Developmental Dysplasia Of The Hip Model Research Articles

Impact of endoplasmic reticulum stress on chondrocyte apoptosis in rat model of DDH.

Developmental dysplasia of the hip (DDH) is a developmental disorder characterized by acetabular dysplasia leading to early osteoarthritis. This study examines the role of endoplasmic reticulum stress (ERS) in chondrocyte apoptosis and cartilage degeneration within a DDH model. In the rat model of DDH, created using a swaddling technique, significant deformities in the femoral head and acetabulum were observed, alongside an upregulation of matrix metalloproteinase-13 in acetabular cartilage. We also noted increased levels of apoptosis and ERS-related factors in the acetabular cartilage of DDH models. Additionally, rat chondrocytes exposed to high-magnitude cyclic tensile strain (CTS, 1 Hz, 10% equibiaxial strain) in vitro exhibited elevated ERS and increased apoptosis. Importantly, treatment with the ERS inhibitor 4-phenylbutyric acid effectively suppressed apoptosis induced by CTS in chondrocytes. Our findings suggest that ERS contributes to the upregulation of apoptosis-related factors in chondrocytes within the DDH model, indicating the potential of ERS modulation as a therapeutic approach for DDH-related cartilage degeneration.

EFFECT AND LIMITATION OF PERIACETABULAR OSTEOTOMY FOR ACETABULAR DYSPLASIA CLARIFIED BY 3D FINITE ELEMENT ANALYSIS

Finite element analysis (FEA) has been applied for the biomechanical analysis of acetabular dysplasia, but not for biomechanical studies of periacetabular osteotomy (PAO) or those performing analysis taking into consideration the severity of acetabular dysplasia. This study aimed to perform biomechanical evaluation of changes in stress distribution following PAO and to determine the effect of the severity of developmental dysplasia of the hip (DDH) using three-dimensional FEA.A normal model was designed with a 25° center-edge (CE) angle and a 25° vertical-center-anterior margin (VCA) angle. DDH models were designed with CE and VCA angles each of 10, 0, or −10°. Post-PAO models were created by separating each DDH model and rotating the acetabular bone fragment in the anterolateral direction so that the femoral head was covered by the acetabular bone fragment, with CE and VCA angles each at 25°.Compared to the normal hip joint model, the DDH models showed stress concentration in the acetabular edge and contacting femoral head, and higher stress values; stress increased with decreasing CE and VCA angles. Compared to the DDH models, the post-PAO models showed near-normal patterns of stress distribution in the acetabulum and femoral head, with stress concentration areas shifted from the lateral to medial sides. Stress dispersion was especially apparent in the severe acetabular dysplasia models. PAO provided greater decreases in the maximum values of von Mises stress in the load-bearing area of the acetabulum and femoral head when applied to the DDH models of higher degrees of severity, although the values increased with increasing severity of DDH. PAO is expected to provide biomechanical improvement of the hip joint, although the results also suggest a limitation in the applicability of PAO for the patients with severe acetabular dysplasia.

Reduced dynamic loads due to hip dislocation induce acetabular cartilage degeneration by IL-6 and MMP3 via the STAT3/periostin/NF-κB axis

Developmental dysplasia of the hip (DDH) is characterized by anatomical abnormalities of the hip joint, ranging from mild acetabular dysplasia to hip subluxation and eventually dislocation. The mechanism underlying the cartilage degeneration of the hip joints exposed to reduced dynamic loads due to hip dislocation remains unknown. We established a rodent hip dislocation (disarticulation; DA) model of DDH (DA-DDH rats and mice) by swaddling. Expression levels of periostin (Postn) and catabolic factors, such as interleukin-6 (IL-6) and matrix metalloproteinase 3 (Mmp3), increased and those of chondrogenic markers decreased in the acetabular cartilage of the DA-DDH models. Postn induced IL-6 and Mmp3 expression in chondrocytes through integrin αVβ3, focal adhesion kinase, Src, and nuclear factor-κB (NF-κB) signaling. The microgravity environment created by a random positioning machine induced Postn expression in chondrocytes through signal transducer and activator of transcription 3 (STAT3) signaling. IL-6 stimulated Postn expression via STAT3 signaling. Furthermore, cartilage degeneration was suppressed in the acetabulum of Postn−/− DA-DDH mice compared with that in the acetabulum of wild type DA-DDH mice. In summary, reduced dynamic loads due to hip dislocation induced acetabular cartilage degeneration via IL-6 and MMP3 through STAT3/periostin/NF-κB signaling in the rodent DA-DDH models.

