Clinical Application of a Specific Simulation Software for 3-Dimensional Orbital Volume Modeling for Orbital Wall Reconstruction.

In this study, we developed a new 3-dimensional (3D) preoperative planning software and investigated its effectiveness by measuring orbital volume change. Fifty-six patients who underwent unilateral orbital wall reconstruction between December 2015 and February 2018 in our institute were recruited. We developed an orbit-specific automatic 3D modeling software (Osteopore CMF 3D) that can automatically detect orbital fracture site, process mirroring, measure volume, and then produce a customized orbital implant. Using preoperative and postoperative 3D computed tomography, orbital volume was measured and compared between Osteopore CMF 3D and the widely used Mimics. Intraclass correlation coefficient between Osteopore CMF 3D and Mimics showed 90.6% concordance for preoperative injured site orbital volume and 86.7% concordance for postoperative orbital volume, both being statistically significant (preoperative intraclass correlation coefficient, 0.906 [confidence interval {CI}, 0.840-0.945; P < 0.000]; postoperative intraclass correlation coefficient, 0.867 [CI, 0.773-0.922; P < 0.000]). For uninjured sites, the 2 software packages showed 76.6% preoperative concordance and 83.9% postoperative concordance, both being statistically significant (preoperative intraclass correlation coefficient, 0.766 [CI, 0.600-0.863; P < 0.000]; postoperative intraclass correlation coefficient, 0.839 [CI, 0.725-0.905; P < 0.000]). This study introduces our newly developed 3D surgical planning software specialized for orbital wall reconstruction and demonstrates its performance to be comparable with that of a widely used software.

Clinical advantages of image-free navigation system using surface-based registration in anatomical anterior cruciate ligament reconstruction.

PURPOSE: To evaluate the clinical advantages of a navigation system developed with an emphasis on attaining an appropriate femoral tunnel length and posterior wall margin with no posterior wall blowout, as well as having accurate tunnel positioning, in anatomical anterior cruciate ligament reconstruction (ACLR). METHODS: Ten freshly frozen human knees were transected at mid-femur and mid-tibia. Each knee specimen underwent arthroscopic single-bundle anterior cruciate ligament reconstruction using the outside-in technique, with two knees by manual ACLR (control group) and another eight knees by only the navigational ACLR without arthroscopic assistance (experimental group). The position/orientation information of tunnel entry point, tunnel length, and posterior wall distance of pre-, intra-, and postoperative tunnel were recorded, and the reliability and errors among them were evaluated. RESULTS: From comparison of the 3D models for preoperative planning and postoperative reconstruction, the mean differences for navigational femoral tunnelling and arthroscopic-assisted femoral tunnelling were recorded, respectively: (1) tunnel entry position, 1.4 mm (SD 0.3) versus 4.9 mm; (2) tunnel length, 0.7 mm (SD 0.2), similar to 0.6 mm in arthroscopic-assisted femoral tunnelling, and (3) posterior wall distance, 0.5 mm (SD 0.2), much smaller than 4.7 mm for arthroscopic-assisted femoral tunnelling. The intraclass correlation coefficients, calculated to determine the accuracy and reliability of navigational femoral tunnelling, showed excellent internal consistency that ranged from 0.965 to 0.989 for tunnel length and from 0.810 to 0.953 for posterior wall distance. CONCLUSION: Navigation systems with enhancement of the registration accuracy by the developed system are feasible in anatomical ACLR, in reducing surgical failures such as short tunnel length or posterior wall breakage of distal femur. The present study revealed that computer navigation could aid in avoiding major mistakes in exact positioning and posterior wall blowout and help in attaining appropriate length for femoral tunnelling in anatomical ACLR.

Relationship of severity of hyponatremia and adverse outcomes in children visiting the emergency department.

