Importance of posterior tibial slope in joint kinematics with an anterior cruciate ligament-deficient knee.

AIMS: To fully quantify the effect of posterior tibial slope (PTS) angles on joint kinematics and contact mechanics of intact and anterior cruciate ligament-deficient (ACLD) knees during the gait cycle. METHODS: In this controlled laboratory study, we developed an original multiscale subject-specific finite element musculoskeletal framework model and integrated it with the tibiofemoral and patellofemoral joints with high-fidelity joint motion representations, to investigate the effects of 2.5° increases in PTS angles on joint dynamics and contact mechanics during the gait cycle. RESULTS: The ACL tensile force in the intact knee was significantly affected with increasing PTS angle. Considerable differences were observed in kinematics and initial posterior femoral translation between the intact and ACLD joints as the PTS angles increased by more than 2.5° (beyond 11.4°). Additionally, a higher contact stress was detected in the peripheral posterior horn areas of the menisci with increasing PTS angle during the gait cycle. The maximum tensile force on the horn of the medial meniscus increased from 73.9 N to 172.4 N in the ACLD joint with increasing PTS angles. CONCLUSION: Knee joint instability and larger loading on the medial meniscus were found on the ACLD knee even at a 2.5° increase in PTS angle (larger than 11.4°). Our biomechanical findings support recent clinical evidence of a high risk of failure of ACL reconstruction with steeper PTS and the necessity of ACL reconstruction, which would prevent meniscus tear and thus the development or progression of osteoarthritis.Cite this article: Bone Joint Res 2022;11(10):708-719.

Histologic Evaluation of Tibial Attachment in 11-Year Double-Bundle ACL Reconstruction with Hamstring Tendons: A Case Report.

CASE: A 49-year-old woman injured her anterior cruciate ligament (ACL) as a high-school student. Double-bundle ACL reconstruction (DBACLR) using hamstring tendon grafts was performed because of recurrent instability. Eleven years after DBACLR, total knee arthroplasty was performed because of osteoarthritis progression. Histologic analysis was completed to observe the osteointegration of the tendon in the obtained proximal tibia. The tibial tunnel showed Sharpey-like fibers anteriorly, connecting the tendon graft and lamellar bone, whereas the intraarticular exit revealed well-aligned chondrocytes posteriorly, indicating chondral metaplasia of the tendon graft. CONCLUSION: Tendon-bone healing can regenerate both fibrous insertion and chondral metaplasia in DBACLR.

An Analysis of the Characteristics and Improved Use of Newly Developed CT-based Navigation System in Total Hip Arthroplasty.

We developed a surface matching-type computed tomography (CT)-based navigation system for total hip arthroplasty (the N-navi; TEIJIN NAKASHIMA MEDICAL, Okayama, Japan). In the registration step, surface matching was performed with digitizing points on the pelvic bone surface after coarse paired matching. In the present study, we made model bones from the CT data of patients whose acetabular shapes had various deformities. We measured the distances and angles after surface matching from the fiducial points and evaluated the ability to correct surface-matching registration on each pelvic form, using several areas and numbers of points. When the surface-matching points were taken on the superior area of the acetabulum, the correction was easy for the external direction, but it was difficult to correct for the anterior and proximal directions. The correction was difficult for external and proximal directions on the posterior area. Each area of surface-matching points has particular directions that are easily corrected and other directions that are difficult to correct. The shape of the pelvis also affected the correction ability. Our present findings suggest that checking the position after coarse paired matching and choosing the surface-matching area and points that are optimal to correct will improve the accuracy of total hip arthroplasty and reduce surgical times.

Verification of Implant Surface Modification by a Novel Processing Method.

Metals have been used clinically as biomaterials, especially in the orthopaedic and dental fields. Metals used as implants wear at contact surfaces, producing metal particles and metal ions that may be harmful. Newly developed metal implants and methods of implant surface modification are currently under scrutiny. We evaluated the use of electrolytic in-process dressing (ELID) as a surface finishing method for metal implants. Metal implants processed using the ELID method (ELID group) or not processed (Non-ELID group) were inserted surgically into rabbit femurs. The rabbits were sacrificed postoperatively over a 24-week period. We assessed the concentrations of the cytokines, interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α, the resistance to implant pull-out, and histopathology at the implant site. There was no significant difference between the groups regarding the cytokine concentrations or implant pull-out resistance. Many particles indicating wear around the implant were noted in the Non-ELID group (n=10) but not the ELID group (n=13), while a fibrous membrane adhering to the every implant was noted in the ELID group. The formation of a fibrous membrane rather than metal particles in the ELID group may indicate improved biocompatibility, and it suggests that ELID may prevent corrosion in the areas of contact.