Early Emergent and Progressive Aberrant Subchondral Bone Remodeling Coupled with Aggravated Cartilage Degeneration in Developmental Dysplasia of the Hip.

ObjectiveDevelopmental dysplasia of the hip (DDH) is the most common skeletal development in children and could result in secondary osteoarthritis. This study aims to clarify the alternations of subchondral trabecular bone remodeling and microstructural properties during the development of DDH, and the potential influence of these alternations on the overlying cartilage degeneration and DDH progression.DesignTraditional straight-leg swaddling method was adopted to establish DDH model in newborn Sprague Dawley rats. Hip joint specimens from normal or DDH rats were used. Typical features of DDH in radiological examination were observed by x-ray analysis. Micro–computed tomography analysis was applied to evaluate the microstructural properties of subchondral bone at postnatal weeks 2, 4, and 6. Histological and immunohistochemical analyses were adopted to appraise subchondral bone remodeling activity and cartilage degeneration. The associations among subchondral bone, articular cartilage, and DDH severity were analyzed via multiple linear regression analysis.ResultsCompared with control group, the subchondral bone in DDH group displayed a gradual trend of deteriorated microstructure and worsening biomechanical properties along with aberrant bone remodeling, which might be responsible for the inhibition of stress transmission from the articular cartilage to the subchondral bone and thus leading to the cartilage degeneration and accelerated DDH progression.ConclusionsOur findings indicate that alternations of subchondral trabecular bone in a time-dependent manner could contribute to the DDH progression and the amelioration on subchondral bone might be a favorable therapeutic candidate for DDH.

The long noncoding RNA H19 attenuates force-driven cartilage degeneration via miR-483–5p/Dusp5

Developmental dysplasia of the hip (DDH) is a common hip disease characterized by abnormal development of the acetabulum and femoral head. In most cases, DDH ultimately leads to osteoarthritis. Anomalous biomechanical force plays an important role in cartilage degeneration in DDH. However, in addition to mechanical wear, the underlying molecular mechanisms in cartilage degeneration in DDH remain unclear. This study analyzed the effect of long noncoding RNA (lncRNA)-H19 on DDH cartilage degradation. To elucidate the specific role of lncRNA H19, we established an intermittent cyclic mechanical stress (ICMS) cell force model to simulate abnormal biomechanical environment in vitro. Then, the roles of lncRNA-H19 were also determined in vivo by establishing a model of swaddling DDH. We observed that patients with DDH possessed low levels of lncRNA-H19, COL2A1, and Aggrecan but high levels of MMP3 and Adamts5. The same results were also obtained in a DDH rat model. Furthermore, the data suggested that ICMS promoted cartilage degeneration and caused reorientation of the cytoskeleton, and lncRNA H19 helped inhibit cartilage degeneration. Bioinformatics analysis and lncRNA sequencing were performed, and luciferase assays showed that lncRNA H19 and Dusp5 are both direct targets of miR-483–5p. Moreover, Dups5 plays a negative role in ICMS-induced cartilage degradation by activating the Erk and p38 pathways. In vivo, lncRNA H19 had protective effects on the swaddling DDH model. These findings indicate that lncRNA-H19 played a positive role in cartilage degradation in DDH through the lncRNA H19/miR-483–5p/Dusp5 axis.

Chondrocyte suppression is mediated by miR-129-5p via GDF11/SMAD3 signaling in developmental dysplasia of the hip.

