Clinical study of preoperative skeletal muscle mass as a predictor of physical performance recovery following palliative surgery for spinal metastases.

BACKGROUND: Surgical treatment of spinal metastases has been associated with high morbidity and mortality in patients with sarcopenia based on low skeletal muscle mass. We assessed physical performance using the Eastern Cooperative Oncology Group performance status scale and the Barthel Index on the 30th day after palliative surgery for spinal metastases and investigated the effectiveness of surgery according to sarcopenia assessed by skeletal muscle mass. METHODS: We retrospectively analyzed 78 consecutive patients with thoracic and lumbar spinal metastases who underwent palliative surgery. The value of the area of the psoas major muscle at the L3 level normalized by the vertebral area was divided into first, middle, and third tertiles. Clinical variables were compared by tertile. Variables affecting the 30-day good performance status were investigated with univariate and multivariate analyses. RESULTS: The 30-day morbidity rates were 50%, 38.5%, and 15.4% by tertile. The 30-day mortality rate was 2%; all were in the first tertile. Good preoperative performance status scores were seen in 15.4% of first and 50% of third tertile patients. Postoperatively, the performance status improved in all groups, with 30.8%, 65.4%, and 92.3% by tertile. Multivariate regression analysis revealed that a good preoperative performance status (OR: 15.50, 95% CI: 1.610-149.00, P < 0.05) and the value of the area of the psoas major muscle at the L3 level normalized by the vertebral area not in the first tertile (OR: 0.22, 95% CI: 0.06-0.82, P < 0.05) were significant predictors of a good postoperative 30-day performance status. CONCLUSIONS: A good preoperative performance status and exclusion from the first tertile were clinical factors predicting a good postoperative 30-day performance status. In patients with large psoas muscle mass (third tertile), a good 30-day performance status can be expected after surgery, suggesting that surgery in this population should be pursued aggressively.

Time course of symptomatic improvement after open-door laminoplasty for cervical spondylotic myelopathy.

BACKGROUND: Postoperative outcomes following cervical spondylotic myelopathy (CSM) are excellent overall, but there are few prospective studies using patient-reported outcome measures that have explored how long improvement can be expected after surgery. The aim of this prospective study was to investigate from when and until when symptoms improve after open-door laminoplasty in patients with CSM and the factors that affect surgical outcomes. METHODS: The subjects were 115 CSM patients who underwent laminoplasty. The Japanese Orthopaedic Association Cervical Myelopathy Evaluation Questionnaire (JOACMEQ) was used for longitudinal evaluation, and surveys on cervical spine function, upper extremity function, lower extremity function, bladder function, and quality of life were conducted before surgery and at 3, 6, 12, 18, and 24 months after surgery. The patients were classified into 'effective' and 'ineffective' groups based on the points obtained for each domain, and the factors affecting the outcome of surgery were examined by multivariate analysis. RESULTS: JOACMEQ scores improved significantly at 3 months postoperatively compared to preoperatively for all four domains except cervical spine function, but there was no significant improvement after 3 months postoperatively. Multivariate analysis showed that the preoperative score (cervical spine function and upper extremity function) and the points obtained at 3 months postoperatively (upper and lower extremity function, bladder function, and quality of life) were the factors associated with membership of the effective group at 24 months postoperatively. CONCLUSIONS: Although upper and lower extremity function, bladder function, and quality of life domains improved in the relatively early postoperative period, improvement after 3 months postoperatively was limited. The results suggest that the preoperative score and the superiority of the improvement obtained up to 3 months after surgery may be indicators of postoperative outcomes, and this may provide new insights into the selection of surgical indications and patient explanations.

The position of the vertebral artery V1 segment relative to the C7 vertebra.

