Effect of Chitosan on Synovial Membrane Derived Cells and Anterior Cruciate Ligament Fibroblasts.

Previously, chitosan reduces the senescence-related phenotypes in human foreskin fibroblasts through the transforming growth factor beta (TGF-ß) pathway, and enhances the proliferation and migration capabilities of these cells are demonstrated. In this study, we examined whether the senescence-delaying effect of chitosan could be applied to primary knee-related fibroblasts, such as human synovial membrane derived cells (SCs) and anterior cruciate ligament fibroblasts (ACLs). These two types of cells were obtained from donors who needed ACL reconstruction or knee replacement. We found that chitosan treatment effectively reduced aging-associated ß-galactosidase (SA-ß-gal)-positive cells, downregulated the expression of senescence-related proteins pRB and p53, and enhanced the 5-bromo-2'-deoxyuridine (BrdU) incorporation ability of SCs and ACLs. Moreover, chitosan could make SCs secret more glycosaminoglycans (GAGs) and produce type I collagen. The ability of ACLs to close the wound was also enhanced, and the TGF-ß and alpha smooth muscle actin (αSMA) protein expression decreased after chitosan treatment. In summary, chitosan not only delayed the senescence but also enhanced the functions of SCs and ACLs, which is beneficial to the application of chitosan in cell expansion in vitro and cell therapy.

The Outcome of under 10 mm Single-Incision Surgery Using a Non-Specialized Volar Plate in Distal Radius Fractures: A Retrospective Comparative Study.

BACKGROUND: The distal radius fracture is a common orthopedic injury. We aimed to share the surgical steps and investigate the outcomes of treating distal radius fractures with wounds ≤10 mm using a globally accessible locking plate. METHODS: We collected 46 patients who underwent surgery via a <10 mm wound, with a control group consisting of 40 patients who underwent conventional procedures. Both groups were treated using the same volar plate. We compared the radiographic reduction quality, including volar tilt angle, radial inclination angle, and ulna variance. Additionally, clinical outcomes, such as pain assessed using VAS, Q-Dash score, and PRWE, were evaluated. Patient satisfaction with the wound was also analyzed. The follow-up time for the clinical outcomes was 24.2 ± 13.47 months. RESULTS: There were no differences in the quality of reduction in parameters such as the volar tilt angle (p = 0.762), radial inclination angle (p = 0.986), and ulna variance (p = 0.166). Both groups exhibited comparable results in pain VAS (p = 0.684), Q-Dash score (p = 0.08), and PRWE (p = 0.134). The ≤10 mm incision group displayed an increase in satisfaction with the wound (p < 0.001). CONCLUSIONS: Treating distal radius fractures with a <10 mm wound using a non-specialized locking plate is a feasible approach. It does not compromise the quality of fracture reduction or functional scores and improves wound satisfaction.

IL-6 induces periostin production in human ACL remnants: a possible mechanism causing post-traumatic osteoarthritis.

OBJECTIVE: Perostin (POSTN) and IL-6 consistently elevated after ACL injury, and ACL has been proposed as the major source of POSTN. However, there is a lack of evidence whether IL-6 induces ACL remnants to produce POSTN. This study aimed to investigate the effect of IL-6 on POSTN production in ACL fibroblasts, which may help us understand more about the mechanism of PTOA after ACL injury and ACL reconstruction. METHODS: ACL remnants were harvested from 27 patients undergoing ACL reconstruction. Quantitative real-time polymerase chain reaction (PCR) was performed to examine the POSTN gene expression of ACL fibroblasts after treatment of different concentrations of IL-6. The POSTN protein production of ACL fibroblasts was determined using western blot analysis. The blockers of possible signaling pathways, including PI3K/Akt, Ras/MAPK, and JAK/STAT pathways, were added to test whether the effect of IL-6 on ACL fibroblast could be attenuated. ACL fibroblast and chondrocyte co-culture was carried out to determine the influence of ACL and IL-6 on chondrocytes. RESULTS: Quantitative real-time PCR showed that IL-6 time-dependently and dose-dependently increased POSTN gene expression of ACL fibroblast. Western blot analysis also revealed that IL-6 dose-dependently induced POSTN protein production. Regarding the chronicity of ACL injury, the POSTN protein production was comparable between ACL remnants which were derived within 3 months of injury and at least 6 months after injury. PI3K/Akt blockers could attenuate the effect of IL-6 on ACL remnants, whereas Ras/MAPK and JAK/STAT did not decrease POSTN production. The coexistence of ACL and IL-6 induced more MMP-13 and ADAMTS-4 by chondrocytes. CONCLUSIONS: IL-6 induced ACL remnants to produce POSTN. This effect could be attenuated by the PI3K/Akt blocker. Coexistence of IL-6 and ACL remnants may accelerate post-traumatic arthritis.