Recent studies have shown that developmental dysplasia of the hip (DDH) during childhood and in animal models is associated with impaired endochondral ossification of the roof of the acetabulum, yet the molecular mechanism of this pathology remains unknown. To address this, an animal model of DDH was established in 4-week-old New Zealand white rabbits by cast immobilization of knee extension. Fifty-six rabbits of DDH were involved in this study, including 21 male rabbits and 25 female rabbits. High-throughput RNA sequencing identified 18 differentially expressed microRNAs; miR-129-5p downregulation was further confirmed by quantitative polymerase chain reaction. Bioinformatics and luciferase reporter assay identified growth differentiation factor 11 (GDF11) as the target gene of miR-129-5p in vitro. miR-129-5p downregulation increased GDF11 expression, which induced the phosphorylation of SMAD family member 3. As a result, the expression of runt-related transcription factor 2, Indian hedgehog homolog, and collagen type X was inhibited in vitro. Meanwhile, Alizarin Red S and Von Kossa staining revealed reduced formation of mineralized nodules by chondrocytes after miR-129-5P downregulation compared with the control. Additionally, proliferation assays and flow cytometry confirmed the suppression of chondrocyte proliferation and G1 cell cycle arrest following miR-129-5p downregulation. These findings indicate that miR-129-5p is able to suppress chondrocyte proliferation and hypertrophic differentiation and decrease mineralization via the miR-129-5p/GDF11/SMAD3 axis. This could present the underlying cause for the observed DDH-associated ossification impairment of the acetabular roof.

WISP-2, an upregulated gene in hip cartilage from the DDH model rats, induces chondrocyte apoptosis through PPARγ in vitro.

Chondrocyte apoptosis plays an important role in the developmental dysplasia of the hip (DDH) development. It has been found that WNT1 inducible signaling pathway protein 2 (WISP-2) and peroxisome proliferator-activated receptor γ (PPARγ) are involved in cell apoptosis. In this study, we performed the straight-leg swaddling DDH rat model and we found that cartilage degradation and chondrocyte apoptosis were remarkably increased in DDH rats in vivo. Moreover, we found that WISP-2 was upregulated in hip acetabular cartilage of DDH rats compared to control rats. Next, the effects of WISP-2 on chondrocyte apoptosis and its possible underlying mechanism were examined in vitro. The lentivirus-mediated gain- and loss-of-function experiments of WISP-2 and peroxisome proliferator-activated receptor γ (PPARγ) for cell viability and apoptosis were performed in primary rat chondrocytes. The results showed that the overexpression of WISP-2 induced chondrocyte apoptosis, and knockdown of WISP-2 could suppress the chondrocyte apoptosis induced by advanced glycation end products (AGE). Additionally, WISP-2 could negatively regulate the expression of PPARγ in chondrocytes. Moreover, the knockdown of PPARγ promoted chondrocyte apoptosis and overexpression of PPARγ abated the increased apoptosis and decreased cell viability of chondrocytes induced by WISP-2. This study demonstrated that WISP-2 might contribute to chondrocyte apoptosis of hip acetabular cartilage through regulating PPARγ expression and activation, which may play an important role in the development of DDH.

Expression of PAPP-A2 and IGF Pathway-Related Proteins in the Hip Joint of Normal Rat and Those with Developmental Dysplasia of the Hip.

Developmental dysplasia of the hip (DDH) is one of the major causes of child disability and early osteoarthritis. Genetic factors play an important role, but which still remain unclear. Pregnancy-associated plasma protein-A2 (PAPP-A2), a special hydrolase of insulin-like growth factor binding protein-5 (IGFBP-5), has been confirmed to be associated with DDH by previous studies. The aim of this study was firstly, to investigate the expression of PAPP-A2 and insulin-like growth factor (IGF) pathway-related proteins in normal rat's hip joints; secondly, to compare the variations of those proteins between DDH model rats and normal ones. The DDH model was established by swaddling the rat's hind legs in hip adduction and extension position. The hip joints were collected for expression study of fetal rats, normal newborn rats, and DDH model rats. Positive expression of PAPP-A2 and IGF pathway-related proteins was observed in all the hip joints of growing-stage rats. Ultimately, IGF1 was downregulated; insulin-like growth factor 1 receptor (IGF1R) showed an opposite trend in DDH rats when compared with normal group. The PAPP-A2 and IGF pathway-associated proteins may also be involved in the development of the rat's hip joint, which bring the foundation for further revealing the pathogenic mechanism of DDH.