BACKGROUND: The vertebral artery (VA) usually enters the transverse foramen at the C6 level. Thus, surgeons prefer to insert pedicle screws (PSs) at C7, but this does not eliminate the risk of VA injury. We aimed to clarify anatomical features of the VA V1 segment at the C7 pedicle level, based on computed tomographic angiography (CTA) of 81 consecutive patients. METHODS: We examined the course of the VA V1 segment on axial CTA images. VA position was classified according to its alignment with the anterior (A), middle (M), or posterior (P) third of the C7 vertebral body at the pedicle level. We also assessed the prevalence of hypoplastic VA (HVA). We measured the distance (VED) from the optimum C7 PS entry point (Ep) to the center of the VA. We also measured the angles formed by the vertebral midline and a line from the inner edge of the VA to the Ep (the VEA), and by the vertebral midline and a line from the inner edge of the pedicle to the Ep (the PEA). RESULTS: The variant location of the VA to the C7 vertebra was A in 13 courses (8.1%), M in 123 (76.9%), and P in 20 (12.5%). HVA was present in the contralateral side in 7 of 20 courses (35%) in the P group, and in 8 of 127 courses (6.3%) in the M group (p < 0.05). The mean VED was 20.2 mm, the mean VEA 6.9°, and the mean PEA angle was 36.3°. CONCLUSION: The 20 VA courses in the P group (12.5% of the total VA courses) were relatively close to the C7 Ep. HVA was present contralateral to the VA in 7 of 20 courses in the P group. CTA should be considered before proceeding with, even if, C7 PS instrumentation, to avoid unexpected pitfall.

Non-invasive skin autofluorescence, blood and urine assays of the advanced glycation end product (AGE) pentosidine as an indirect indicator of AGE content in human bone.

BACKGROUND: Bone mineral density (BMD) measurements are widely used to assess fracture risk. However, the finding that some fracture patients had high BMD together with the low contribution of drugs to osteoporosis suggests that bone strength factors other than BMD contribute to bone quality. We evaluated the amount of advanced glycation end products (AGEs) by non-invasive assays of serum and urine as well as by skin autofluorescence to measure the levels of a representative AGE, pentosidine, to investigate whether pentosidine can serve as an indirect indicator of AGEs formation in bone collagen. METHODS: A total of 100 spinal surgery patients without fragility fracture (54 males and 46 females) treated at our hospital were enrolled. The amount of pentosidine in blood, urine, skin and bone (lumbar lamina) samples from these patients was measured. AGE accumulation was assessed by measuring skin autofluorescence. We examined the correlation between pentosidine content in tissues and body fluid, as well as skin AGEs with age, height, body weight, BMI, and estimated glomerular filtration rate (eGFR). RESULTS: A significant age-related increase in pentosidine levels in tissues was observed, while there was a significant negative correlation between tissue pentosidine and eGFR. The amount of skin pentosidine was significantly and positively correlated with pentosidine content of the bone in those under 50 years of age. Urine pentosidine also correlated positively with bone pentosidine and skin pentosidine, but only in females. The total amount of AGEs in skin did not correlate with bone pentosidine. CONCLUSION: In this study, the strong correlation between the pentosidine content in each sample and eGFR may indicate that renal dysfunction with advancing age increases oxidative stress and induces AGEs formation in collagen-containing tissues. The correlation of skin pentosidine concentration and eGFR, with AGEs formation in bone collagen suggests that pentosidine would be a useful indirect index of decreased bone quality. Skin AGEs estimated by autofluorescence in clinical situations may not be suitable as an indirect assessment of bone quality. Because urine pentosidine correlated positively with bone pentosidine and skin pentosidine in females, urine pentosidine may be a candidate for an indirect assessment of bone quality.

Sciatica Caused by Perineural Spread of Prostate Cancer.

An 81-year-old man with a history of prostate cancer developed sciatica and underwent L4/5 laminectomy followed by L5/S1 transforaminal lumbar interbody fusion. Postoperatively, pain improved temporarily, then deteriorated. Tumor resection was performed after enhanced magnetic resonance imaging showed a mass distal to the left greater sciatic foramen. Histopathological examination showed the perineural spread of prostate cancer to the sciatic nerve. Developments in diagnostic imaging have revealed that prostate cancer can undergo perineural spread. Imaging studies are essential when sciatica is diagnosed in patients with a history of prostate cancer.

Efficacy of the PainVision apparatus for assessment of axial neck pain after cervical laminoplasty: a prospective study.

BACKGROUND: Axial neck pain is one of the complications of posterior cervical surgeries such as laminoplasty. This study aimed to investigate the efficiency of the PainVision apparatus for axial neck pain assessment by comparing it with other methods. METHODS: This prospective study included 118 patients (90 men and 28 women; average age: 66.9 (32-86) years) with cervical myelopathy who underwent open-door laminoplasty at our medical center between April 2009 and August 2019. Pain degree (PD) measured by PainVision, visual analog scale (VAS), and bodily pain (BP), a subitem of the MOS 36-Item Short-Form Health Survey (SF36), were used to evaluate axial neck pain, which was investigated preoperatively and at 3, 6, 12, 18, and 24 months postoperatively. RESULTS: Comparison of the scores at each evaluation time point found significant improvement between the pre- and post-operative values for all assessment methods. Further, on comparing the amounts of change between pre- and post-operative scores in each pain assessment method, we found significant differences in PD and VAS but not in BP. We also found significant positive correlations between PD and VAS at each time point (all p < 0.001) and significant negative correlations between PD and BP (all p < 0.05) and between VAS and BP (all p < 0.01) at each time point. CONCLUSIONS: In this study, we demonstrated that PD and VAS are more sensitive indicators of changes in axial neck pain than BP and also that PD has an excellent correlation with VAS. These results suggest that the PainVision apparatus may be an effective instrument for quantifying axial neck pain after cervical laminoplasty, though its superiority over VAS needs to be verified in future studies.

