Femoral Growth Plate Research Articles

Functional analysis of a novel pathogenic variant in CREBBP associated with bone development.

CREBBP has been extensively studied in syndromic diseases associated with skeletal dysplasia. However, there is limited research on the molecular mechanisms through which CREBBP may impact bone development. We identified a novel pathogenic CREBBP variant (c.C3862T/p.R1288W, which is orthologous to mouse c.3789 C > T/p.R1289W) in a patient with non-syndromic polydactyly. We created a homozygous Crebbp p.R1289W mouse model and compared their skeletal phenotypes to wild-type (WT) animals. Bone marrow stem cells (BMSCs) were isolated and assessed for their proliferative capacity, proportion of apoptotic cells in culture, and differentiation to chondrocytes and osteocytes. We observed a significant decrease in body length in 8-week-old homozygous Crebbp p.R1289W mice. The relative length of cartilage of the digits of Crebbp p.R1289W mice was significantly increased compared to WT mice. BMSCs derived from Crebbp p.R1289W mice had significantly decreased cell proliferation and an elevated rate of apoptosis. Consistently, cell proliferative capacity was decreased and the proportion of apoptotic cells was increased in the distal femoral growth plate of Crebbp p.R1289W compared to WT mice. Chemical induction of BMSCs indicated that Crebbp p.R1289W may promote chondrocyte differentiation. The Crebbp p.R1289W variant plays a pathogenic role in skeletal development in mice. CREBBP has been extensively studied in syndromic diseases characterized by skeletal dysplasia. There is limited research regarding the molecular mechanism through which CREBBP may affect bone development. To our knowledge, we generated the first animal model of a novel Crebbp variant, which is predicted to be pathogenic for skeletal diseases. Certain pathogenic variants, such as Crebbp p.R1289W, can independently lead to variant-specific non-syndromic skeletal dysplasia.

Intraosseous contrast administration for coelomic computed tomography angiography in green iguanas (Iguana iguana): Preliminary findings show promise as an alternative to the intravenous route.

Computed tomography is commonly used to evaluate the reptile coelom. One challenge in the acquisition of contrast-enhanced CT studies in reptiles is the difficulty in placing intravenous catheters due to the poor conspicuity of peripheral vessels. To overcome these limitations, intraosseous catheterization of the long bones (humerus, femur, and tibia) can be used. The purpose of this prospective study was to assess the feasibility and diagnostic value of contrast-enhanced CT of the coelom using a femoral intraosseous route for contrast administration. Twelve healthy juvenile green iguanas were enrolled. In 9 of 12 iguanas, vascular phases were obtained, providing strong, homogeneous enhancement of the arterial and venous tree concurrently. In these nine cases, Phase #1, acquired 60s after beginning the injection of contrast medium, provided maximal contrast between the hepatic parenchyma and hepatic vasculature. In the remaining three cases, injection failure resulted from intracortical placement of the catheter tip. Histological examination of the liver, kidneys, and femur was performed to determine the pathomorphological correlates of the CT findings; in a third of patients, some degree of contrast-induced renal toxicity was documented, which in mammalians shows potential for reversibility. Four iguanas had iatrogenic structural damage to the femoral growth plates attributed to intraosseous catheter placement. A femoral intraosseous route can be used in green iguanas for iodinated contrast medium administration for CT angiography. For the greatest contrast between coelomic organs and vascular structures, at least one postcontrast acquisition 60s after initiation of contrast administration is recommended when using this technique.

ESMAC Best Paper Award 2023: Increased knee flexion in participants with cerebral palsy results in altered stresses at the distal femoral growth plate compared to a typically developing cohort