Validation of the Efficacy of Ultrasound Speckle Tracking in Measuring Tendon Gliding After Finger Flexor Tendon Repair.

OBJECTIVE: Restricted tendon gliding is commonly observed in patients after finger flexor tendon (FFT) repair. The study described here was aimed at quantifying the amount of FFT gliding to evaluate the recovery of post-operative tendons using a 2-D radiofrequency (RF)-based ultrasound speckle tracking algorithm (UST). METHODS: Ex vivo uniaxial tensile testing of porcine flexor tendons and in vivo isometric testing of human FFT were implemented to verify the efficacy of UST beforehand. The verified UST was then applied to the patients after FFT repair to compare tendon gliding between affected and healthy sides and to investigate its correlation with the joint range of motion (ROM). RESULTS: Excellent validity was confirmed with the average R2 value of 0.98, mean absolute error of 0.15 ± 0.08 mm and mean absolute percentage error of 5.19 ± 2.43% between results from UST and ex vivo testing. The test-retest reliability was verified with good agreement of ICC (0.90). The affected side exhibited less gliding (p = 0.001) and smaller active ROM (p = 0.002) than the healthy side. Meanwhile, a significant correlation between tendon gliding and passive ROM was found only on the healthy side (ρ = 0.711, p = 0.009). CONCLUSION: The present study provides a promising protocol to evaluate post-operative tendon recovery by quantifying the amount of FFT gliding with a validated UST. FFT gliding in patients with different levels of ROM restriction should be further explored for categorizing the severity of tendon adhesion.

Novel Modular Spine Blocks Affect the Lumbar Spine on Finite Element Analysis.

Introduction: There are various surgical interventions to manage osteoporotic vertebral compression fracture. Modular spine block (MSB) is a novel intravertebral fixator that can be assembled. This study aimed to quantitatively investigate the force distribution in vertebrae with the various structural designs and implantation methods by finite element analysis (FEA). Methods: A three-dimensional nonlinear FEA of the L3 implanted with MSB was constructed. Different structural designs (solid vs. hollow) and implantation methods (three-layered vs. six-layered and unilateral vs. bilateral) were studied. The model was preloaded to 150 N-m before the effects of flexion, extension, torsion, and lateral bending were analyzed at the controlled ranges of motion of 20°, 15°, 8°, and 20°, respectively. The resultant intervertebral range of motion (ROM) and disk stress as well as intravertebral force distribution were analyzed at the adjacent segments. Results: The different layers of MSB provided similar stability at the adjacent segments regarding the intervertebral ROM and disk stress. Under stress tests, the force of the solid MSB was shown to be evenly distributed within the vertebrae. The maximum stress value of the unilaterally three-layered hollow MSB was generally lower than that of the bilaterally six-layered solid MSB. Conclusions: The MSB has little stress shielding effect on the intervertebral ROM and creates no additional loading to the adjacent disks. The surgeon can choose the appropriate numbers of MSB to fix vertebrae without worrying about poly(methyl methacrylate) extravasation, implant failure, or adjacent segment disease.

A simple method to improve the antibiotic elution profiles from polymethylmethacrylate bone cement spacers by using rapid absorbable sutures.