Mild hyponatremia is often found in patients visiting pediatric emergency departments (PEDs), but there are few large-scale studies on its association with adverse outcomes, including mortality. We conducted this study to identify the association of mild hyponatremia with adverse outcome. This retrospective observational study included children under 18 years of age visiting the PED at a tertiary hospital. We used electronic medical record data from January 1, 2009 to December 31, 2020. Clinical outcomes, including ward admission, vasopressor administration, pediatric intensive care unit (PICU) admission, and mortality, were assessed for the total of 44,147 patients. Among these, 1,639 (3.7%) were in the hyponatremia group, with 1,521 (3.4%) exhibiting mild hyponatremia. Mild hyponatremia was more prevalent in younger patients, particularly in the 1-3 years age group, and less common in females. Patients with mild hyponatremia had a significantly prolonged median length of stay in the PED compared to normonatremic patients (5.8 h vs. 4.4 h, p < 0.001). Moreover, they showed significantly higher rates of ward admission (51.1% vs. 35.6%, p < 0.001), vasopressor administration (1.1% vs. 0.6%, p = 0.014), PICU admission (2.4% vs. 1.0%, p < 0.001), and mortality (1.5% vs. 0.3%, p < 0.001). Compared with the normonatremia group, the odds ratios (95% CI) for ward admission, vasopressor administration, PICU admission, and mortality in the mild hyponatremia group were 1.90 (1.71-2.10), 1.91 (1.17-3.13), 2.62 (1.86-3.68), and 5.56 (3.51-8.80), respectively. Furthermore, our findings demonstrate a notable upward trend in adverse outcomes, including vasopressor administration, PICU admission, and mortality, from mild hyponatremia to severe hyponatremia. In conclusion, we found that adverse outcomes increase with the severity of hyponatremia in children presenting to the PED, highlighting the importance of immediate intervention alongside the identification of the underlying cause.

Association among basal serum BDNF, cardiorespiratory fitness and cardiovascular disease risk factors in untrained healthy Korean men.

Evidence suggests that serum brain-derived neurotrophic factor (serum BDNF) can be affected by cardiorespiratory fitness (CRF), but this relationship is far from clear. Recent reports show an inverse relationship between serum BDNF and CRF in healthy individuals, and other studies suggest a possible association between serum BDNF and cardiovascular disease. However, the possible interaction between serum BDNF, CRF, and cardiovascular disease risk has not been studied. The purpose of this study was to examine the association among serum BDNF, CRF, and cardiovascular disease risk factors in healthy men. The investigation involved a large sample of men (n = 995, age range: 20-76 years) who live in the central area of South Korea and were recruited into the Preventive Health Study. Our study showed a significant inverse relationship between serum BDNF and relative VO(2)max (r = -0.412, p < 0.0001) and heart rate reserve (r = -0.194, p < 0.0001). Serum BDNF was positively correlated with body mass index (r = 0.80, p < 0.0001), total cholesterol (r = 0.185, p < 0.0001), and triglyceride (r = 0.320, p < 0.0001). Our data suggest that serum BDNF may be associated with effects of increased CRF on cardiovascular disease. However, more research is clearly needed before a determination of whether, and to what extent, serum BDNF may be responsible for some of the health benefits associated with CRF.

Influence of performance level on anaerobic power and body composition in elite male judoists.

This study examined the relationship between 30-second anaerobic power and body composition by performance level in elite Judoists. During a 3-month period, 10 male Korean Judo national team athletes (NT), 26 male university varsity team athletes (VT), and 28 male junior varsity team athletes (JT) were assessed for 30-second anaerobic power and body composition at the Youngin University. Anaerobic power was measured using a 30-second Wingate test. Body composition was assessed via bioelectric impedance analysis in standardized conditions using BioSpace (Korean)-specific prediction formulas. All testing occurred at the beginning of the winter nonseason period but excluded a brief weight-loss period before the competition phase. Anaerobic power measures were significantly greater (p < 0.05) in NT and VT than in JT. Fat-free mass (FFM), muscle mass (MM), and total body water in JT were also greater than in VT and JT (p < 0.05). Muscle mass in VT was significantly lower than in NT (p < 0.05). Fat-free mass in NT was strongly correlated to mean and peak anaerobic power (r = 0.77, p = 0.009; r = 0.87, p < 0.001, respectively). Varsity team athletes also indicated a moderate association between FFM and peak and mean anaerobic power (r = 0.63, p < 0.001; r = 0.48, p = 0.013, respectively). However, relationship between FFM and anaerobic power was not statistically significantly correlated in JT (r = 0.14, p = 0.470; r = 0.23, p = 0.232, separately). In conclusion, our data indicated that anaerobic power is closely correlated with increase in FFM and MM and was different dependent among performance levels. Further research in the field is warranted to elucidate the Judo-specific relationship between FFM and performance.

Decreased Maximal Tongue Protrusion Length May Predict the Presence of Dysphagia in Stroke Patients.