IntroductionFemoral deformities are highly prevalent in children with cerebral palsy (CP) and can have a severe impact on patients’ gait abilities. While the mechanical stress regime within the distal femoral growth plate remains underexplored, understanding it is crucial given bone's adaptive response to mechanical stimuli. We quantified stresses at the distal femoral growth plate to deepen our understanding of the relationship between healthy and pathological gait patterns, internal loading, and femoral growth patterns. MethodsThis study included three-dimensional motion capture data and magnetic resonance images of 13 typically developing children and twelve participants with cerebral palsy. Employing a multi-scale mechanobiological approach, integrating musculoskeletal simulations and subject-specific finite element analysis, we investigated the orientation of the distal femoral growth plate and the stresses within it. Limbs of participants with CP were grouped depending on their knee flexion kinematics during stance phase as this potentially changes the stresses induced by knee and patellofemoral joint contact forces. ResultsDespite similar growth plate orientation across groups, significant differences were observed in the shape and distribution of growth values. Higher growth rates were noted in the anterior compartment in CP limbs with high knee flexion while CP limbs with normal knee flexion showed high similarity to the group of healthy participants. DiscussionResults indicate that the knee flexion angle during the stance phase is of high relevance for typical bone growth at the distal femur. The evaluated growth rates reveal plausible results, as long-term promoted growth in the anterior compartment leads to anterior bending of the femur which was confirmed for the group with high knee flexion through analyses of the femoral geometry. The framework for these multi-scale simulations has been made accessible on GitHub, empowering peers to conduct similar mechanobiological studies. Advancing our understanding of femoral bone development could ultimately support clinical decision-making.

Does spinopelvic alignment affect femoral head cartilage and the proximal femoral physis in slipped capital femoral epiphysis? A finite element analysis

BackgroundSlipped capital femoral epiphysis is a prevalent pediatric hip disorder. Recent studies suggest the spine's sagittal profile may influence the proximal femoral growth plate's slippage, an aspect not extensively explored. This study utilizes finite element analysis to investigate how various spinopelvic alignments affect shear stress and growth plate slip. MethodsA finite element model was developed from CT scans of a healthy adult male lumbar spine, pelvis, and femurs. The model was subjected to various sagittal alignments through reorientation. Simulations of two-leg stance, one-leg stance, walking heel strike, ascending stairs heel strike, and descending stairs heel strike were conducted. Parameters measured included hip joint contact area, stress, and maximum growth plate Tresca (shear) stress. FindingsPosterior pelvic tilt cases indicated larger shear stresses compared to the anterior pelvic tilt variants except in two leg stance. Two leg stance resulted in decreases in the posterior tilted pelvi variants hip contact and growth plate Tresca stress compared to anterior tilted pelvi, however a combination of posterior pelvic tilt and high pelvic incidence indicated larger shear stresses on the growth plate. One leg stance and heal strike resulted in higher shear stress on the growth plate in posterior pelvic tilt variants compared to anterior pelvic tilt, with a combination of posterior pelvic tilt and high pelvic incidence resulting in the largest shear. InterpretationOur findings suggest that posterior pelvic tilt and high pelvic incidence may lead to increased shear stress at the growth plate. Activities performed in patients with these alignments may predispose to biomechanical loading that shears the growth plate, potentially leading to slip.

Calcified Cartilage-Guided Identification of Osteogenic Molecules and Geometries.

Calcified cartilage digested by chondroclasts provides an excellent scaffold to initiate bone formation. We analyzed bioactive proteins and microarchitecture of calcified cartilage either separately or in combination and evaluated biomimetic osteogenic culture conditions of surface-coated micropatterning. To do so, we prepared a crude extract from porcine femoral growth plates, which enhanced in vitro mineralization when coated on flat-bottom culture dishes, and identified four candidate proteins by fractionation and mass spectrometry. Murine homologues of two candidates, desmoglein 4 (DSG4) and peroxiredoxin 6 (PRDX6), significantly promoted osteogenic activity based on in vitro mineralization and osteoblast differentiation. Moreover, we observed DSG4 and PRDX6 protein expression in mouse femur. In addition, we designed circular, triangular, and honeycomb micropatterns with 30 or 50 μm units, either isolated or connected, to mimic hypertrophic chondrocyte-sized compartments. Isolated, larger honeycomb patterns particularly enhanced osteogenesis in vitro. Mineralization on micropatterns was positively correlated with the reduction of osteoblast migration distance in live cell imaging. Finally, we evaluated possible combinatorial effects of coat proteins and micropatterns and observed an additive effect of DSG4 or PRDX6 coating with micropatterns. These data suggest that combining a bioactive surface coating with osteogenic micropatterns may recapitulate initiation of bone formation during endochondral ossification.