Contrast-enhanced Computed Tomography Screening Is Effective for Detecting Venous Thromboembolism not Prevented by Prophylaxis after Total Knee Arthroplasty.

Venous thromboembolism (VTE) is a potential complication occurring after total knee arthroplasty (TKA). We investigated the incidence of VTE after TKA using contrast-enhanced computed tomography (CT), and assessed the efficacy of VTE prophylaxis (fondaparinux and enoxaparin). At our hospital, 189 patients (225 knees) underwent TKA between April 2007 and October 2011. The 225 knees were divided into a control group with no VTE prophylaxis (31 cases), a fondaparinux group (107 cases), and an enoxaparin group (87 cases). Contrast-enhanced CT screening for VTE was performed in all cases on day 5 or 6 after TKA. D-dimer levels were measured on day 5 after TKA, and were significantly lower in the fondaparinux (9.8 ± 3.8) and enoxaparin groups (9.4 ± 4.9) than in the control group (15.6 ± 9.8) (p ＜ 0.001). However, no statistically significant difference in the incidence of VTE was observed among the groups (control, 61.3%;fondaparinux, 49.5%;enoxaparin, 50.6%). Prophylaxis was not effective for the prevention of VTE as detected by contrast-enhanced CT after TKA. CT should be performed after TKA, even when VTE prophylaxis is used.

Modification of the cell adhesion and hydrophilic characteristics of poly(ether-ether-ketone) by 172-nm xenon excimer radiation.

BACKGROUND: Poly-ether-ether-ketone (PEEK) has biomechanical and chemical properties that are excellent for biomedical applications; however, PEEK adhesion to bone or chondral tissue proceeds slowly due to poor hydrophilicity and other surface characteristics. OBJECTIVE: We investigated the structural change, hydrophilicity, and cytocompatibility of a PEEK surface after 172-nm xenon excimer UV-irradiation. METHODS: The surface characteristics before and after irradiation were evaluated by contact angle and ATR-FTIR measurements. Mouse osteoblast-like cells (MC3T3-E1) were cultured on PEEK plates and collected after 6, 12 and 24 h for cell adhesion analysis by crystal violet staining (CVS) and scanning electron microscopy (SEM). RESULTS: UV-irradiation improved PEEK surface hydrophilicity, as indicated by a significant drop in water contact angle (p<0.05). Irradiated PEEK showed additional peaks around 3370 cm-1 and 1720 cm-1, highlighting the generation of hydroxyl and carbonyl groups. CVS and SEM revealed improved adhesion to the PEEK surface after UV-irradiation. CONCLUSION: Our results suggest that 172-nm UV-irradiated PEEK may be used in biomedical applications that require good cell adhesion.

Intercondylar notch size influences cyclops formation after anterior cruciate ligament reconstruction.

PURPOSE: The purpose of this study was to investigate the incidence of cyclops lesions and its relationship with the cross-sectional area of the intercondylar notch. METHODS: For this study, 55 patients (24 male and 31 female) underwent follow-up arthroscopy after bi-socket anterior cruciate ligament reconstruction with hamstring tendon grafts were included. All patients underwent magnetic resonance imaging measurements of intercondylar notch dimensions. We compared the femoral intercondylar notch sizes and bone tunnel sizes between knees with cyclops lesions (cyclops group) and those without cyclops lesions (no-cyclops group). The mean percentage of the tunnel size to the cross-sectional area of the femoral intercondylar notch was also compared between the groups. The median follow-up duration was 3.8 years. RESULTS: Cyclops lesions were found in 15 of the 55 knees (27.3 %) on second-look arthroscopy (cyclops group). Only 6 of the 55 knees (10.9 %) had extension loss (cyclops syndrome). The cyclops group included 3 men and 12 women. The two groups showed a statistical difference in sex variation (P = 0.04). No significant differences were found in the femoral and tibial tunnel sizes between the two groups. The cross-sectional area of the femoral intercondylar notch was significantly smaller in the cyclops group (251.7 ± 63.2 mm(2)) than in the no-cyclops group (335.6 ± 77.6 mm(2)) (P < 0.001). The percentage of the total femoral tunnel size to the cross-sectional area of the femoral intercondylar notch was significantly higher in the cyclops group (18.6 ± 5.3 %) than in the no-cyclops group (13.2 ± 3.6 %) (P = 0.02). CONCLUSIONS: A smaller intercondylar notch size may be a potential risk factor for cyclops lesion formation. LEVEL OF EVIDENCE: Case-control study, Level IV.