OBJECTIVE: Antibiotic-loaded bone cement beads and spacers have been widely used for orthopaedic infection. Poor antibiotic elution is not capable of eradicating microbial pathogens and could lead to treatment failure. The elution profiles differ among different cement formulations. Although Simplex P cement has the least release amount, it is widely used due to its ready availability. Previous methods aiming to improve the elution profiles were not translated well to clinical practice. We sought to address this by using easily available materials to improve the elution profile of antibiotics from PMMA, which allows clinicians to implement the method intraoperatively. METHODS: Vancomycin was mixed with Simplex P cement. We used Vicryl Rapide sutures to fabricate sustained-release cement beads by repetitively passing the sutures through the beads and/or mixing suture segments into the cement formulation. Vancomycin elution was measured for 49 days. The mechanism of antibiotic release was observed with gross appearance and scanning electron microscopic images. The antimicrobial activities against MRSA were tested using an agar disk diffusion bioassay. RESULTS: Passing Vicryl Rapide sutures through cement beads significantly improved the elution profiles in the 7-week period. The increased ratios were 9.0% on the first day and 118.0% from the 2nd day to the 49th day. Addition of suture segments did not increase release amount. The Vicryl Rapide sutures completely degraded at the periphery and partially degraded at the center. The antibiotic particles were released around the suture, while antibiotic particles kept densely entrapped in the control group. The antimicrobial activities were stronger in passing suture groups. CONCLUSION: Passing fast absorbable sutures through PMMA cement is a feasible method to fabricate sustained-release antibiotic bone cement. Intra-cement tunnels can be formed, and the effect can last for at least 7 weeks. It is suitable for a temporary spacer between two stages of a revision surgery.

Finite element analysis after rod fracture of the spinal hybrid elastic rod system.

BACKGROUND: The spinal hybrid elastic (SHE) rod dynamic stabilization system can provide sufficient spine support and less adjacent segment stress. This study aimed to investigate the biomechanical effects after the internal fracture of SHE rods using finite element analysis. METHODS: A three-dimensional nonlinear finite element model was developed. The SHE rod comprises an inner nitinol stick (NS) and an outer polycarbonate urethane (PCU) shell (PS). The fracture was set at the caudal third portion of the NS, where the maximum stress occurred. The resultant intervertebral range of motion (ROM), intervertebral disc stress, facet joint contact force, screw stress, NS stress, and PCU stress were analyzed. RESULTS: When compared with the intact spine model, the overall trend was that the ROM, intervertebral disc stress, and facet joint force decreased in the implanted level and increased in the adjacent level. When compared with the Ns-I, the trend in the Ns-F decreased and remained nearly half effect. Except for torsion, the PS stress of the Ns-F increased because of the sharing of NS stress after the NS fracture. CONCLUSIONS: The study concluded the biomechanical effects still afford nearly sufficient spine support and gentle adjacent segment stress after rod fracture in a worst-case scenario of the thinnest PS of the SHE rod system.

Knee-ankle joint line angle: a significant contributor to high-degree knee joint line obliquity in medial opening wedge high tibial osteotomy.

BACKGROUND: Medial opening wedge high tibial osteotomy (MOWHTO) changes the knee joint inclination in the coronal plane, which can be compensated by the ankle joint. Once there is a decompensated knee joint obliquity, it can induce excessive shear force on the articular cartilage. This study aimed to investigate the capacity of the compensation by analyzing the correlation of the knee-ankle joint line angle (KAJA) and the knee joint line obliquity (KJLO). PATIENTS AND METHODS: Ninety-six patients undergoing MOWHTO were included. We measured potential predictors including preoperative or postoperative body mass index (BMI), weight-bearing line (WBL) ratio/correction amount, knee-ankle joint line angle(KAJA), mechanical lateral distal femoral angle (mLDFA), medial proximal tibia angle (MPTA), ankle joint line obliquity (AJLO), mechanical hip-knee-ankle angle (mHKA) and joint line convergence angle (JLCA). The correlations of these predictors and postoperative KJLO were determined using Pearson correlation coefficient. The contribution of significant predictors was further analyzed using multiple linear regression. Finally, the cutoff value of the most contributing factor resulting in decompensated KJLO was derived with receiver operating characteristic (ROC) curve analysis. RESULTS: Preoperative AJLO, JLCA, MPTA, mHKA and KJLO and postoperative KAJA and MPTA correlated with postoperative KJLO. After multiple linear regression, only preoperative AJLO and JLCA and postoperative KAJA still showed significant contribution to postoperative KJLO. Postoperative KAJA made the greatest contribution. The cutoff value of postoperative KAJA was at 9.6° after ROC analysis. The incidence rate of high-grade KJLO was 69.6% when postoperative KAJA exceeded 9.6°. CONCLUSIONS: Postoperative KAJA is a significant contributor to high-grade KJLO after MOWHTO. The incidence was increased at angles greater than 9.6°. The results suggest that KAJA should be carefully assessed during preoperative planning or intraoperative evaluation. Postoperative KAJA < 9.6° can lower the rate of early high-degree KJLO.

