progressiva (FOP) is a rare genetic disease characterized by extraskeletal bone formation through endochondral ossification. FOP patients harbor point mutations in ACVR1 (also known as ALK2), a type I receptor for bone morphogenetic protein (BMP). Two mechanisms of mutated ACVR1 (FOP-ACVR1) have been proposed: ligand-independent constitutive activity and ligand-dependent hyperactivity in BMP signaling. Here, by using FOP patient-derived induced pluripotent stem cells (FOP-iPSCs), we report a third mechanism, where FOP-ACVR1 abnormally transduces BMP signaling in response to Activin-A, a molecule that normally transduces TGF-ß signaling but not BMP signaling. Activin-A enhanced the chondrogenesis of induced mesenchymal stromal cells derived from FOP-iPSCs (FOP-iMSCs) via aberrant activation of BMP signaling in addition to the normal activation of TGF-ß signaling in vitro, and induced endochondral ossification of FOP-iMSCs in vivo. These results uncover a novel mechanism of extraskeletal bone formation in FOP and provide a potential new therapeutic strategy for FOP.

Differences in impact on adjacent compartments in medial unicompartmental knee arthroplasty versus high tibial osteotomy with identical valgus alignment.

BACKGROUND: It is unclear why medial unicompartmental knee arthroplasty (UKA) with postoperative valgus alignment causes adjacent compartment osteoarthritis more often than high tibial osteotomy (HTO) for moderate medial osteoarthritis of the knee with varus deformity. This study used a computer simulation to evaluate differences in knee conditions between UKA and HTO with identical valgus alignment. METHODS: Dynamic musculoskeletal computer analyses of gait were performed. The hip-knee-ankle angle in fixed-bearing UKA was changed from neutral to 7° valgus by changing the tibial insert thickness. The hip-knee-ankle angle in open-wedge HTO was also changed from neutral to 7° valgus by opening the osteotomy gap. RESULTS: The lateral tibiofemoral contact forces in HTO were larger than those in UKA until moderate valgus alignments. However, the impact of valgus alignment on increasing lateral forces was more pronounced in UKA, which ultimately demonstrated a larger lateral force than HTO. Valgus alignment in UKA caused progressive ligamentous tightness, including that of the anterior cruciate ligament, resulting in compression of the lateral tibiofemoral compartment. Simultaneously, patellofemoral shear forces were slightly increased and excessive external femoral rotation against the tibia occurred due to the flat medial tibial insert surface and decreased lateral compartment congruency. By contrast, only lateral femoral slide against the tibia occurred in excessively valgus-aligned HTO. CONCLUSIONS: In contrast to extra-articular correction in HTO, which results from opening the osteotomy gap, intra-articular valgus correction in UKA with thicker tibial inserts caused progressive ligamentous tightness and kinematic abnormalities, resulting in early osteoarthritis progression into adjacent compartments.

Varus alignment after total knee arthroplasty results in greater axial rotation during deep knee bend activity.

BACKGROUNDS: The correlation between in vivo knee kinematics and alignment has not been fully elucidated. Recently, similar or better clinical outcomes have been reported by restoration of mild varus alignment after total knee arthroplasty for preoperative varus knees. The aim of this study was to evaluate the effect of postoperative alignment on knee kinematics during a deep knee bend activity. METHODS: In vivo knee kinematics of 36 knees (25 patients) implanted with tri-condylar total knee arthroplasty were analyzed with a three dimensional model fitting approach using fluoroscopy. Under fluoroscopic surveillance, individual video frames were digitized at 30° increments from full extension to maximum flexion. Postoperative coronal and sagittal alignments were assessed using radiographs, and rotational alignment was assessed with computed tomography. Pearson correlation coefficients were calculated to determine the correlations between the alignment data and kinematic factors. FINDINGS: Correlation analysis showed that coronal alignment was significantly correlated with knee kinematics. The varus alignment of the limb and tibial component led to a greater axial rotation from full extension to maximum flexion and more rotated position in the mid to deep flexion range. Neither the rotational alignment of the femoral nor tibial components showed significant correlation with axial rotation from full extension to maximum flexion. INTERPRETATION: Varus alignment resulted in greater axial rotation, which could represent near-normal knee kinematics. The current study can be a kinematic rationale reporting similar or better clinical and functional outcomes for the total knee arthroplasty with residual varus alignment.

Abnormal knee kinematics caused by mechanical alignment in symmetric bicruciate-retaining total knee arthroplasty are alleviated by kinematic alignment.