Shear Wave Velocity to Evaluate the Effect of Botulinum Toxin on Post-Stroke Spasticity of the Lower Limb.

(1) Background: The evaluation of muscles with spasticity using ultrasound elastography has attracted attention recently, and the shear wave velocity (SWV) technique can measure the mechanical properties of tissues objectively and quantitatively. The purpose of this study was to evaluate the effect of using SWV to assess the effect of Botulinum toxin type A (BoNT-A) treatment in adult patients with post-stroke lower limb spasticity. (2) Methods: We assessed the modified Ashworth Scale, the modified Tardieu Scale, and SWV at rest and after stretching before and at 1 month after BoNT-A treatment in 10 adult participants with post-stroke lower limb spasticity. (3) Results: Significant changes in SWV of the ankle joint in maximum dorsiflexion to the extent possible (SWV stretched) were observed after BoNT-A treatment. SWV stretched was positively correlated with joint range of motion. Participants whose joint range of motion did not improve (i.e., gastrocnemius medialis muscle (GCM) extension distance did not change) had significantly more reductions in SWV stretched after BoNT-A treatment. (4) Conclusions: Our results suggest that the SWV measurements may serve as a quantitative assessment to determine the effect of the BoNT-A treatment in adult stroke patients. SWV measurements to assess GCM spasticity should consider the effects of tension, material properties and activation level of muscles. The challenge is to measure SWV with matching limb positions in patients without contractures.

Efficacy of Laminoplasty in Improving Sensory Disturbances in Patients with Cervical Spondylotic Myelopathy: A Prospective Study.

OBJECTIVE: Upper extremity sensory disturbances are primary symptoms that affect the quality of life (QOL) of patients with cervical spondylotic myelopathy. Although laminoplasty is 1 of the surgical options, its effects on sensory disturbances have remained unclear. We aimed to determine whether surgical intervention would improve the sensory disturbances of patients with cervical spondylotic myelopathy. METHODS: We conducted a prospective clinical trial of 101 patients who had undergone open door laminoplasty. For an objective sensory assessment, we measured the current perception thresholds (CPTs) in the patients' forearms and palms using PainVision PS-2100. For a subjective sensory assessment, numbness in the upper extremities was rated using a visual analog scale (VAS). Using the VAS scores, the patients were divided into those with improvement and without improvement. Their self-reported 36-item short-form health survey and Japanese Orthopaedic Association cervical myelopathy evaluation questionnaire scores were compared. RESULTS: The postoperative CPTs in relationship to the preoperative CPTs at 3, 6, and 12 months was 99.3%, 98.1%, and 93.8% in the forearm and 93.6%, 90.6%, and 87.8% in the palm, respectively. The corresponding postoperative numbness VAS scores were 63.8%, 50.5%, and 48.0%. At 12 months postoperatively, the 36-item short-form health survey physical and role component summary scores, cervical spine function effectiveness rates, upper and lower extremity function, and QOL items in the Japanese Orthopaedic Association cervical myelopathy evaluation questionnaire were significantly higher in the improvement group. CONCLUSIONS: Our findings have indicated that improvement in postoperative subjective sensory disturbances will occur relatively earlier and will be significantly greater than the improvement in objective sensory disturbances. Furthermore, improvement in the subjective sensory disturbances contributes to functional spinal cord recovery and patients' health-related QOL.

New Method for the Quantitative Assessment of Sensory Disturbances in Cervical Myelopathy: Application for Neurological Level Diagnosis.