Safety and Efficacy of Kartigen® in Treating Cartilage Defects: A Randomized, Controlled, Phase I Trial.

Here, we aimed to investigate the safety and preliminary efficacy of Kartigen®, a matrix with autologous bone marrow mesenchymal stem cell-derived chondrocyte precursors embedded in atelocollagen. As a surgical graft, Kartigen® was implanted onto the cartilage defects at the weight-bearing site of the medial femoral condyle of the knee. Fifteen patients were enrolled and stratified into two groups, undergoing either Kartigen® implantation (n = 10) or microfracture (control group, n = 5). The primary endpoint was to evaluate the safety of Kartigen® by monitoring the occurrence of adverse events through physician queries, physical examinations, laboratory tests, and radiological analyses for 2 years. There were no infections, inflammations, adhesions, loose body, or tumor formations in the Kartigen®-implanted knees. The preliminary efficacy was assessed using the International Knee Documentation Committee (IKDC) score, visual analog scale, and second-look arthroscopy. The postoperative IKDC scores of the Kartigen® group significantly improved in the 16th week (IKDC = 62.1 ± 12.8, p = 0.025), kept increasing in the first year (IKDC = 78.2 ± 15.4, p < 0.005), and remained satisfactory in the second year (IKDC = 73.6 ± 13.8, p < 0.005), compared to the preoperative condition (IKDC = 47.1 ± 17.0), while the postoperative IKDC scores of the control group also achieved significant improvement in the 28th week (IKDC = 68.5 ± 6.1, p = 0.032) versus preoperative state (IKDC = 54.0 ± 9.1). However, the IKDC scores decreased in the first year (IKDC = 63.5 ± 11.6) as well as in the second year (IKDC = 52.6 ± 16.4). Thirteen patients underwent second-look arthroscopy and biopsy one year after the operation. The Kartigen® group exhibited integration between Kartigen® and host tissue with a smooth appearance at the recipient site, whereas the microfracture group showed fibrillated surfaces. The histological and immunohistochemical analyses of biopsy specimens demonstrated the columnar structure of articular cartilage and existence of collagen type II and glycosaminoglycan mimic hyaline cartilage. This study indicates that Kartigen® is safe and effective in treating cartilage defects.

Focal proximal fibular angle: A potential indicator of the tibial mechanical axis in opening-wedge high tibial osteotomy.

BACKGROUND: For opening-wedge high tibial osteotomy, correct alignment is essential for a better prognosis. It is difficult to evaluate the mechanical axis of the lower extremity or tibia using a single fluoroscopic image. This study aimed to discuss the use of focal proximal fibular angle (FPFA), which can be assessed by a single fluoroscopic image, as an intraoperative indicator. METHODS: Eligible for analyses were 111 consecutively treated patients; for the final analyses 96 patients were included. The preoperative and postoperative medial proximal tibial angle (MPTA) and FPFA were measured. The relationship between these two angles, correction amount, weight-bearing line ratio and patient characteristics were analyzed. RESULTS: The preoperative FPFA and MPTA were 96.5 ± 3.8° (mean ± standard deviation, SD) and 84.8 ± 3.0°, while the postoperative FPFA and MPTA were 87.6 ± 4.1° and 94.0 ± 3.5°, respectively. The preoperative and postoperative sums of the MPTA and FPFA were constant. The discrepancy was less than 3° in all knees, less than 2° in 92.7% knees and less than 1° in 68.8% knees. It was not correlated with age, sex, weight-bearing line ratio, or correction amount. CONCLUSION: The study findings confirmed the constancy of the sum of the MPTA and FPFA. The FPFA can be easily evaluated on a single fluoroscopic image of the knee. Use of the FPFA as guidance may simplify the procedure of opening-wedge high tibial osteotomy and approximately predict the tibial mechanical axis.