A new navigation system for minimally invasive total knee arthroplasty.

A computer-assisted navigation system to be used for total knee arthroplasties (TKAs) was reported to improve the accuracy of bone resection and result in precise implant placement, but the concomitant surgical invasion and time consumption are clinical problems. We developed a computed tomography (CT)-based navigation system (NNS) to be used for minimally invasive TKA. It requires only the reference points from a small limited area of the medial femoral condyle and proximal tibia through a skin incision to obtain optical images. Here we evaluated the usefulness and accuracy of the NNS in comparison with the commercially available BrainLAB image-free navigation system (BLS). In a clinical experiment, the registration times obtained with the NNS tended to be shorter than those obtained with the BLS, but not significantly so. The NNS group tended to be in the extended position in the sagittal plane of the distal femur within 3 degrees, and the BLS group showed rather flexed deviation in the sagittal plane of the anterior femur.

Histological analysis of failed cartilage repair after marrow stimulation for the treatment of large cartilage defect in medial compartmental osteoarthritis of the knee.

Bone marrow-stimulating techniques such as microfracture and subchondral drilling are valuable treatments for full-thickness cartilage defects. However, marrow stimulation-derived reparative tissues are not histologically well-documented in human osteoarthritis. We retrospectively investigated cartilage repairs after marrow stimulation for the treatment of large cartilage defects in osteoarthritic knees. Tissues were obtained from patients who underwent total knee arthroplasty (TKA) after arthroscopic marrow stimulation in medial compartmental osteoarthritis. Clinical findings and cartilage repair were assessed. Sections of medial femoral condyles were histologically investigated by safranin O staining and anti-type II collagen antibody. Marrow stimulation decreased the knee pain in the short term. However, varus leg alignment gradually progressed, and TKA conversions were required. The grade of cartilage repair was not improved. Marrow stimulations resulted in insufficient cartilage regeneration on medial femoral condyles. Safranin O-stained proteoglycans and type II collagen were observed in the deep zone of marrow-stimulated holes. This study demonstrated that marrow stimulation resulted in failed cartilage repair for the treatment of large cartilage defects in osteoarthritic knees. Our results suggest that arthroscopic marrow stimulation might not improve clinical symptoms for the long term in patients suffering large osteoarthritic cartilage defects.

Chondromodulin-I derived from the inner meniscus prevents endothelial cell proliferation.

The meniscus is a fibrocartilaginous tissue that plays an important role in controlling complex biomechanics of the knee. A perimeniscal capillary plexus supplies the outer meniscus, whereas the inner meniscus is composed of avascular tissue. Anti-angiogenic molecules, such as chondromodulin-I (ChM-I) and endostatin, have pivotal roles in preserving the avascularity of cartilage. However, the anti-angiogenic role of ChM-I is unclear in the meniscus. We hypothesized that the inner meniscus might maintain its avascular feature by expressing ChM-I. Immunohistochemical analyses revealed that ChM-I was mainly detected in the inner and superficial zones of the meniscus. On the other hand, endostatin distribution was similar between the inner and outer meniscus. In Western blot, ChM-I was detected only in the inner meniscus, whereas endostatin was equally observed in both inner and outer menisci. In addition, ChM-I concentration of the inner meniscus-derived conditioned medium was higher than that of the outer meniscus-derived medium. ChM-I removal from the inner meniscus-derived medium and functional blocking of ChM-I significantly increased endothelial cell proliferation. In this study, we demonstrated that the inner meniscus contained larger amounts of ChM-I, and that the inner meniscus-derived ChM-I inhibited endothelial cell proliferation. Our results suggest that ChM-I may be a key anti-angiogenic factor for maintaining the avascularity of the inner meniscus.

Comparison between loose fragment chondrocytes and condyle fibrochondrocytes in cellular proliferation and redifferentiation.