Altered Sox9 and FGF signaling gene expression in Aga2 OI mice negatively affects linear growth.

Osteogenesis imperfecta (OI), or brittle bone disease, is a disorder characterized by bone fragility and increased fracture incidence. All forms of OI also feature short stature, implying an effect on endochondral ossification. Using the Aga2+/- mouse, which has a mutation in type I collagen, we show an affected growth plate primarily due to a shortened proliferative zone. We used single-cell RNA-Seq analysis of tibial and femoral growth plate tissues to understand transcriptional consequences on growth plate cell types. We show that perichondrial cells, which express abundant type I procollagen, and growth plate chondrocytes, which were found to express low amounts of type I procollagen, had ER stress and dysregulation of the same unfolded protein response pathway as previously demonstrated in osteoblasts. Aga2+/- proliferating chondrocytes showed increased FGF and MAPK signaling, findings consistent with accelerated differentiation. There was also increased Sox9 expression throughout the growth plate, which is expected to accelerate early chondrocyte differentiation but reduce late hypertrophic differentiation. These data reveal that mutant type I collagen expression in OI has an impact on the cartilage growth plate. These effects on endochondral ossification indicate that OI is a biologically complex phenotype going beyond its known impacts on bone to negatively affect linear growth.

Effect of hip dysplasia on the development of the femoral head growth plate.

The purpose of this study was to observe whether developmental dysplasia of the hip (DDH) affects the development of the femoral head growth plate and to analyze the risk factors. We selected female patients aged between 11 and 20 years with unilateral DDH and unclosed femoral head growth plate (s). The selected patients underwent anteroposterior radiography of the hip joint to compare the degree of development of the femoral head growth plate on both sides and to identify risk factors that affect the development of the growth plate in the femoral head. We included 48 female patients with unilateral DDH, with an average age of 14 years (range: 11.1-18.5 years) and an average BMI of 20.4 kg/m² (range: 15.5 kg/m²-27.9 kg/m²). Among them, 23 patients had earlier development of the femoral head growth plate on the affected side than on the healthy side, while the degree of development of the femoral head growth plate in 25 patients was the same as that on the contralateral side. When the Tönnis angle was greater than 29.5°C and/or the Reimers migration index was greater than 48.5%, there was a statistically significant difference in the acceleration of femoral head growth plate development. An abnormal relative position of the acetabulum-femoral head caused by DDH can accelerate closure of the femoral head growth plate in immature female patients. The risk factors are a Tönnis angle greater than 29.5°C and/or Reimers migration index greater than 48.5%.

The Risk of Growth Disturbance Is Low After Pediatric Anterior Cruciate Ligament Reconstruction With a Femoral Growth Plate Sparing Technique

To evaluate radiological tibial and femoral length and axis growth disturbances, as well as clinical outcome in skeletal immature anterior cruciate ligament reconstruction (ACLR) patients treated with a femoral growth plate-sparing ACLR technique. Skeletally immature patients who underwent operation between 2013 to 2019 with ALCR using the femoral growth plate-sparing technique were investigated with follow-up after growth plate closure. The inclusion criteria were isolated ACL rupture in patients with open physis in the distal femur and proximal tibia seen at plain radiography. The minimum follow-up time was 29 months. Patients were evaluated with full extremity radiographs measuring limb length discrepancy and coronal knee alignment compared to contralateral leg, as well as clinical evaluation with Rolimeter measurements and the Knee Osteoarthritis Outcome Score (KOOS), the International Knee Documentation Committee subjective knee form (IKDC), and Tegner Activity Scale scores. Sixty-five patients were examined with radiography, and 52 patients were assessed with clinical examination. The mean follow-up time was 68 (range, 29-148) months. No limb-length discrepancy (-0.65 mm [confidence interval {CI},-2.21 to 0.92]) or angular deformity at tibia (-0.25° [CI,-0.78° to 0.28°]) was found. There was a small but statistically significant different angular deformity at the distal femur compared to the contralateral leg (-1.51° [CI,-2.31 to-0.72]) at follow-up. The side-to-side difference in knee laxity at follow-up was 2.4 mm. At follow-up the KOOS Sport, KOOS Quality of Life (QoL), IKDC, and Tegner scores were 80, 75, 86, and 5, respectively. Sixty-seven percent of the patients met the Patient Acceptable Symptom State, and 52% reported results exceeding the KOOS Sport MCID Level and 69% the KOOS QoL level. Femoral physis-sparing ALCR is associated with a low risk of alignment and length disturbances. The technique provides otherwise good subjective clinical outcome and knee stability. Level IV, therapeutic case series.