Cellular and molecular changes to chondrocytes in an invitro model of developmental dysplasia of the hip‑an experimental model of DDH with swaddling position.

The aim of the present study was to assess the cellular and molecular changes to chondrocytes in a developmental dysplasia of the hip (DDH) model and to investigate the early metabolism of chondrocytes in DDH. Neonatal Wistar rats were used for the DDH model with swaddling position. Primary cultures of chondrocytes were prepared at serial interval stages (2, 4, 6 and 8 weeks) to investigate cellular proliferation. The expression of collagen II and aggrecan mRNA was detected to assess the anabolic ability of chondrocytes. The expression of matrix metallopeptidase (MMP)-13 and ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS-5) mRNA was measured to investigate the degradation of collagen II and aggrecan, respectively. Morphological changes were observed in coronal dissection samples after the removal of fixation. Primary chondrocytes at serial intervals were assessed using a Cell Counting Kit-8 assay and the results revealed that DDH chondrocytes had more proliferative activity. The expression of collagen II mRNA was upregulated at 2 weeks and was more sensitive to mechanical loading compared with aggrecan. Similar changes occurred at 6 weeks. Furthermore, MMP-13 and ADAMTS-5 mRNA expression levels were upregulated at 2 weeks. It was also demonstrated that DDH chondrocytes exhibited high proliferative activity at the early stages and degeneration later.

Canine hip dysplasia: A natural animal model for human developmental dysplasia of the hip.

Developmental dysplasia of the hip (DDH) in humans is a common condition that is associated with hip pain, functional limitations, and secondary osteoarthritis (OA). Surgical treatment of DDH has improved in the last decade, allowing excellent outcomes at short- and mid-term follow-up. Still, the etiology, mechanobiology, and pathology underlying this disease are not well understood. A pre-clinical animal model of DDH could help advance the field with a deeper understanding of specific pathways that initiate hip joint degeneration secondary to abnormal biomechanics. An animal model would also facilitate different interventional treatments that could be tested in a rigorous and controlled environment. The dog model exhibits several important characteristics that make it valuable as a pre-clinical animal model for human DDH. Dogs are naturally prone to develop canine hip dysplasia (CHD), which is treated in a similar manner as in humans. Comparable to human DDH, CHD is considered a pre-OA disease; if left untreated it will progress to OA. However, progression to OA is significantly faster in dogs than humans, with progression to OA within 1-2 years of age, associated with their shorter life span compared to humans. Animal studies could potentially reveal the underlying biochemical pathway(s), which can inform refined treatment modalities and provide opportunities for new treatment and prevention targets. Herein, we review the similarities and differences between the two species and outline the argument supporting CHD as an appropriate pre-clinical model of human DDH. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1807-1817, 2018.

Guided bone regeneration using a bone tissue engineering complex consisting of a poly-dl-lactide membrane and bone mesenchymal stem cells.

Developmental dysplasia of the hip (DDH) is one of the most common diseases encountered in pediatric orthopedic departments. Current treatment strategies seek to improve acetabular coverage, the principal defect of acetabular dysplasia, but are not very successful. We developed a guided bone regeneration (GBR) strategy to improve acetabular coverage via bone tissue engineering (BTE). Poly-dl-lactide (PDLLA) membranes were seeded with bone marrow mesenchymal stem cells (BMSCs) to form a BTE complex, which was then implanted into the superior margin of the acetabulum in a rabbit DDH model. Twelve weeks later, a small amount of high-density shadowing was evident on X-rays of the superior margin of the acetabulum, specimens of which exhibited new bone formation. Micro-computed tomography yielding three-dimensional images revealed that new bone had formed in the superior acetabulum, the basal part of which had fused with (and thus reconstructed) the autogenous bone, and new trabecular bone featuring transverse interlacing was evident in the interior of the hip. No clear evidence of bone formation was observed in rabbits that underwent sham operations or that were implanted with PDLLA only. Thus, it may be possible to improve acetabular coverage via BTE-based bone regeneration.

What is the Best Material for Molding Casts in Children?