BACKGROUND: Loose fragments in spontaneous osteonecrosis of the knee (SONK) are usually removed by surgical treatment. However, the healing potential of osteonecrotic loose fragments and their clinical availability, for example as a cell source for cartilage repair and tissue engineering, have not been investigated. The objective of this study was to evaluate the cellular proliferation and redifferentiation ability of loose fragment chondrocytes for the treatment of SONK. METHODS: Cells were obtained from the remaining cartilage of chondral loose fragments or fibrocartilaginous tissue under the affected femoral condyle in SONK. The proliferation activity of loose fragment-derived chondrocytes and condyle-derived fibrochondrocytes was evaluated. In-vitro differentiation ability was assessed by PCR and histological analysis. RESULTS: The deposition of proteoglycans and type II collagen were maintained in loose fragments. However, loose fragment-derived chondrocytes had lower proliferating activity than condyle-derived fibrochondrocytes. Chondrogenic redifferentiation ability was lower in loose fragment chondrocytes than in condyle fibrochondrocytes. Differentiation towards adipogenic and osteogenic lineages was not observed in loose fragment chondrocytes. On the other hand, lipid vacuoles were detected in fibrochondrocytes after adipogenic treatment. CONCLUSIONS: This study demonstrated that loose fragment-derived chondrocytes in SONK had lower potential than fibrochondrocytes in cellular proliferation and redifferentiation. Our experimental results suggest that osteonecrotic loose fragments might have restricted cellular properties in the healing of SONK-related osteochondral defects.

Inner meniscus cells maintain higher chondrogenic phenotype compared with outer meniscus cells.

Meniscus cells have several distinct properties in cellular morphology and extracellular matrix production. Inner meniscus cells are considered to have more chondrocytic phenotype compared with outer meniscus cells. However, the chondrogenic property of each meniscus cell has not been elucidated in detail. In this study, we investigated the difference between human inner and outer meniscus-derived cells in extracellular matrix deposition and chondrogenic potential. Monolayer-cultured inner meniscus cells showed small and ovoid shapes though slender and fibroblastic cells were obtained from outer half of human meniscus. The syntheses of type II collagen and safranin O-stained proteoglycans were increased in chondrogenic pellets derived from inner meniscus cells, rather than in outer meniscus cell-derived pellets. On the other hand, adipogenic lipid vacuoles were equally accumulated in both inner and outer meniscus cells after adipogenic treatment. Chondrogenic treatments also enhanced the expression of chondrogenic marker genes, such as Sry-type HMG box (SOX) 9, Scleraxis, and alpha1(II) collagen, in inner meniscus cells. However, SOX9 expression was not increased in outer meniscus cells even after chondrogenic treatment. This study demonstrated that inner meniscus cells maintained higher chondrogenic potential compared with outer meniscus cells. Our results suggest that the difference between inner and outer meniscus cells in chondrogenic property might have an essential role in preserving a zone-specific meniscal feature.

Combined use of bFGF and GDF-5 enhances the healing of medial collateral ligament injury.

Basic fibroblast growth factor (bFGF) and growth and differentiation factor (GDF)-5 stimulate the healing of medial collateral ligament (MCL) injury. However, the effect of isolated and combined use of bFGF/GDF-5 remains still unclear. We investigated cellular proliferation and migration responding to bFGF/GDF-5 using rabbit MCL fibroblasts. Rabbit MCL injury was treated by bFGF and/or GDF-5 with peptide hydrogels. Gene expression and deposition of collagens in healing tissues were evaluated. bFGF/GDF-5 treatment additively enhanced cell proliferation and migration. bFGF/GDF-5 hydrogels stimulated Col1a1 expression without increasing Col3a1 expression. Combined use of bFGF/GDF-5 stimulated type I collagen deposition and the reorganization of fiber alignment, and induced better morphology of fibroblasts in healing MCLs. Our study indicates that combined use of bFGF/GDF-5 might enhance MCL healing by increasing proliferation and migration of MCL fibroblasts, and by regulating collagen synthesis and connective fiber alignment.

GDF-5/7 and bFGF activate integrin alpha2-mediated cellular migration in rabbit ligament fibroblasts.