Femoral growth plate stresses in children with cerebral palsy compared to typically developing children

Femoral growth plate stresses in children with cerebral palsy compared to typically developing children

Influence of tension-band plates on the mechanical loading of the femoral growth plate during guided growth due to coronal plane deformities.

Introduction: Correction of knee malalignment by guided growth using a tension-band plate is a common therapy to prevent knee osteoarthritis among other things. This approach is based on the Hueter-Volkmann law stating that the length growth of bones is inhibited by compression and stimulated by tension. How the locally varying mechanical loading of the growth plate is influenced by the implant has not yet been investigated. This study combines load cases from the gait cycle with personalized geometry in order to investigate the mechanical influence of the tension-band plates. Methods: Personalized finite element models of four distal femoral epiphyses of three individuals, that had undergone guided growth, were generated. Load cases from the gait cycles and musculoskeletal modelling were simulated with and without implant. Morphological features of the growth plates were obtained from radiographs. 3D geometries were completed using non-individual Magnetic Resonance Images of age-matched individuals. Boundary conditions for the models were obtained from instrumented gait analyses. Results: The stress distribution in the growth plate was heterogenous and depended on the geometry. In the insertion region, the implants locally induced static stress and reduced the cyclic loading and unloading. Both factors that reduce the growth rate. On the contralateral side of the growth plate, increased tension stress was observed, which stimulates growth. Discussion: Personalized finite element models are able to estimate the changes of local static and cyclic loading of the growth plate induced by the implant. In future, this knowledge can help to better control growth modulation and avoid the return of the malalignment after the treatment. However, this requires models that are completely participant-specific in terms of load cases and 3D geometry.

Knee valgus and patellofemoral instability after pediatric anterior cruciate ligament reconstruction: a case report and review of the literature

BackgroundPediatric athletes who undergo anterior cruciate ligament reconstruction are at risk for a growth deformity if the surgery violates the physes.CaseA 12-year-old African American boy underwent anterior cruciate ligament reconstruction using a hamstring autograft. The procedure violated the distal femoral growth plate and the perichondrial ring of LaCroix, resulting in a distal femoral lateral physeal growth arrest. Three years later, he had developed a 15° valgus deformity, an increased quadriceps angle and patellofemoral instability. He was able to return to sports after undergoing a distal femoral osteotomy to correct the valgus and medial patellofemoral ligament reconstruction to stabilize the patella.ConclusionAnterior cruciate ligament reconstruction in athletes with open physes has the potential to cause distal femoral valgus deformity, an increased quadriceps angle, and subsequent patellofemoral instability.

Double-Bundle Quadriceps Tendon Autograft for Reconstruction of the Medial Patellofemoral Complex to Manage Recurrent Patellar Dislocation in Patients With Open Physes

Management of chronic patellar instability in patients with open physis requires special reconstruction techniques to minimize the risks of femoral growth plate injury due to the close proximity of the open physis to the native femoral origin of the medial patellofemoral ligament (MPFL). Children and adolescents have a relatively smaller patella than the adult group, so, there is a higher risk of patellar fracture when tunnels are performed in the patella. It is wise to mimic the normal anatomy of the medial patellofemoral complex (MPFC) by reconstruction of both of the medial quadriceps tendon femoral ligament (MQTFL) and MPFL, so as to restore the normal fan-shaped MPFC, with its wide anterior attachment to both of the patella and quadriceps tendon (QT). This article describes a simple, safe, reproducible, and cost effective technique for surgical management of chronic patellar instability in patients with open physis by reconstruction of the MPFC using a double-bundle QT autograft.