Atelocollagen-Embedded Chondrocyte Precursors as a Treatment for Grade-4 Cartilage Defects of the Femoral Condyle: A Case Series with up to 9-Year Follow-Up.

We demonstrated the safety and efficacy of autologous chondrocyte precursor (CP) cell therapy in repairing Grade 4 cartilage defects of medial femoral condyles. The autologous bone marrow mesenchymal stem cells of each participant were isolated, amplified, and then differentiated into CPs in atelocollagen. Neotissues made of CPs were implanted into cartilage defects with an average cell density of 4.9 ± 2.1 × 106 cells/cm2 through arthrotomy. The knee function was evaluated with the International Knee Documentation Committee (IKDC) subjective knee form. Patients' knee functions significantly improved by the 28th week (IKDC score = 68.3 ± 12.1), relative to the initial functionality before the CP therapy (IKDC score = 46.1 ± 16.4, p-value = 0.0014). Nine of these twelve patients maintained good knee functions for 9 years post-implantation (IKDC score = 69.8 ± 12.3) at levels higher than the pre-implantation values (p-value = 0.0018). Patients were evaluated with MRI and arthroscopy, and the defective sites exhibited a smooth surface without a gap between the implant and host tissue. This study demonstrates that autologous CPs successfully engraft into the host tissue and result in the re-formation of hyaline-like cartilage, thereby improving the impaired knee functions. Most importantly, no adverse event was reported during this long-term follow-up period.

Medial joint space width and convergence angle change with time after medial opening-wedge high tibial osteotomy.

BACKGROUND: Although the medial joint space width (MJSW) is commonly used for radiographic evaluation of knee osteoarthritis, the changes in knee joint space width (JSW) during weight bearing after medial opening-wedge high tibial osteotomy (MOWHTO) remain unclear. This study aimed to depict how medial and lateral JSWs and convergence angles change gradually after MOWHTO. METHODS: We retrospectively followed up 81 MOWHTO cases for over 45 months on average. Pre- and postoperative mechanical axes were recorded. The JSWs and convergence angles were measured preoperatively, immediately postoperatively, and 3-6, 9-12, and 21-24 months postoperatively. Patient-reported outcomes were measured using a visual analogue scale (VAS). RESULTS: The mean mechanical femoral-tibial angle improved from 8.1° varus to 2.4° valgus. At the aforementioned times, the respective mean values of MJSW were 2.6, 3.5, 3.8, 4.0, and 4.2 mm; mean convergence angles were 4.8°, 2.9°, 2.2°, 2.1°, and 1.9°; and the mean VAS scores were 7.2, 7.8, 4.8, 1.4, and 1.3. The MJSW continued to increase significantly in the first year postoperatively and then plateaued for a minimum of 2 years follow up after MOWHTO. The convergence angle decreased significantly in the first 6 months postoperatively and was then maintained. CONCLUSIONS: The MJSW, convergence angle, and VAS scores continued to improve through weight bearing during the first year after MOWHTO and were maintained for at least 2 years. Thus, JSW measurement may be an easy and representative way of radiographically monitoring the effect of MOWHTO.

Label-free platform on pH-responsive chitosan: Adhesive heterogeneity for cancer stem-like cell isolation from A549 cells via integrin ß4.