INTRODUCTION: Cervical myelopathy frequently manifests as sensory disturbances, including numbness, and their distribution pattern aids in neurological level diagnosis. However, the objective assessment of sensory disturbances is challenging. In this study, we attempted to quantitatively evaluate sensory symptoms in patients with cervical myelopathy according to lesion level using PainVisionⓇ. METHODS: Dermal sensations were evaluated in patients (n = 158) and healthy volunteers (n = 100) using PainVisionⓇ PS-2100, which measured the current perception threshold (CPT). The results were analyzed for their correlation with magnetic resonance imaging (MRI) data, visual analog scale (VAS) scores, and patient functional status assessed by the Japanese Orthopaedic Association (JOA) and JOA Cervical Myelopathy Evaluation Questionnaire (JOACMEQ) scores. RESULTS: Forearm and palm CPT values were significantly higher in patients with cervical myelopathy (both sites, P < 0.001) and were negatively correlated with the JOA score (forearm, r = -0.33; palm, r = -0.35; P < 0.001) and the JOACMEQ scores for upper extremity function (forearm, r = -0.37; palm, r = -0.39; P < 0.001), lower extremity function (forearm, r = -0.39; palm, r = -0.40; P < 0.001), and quality of life (forearm r = -0.27, P = 0.0025); however, no correlation was observed with the VAS score. Stratification of patients according to their lesion levels determined by MRI revealed that the C3/C4 subgroup had significantly higher forearm CPT values than the C4/C5 (P = 0.024) and C5/C6 (P = 0.0013) subgroups and higher palm CPT values than the C5/C6 subgroup (P = 0.009). CONCLUSIONS: Quantitative measurements of sensory disturbances using the PainVisionⓇ device correspond to the degree of patient functional disability and the lesion level. This indicates that both the distribution and intensity of sensory abnormalities are important for neurological level diagnosis in patients with cervical myelopathy.

Dietary Habits Had No Relationship with Adolescent Idiopathic Scoliosis: Analysis Utilizing Quantitative Data about Dietary Intakes.

Although several genetic and environmental factors have been identified as risk factors of adolescent idiopathic scoliosis (AIS), the influence of dietary intake has not been elucidated. We evaluated the association between AIS and dietary habits among female students. Junior high school girls aged 12 to 15 years in the Tokyo metropolitan area who underwent a second school screening for scoliosis were recruited. AIS was diagnosed by orthopedic surgeons specializing in scoliosis, using standing whole spine radiography. Students with a Cobb angle of ≥15° were classified into the AIS group, and others were considered healthy controls. Dietary assessment was performed using a validated diet history questionnaire. Dietary intakes were categorized into quintiles based on distribution, and crude and multivariable odds ratios and 95% confidence intervals for AIS for each quintile category of dietary variable were calculated, with the lowest quintile category used as a reference. In total, 2431 subjects were included in the analysis, and 47.8% of them were diagnosed with AIS. None of the selected nutrients or food groups was significantly associated with AIS. In conclusion, dietary habits may not be associated with AIS.

Insertional Torque in Cervical Vertebrae Lateral Mass Screw Fixation: Magerl Technique Versus Roy-Camille Technique.

STUDY DESIGN: This is a prospective multicenter comparison study. OBJECTIVE: To measure the insertional torque (IT) for cervical vertebra lateral mass screw (LMS) fixation using the Magerl and Roy-Camille methods, at the discretion of the surgeon. SUMMARY OF BACKGROUND DATA: Current fixation methods for patients with preoperative cervical spine instability use LMSs; however, few studies have closely examined intraoperative IT. Patients' bone quality was not measured. METHODS: A total of 637 posterior cervical multiaxial screws were inserted for LMS (C3-C6) (Magerl technique, 423; Roy-Camille technique, 214) in 107 cases. Patients' mean age was 66 years. Patients treated with the Magerl method were divided into 2 groups, with the men in the MM group and the women in the MF group. Similarly, patients treated with the Roy-Camille method were divided into 2 groups, with the men in the RM group and the women in the RF group. The contralateral cortex was penetrated, and the IT at cerclage was measured at the last time. RESULTS: IT of the lateral mass screw was 53.8±22.4, 45.4±21, 45.5±16.9, and 34±16.4 cN.m in the MM group (258), MF group (165), RM group (163), and RF group (51), respectively. The MM group had a significantly higher IT than the other 3 groups, and the RM group had a significantly higher IT than the RF group. When the correlation between screw length and IT was evaluated, IT was significantly (P<0.05) higher with a longer screw by the Magerl method. CONCLUSIONS: The IT was higher with the Magerl method with a longer screw. Screw length and IT were not correlated with the Roy-Camille method; furthermore, the Roy-Camille method went through bilateral cortical bone perpendicularly, so that IT was determined by the fixation power in the cortical part of the bone, which was not thought to be affected by screw length. LEVEL OF EVIDENCE: Level III.