DTI assessment of the maturing growth plate of the knee in adolescents and young adults

PurposeTo assess the growth plates of the knee in a healthy population of young adults and adolescents using DTI, and to correlate the findings with chronological age and skeletal maturation. MethodsA prospective, cross-sectional study to assess the tibial and femoral growth plates with DTI in 155 healthy volunteers aged between 14.0 and 21 years old. Echo-planar DTI with 15 directions and b value of 0 and 600 s/mm2 was performed on a 3 T whole-body scanner. ResultsA relationship was observed between chronological age and most DTI metrics (fractional anisotropy, mean diffusivity, and radial diffusivity), tract length and volume. (No significant relationship could be seen for axonal diffusivity and tract length.) Subdivision according to skeletal maturation showed the greatest tract lengths and volumes seen in stage 4b and not 4a. The intra-observer agreement was significant (P = 0.01) for all the measured variables, but agreement varied (femur 0.53 – 0.98; tibia 0.58 – 0.98). Spearman’s correlation showed a significant correlation for age (P = 0.05; P = 0.01) as well as for the fractional anisotropy value within all variables in both femur and tibia. Tract number and volume had a similar correlation with most variables, especially the DTI metrics, and would seem to be interchangeable. ConclusionThe current study indicates that DTI metrics could be a tool to assess the skeletal maturation process of the growth plate and its activity. Tractography seems promising to assess the activity of the growth plate in a younger population but must be used with caution in the more mature growth plate.

Intra- and inter-subject variability of femoral growth plate stresses in typically developing children and children with cerebral palsy.

Little is known about the influence of mechanical loading on growth plate stresses and femoral growth. A multi-scale workflow based on musculoskeletal simulations and mechanobiological finite element (FE) analysis can be used to estimate growth plate loading and femoral growth trends. Personalizing the model in this workflow is time-consuming and therefore previous studies included small sample sizes (N < 4) or generic finite element models. The aim of this study was to develop a semi-automated toolbox to perform this workflow and to quantify intra-subject variability in growth plate stresses in 13 typically developing (TD) children and 12 children with cerebral palsy (CP). Additionally, we investigated the influence of the musculoskeletal model and the chosen material properties on the simulation results. Intra-subject variability in growth plate stresses was higher in cerebral palsy than in typically developing children. The highest osteogenic index (OI) was observed in the posterior region in 62% of the TD femurs while in children with CP the lateral region was the most common (50%). A representative reference osteogenic index distribution heatmap generated from data of 26 TD children's femurs showed a ring shape with low values in the center region and high values at the border of the growth plate. Our simulation results can be used as reference values for further investigations. Furthermore, the code of the developed GP-Tool ("Growth Prediction-Tool") is freely available on GitHub (https://github.com/WilliKoller/GP-Tool) to enable peers to conduct mechanobiological growth studies with larger sample sizes to improve our understanding of femoral growth and to support clinical decision making in the near future.

How does hemiepiphysiodesis by use of tension band implants change the stress distributions in femoral growth plate tissue during guided growth?

How does hemiepiphysiodesis by use of tension band implants change the stress distributions in femoral growth plate tissue during guided growth?

Effects of acute femoral head ischemia on the growth plate and metaphysis in a piglet model of Legg-Calvé-Perthes disease