BACKGROUND: Bicruciate-retaining total knee arthroplasty (BCR-TKA) was developed to maintain anterior cruciate ligament function and thus reproduce natural knee kinematics postoperatively. Traditional surgical techniques, however, may cause several complications secondary to kinematic conflict and ligament overtension. The objective of this study was to use a computer simulation of symmetric BCR-TKA to evaluate the effects of alternative surgical techniques on knee kinematics and ligaments. METHODS: A musculoskeletal computer model of a healthy knee was constructed and was used to simulate a BCR model with mechanical alignment (MA). Five adjusted models were investigated, characterized, respectively, by kinematic alignment (KA), two degrees increased tibial slope, two-millimeter distal setting of the tibial component, and an undersized femoral component with either MA or KA. RESULTS: All models exhibited a normal femoral position against the tibia at knee extension, with no anterior paradoxical motion during mid-flexion. The healthy knee model showed medial pivot motion and rollback. In contrast, the BCR MA model demonstrated abnormal bi-condylar rollback with excessive tensions of the lateral collateral ligament and posterior cruciate ligament during knee flexion, whereas the undersized femoral model with MA partly reduced both tensions. The BCR KA model retained relatively physiological kinematics and suppressed excessive ligament tensions. However, no adjusted model completely reproduced healthy knee conditions. CONCLUSIONS: The BCR MA model showed abnormal biomechanics due to kinematic conflict between the retained ligaments and the replaced joint surface. Surgeons using symmetric BCR-TKA should consider using the KA method to achieve sufficient ligament laxity throughout knee flexion.

Large medial proximal tibial angles cause excessively medial tibiofemoral contact forces and abnormal knee kinematics following open-wedge high tibial osteotomy.

BACKGROUND: Recurrent varus deformity and poor outcome sometimes occur following open-wedge high tibial osteotomy, but the mechanism remains unclear. The hypothesis of this study was that an excessively large medial proximal tibial angle with lateral joint surface inclination can worsen postoperative knee biomechanics. METHODS: A computer-simulated knee model was validated based on a volunteer knee. Osteotomy models with medial proximal tibial angles ranging from 90° to 97° in 1° increments were developed. Varus alignment correction of the distal femur was performed in each model to maintain identical coronal alignment passing through a point 62.5% lateral to the tibial plateau. The peak tibiofemoral contact forces and knee kinematics were compared in each model during walking and squatting. FINDINGS: All the osteotomy models demonstrated higher peak contact forces on the lateral tibiofemoral joints than on the medial tibiofemoral joints during walking. However, larger medial proximal tibial angles caused excessive increases in medial tibiofemoral contact forces, and the dominant tibiofemoral contact forces shifted to the medial side. Increased medial proximal tibial angles also caused progressive medial collateral ligament tension in knee flexion, but partial medial collateral ligament release effectively reduced medial tibiofemoral contact forces. Models with large medial proximal tibial angles showed nonphysiological roll-forward of the lateral femoral condyle during squatting and no screw-home movement around knee extension. INTERPRETATION: Excessively large medial proximal tibial angles following open-wedge high tibial osteotomy resulted in increased medial tibiofemoral contact forces and abnormal knee kinematics during knee flexion due to medial joint line elevation and ligament imbalance.

Impact of intraoperative adjustment method for increased flexion gap on knee kinematics after posterior cruciate ligament-sacrificing total knee arthroplasty.

BACKGROUND: In general, the flexion gap is larger than the extension gap with posterior cruciate ligament-sacrificing total knee arthroplasty. Several methods compensate for an excessive flexion gap, but their effects are unknown. The purpose of this study was to compare three methods to compensate for an increased flexion gap. METHODS: In this study, squatting in knees with excessive (4 mm) and moderate (2 mm) flexion gaps was simulated in a computer model. Differences in knee kinematics and kinetics with joint line elevation, setting the femoral component in flexion, and using a larger femoral component as compensatory methods were investigated. FINDINGS: The rotational kinematics during flexion with setting the femoral component in flexion were opposite to those in the other models. Using a larger femoral component resulted in the most physiological motion. The peak anterior translation was 10 mm in the joint line elevation model compared with approximately 6 mm in the other models. In the joint line elevation model, patellofemoral contact stress was excessively increased at 90° of knee flexion. In contrast, tibiofemoral contact stress was higher during knee extension with setting the femoral component in flexion due to anterior impingement. There were few differences in the effect of the three compensatory methods with a moderate flexion gap. INTERPRETATION: A larger femoral component should be used to compensate for an excessive flexion gap because it has less negative impact on posterior cruciate ligament-sacrificing total knee arthroplasty, whereas any compensation method might be acceptable for a moderate flexion gap.