Physical Activities and Lifestyle Factors Related to Adolescent Idiopathic Scoliosis.

BACKGROUND: In addition to genetic factors, environmental and lifestyle factors are thought to play an important role in the onset of adolescent idiopathic scoliosis (AIS). This cross-sectional study was conducted to explore lifestyle factors related to AIS. METHODS: This study included 2,759 Japanese female junior high school students who planned a secondary screening after an initial moiré topography screening indicated possible scoliosis. The students and their mothers, or guardians, were asked to fill out a questionnaire consisting of 38 questions about demographic factors, lifestyle-related factors, social factors, household environment, participation in sports, health status, and factors related to the mother's pregnancy and delivery. The questionnaire was completed by 2,747 students (a 99.6% response rate). After excluding students with heart disease, neurological disease, or a congenital vertebral anomaly, 2,600 students were eligible for assessment. After undergoing a secondary screening with standing radiographs of the spine, students were assigned to the normal (control) group if radiographs showed a curve of <15° or to the scoliosis group if they had a curve of ≥15°. The odds ratios (ORs) for AIS in relation to the possible risk or preventive factors were estimated by logistic regression analyses. RESULTS: No lifestyle-related factor was significantly associated with AIS. However, AIS was associated with classical ballet training (OR, 1.38; 95% confidence interval [CI], 1.09 to 1.75); the odds of AIS developing increased as the child's frequency of training, number of years of experience, and duration of training in ballet increased. The OR for AIS was 1.5 times higher for participants whose mothers had scoliosis. AIS was also associated with a low body mass index (BMI). These associations remained even after mutual adjustment was performed. CONCLUSIONS: No association was found between AIS and lifestyle-related factors. However, classical ballet training, a family history of scoliosis, and low BMI may be associated with AIS. LEVEL OF EVIDENCE: Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.

Anatomical considerations for cervical pedicle screw insertion: the use of multiplanar computerized tomography reconstruction measurements.

OBJECT: The purpose of this study was to evaluate the linear and angular parameters of the vertebral body (VB) required for cervical pedicle screw (CPS) insertion by using multiplanar computerized tomography (CT) reconstructions. METHODS: Three hundred fifteen vertebrae from C-3 to C-7 in 63 patients were studied. Pedicle dimensions such as pedicle transverse angle (PTA), pedicle sagittal angle (PSA), and pedicle outer width (POW) were measured on axial CT reconstructions, as were linear parameters including the lateral mass thickness (LMT), the anteroposterior (AP) and mediolateral distances between spinal canal and transverse foramen, and spinal canal longitudinal and transverse diameter. In addition, the correlations between PTA and other parameters were calculated using univariate linear regression analysis. The overall mean LMT ranged from 10.7 to 12.6 mm. The smallest mean AP spinal canal-transverse foramen distance was found at C-7 (1.1 mm),whereas the largest mean distance was at C-4 (3.1 mm). The smallest mean mediolateral spinal canal-transverse foramen distance was found at C-4 (1.2 mm), whereas the largest mean distance was at C-7 (4.7 mm). There were significant intergroup differences between male and female patients except for PTA and spinal canal longitudinal diameter. The PTA had a direct linear correlation with AP and mediolateral spinal canal-transverse foramen distances. The largest Pearson coefficient was 0.71 between the PTA and AP spinal canal-transverse foramen distance and the inverse one was -0.73 between the PTA and mediolateral spinal canal-transverse foramen distance. CONCLUSIONS: Analysis of the data obtained in this study suggests that not only pedicle dimensions but also linear and angular parameters of the VB can be useful data when inserting a CPS.

Effect of hyper- and microgravity on collagen post-translational controls of MC3T3-E1 osteoblasts.