OBJECTIVE: To investigate the relationship between maximal tongue protrusion length (MTPL) and dysphagia in post-stroke patients. METHODS: Free tongue length (FTL) was measured using the quick tongue-tie assessment tool and MTPL was measured using a transparent plastic ruler in 47 post-stroke patients. The MTPL-to-FTL (RMF) ratio was calculated. Swallowing function in all patients was evaluated via videofluoroscopic swallowing study (VFSS), PenetrationAspiration Scale (PAS), Functional Oral Intake Scale (FOIS), and Videofluoroscopic Dysphagia Scale (VDS). RESULTS: The MTPL and RMF values were significantly higher in the non-aspirator group than in the aspirator group (MTPL, p=0.0049; RMF, p<0.001). MTPL and RMF showed significant correlations with PAS, FOIS and VDS scores. The cut-off value in RMF for the prediction of aspiration was 1.56, with a sensitivity of 84% and a specificity of 86%. CONCLUSION: There is a relationship between MTPL and dysphagia in post-stroke patients. MTPL and RMF can be useful for detecting aspiration in post-stroke patients.

Optimal Ultrasonographic Measurements for Diagnosing Carpal Tunnel Syndrome in Patients With Diabetic Sensorimotor Polyneuropathy: A Case-Control Study.

OBJECTIVE: To investigate the optimal sonographic method for diagnosing carpal tunnel syndrome (CTS) in patients with diabetic sensorimotor polyneuropathy (DSP). METHODS: A total of 190 participants were divided into four groups based on DSP history and electrodiagnostic results of CTS. The absolute parameters were measured at baseline and the relative values were calculated: maximal cross-sectional area (CSA) of the median nerve throughout the carpal tunnel (Mmax), median nerve CSA at the forearm level (Mf), ulnar nerve CSA at the pisiform level (Upi), difference between Mmax and Mf (∆MM), and difference between Mmax and Upi (∆MU). Then, the optimal ultrasonographic parameters for diagnosing CTS, according to the presence of DSP, using absolute and relative cutoff values were analyzed. RESULTS: Median and ulnar nerve CSAs were significantly larger in the DSP group than in the control group. In the DSP participants, the mean Mmax, ∆MM, and ∆MU values were significantly larger in patients with both DSP and CTS than in patients with DSP only. The Mmax thresholds of 9.5 mm2 in the control group and 11.5 mm2 in the DSP group showed the greatest sensitivity and specificity for diagnosing CTS. The ∆MM thresholds of 2.5 mm2 and ∆MU thresholds of 4.5 mm2 had the greatest sensitivity and specificity in both the DSP and control groups. CONCLUSION: Measurement of Mmax, ∆MM and ∆MU is an optimal ultrasonographic evaluation method for diagnosing CTS in patients with DSP.

Three-dimensional orbital wall modeling using paranasal sinus segmentation.

PURPOSE: Three-dimensional orbital wall modeling is a time-consuming process because of the presence of pseudoforamina. We developed an automated three-dimensional modeling software to characterize the orbital wall, and evaluated it using data from fracture patients. METHODS: We first characterized the air and face regions using multiphase segmentation; the sinuses were segmented by applying morphological operations to air regions. Pseudoforamina of the orbital wall were offset with the segmented sinuses. Finally, the three-dimensional facial bone model, with orbital wall, was reconstructed from the segmented images. RESULTS: Ten computed tomography data sets were used to evaluate the proposed method. Results were compared with those obtained using the active contour model and manual segmentation. The process took 31.7 ± 8.0 s, which was 30-60 times faster than other methods. The average distances between surfaces obtained with the proposed method and those obtained with manual segmentation (normal side: 0.20 ± 0.06 mm; fractured side: 0.28 ± 0.10 mm) were approximately half those obtained using the active contour model. CONCLUSIONS: Three-dimensional orbital wall models, which were very similar to the manually segmented models, were archived within 1 min using the developed software, regardless of fracture presence. The proposed method might improve the safety and accuracy of surgical procedures.

Registration accuracy enhancement of a surgical navigation system for anterior cruciate ligament reconstruction: A phantom and cadaveric study.