To determine the effects of acute (≤7 days) femoral head ischemia on the proximal femoral growth plate and metaphysis in a piglet model of Legg-Calvé-Perthes disease (LCPD). We hypothesized that qualitative and quantitative histological assessment would identify effects of ischemia on endochondral ossification. Unilateral femoral head ischemia was surgically induced in piglets, and femurs were collected for histological assessment at 2 (n=7) or 7 (n=5) days post-ischemia. Samples were assessed qualitatively, and histomorphometry of the growth plate zones and primary spongiosa was performed. In a subset of samples at 7 days, hypertrophic chondrocytes were quantitatively assessed and immunohistochemistry for TGFβ1 and Indian hedgehog was performed. By 2 days post-ischemia, there was significant thinning of the proliferative and hypertrophic zones, by 63μm (95% CI-103,-22) and-19 μm (95% CI-33,-5), respectively. This thinning persisted at 7 days post-ischemia. Likewise, at 7 days post-ischemia, the primary spongiosa was thinned to absent by an average of 311μm (95% CI-542,-82) in all ischemic samples. TGFβ1 expression was increased in the hypertrophic zone at 7 days post-ischemia. Alterations to the growth plate zones and metaphysis occurred by 2 days post-ischemia and persisted at 7 days post-ischemia. Our findings suggest that endochondral ossification may be disrupted at an earlier time point than previously reported and that growth disruption may occur in the piglet model as occurs in some children with LCPD.

Isolated MPFL reconstruction with soft tissue femoral fixation technique in 54 skeletally immature patients: Clinical outcomes at 2 years follow-up. A French multicenter retrospective study

BackgroundMedial patello-femoral ligament (MPFL) reconstruction is one of the therapeutic options to treat patellofemoral instability. Classically, a à la carte treatment of skeletal and ligament abnormalities is described. This option is difficult to achieve in children because bony procedures can damage the femoral and/or tibial growth plate. The objective was to evaluate a strategy for isolated reconstruction of the MPFL in the treatment of objective patellar instabilities in children, in a large cohort. The return to sport, knee function and pain or discomfort were studied as secondary endpoints. MethodsThis French multicenter retrospective study included 54 pediatric patients with objective patellofemoral instability. Patients were included if they had presented at least 2 episodes of objective patella dislocation. A Deie-like technique with gracilis tendon graft, soft tissue femoral fixation and patellar bone tunnels for patellar fixation was used. Recurrence of dislocation was studied as the primary endpoint, and the recurrence rate was compared with the literature. A comparison of functional scores (Kujala, Lille femoro-patellar instability score or LFPI Score and Tegner activity score) and NRS between pre- and postoperative was studied as a secondary objective. ResultsA recurrence of femoro-patellar instability was observed for five patients within 2 years follow up (9%). A significant improvement of the Kujala, LFPI score, Tegner and NRS scores was observed (p<0.001). ConclusionIsolated reconstruction of the MPFL presents a risk of recurrence of 9% at 2years follow-up. This technique significantly improves the functional scores of the knee. This modified Deie technique provides good clinical and functional results, allowing return to sports with an acceptable risk of recurrence of patellar dislocation, similar to those observed in the literature. Isolated MPFL reconstruction as a first-line treatment appears to be a reliable and effective technique in terms of recurrence of dislocation and functional scores. It allows early recovery and rehabilitation and has lower morbidity than procedures requiring bone gestures. Level of evidenceIII, retrospective comparative study.

Bone wax in the treatment of partial epiphysiodesis of distal femoral growth plate: Case report at 10-year follow-up

The growth plate is the weakest structure in the skeleton of a child and a frequent site of injury or fracture; physeal injuries represent 15%-30% of all fractures in children. Of all growth plate fractures, the incidence of growth arrest and disorders is around 15%. Here, we discuss a female patient who, at the age of 5 years, was treated for a polytrauma that involved a complex lesion of the growth plates of the knee. Four days after trauma, she underwent closed reduction surgery and internal fixation with cannulated screws for femoral and tibial fractures of the growth plate. A 20° valgus deviation of the left knee was found at 3-month postoperative clinical check-up likely as a result of a growth disorder of the femur. She was diagnosed with valgus knee secondary to epiphysiodesis of the lateral portion of the femoral physis and she was readmitted to the hospital. In the operating theater, an open femoral de-epiphysiodesis was performed with a burr; the drilled hole was then filled with bone wax. At 20-month post-trauma follow-up, the left knee was still valgus about 20° relative to the other side. During follow-up, a slow but progressive improvement in the axis of the lower limbs was noted. Clinical and radiographic control 10 years after the trauma showed a complete recovery of the axis of the lower limbs. In the initial stages, the presence of bone wax in the area of de-epiphysiodesis allowed for stabilization of the deformity on the 20° of preoperative valgus. The interpretation of the growth cartilage activity occurred in an asymmetrical way such as to realign the femoral load axis, it can be based on the different mechanical stimulus on the two knee areas due to the preexisting deformity. There is no unanimous evidence in the literature in terms of management of growth disorders resulting from this type of injury. Bone wax resulted in effectively filling the hole of de-epiphysiodesis in the distal femoral growth plate and allowed us to obtain the response of the growth plate and to improve the recovery time in young children.