UNLABELLED: We attempted to study the effects of microgravity (by clinostat) and hypergravity (using centrifugation) on collagen metabolism using murine MC3T3-E1 osteoblasts, especially focusing on collagen cross-link formation. We found that altered gravitational load affected the post-translational modification of collagen, particularly the collagen maturation pathway, through altered expression of enzymes involved in cross-link formation. INTRODUCTION: Gravitational loading plays important roles in the stimulation of differentiated osteoblast function and in the maintenance of skeletal tissues, whereas microgravity seems to result in osteopenia caused by impaired osteoblast differentiation. The aim of our study was to clarify the effects of altered gravitational environments on collagen metabolism, particularly the relationship between post-translational collagen quality and enzymes involved in cross-link formation, using murine osteoblastic MC3T3-E1 cells. MATERIALS AND METHODS: Cells were cultured under vector-averaged microgravity (1 x 10(-3) g) using a clinostat or under conventional centrifugation techniques to generate hypergravity (20 g and 40 g) for 72 h. We then examined the expression patterns of lysyl oxidase and the two lysyl hydroxylase isoforms telopeptidyl lysyl hydroxylase (TLH; procollagen-lysine, 2-oxyglutarate, 5-dioxigenase 2 [PLOD2]) and helical lysyl hydroxylase (HLH; [PLOD1]) by quantitative real time polymerase chain reaction (PCR) analysis. Quantitative analysis of reducible immature (dihydroxylysinonorleucine, hydroxylysinonorleucine, and lysinonorleucine) and nonreducible mature (pyridinoline and deoxypyridinoline) cross-links, and maturation rate analysis of immature to mature cross-links by conventional metabolic labeling using tritium lysine were also performed. RESULTS: Hypergravity upregulated both TLH mRNA expression and enzyme activity compared with stationary cultures, whereas microgravity stimulated both HLH mRNA expression and enzyme activity. These results were consistent with increased relative occupancy rates of telopeptidyl hydroxylysine-derived cross-links and helical hydroxylysine-derived forms observed under hypergravity and microgravity, respectively. Hypergravity stimulated not only lysyl oxidase mRNA expression but also increased enzyme activity and the sum of immature and mature cross-links. Furthermore, the conversion rate of immature cross-links to mature compounds was markedly increased under hypergravity but decreased under microgravity. CONCLUSION: Altered gravitational loading may affect the post-translational modification of collagen through altered expression of enzymes involved in cross-link formation. These observations may be important in elucidating the mechanisms of osteopenia during space flight.

Intensity-related differences in collagen post-translational modification in MC3T3-E1 osteoblasts after exposure to low- and high-intensity pulsed ultrasound.

Low-intensity pulsed ultrasound (LIPUS) has distinct effects on biologic mineralization at intensities of <100 mW/cm2. Intensity-dependent differences in the pattern of accelerated mineralization may be due to different alterations in regulation of collagenous matrix formation. However, little is known about the influence of LIPUS on collagen metabolism in the context of mineralization processes. Therefore, we attempted to evaluate differential effects of two intensities of pulsed ultrasound (30 vs. 120 mW/cm2) on collagen post-translational modification and mineralization in osteoblastic MC3T3-E1 cells. Murine osteoblastic MC3T3-E1 cells were exposed to pulsed ultrasound (1.5-MHz, 200-ms burst sine wave at 1.0-kHz frequency, either 30 or 120 mW/cm2 SATA, for 20 min/day from Day 14 to Day 35 postconfluence). Expression patterns of lysyl oxidase (LO), procollagen-lysine, 2-oxyglutarate, 5-dioxigenase 1 (PLOD1, LH1), and 2 (PLOD2, LH2) was examined using quantitative PCR. Quantitative analysis of reducible immature cross-links (dihydroxylysinonorleucine, hydroxylysinonorleucine, and lysinonorleucine) and nonreducible mature cross-links (pyridinoline and deoxypyridinoline) as well as analysis of the maturation of immature to mature cross-links were performed. Exposure to 30 mW/cm2 LIPUS upregulated LH2 mRNA expression and enzyme activity compared to controls. It was associated with increased relative amounts of telopeptidyl hydroxylysine (Hyl)-derived cross-links beginning on Day 14, upregulated LO mRNA expression, increased total reducible and nonreducible cross-links, and increased ratios of newly formed nonreducible to reducible cross-links. Similarities in the pattern of cross-link formation and calcium deposition in matrices between 30 mW/cm2 LIPUS-treated MC3T3-E1 cultures and bone suggest that 30 mW/cm2 LIPUS may promote the maturation of collagenous matrix as a scaffold for calcification. In contrast, exposure to 120 mW/cm2 ultrasound increased calcium accumulation compared to control at Day 35, but increases were delayed until Day 25. No differences in the extent and pattern of cross-links were observed compared to controls. These results suggest that the promotion of mineralization induced by 120 mW/cm2 may be attributed to other factors involved in mineralization process rather than cross-link pattern. Our results demonstrated the existence of differential effects of lower versus higher intensities of ultrasound on mineralization processes in vitro.