Casts are used to treat clubfeet, developmental dysplasia of the hip (DDH), forearm fractures, and femur fractures. The ability of a cast to maintain a desired shape is termed moldability. Clinicians use plaster, fiberglass, and soft casts. To our knowledge the biomechanical molding characteristics of these 3 materials have never been reported. We hypothesized that moldability of plaster would be better than fiberglass and fiberglass would be better than soft cast. We compared 12.7 cm wide casts of plaster, fiberglass, and soft cast. Casts were 5 layers thick, prepared in 40°C water, and placed over 2 layers of cotton padding on 5.1 cm and 15.2 cm diameter foam cylinders. A loading device simulated loads applied by clinicians when molding casts for 4 conditions: clubfoot (thumb-shaped 50 N load on 5.1 cm model), DDH (thumb-shaped 100 N load on 15.2 cm model), forearm fracture (palm-shaped 50 N load on 5.1 cm model), and femur fracture (palm-shaped 100 N load on 15.2 cm model). The loading device applied molding for 7 minutes. Five casts of each material were made for each model. Casts were removed, photographed, and the area of maximal deformation was compared with an unmolded cast. A large area of maximal deformation meant that the deformation was spread out over a large area, less precise molding. In the clubfoot model, plaster was more precise than fiberglass (P=0.002) and soft cast (P<0.0001). In the DDH model, plaster was more precise than fiberglass (P<0.0001) and soft cast (P<0.0001) and fiberglass was more precise than soft cast (P<0.0001).In the femur fracture model, plaster was more precise than fiberglass (P=0.001) and soft cast (P=0.001). The moldability of plaster is better than fiberglass and soft cast and fiberglass is better than soft cast. If precise molding is required, plaster has the best moldability. In cases not requiring precise molding, fiberglass and soft cast are lightweight, waterproof, and available in child-friendly colors.

Early Cartilage Degeneration in a Rat Experimental Model of Developmental Dysplasia of the Hip

Osteoarthritis (OA) is a common long-term complication of developmental dysplasia of the hip (DDH) that is associated with a higher incidence of OA. In addition, the age of onset of OA in DDH patients is significantly younger than in the general population. In order to investigate the early degeneration in DDH cartilage, we used a rat DDH model that was established by the straight-leg swaddling position. The hips were isolated from the DDH model rats and an untreated control group at postnatal weeks 2, 4, 6, and 8. Histology and proteoglycan levels were observed in articular cartilage using Safranin O staining. Biomarkers of cartilage degeneration, including type X collagen and matrix metalloproteinase (MMP)-13, were assessed using immunohistochemistry and quantitative real-time polymerase chain reaction. In addition, expressions of ADAMTS-4 and ADAMTS-5 were studied using quantitative real-time polymerase chain reaction at different ages. DDH rats showed decreased proteoglycans and derangement of chondrocytes when compared with the control group. Collagen X and MMP-13 expressions were higher in the superficial zone of DDH rats than in that of controls (p < 0.05), and the increase was age-dependent. mRNA expression of Collagen X and MMP-13 showed similar results (p < 0.05). A significant increase in mRNA expression of ADAMTS-5 was found in the DDH model cartilage at 8 weeks (p < 0.05). However, no change was observed in ADAMTS-4 expression. This study shows that degenerative cartilage changes occur at an early stage in the rat DDH model and become aggravated with age.

Femoral Head Shape Differences During Development May Identify Hips at Risk of Degeneration

Developmental dysplasia of the hip (DDH) is a common cause of elevated contact stress and early onset osteoarthritis (OA). We hypothesized that adaptation to focal loading during postnatal development would result in signature changes to the shape of the femoral head secondary center of ossification (SCO). SCO shape was evaluated in a canine model of DDH at ages 14 and 32 weeks. The evolving 3D morphology of the SCO was captured using serial quantitative computed tomography. A discrete medial representation shape model was fit to each SCO and served as the basis for quantitative thickness and bending measurements. Shape measurements were tested for associations with hip subluxation and degeneration. At 32 weeks, the SCO was thinner (flatter) in the perifoveal region, the site of focal loading; a greater bend to the SCO was present lateral to the site of thinning; SCO thinning and bending were associated with less femoral head coverage and with a higher probability of degeneration. Shape changes were not detected at 14 weeks. Measurement and visualization of SCO shape changes due to altered loading may provide a basis for identifying hips at risk of early onset OA and a tool for surgical planning of hip restructuring.