BACKGROUND: Recently, surgical navigation systems have been widely used to improve the results of various orthopaedic surgeries. However, surgical navigation has not been successful in anterior cruciate ligament reconstruction, owing to its inaccuracy and inconvenience. This study investigated the registration of preoperative and intraoperative data, which are the key components in improving accuracy of the navigation system. METHODS: An accurate registration method was proposed using new optical tracking markers and landmark retake. A surgical planning and navigation system for anterior cruciate ligament reconstruction was developed and implemented. The accuracy of the proposed system has been evaluated using phantoms and eight cadaveric knees. The present study investigated only the registration accuracy excluding the errors of optical tracking hardware and surgeon. RESULTS: The target registration errors of femoral tunnelling for anterior cruciate ligament reconstruction in phantoms were found to be 0.24±0.03mm and 0.19±0.10° for the tunnel entry position and tunnel direction, respectively. The target registration errors measured using cadavers were 0.9mm and 1.94°, respectively. CONCLUSIONS: The preclinical experimental results showed that the proposed methods enhanced the registration accuracy of the developed system. As the system becomes more accurate, surgeons could more precisely position and orient the femoral and tibial tunnels to their original anatomical locations.

Blockage of Autophagy Rescues the Dual PI3K/mTOR Inhibitor BEZ235-induced Growth Inhibition of Colorectal Cancer Cells.

Molecular targeting for the altered signaling pathways has been proven to be effective for the treatment ofmany types of human cancer, including colorectal cancer (CRC). The dual phosphatidylinositol-3-kinase (PI3K) and mammalian target of rapamycin (mTOR) inhibitor BEZ235 has shown to exhibit potent antitumor activity against solid tumors. Autophagy is a cellular lysosomal catabolic process to maintain metabolic homeostasis, which has been known to be induced in response to many therapeutic agents in cancer cells. This process is negatively regulated by mTOR and often acts as prosurvival or prodeath mechanism following cancer therapeutics. The current study was designed to investigate the antiproliferation activity of BEZ235 and to evaluate the role of autophagy induced by BEZ235 using HCT15 CRC cells bearing ras oncogene mutation. We found that BEZ235 decreases cell viability, which was mostly dependent on G1 arrest of cell cycle via suppression of cyclin A expression. BEZ235 affects PI3K/Akt/mTOR signaling pathway by increasing the phosphorylation of AKT at Ser(473) and RAS/RAF/MEK/ERK pathway by decreasing the phosphorylation of ERK at Tyr(204). BEZ235 also stimulated autophagy induction as evidenced by the increased expression of LC3-II and abundant acidic vesicular organelles (AVOs) in the cytoplasm. In addition, the combination of BEZ235 with autophagy inhibitor chloroquine, a known antagonist of autophagy, counteracted the antiproliferation effect of BEZ235. Thus, our study indicates that autophagy induced in response to BEZ235 treatment appears to act as cell death mechanism in HCT15 CRC cells.

The Effect of Optical Marker Configuration on Tracking Accuracy in Image Guided Surgery.

In this study, the effect of the geometrical configuration of retroreflective markers on the maximum tracking uncertainty of target points during a medical navigation is analyzed. Methods that help users select and set up an optimal configuration for minimizing the navigation uncertainty are proposed. The methods are evaluated by simulating surgical navigation environment using an optical tracking system. The result shows that the maximum uncertainty of tracking the target points lying outside the marker region is a function of the configuration of some of the associated markers and the precision of the optical tracking system used. We also show that the use of flexible rigid bodies minimizes target tracking uncertainty by enabling a customized reconfiguration of markers for optimal pose with respect to individual surgical target position during preoperative planning phase.

Markerless surgical robotic system for intracerebral hemorrhage surgery.

Conventional intracerebral hemorrhage (ICH) surgery uses a stereotactic frame to access an intracerebral hematoma. Using a stereotactic frame for ICH surgery requires a long preparation time. In order to resolve this problem, we propose a markerless surgical robotic system. This system uses weighted iterative closest point technology for surface registration, hand-eye calibration for needle insertion, and 3D surface scanning for registration. We need calibration to integrate the technologies: calibration of robot and needle coordinates and calibration of 3D surface scanning and needle coordinates. These calibrations are essential elements of the markerless surgical robotic system. This system has the advantages of being non-invasive, a short total operation time, and low radiation exposure compared to conventional ICH surgery.

Effects of potato and lotus leaf extract intake on body composition and blood lipid concentration.