Cranial Base Synchondrosis Lacks PTHrP-Expressing Column-Forming Chondrocytes.

The cranial base contains a special type of growth plate termed the synchondrosis, which functions as the growth center of the skull. The synchondrosis is composed of bidirectional opposite-facing layers of resting, proliferating, and hypertrophic chondrocytes, and lacks the secondary ossification center. In long bones, the resting zone of the epiphyseal growth plate houses a population of parathyroid hormone-related protein (PTHrP)-expressing chondrocytes that contribute to the formation of columnar chondrocytes. Whether PTHrP+ chondrocytes in the synchondrosis possess similar functions remains undefined. Using Pthrp-mCherry knock-in mice, we found that PTHrP+ chondrocytes predominantly occupied the lateral wedge-shaped area of the synchondrosis, unlike those in the femoral growth plate that reside in the resting zone within the epiphysis. In vivo cell-lineage analyses using a tamoxifen-inducible Pthrp-creER line revealed that PTHrP+ chondrocytes failed to establish columnar chondrocytes in the synchondrosis. Therefore, PTHrP+ chondrocytes in the synchondrosis do not possess column-forming capabilities, unlike those in the resting zone of the long bone growth plate. These findings support the importance of the secondary ossification center within the long bone epiphysis in establishing the stem cell niche for PTHrP+ chondrocytes, the absence of which may explain the lack of column-forming capabilities of PTHrP+ chondrocytes in the cranial base synchondrosis.

MODL-03. Establishment of intraventricular Shh inhibition as a therapeutic option for young patients with medulloblastoma

The prognosis of pediatric medulloblastoma is still dissatisfying today and tumor survivors often suffer from severe treatment-related morbidities. This poses an urgent need for more efficient therapies. Shh medulloblastoma is characterized by mutations in the Sonic Hedgehog (Shh) pathway, providing an elegant way of targeted therapy. The small molecule Vismodegib allosterically inhibits Smoothened (SMO), an upstream activator of Shh, and shows promising anti-tumor effects against Shh medulloblastoma. Unfortunately, Vismodegib caused severe bone deformities in preclinical studies and clinical trials, preventing its systemic application in children. In a mouse model, we established an intraventricular therapy with Vismodegib combining the benefits of targeted drug delivery and minimal systemic side effects. We compare intraventricular, oral, and placebo treatment regarding effects on survival, tumor biology, and bone morphology.Math1-cre::Ptch1Fl/Fl mice show a homozygous loss of Ptch1 in Math1-positive cells, resulting in Shh pathway overactivation and development of Shh medulloblastomas. At postnatal day 11-13, Math1-cre::Ptch1Fl/Fl mice were randomized in four treatment arms: Group A (n=14) received intraventricular placebo, B (n=12) received 200 mg/kd/d oral Vismodegib, C (n=16) received 0.2 mg/kg/d intraventricular Vismodegib, and D (n=9) received 1.6 mg/kg/d intraventricular Vismodegib. Kaplan-Meier survival curves show a significant survival benefit of 1.6 mg/kg/d intraventricular Vismodegib over placebo (p=0.003). While all intraventricular treated animals develop proliferative tumors at end of observation, investigations at an early time point after completed treatment show promising anti-tumor effects with reduced or absent proliferation in the cerebellum compared to placebo. Bone histology and X-ray analysis of intraventricular treated mice show intact femoral and tibial growth plates, in contrast to orally treated mice that develop severe skeletal malformations. Based on these preliminary experimental results, we conclude that intraventricular application of a SMO-inhibitor might evolve as a promising new way of targeted treatment of Shh medulloblastoma in children.