Cellular activities responding to growth factors are important in ligament healing. The anterior cruciate ligament (ACL) has poor healing potential compared to the medial collateral ligament (MCL). To assess the differences, we investigated the proliferation, migration, adhesion, and matrix synthesis responding to growth factors in rabbit ACL and MCL fibroblasts. ACL cell proliferation to basic fibroblast growth factor (bFGF), bone morphogenetic protein-2, growth and differentiation factor (GDF)-5, and GDF-7 treatment was similar to that of MCL cells. GDF-5 enhanced Col1a1 expression in ACL and MCL fibroblasts up to 4.7- and 17-fold levels of control, respectively. MCL fibroblasts showed stronger migration activities in response to bFGF and GDF-5 than ACL cells. GDF-5/7 and bFGF also changed the stress fiber formation and cellular adhesion by modulating the distribution of integrin alpha2. Functional blocking analyses using anti-integrin alpha2 antibodies revealed that cellular migration responding to growth factors depended on the integrin alpha2-mediated adhesion on type I collagen. The expression of integrin alpha2 was also increased by growth factors in both cells. Our results demonstrate that GDF-5/7 and bFGF stimulate cellular migration by modulating integrin alpha2 expression and integrin alpha2-dependent adhesion, especially in MCL fibroblasts. These findings suggest that the different healing potential between ACL and MCL may be caused by different cellular behavior in the integrin alpha2-mediated cellular migration in response to growth factors.

Mechanical stretch stimulates integrin alphaVbeta3-mediated collagen expression in human anterior cruciate ligament cells.

Biomechanical stimuli have fundamental roles in the maintenance and remodeling of ligaments including collagen gene expressions. Mechanical stretching signals are mainly transduced by cell adhesion molecules such as integrins. However, the relationships between stress-induced collagen expressions and integrin-mediated cellular behaviors are still unclear in anterior cruciate ligament cells. Here, we focused on the stretch-related responses of different cells derived from the ligament-to-bone interface and midsubstance regions of human anterior cruciate ligaments. Chondroblastic interface cells easily lost their potential to produce collagen genes in non-stretched conditions, rather than fibroblastic midsubstance cells. Uni-axial mechanical stretches increased the type I collagen gene expression of interface and midsubstance cells up to 14- and 6-fold levels of each non-stretched control, respectively. Mechanical stretches also activated the stress fiber formation by shifting the distribution of integrin alphaVbeta3 to the peripheral edges in both interface and midsubstance cells. In addition, integrin alphaVbeta3 colocalized with phosphorylated focal adhesion kinase in stretched cells. Functional blocking analyses using anti-integrin antibodies revealed that the stretch-activated collagen gene expressions on fibronectin were dependent on integrin alphaVbeta3-mediated cellular adhesions in the interface and midsubstance cells. These findings suggest that the integrin alphaVbeta3-mediated stretch signal transduction might have a key role to stimulate collagen gene expression in human anterior cruciate ligament, especially in the ligament-to-bone interface.

Novel magnetic resonance imaging evaluation for valgus instability of the knee caused by medial collateral ligament injury.

Instability of the knee after the medial collateral ligament (MCL) injury is usually assessed with the manual valgus stress test, even though, in recent years, it has become possible to apply magnetic resonance imaging (MRI) to the assessment of the damage of the ligament. The valgus instability of 24 patients (12 isolated injuries and 12 multiple ligament injuries) who suffered MCL injury between 1993 and 1998 was evaluated with the Hughston and Eilers classification, which involves radiographic assessment under manual valgus stress to the injured knees. We developed a novel system for classifying the degree of injury to the MCL by calculating the percentage of injured area based on MRI and investigated the relationship between this novel MRI classification and the magnitude of valgus instability by the Hughston and Eilers classification. There was a significant correlation between the 2 classifications (p=0.0006). On the other hand, the results using other MRI based classification systems, such as the Mink and Deutsch classification and the Petermann classification, were not correlated with the findings by the Hughston and Eilers classification in these cases (p0.05). Since MRI is capable of assessing the injured ligament in clinical practice, this novel classification system would be useful for evaluating the stability of the knee and choosing an appropriate treatment following MCL injury.

Disease-specific screening for deep venous thrombosis and pulmonary thromboembolism using plasma D-dimer values after total knee arthroplasty.