Great efforts have been paid to develop methodologies for cancer stem-like cell (CSLC) isolation in anti-cancer research. The major obstacle lies in the lack of generic biomarkers for different cancer types and the requirement of complicated immuno-labeling procedures. The purpose of this study is to establish a label-free platform for CSLC isolation using pH-responsive chitosan. Based on the adhesive heterogeneity, 15.7 ± 1.9 % of human non-small cell lung cancer (NSCLC) cell line A549 detached from the chitosan substrate following medium pH elevation from 6.99 to 7.65 within 1 h. As a result, this subpopulation of cells with low adhesiveness exhibited superior CSLC hallmarks, including self-renewal, invasive and metastatic potential, therapeutic-resistance, colony formation in vitro, as well as nude mice xenograft in vivo for tumorigenesis, in comparison with their high-adhesive counterpart. Furthermore, integrin ß4 is decisive in controlling CSLC detachment of NSCLC. Conclusively, this pH-dependent isolation provides new insights into biomaterial-based CSLC isolation.

Whole body balance control in Lenke 1 thoracic adolescent idiopathic scoliosis during level walking.

INTRODUCTION: Altered trunk shape and body alignment in Lenke 1 thoracic adolescent idiopathic scoliosis (AIS) may affect the body's balance control during activities. The current study aimed to identify the effects of Lenke 1 thoracic AIS on the balance control during level walking in terms of the inclination angles (IA) of the center of mass (COM) relative to the center of pressure (COP), the rate of change of IA (RCIA), and the jerk index of IA. The association between the Cobb angle, IA and RCIA was also evaluated. MATERIALS AND METHODS: Sixteen adolescents with AIS (age: 14.0±1.8 years, height: 154.8±4.7 cm, mass: 42.0±7.5 kg) and sixteen healthy controls (age: 14.4±2.0 years, height: 158.4±6.2 cm, body mass: 48.6±8.9 kg) performed level walking in a gait laboratory. The kinematic and ground reaction force data were measured for both concave-side and convex-side limb cycles, and used to calculate the IA and RCIA, the jerk index of IA, and the temporal-spatial parameters. Correlations between the Cobb angle, IA and RCIA were quantified using Pearson's correlation coefficients (r). RESULTS: The patients showed less smooth COM-COP motion with increased jerk index of IA in the sagittal plane during single limb support (SLS) of the concave-limb (p = 0.05) and in the frontal plane during double limb support (DLS) (p < 0.05). The patients also showed significantly increased posterior RCIA on both the concave and convex side during initial (p = 0.04, p = 0.03) and terminal (p = 0.04, p = 0.03) DLS when compared to healthy controls. In the frontal plane, the patients walking on the concave-side limb showed decreased IA over SLS (p = 0.01), and at contralateral toe-off (p<0.01) and contralateral heel-strike (p = 0.02), but increased mean IA magnitude over terminal DLS (p = 0.01). The frontal IA at contralateral toe-off and SLS for AIS-A showed a moderate to strong correlation with Cobb angles (r = -0.46 and -0.61), and the sagittal RCIA over the initial DLS for AIS-A also showed a significant, strong correlation with Cobb angles (r = -0.50). CONCLUSIONS: The patients with Lenke 1 thoracic scoliosis in the current study showed altered and jerkier COM-COP control during level walking when compared to healthy controls. During DLS, the patients increased the posterior RCIA in the sagittal plane with increased IA jerk index in the frontal plane for both the concave- and the convex-side limb, indicating their difficulty in maintaining a smooth transfer of the body weight. During SLS of the concave-side limb, the patients adopted a conservative COM-COP control strategy, as indicated by a decreased IA in the frontal plane, but showed a jerky COM-COP control in the sagittal plane. The COM-COP control of the patients was associated with the severity of the spinal deformity. The current results suggest that this patient group should be monitored for signs of an increased risk of loss of balance during weight transfer on the concave-side limb.

Altered balance control in thoracic adolescent idiopathic scoliosis during obstructed gait.