PURPOSE: The purpose of this study was to investigate the effects of potato and lotus leaf extract intake on body composition, abdominal fat, and blood lipid concentration in female university students. METHODS: A total of 19 female university students participated in this 8-week study, and they were randomly assigned into 2 groups; potato and lotus leaf extract (skinny-line) administered group (SKG, n =9) and placebo group (PG, n = 10). The main results of the present study are presented below. RESULTS: 1) Body mass index, and percent body fat and abdominal fat in students of the SKG showed a decreasing tendency without significant interaction, 2) total cholesterol (TC), triglyceride (TG), and low density lipoprotein (LDL-C) in students of the SKG showed an averagely decreasing tendency and there was a significant interaction of TC only, 3) high density lipoprotein (HDL-C) in students of the SKG showed an increasing tendency without significant interaction, and 4) Z-score of fatness testing interaction in group × repetition did not show a significant interaction; however, there was a significant interaction of TC in group × repetition. Based on these results, 8-week intake of potato and lotus leaf extract had a positive effect of lowering TC. On the other hand, it had no significant effect on other types of lipids and percent body fat changes. CONCLUSION: There was a positive tendency of blood lipids in students of the SKG and it seems that potato and lotus leaf extract intake might prevent obesity and improve obesity related syndromes.

Effects of the controlled-released TGF-beta 1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold.

The objectives of this study were (1) to develop a three-dimensional collagen/chitosan/glycosaminoglycan (GAG) scaffold in combination with transforming growth factor-beta1 (TGF-beta 1)-loaded chitosan microspheres, and (2) to evaluate the effect of released TGF-beta 1 on the chondrogenic potential of rabbit chondrocytes in such scaffolds. TGF-beta 1 was loaded into chitosan microspheres using an emulsion-crosslinking method. The controlled release of TGF-beta 1, as measured by enzyme-linked immunosorbent assay (ELISA), was monitored for 7 days. The porous scaffolds containing collagen and chitosan were fabricated by using a freeze drying technique and crosslinked using 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide (EDC) in the presence of chondroitin sulfate (CS), as a GAG component. The TGF-beta 1 microspheres were encapsulated into the scaffold at a concentration of 10 ng TGF-beta 1/scaffold and then chondrocytes were seeded in the scaffold and incubated in vitro for 3 weeks. Both proliferation rate and glycosaminoglycan (GAG) production were significantly higher in the TGF-beta 1 microsphere-incorporated scaffolds than in the control scaffolds without microspheres. Extracellular matrix staining by Safranin O and immunohistochemistry for type II collagen were elevated in the scaffold with TGF-beta 1 microspheres. These results suggest that TGF-beta 1 microspheres when incorporated into a scaffold have the potential to enhance cartilage formation.

The concentrations of serum, plasma and platelet BDNF are all increased by treadmill VO2max performance in healthy college men.

The most current human-based studies in which brain-derived neurotrophic factor (BDNF) levels in the peripheral blood system are analyzed use it as an indicator that represents BDNF levels in the CNS. However, whether circulating BDNF (serum and plasma) is positively or inversely associated with cardiorespiratory fitness levels (VO(2max)) is still controversial, and no study has done to investigate exercise effects on the concentration of BDNF stored in circulating platelets which, in fact, store a large amount of circulating BDNF. Thus, the purpose of this study was to determine the relation between VO(2max) and all circulating BDNF levels (serum, plasma and platelets) in college male students (N=18; age, 19 ± 1 years; height, 173.22 ± 7.65 cm; weight, 78.25 ± 14.25 kg; body fat percent, 13.82 ± 5.68%). Dual X-ray energy absorptiometry whole body scan was used to measure their body composition. After the overnight fast, all participants were performed VO(2max) test, and their blood was collected at rest and immediately after the exercise. Our data resulted in significant increases in platelet counts and serum, plasma and platelet BDNF levels immediately after the exercise (p<0.01). VO(2max) had a significant negative correlation with serum BDNF, plasma BDNF and platelet BDNF at rest (p<0.05) but a significant positive correlation with serum, plasma BDNF, and platelet BDNF immediately after the exercise (p<0.01). However, our data show no correlation between VO(2max) and platelet count both at rest and immediately after the exercise. In conclusion, this is the first study showing that basal BDNF levels are inversely correlated with cardiorespiratory fitness levels but that the inverse correlations turn into positive correlations with all circulating BDNF levels immediately after the exercise. Moreover, it is the first time to provide evidence that platelet BDNF levels are also positively affected by the exercise. However, future studies will be needed to investigate what tissues provide BDNF into the circulating system and to elucidate the role of circulating BDNF.