We prospectively evaluated the disease-specific features of the early postoperative plasma D: -dimer value and the relationship with deep venous thrombosis and/or pulmonary thromboembolism (DVT/PE) in 95 patients following total knee arthroplasty. Patients in whom DVT/PE was highly suspected were diagnosed by high-resolution multi-detector row computed tomography scanning (MDCT). Forty-nine knees in 46 patients with rheumatoid arthritis (RA, 24 knees) or osteoarthritis (OA, 25 knees) were finally recruited. DVT/PE was detected in 28 (57.1%) of the 49 cases examined by diagnostic MDCT: 12 (50.0%) of the 24 cases of RA, and 16 (64.0%) of the 25 cases of OA. Of these, PE was found in 11 cases (39.2%), but none of them showed clinical symptomatic signs of dyspnea or chest pain. In both RA and OA cases, there were statistically significant differences in the D: -dimer value on postoperative day 3 (P = 0.027) and after day 28 (P = 0.037) between the groups with and without DVT/PE. In OA cases, there were significant differences between the two groups on postoperative days 1 (P = 0.034), 3 (P = 0.020), 5 (P = 0.005), and 7 (P = 0.045), respectively. At the baseline, perioperative D: -dimer levels in the RA group without DVT/PE were higher than in the OA group. However, multivariate logistic regression analysis showed that RA was not a significant risk factor of DVT/PE in comparison with OA. In conclusion, individual evaluation of the D: -dimer level between RA and OA should provide a more precise predictive indicator of early postoperative DVT/PE.

Cutting tool system to minimize soft tissue damage for robot-assisted minimally invasive orthopedic surgery.

Minimally invasive surgery in orthopedic field is considered to be a challenging problem with a milling robot. One objective of this study is to minimize collision of the cutting tool with soft tissue. The authors have developed a robot with redundant axis to avoid the collision so far. Some important components are modeled based on physical requirements, and a geometric optimization approach based on the model has been also proposed to improve performance. In this paper, a protective mechanism to cover the non-working part of the cutting edge is proposed to avoid soft tissue damage. Hardware and software have been developed for this application and the effectiveness of this technique was evaluated with urethane bone.

New approach for assessing vascular distribution within bone tumors using dynamic contrast-enhanced MRI.

PURPOSE: To differentiate benign from malignant bone tumors by analyzing the vascular distribution within bone tumors with dynamic contrast-enhanced MRI. METHODS: We studied dynamic contrast-enhanced MRI for 49 bone tumors (22 malignant and 27 benign tumors). Seven small regions of interest (ROI) were set inside the largest portion of each tumor. Four ROI were placed evenly on the periphery and three ROI were placed evenly on the line of the longest breadth within the tumor. The slope of the curve (%Slope) was calculated on the time-intensity curves of the whole tumor and of each ROI. The variance values for the %Slope of the ROI were calculated to assess the dispersion of the intensity change at each ROI within the tumor. RESULTS: Mean value of the %Slopes of whole tumor regions for malignant bone tumors (70.4 +/- 60.3%) was significantly higher than that for benign bone tumors (37.6 +/- 52.9%) (P = 0.015), although giant cell tumor (GCT), a locally aggressive tumor, had a relatively higher %Slope. Mean value of the variance of %Slopes for malignant bone tumors (3485.9 +/- 5942.5) was significantly higher than that for all benign tumors (470.4 +/- 583.9) (P = 0.012), indicating that the %Slope values of seven ROI within malignant bone tumors varied more widely compared with the ROI inside benign bone tumors. GCT also demonstrated a lower value. CONCLUSION: Our method of analyzing the signal intensity change at seven separate regions that evaluates the vascular distribution within a tumor could be a useful tool for differentiating between benign and malignant bone tumors.

Decreased levels of insulin-like growth factor-1 and vascular endothelial growth factor relevant to the ossification disturbance in femoral heads spontaneous hypertensive rats.

Ossification disturbance in femoral head reportedly is seen in the Spontaneously Hypertensive rats (SHR) between ages of 10 and 20 weeks. We investigated serum and tissue levels of insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF) in SHR relevant to the ossification disturbance and osteonecrosis of the femoral head. Serum levels of IGF-1 and VEGF were significantly lower in SHR than in Wistar Kyoto rats (WKY) at weeks 5, 10, 15 and 20 (p<0.005). The incidence of histological ossification disturbance of the femoral head was higher in SHR (59%) than in WKY (40%) at week 20. Lower serum and local levels of VEGF in SHR appeared to be related to the incomplete ossification of the femoral heads. Immunohistochemical study showed significantly lower numbers of IGF-1 and VEGF positive chondrocytes in the femoral epiphyseal cartilage of SHR than in those of WKY at weeks 10, 15 and 20. Our results suggest that local and/or systemic levels of IGF-1 and VEGF between ages of 5 and 20 weeks might play roles in the pathogenesis of ossifi cation disturbance of the femoral head in SHR.