In vitro bone-like nodules generated from patient-derived iPSCs recapitulate pathological bone phenotypes.

The recapitulation of bone formation via the in vitro generation of bone-like nodules is frequently used to understand bone development. However, current bone-induction techniques are slow and difficult to reproduce. Here, we report the formation of bone-like nodules within ten days, via the use of retinoic acid (RA) to induce the osteogenic differentiation of human induced pluripotent stem cells (hiPSCs) into osteoblast-like and osteocyte-like cells that create human bone tissue when implanted in calvarial defects in mice. We also show that the induction of bone formation depends on cell signalling through the RA receptors RARα and RARß, which simultaneously activate the BMP (bone morphogenetic protein) and Wnt signalling pathways. Moreover, by using patient-derived hiPSCs, the bone-like nodules recapitulated the osteogenesis-imperfecta phenotype, which was rescued via the correction of disease-causing mutations and partially by an mTOR (mechanistic target of rapamycin) inhibitor. The method of inducing bone nodules may serve as a fast and reproducible model for the study of the formation of both healthy and pathological bone.

Cell-type dependent enhancer binding of the EWS/ATF1 fusion gene in clear cell sarcomas.

Clear cell sarcoma (CCS) is a rare soft tissue sarcoma caused by the EWS/ATF1 fusion gene. Here, we established induced pluripotent stem cells (iPSCs) from EWS/ATF1-controllable murine CCS cells harboring sarcoma-associated genetic abnormalities. Sarcoma-iPSC mice develop secondary sarcomas immediately after EWS/ATF1 induction, but only in soft tissue. EWS/ATF1 expression induces oncogene-induced senescence in most cell types in sarcoma-iPSC mice but prevents it in sarcoma cells. We identify Tppp3-expressing cells in peripheral nerves as a cell-of-origin for these sarcomas. We show cell type-specific recruitment of EWS/ATF1 to enhancer regions in CCS cells. Finally, epigenetic silencing at these enhancers induces senescence and inhibits CCS cell growth through altered EWS/ATF1 binding. Together, we propose that distinct responses to premature senescence are the basis for the cell type-specificity of cancer development.

Osteochondroma Presenting as a Calcified Mass in the Sellar Region and Review of the Literature.

Objective Osteochondroma (OC) is the most common benign bone neoplasm. It infrequently occurs in the cranial cavity as a calcified lesion and very rarely presents in the sellar region. The present study summarizes the knowledge about OCs of the sellar region. Methods We searched the literature search for the clinical appearance of OCs and other calcified pathologies occurring in the sellar region. Results A total of 21 English-language articles published from 1961 to 2015 documented cases of calcified lesions in the sellar region including cerebral aneurysm, chondroid chordoma, chondroma, craniopharyngioma, OC, odontome, osteoma, pituitary adenoma, pituitary stone, Rathke cleft cyst, retinoblastoma, schwannoma, and xanthogranuloma. Among them, six were OC cases: three in the parasellar region and three in the sellar-suprasellar region. Patients with sellar-suprasellar OCs presented with visual loss and hypopituitarism; patients with parasellar OCs did not show these symptoms. OCs appeared as irregular and multilobulated calcifications on X-ray and computed tomography. On magnetic resonance imaging, OCs showed variable intensity on T1-weighted sequences and consistently heterogenous intensity on T2. Four patients underwent transcranial tumor resection, and the transsphenoidal route was selected for one. Five of the six resulted in a partial resection or internal decompression with a satisfactory outcome. Conclusions Calcified tumors occurring in the sellar region may be OCs, especially if they appear as irregular multilobulated calcification.

Analysis of neural crest cells from Charcot-Marie-Tooth disease patients demonstrates disease-relevant molecular signature.

Charcot-Marie-Tooth disease (CMT) is the most common inherited neuropathy. The majority of CMT is demyelinating type (demyelinating CMT) caused by Schwann cell involvement. Although a large number of genes responsible for demyelinating CMT have been found, the common molecular target of the pathophysiology caused by these different genes in demyelinating CMT is still unknown. We generated induced pluripotent stem cells (iPSCs) from healthy controls and patients with demyelinating CMT caused by duplication in peripheral myelin protein 22 kDa (PMP22) or point mutations in myelin protein zero (MPZ) or early growth response 2 (EGR2). iPSCs were differentiated into neural crest cells, progenitors of Schwann cells, followed by purification using the neural crest cell markers p75 and human natural killer-1. To identify a disease-relevant molecular signature at the early stage of demyelinating CMT, we conducted global gene expression analysis of iPSC-derived neural crest cells and found that a glutathione-mediated detoxification pathway was one of the related pathways in demyelinating CMT. mRNA expression of glutathione S-transferase theta 2 (GSTT2), encoding an important enzyme for glutathione-mediated detoxification, and production of reactive oxygen species were increased in demyelinating CMT. Our study suggested that patient-iPSC-derived neural crest cells could be a cellular model for investigating genetically heterogeneous disease CMT and might provide a therapeutic target for the disease.