Femoral Head Bone Mineral Density Patterns May Identify Hips at Risk of Degeneration

Developmental dysplasia of the hip (DDH) is a common abnormality that causes elevated contact stress and early onset osteoarthritis (OA). We hypothesized that adaptation of femoral head bone mineral density (BMD) patterns to focal loading during postnatal development could be used to identify hips at risk of degeneration. Evolving BMD patterns of the femoral head secondary center of ossification (SCO) were quantified and tested for differences with hip subluxation and degeneration. BMD was measured using quantitative computed tomography of hips in a canine model of DDH from 4weeks to early skeletal maturity at 32weeks. During body weight and SCO volumetric growth deceleration, SCO mean BMD increased rapidly and local regions of high BMD formed. Greater subluxation was associated with a lower mean BMD up to 14weeks. At 32weeks, greater subluxation was associated with a larger area of high BMD that was more laterally located and had a greater maximum BMD. BMD differences were associated with a higher probability of cartilage degeneration. Measurement and visualization of BMD pattern changes due to altered mechanical loading provide a basis for identifying hips at risk of early onset OA and a tool for surgical planning of contact stress reduction procedures.

Effect of periacetabular osteotomy for acetabular dysplasia clarified by three-dimensional finite element analysis

BackgroundFinite element analysis (FEA) has been applied for the biomechanical analysis of acetabular dysplasia, but not for biomechanical studies of periacetabular osteotomy (PAO) or those performing analysis taking into consideration the severity of acetabular dysplasia. This study aimed to perform biomechanical evaluation of changes in stress distribution following PAO and to determine the effect of the severity of developmental dysplasia of the hip (DDH) using three-dimensional FEA. MethodsA normal model was designed with a 25° center-edge (CE) angle and a 25° vertical-center-anterior margin (VCA) angle. DDH models were designed with CE and VCA angles each of 10, 0, or −10°. Post-PAO models were created by separating each DDH model and rotating the acetabular bone fragment in the anterolateral direction so that the femoral head was covered by the acetabular bone fragment, with CE and VCA angles each at 25°. ResultsCompared to the normal hip joint model, the DDH models showed stress concentration in the acetabular edge and contacting femoral head, and higher stress values; stress increased with decreasing CE and VCA angles. Compared to the DDH models, the post-PAO models showed near-normal patterns of stress distribution in the acetabulum and femoral head, with stress concentration areas shifted from the lateral to medial sides. Stress dispersion was especially apparent in the severe acetabular dysplasia models. PAO provided greater decreases in the maximum values of von Mises stress in the load-bearing area of the acetabulum and femoral head when applied to the DDH models of higher degrees of severity, although the values increased with increasing severity of DDH. ConclusionsPAO is expected to provide biomechanical improvement of the hip joint and to be particularly effective in patients with severe preoperative DDH, although the results also suggested a limitation in the applicability of PAO for these patients.

Effect of Early Postnatal Body Weight on Femoral Head Ossification Onset and Hip Osteoarthritis in a Canine Model of Developmental Dysplasia of the Hip

Developmental dysplasia of the hip (DDH) is a well-known precipitator of hip osteoarthritis. An increase in body weight during the critical early postnatal growth period may alter joint contact, and thus alter hip development and influence joint health in adulthood. The objective of this study was to determine whether early postnatal body weight affected the course of hip development and the onset of osteoarthritis in a canine model of DDH. A longitudinal study, from birth to skeletal maturity, was conducted. Serial body weight, age at femoral head ossification onset, and femoral head coverage at 4 mo were measured. Presence and severity of degeneration at 8 mo were determined using necropsy and cartilage biochemistry. There was a negative association between birth weight and age at femoral head ossification onset; however, the association was likely due to skeletal maturity level rather than body weight per se. Lower birth weight subjects had greater femoral head coverage at 4 mo. Greater birth weight was associated with greater probability of moderate degenerative changes or macroscopic lesions at 8 mo. These results support the hypothesis that increased birth weight is sufficient to alter the course of hip development and result in measurable degenerative changes at adulthood.