Adolescent idiopathic scoliosis (AIS) is the most common spinal deformity during adolescence, leading to altered postural control with compromised stability. To identify the effects of AIS on whole-body balance control during obstacle-crossing, 14 adolescents with Lenke 1 thoracic AIS and 14 healthy controls were compared in terms of the inclination angle (IA) of the body's center of mass (COM) relative to the center of pressure (COP), the rate of change of IA (RCIA) and the jerk index of IA. Between-side comparisons were also performed for the AIS group. The patients showed less smooth COM-COP motion in the sagittal plane with significantly increased anterior RCIA and IA jerk index during crossing with either the concave side (p = 0.001) or the convex side (p = 0.001) leading when compared to healthy controls. In the frontal plane, the patients showed close-to-zero RCIA (p = 0.002) while crossing with the leading limb, with an increased IA magnitude (p = 0.039) only while crossing with the concave-side limb leading. The patients with Lenke 1 thoracic AIS were found to cross obstacles with altered, compromised COM-COP control in both sagittal and frontal planes when compared to healthy controls. The results suggest that the thoracic spinal deformity in Lenke 1 AIS affects the whole-body balance control during obstacle-crossing, which should be monitored for signs of increased risk of loss of balance in the management of such patient groups.

PCL-Blended Chitosan Substrates for Patterning the Heterotypic Cell Distribution in an Epithelial and Mesenchymal Coculture System.

Cell-cell and cell-substrate interactions in coculture systems are very important to the context of biomaterial scaffolds for tissue engineering applications. Understanding the cellular interactions and distribution of epithelial-mesenchymal microtissues on the controllable biomaterial surfaces is useful to study the organoid applications. The aim of the present study is to investigate the effects of chitosan/poly(ε-caprolactone) (PCL)-blended biomaterials on the distribution and spheroid formation of HaCaT and Hs68 cells in a coculture system. In this study, we demonstrated that the cocultured cells gradually changed their pattern from core/shell spheroid to monolayered morphology as the PCL content increased in the blended substrates. This indicates that the chitosan/PCL-blended substrates are able to regulate cell-substrate and cell-cell interactions to modify the distribution of HaCaT and Hs68 cells similar to various mesenchymal-epithelial organizations in biological tissues. Moreover, we also developed a two-dimension lattice model to elaborate the dependence of cell spheroid development on complex cell-cell interactions. This information may be helpful to develop appropriate biomaterials with appropriate properties to the applications of engineered epithelial-mesenchymal organoids.

A self-assembled layer-by-layer surface modification to fabricate the neuron-rich model from neural stem/precursor cells.

BACKGROUND/PURPOSE: In vitro neural cell-based models have been widely used to mimic the in vivo neural tissue environments and quantitatively understand the effects of pharmaceutical molecules on neural diseases. Recently, several biomimetic neural tissue models have been widely developed by using biomaterials or surface modification. However, the complex protocols of material synthesis or surface modification lack an easy execution to fabricate the neuron favorite environment. METHODS: In this study, we utilized a layer-by-layer technique as a surface modification method for regulating the behaviors of neural stem/precursor cells (NSPCs) on material surfaces. Polyelectrolyte multilayers (PEMs) via alternate deposition of poly (allylamine hydrochloride) (PAH) and poly (sodium-4-styrenesulfonate) (PSS) were used to culture NSPCs. After incubation for 7 days, the neuronal differentiation of NSPCs and synapse function of differentiated neurons were identified by immunocytochemistry for lineage specific markers. RESULTS: Compared with the only PAH film, the PSS-ending film (neuron-rich model) was shown to significantly promote differentiation of NSPCs into neurons (more than 50%), form a neuronal network structure; and differentiated neurons exhibiting functional synaptic activity. CONCLUSION: This study shows that the PEMs provided an easily alternative approach to modify the surface properties; and might be a method to obtain a neuron-rich model for the biological/pharmaceutical applications.

The correlation of sagittal osteotomy inclination and the anteroposterior translation in medial open-wedge high tibial osteotomy-one of the causes affecting the patellofemoral joint?