SS18-SSX, the Oncogenic Fusion Protein in Synovial Sarcoma, Is a Cellular Context-Dependent Epigenetic Modifier.

The prevalence and specificity of unique fusion oncogenes are high in a number of soft tissue sarcomas (STSs). The close relationship between fusion genes and clinicopathological features suggests that a correlation may exist between the function of fusion proteins and cellular context of the cell-of-origin of each tumor. However, most STSs are origin-unknown tumors and this issue has not yet been investigated in detail. In the present study, we examined the effects of the cellular context on the function of the synovial sarcoma (SS)-specific fusion protein, SS18-SSX, using human pluripotent stem cells (hPSCs) containing the drug-inducible SS18-SSX gene. We selected the neural crest cell (NCC) lineage for the first trial of this system, induced SS18-SSX at various differentiation stages from PSCs to NCC-derived mesenchymal stromal cells (MSCs), and compared its biological effects on each cell type. We found that the expression of FZD10, identified as an SS-specific gene, was induced by SS18-SSX at the PSC and NCC stages, but not at the MSC stage. This stage-specific induction of FZD10 correlated with stage-specific changes in histone marks associated with the FZD10 locus and also with the loss of the BAF47 protein, a member of the SWI/SNF chromatin-remodeling complex. Furthermore, the global gene expression profile of hPSC-derived NCCs was the closest to that of SS cell lines after the induction of SS18-SSX. These results clearly demonstrated that the cellular context is an important factor in the function of SS18-SSX as an epigenetic modifier.

Derivation of mesenchymal stromal cells from pluripotent stem cells through a neural crest lineage using small molecule compounds with defined media.

Neural crest cells (NCCs) are an embryonic migratory cell population with the ability to differentiate into a wide variety of cell types that contribute to the craniofacial skeleton, cornea, peripheral nervous system, and skin pigmentation. This ability suggests the promising role of NCCs as a source for cell-based therapy. Although several methods have been used to induce human NCCs (hNCCs) from human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), further modifications are required to improve the robustness, efficacy, and simplicity of these methods. Chemically defined medium (CDM) was used as the basal medium in the induction and maintenance steps. By optimizing the culture conditions, the combination of the GSK3ß inhibitor and TGFß inhibitor with a minimum growth factor (insulin) very efficiently induced hNCCs (70-80%) from hPSCs. The induced hNCCs expressed cranial NCC-related genes and stably proliferated in CDM supplemented with EGF and FGF2 up to at least 10 passages without changes being observed in the major gene expression profiles. Differentiation properties were confirmed for peripheral neurons, glia, melanocytes, and corneal endothelial cells. In addition, cells with differentiation characteristics similar to multipotent mesenchymal stromal cells (MSCs) were induced from hNCCs using CDM specific for human MSCs. Our simple and robust induction protocol using small molecule compounds with defined media enabled the generation of hNCCs as an intermediate material producing terminally differentiated cells for cell-based innovative medicine.

Production of 2-phenylethanol in roses as the dominant floral scent compound from L-phenylalanine by two key enzymes, a PLP-dependent decarboxylase and a phenylacetaldehyde reductase.

We investigated the biosynthetic pathway for 2-phenylethanol, the dominant floral scent compound in roses, using enzyme assays. L-[(2)H8] Phenylalanine was converted to [(2)H8] phenylacetaldehyde and [(2)H8]-2-phenylethanol by two enzymes derived from the flower petals of R. 'Hoh-Jun,' these being identified as pyridoxal-5'-phosphate-dependent L-aromatic amino acid decarboxylase (AADC) and phenylacetaldehyde reductase (PAR). The activity of rose petal AADC to yield phenylacetaldehyde was nine times higher toward L-phenylalanine than toward its D-isomer, and this conversion was not inhibited by iproniazid, a specific inhibitor of monoamine oxidase. Under aerobic conditions, rose petal AADC stoichiometrically produced NH3 together with phenylacetaldehyde during the course of decarboxylation and oxidation, followed by the hydrolysis of L-phenylalanine. Phenylacetaldehyde was subsequently converted to 2-phenylethanol by the action of PAR. PAR showed specificity toward several volatile aldehydes.