Independent and joint associations of cardiorespiratory fitness and muscle fitness with metabolic syndrome in Korean men.

BACKGROUND: Higher cardiorespiratory fitness (CRF) and muscle fitness (MF) attenuate metabolic syndrome prevalence in men. However, associations of CRF and MF with metabolic syndrome were poorly understood in South Korean men. Therefore, this study aimed to examine the independent and joint associations of CRF and MF on metabolic syndrome prevalence in Korean men. METHODS: Korean men (n=1,097, mean age=44 years) were recruited from the central areas of South Korea for this Preventive Health Study (PHS) between May, 2007, and October, 2008, and they underwent a health examination, maximal treadmill test, and five different muscle fitness tests for this study. RESULTS: From our data, separate age- and current smoking-adjusted logistic regression models revealed a graded inverse association of metabolic syndrome prevalence with CRF (P<0.0001) and MF (P<0.0001). The association of MF with metabolic syndrome remained significant and became attenuated with further adjustment, for CRF (P=0.02). However, the association of CRF with metabolic syndrome remained unchanged when adjusted for MF (P<0.0001). In the joint analysis using the low-strength and CRF unfit group as the baseline, the odds ratios (95% confidence intervals) for having metabolic syndrome were 0.52 (0.26-1.05) for the high-strength and CRF unfit group, 0.39 (0.24-0.65) for the low-strength and CRF fit group, and 0.28 (0.17-0.46) for the high-strength and CRF fit group. CONCLUSIONS: Both cardiorespiratory fitness and muscle fitness showed independent and joint inverse associations with metabolic syndrome prevalence in Korean adult men.

Struma ovarii and peritoneal strumosis with thyrotoxicosis.

BACKGROUND: Struma ovarii is a highly specialized form of mature ovarian teratoma consisting of thyroid tissue and exhibiting all the histological features of the thyroid gland. Malignant transformation of thyroid tissue in struma ovarii and metastasis are extremely uncommon. In rare cases, benign thyroid tissue may spread to the peritoneal cavity, and pathologic examination of the peritoneal implants shows multiple nodules of varying sizes of mature thyroid tissue similar to struma ovarii. This condition is termed "peritoneal strumosis." SUMMARY: We report a 49-year-old woman with struma ovarii complicated by peritoneal strumosis with thyrotoxicosis. After surgical resection of the struma ovarii and peritoneal strumosis the patient became euthyroid. CONCLUSION: To the best of our knowledge this is the first report of a patient with peritoneal strumosis complicated by thyrotoxicosis. The relative contribution to circulating thyroid hormones by the patient's struma ovarii as compared to the peritoneal strumosis is not known.

Effects of a chitosan scaffold containing TGF-beta1 encapsulated chitosan microspheres on in vitro chondrocyte culture.

The objectives of this study were (1) to develop a three-dimensional chitosan scaffold in combination with transforming growth factor-beta1 (TGF-beta1)-loaded chitosan microspheres and (2) to evaluate the effect of the TGF-beta1 release on the chondrogenic potential of rabbit chondrocytes in the scaffolds. TGF-beta1 was loaded into chitosan microspheres using an emulsion-crosslinking method, resulting in spherical shapes with a size ranging from 0.3 to 1.5 microm. Controlled release of TGF-beta1, as measured by enzyme-linked immunosorbent assay (ELISA), was observed with chitosan microspheres over 7 days. Chitosan solutions (2% and 3%) were fabricated into two types of scaffolds with different pore morphologies and mechanical properties using a freeze-drying technique, with the result that scaffold with higher concentrations showed smaller pores and lower porosity, leading to a much stronger scaffold. The TGF-beta1 microspheres were incorporated into the scaffolds at a concentration of 10 ng TGF-beta1/scaffold and then chondrocytes seeded into each scaffold and incubated in vitro for 2 weeks. The 2% chitosan scaffolds showed higher cell attachment levels than the 3% chitosan scaffolds (P < 0.01), regardless of the TGF-beta1 microspheres. Both the proliferation rate and glycosaminoglycan (GAG) production were significantly higher for scaffolds incorporating TGF-beta1 microspheres than for the control scaffolds without microspheres 10 days after incubation. Extracellular matrix staining by Safranin O and immunohistochemistry for type II collagen both significantly increased in scaffolds containing TGF-beta1 microspheres. These results suggest that the TGF-beta1 microsphere incorporated in scaffolds have the potential to enhance cartilage formation.