PURPOSE: For opening-wedge high tibial osteotomy, previous studies have shown that most osteotomies were anterior-inclined. The purpose of this study was to determine the effect of sagittal osteotomy inclination on the anteroposterior translation of osteomized fragments and discuss its possible impact on the patellofemoral joint. METHODS: We retrospectively measured the angle between the joint line and the sagittal osteotomy line. We also evaluated the anteroposterior translation of osteomized fragments by measuring the distance from the most posterior point of the tibial plateau to the tibial tuberosity and the anterior cortical line. Correlation between the sagittal osteotomy inclination and the anteroposterior translation of fragments was analyzed. RESULTS: The mean sagittal osteotomy inclination was 6.3 ± 8.4° anteriorly to the joint line and 82% of osteotomies were anterior-inclined. The anteroposterior translation of the osteomized fragments was moderately correlated to the sagittal inclination. Anterior-inclined osteotomy tends to result in the anterior translation of the proximal fragment. CONCLUSION: High rates of anterior-inclined osteotomy have been described previously as well as in this study. Anterior-inclined osteotomy tends to result in the anterior translation of the proximal fragment. This may result in increased vertical vector force onto the patellofemoral joint, which further accelerates patellofemoral joint degeneration. Therefore, surgeons should attempt to perform parallel osteotomy or avoid anterior displacement of the proximal fragment if there is concern of anterior-inclined osteotomy.

Core/shell multicellular spheroids on chitosan as in vitro 3D coculture tumor models.

An ideal in vitro drug screening model is important for the drug development. In addition to monoculture systems, 3 dimensional (3D) coculture systems are extensively used to simulate the in vivo tumor microenvironment as cell-cell and cell-extracellular matrix interactions within the tumor tissues can be mimicked. In this study, in vitro 3D suspension coculture multicellular spheroids with core/shell cell distribution were developed on chitosan-coated surfaces. Based on the characteristic of chitosan inhibiting cell adhesion, SW620 (colon cancer cell line), 3A6 (mesenchymal stem-like cell line) and Hs68 (foreskin fibroblast line) cells could aggregate to form 3D coculture spheroids with intimate cell contacts. When cells were cocultured on chitosan, 3A6 and Hs68 cells always located in the core of spheroids and were completely enveloped by SW620 cells due to their N-cadherin protein expression following the differential adhesion hypothesis. The core cells could be the feeder layers to stimulate the shell SW620 cells to enhance their mitochondria activity. Moreover, 3D coculture core/shell multicellular spheroids could enhance the resistance of SW620 cells against the cytotoxicity effect of chemotherapy drugs. To sum up, based on the specificity of the core/shell coculture multicellular spheroids, a novel in vitro tumor model was proposed in this study.

Poly(N-(4-aminobutyl)-acrylamide) as mimetic polylysine for improving survival and differentiation of cerebellar granule neurons.

Poly(N-(4-aminobutyl)-acrylamide) (P4Am) and poly-d-lysine (PDL) possess the same butylamine side chain. The main difference is the peptide structure, which is in the main chain of PDL but in the side chain of P4Am. PDL has been extensively used in the preparation of neuronal cultures. We assumed neurons are sensitive enough to distinguish such structure difference so these two cationic polymers were compared at serial coating concentrations for culturing cerebellar granule neurons from 7-day-old Wistar rats in this study. Cellular viability and morphology assay showed no obvious difference for neurons cultured at high coating concentrations (>0.31 µg/mL) of these two polymers. In contrast, the difference in the peptide structure between P4Am and PDL could be distinguished by neurons at low coating concentrations (< 0.16 µg/mL). P4Am at low coating concentration could keep aggregates with three or four thick processes to support a more complete neural network with higher cellular viability than PDL. This suggests the interaction between neurons and the specific peptide structure of P4Am at low coating concentration was able to improve survival and differentiation of cultured cerebellar granule neurons. Although neural cells exhibited different morphologies and activities on high and low P4Am coating concentrations, immunofluorescence imaging confirmed most of cultured cells were functionally mature neurons, stained by neurofilament, synapsin I, and GAP43. The information should be useful for designing new biomaterials for regeneration of damaged circuits following disease or trauma. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1194-1201, 2018.