Objective To study whether the human trabecular bone-derived cells have human mesenchymal stem cell characteristics.Methods 3 mm×3 mm×3 mm trabecular bones of human ilium and femur were obtained,cut into 3 mm×3 mm×3 mm chippings,rinsed with DMEM medium.The cells were purified by density gradient centrifuge and cuhured in vitro.The attached cells were cultured by the method of limiting dilution.The selected clones were induced into osteoblasts,chondrcytes and cardiomyo- cytes.The characteristic phenotypes of the differentiated cells were tested by morpbologic observation,RT- PCR mRNA analysis,special chemical staining and immunocytochemical staining.Results The trabocular bone-derived stromal cells could readily differentiate into osteoblasts,chondrocytes and cardiomyocytes if subjected to some appropriate chemicals (dexamethasone,β-glycerphosphoric,Vitamin C ),growth factor (transforming growth factor-β) or 5-azacytidine.These induced cells expressed relevant characteristic phe- notypes of the differentiated cells.Conclusion The human trabecular bone-derived stromal cells are e- quivalent to the mesenchymal stem cells (MSCs) isolated from bone marrows and have the biological char- acteristics of adult stem cells.These cells can be used in tissue engineering and gene project.
[Objective]To explicate the biological characters of an immortal human bone marrow stromal cell line MSCxj.[Method]The morphology,growth and proliferation of the cells were observed during continuous culture in vitro and compared with that of human bone marrow stromal cells(BMSCs-2).[Result]The MSCxj cells showed fibroblast-like feature with rich in organelle.The population doubling time(PDT) was 40.8h,it was shorter than the 43.82h of MSCxj's PDT.The rate of clone forming was 10.9%,the BMSC-2 was 11.5%.The MSCxj cell grew and proliferated well and were able to form cell clone in vitro.[Conclusion]The MSCxj remains the biological characters of human bone marrow stromal cells.
The classification of lumbar spondylolysis varies, and there is currently no clear consensus on a standardized system. This study examines the morphological characteristics and parameter differences of the L5 vertebra in patients with isolated versus fused spondylolysis using CT measurements. It also proposes a preliminary classification system based on the separation distance at the fracture site and explores its clinical significance. A total of 117 young male patients with L5 spondylolysis related to high-intensity physical activity were enrolled. Patients with a pars interarticularis separation distance ≥ 2 mm were classified into the isolated group (Group A, 66 patients), while those with a separation distance < 1 mm were classified into the fused group (Group B, 51 patients).Additionally, 117 patients without spondylolysis but experiencing lower back pain were included as the control group (group C). Multislice spiral computed tomography (MSCT) was used to measure the morphological parameters of the L5 vertebra in all three groups, including the sagittal pedicle height (SPH), transverse pedicle width (TPW), transverse pedicle vertical length (TPVL), pedicle screw trajectory length (PSTL), pedicle angle of attack (PAA), frontal vertebral body height (FVH), posterior vertebral body height (PVH), sagittal midline intervertebral space height (SMISH), horizontal vertebral body angle (HVA), and vertical vertebral body angle (VVA). Differences in the morphological imaging parameters of the L5 vertebrae and pedicles among the three groups were compared. There were no significant differences in age, height, weight, body mass index (BMI), or Pfirrmann grade among the three groups. No significant differences were observed in any of the pedicle parameters between the left and right sides within the groups. Group A showed significantly greater TPVL and PSTL values compared to Group B, while TPW and PAA were significantly lower. No significant difference in SPH was observed between Group A and Group B. When compared to Group C, Group A exhibited significant differences in SPH, TPW, TPVL, and PSTL, but not in PAA. Group B, compared to Group C, demonstrated significant differences in SPH and PAA, but no significant differences were observed in TPW, TPVL, or PSTL. Significant differences were also found in FVH, HVA, and VVA between Group A and Group B, with Group A showing a smaller PVH. No significant difference in SMISH was observed between the two groups. Compared to Group C, Group A showed significant differences in PVH, HVA, and VVA, but no significant differences were found in FVH or SMISH. In Group B, significant differences were noted in FVH and HVA compared to Group C, but no differences were observed in PVH, SMISH, or VVA. Differences in the sagittal morphological parameters of the pedicles and vertebral bodies can be observed between the two types of spondylolysis patients. In the isolated spondylolysis pattern, the pedicles exhibit a "thin, long, and contracted" morphology, while the vertebral bodies present a "stuffed bun" shape, both anteriorly, posteriorly, and superiorly. In contrast, the fused type is characterized by "short, thick, and expanded" pedicles, with the vertebral bodies showing a "less pronounced stuffed bun" shape in the anterior–posterior direction. These morphological differences indicate that spondylolysis separation may involve adaptive stress-induced bone remodeling. Surgeons must pay special attention when choosing surgical techniques, as isolated spondylolysis may present a tendency toward slippage. Caution is advised in performing isolated pars repair surgeries, especially during the placement of pedicle screws, where special attention must be given to the length and direction of the screws to avoid additional damage.
Abstract An electrochemically enabled tandem cyclization reaction of unsaturated sulfoximines with diselenides has been well developed, affording metal‐ and external oxidant‐free access to sulfoximidoyl‐based heterocycles containing selenium in moderate to good yields. With n Bu 4 NBr as the role of catalyst and electrolyte, this methodology shows broad substrate scope and functional group tolerance under mild conditions.
Various strategies, each with its own set of limitations, are available for managing lumbar spondylolysis. In response, our department has developed an innovative solution: a V-shaped titanium cable integrated with a pedicle screw internal fixation system specifically designed for lumbar spondylolysis in young adults.
To resolve the problem of the insufficient availability of seed cells and to provide seed cells for tissue engineering research, an immortalized human bone marrow stromal stem cell line (MSCxj cells) was established in our department to investigate the ectopic osteogenesis of MSCxj cells.MSCxjs were grown with a heterogeneous bone scaffold for 48 h. Three groups were included: group A: MSCxjs of 35 PDs were maintained with heterogeneous bone; group B: MSCxjs of 128 PDs were maintained with heterogeneous bone; and group C: heterogeneous bone alone. Tetracycline fluorescence staining, H&E staining, and ponceau staining, immunohistochemistry and bone histomorphometry were performed. At the same time, scanning electron microscopy was conducted to detect the growth of MSCxjs and heterogeneous bone.Scanning electron microscopy showed favorable adherence of MSCxjs to heterogeneous bone. A large number of newly generated filamentous extracellular matrix and fine granular materials were found to cover the cells. The results from staining showed that the osteogenesis was not obvious in group A/B 4 weeks after transplantation. Eight weeks after implantation, osteoid matrix deposition was noted in and around the heterogeneous bone in group A/B. Twelve weeks after implantation, osteogenesis was increased in group A/B. There were no significant differences in the time course for bone formation and the amount of newly generated bone between group A/B.Like primary hBMSCs, MSCxj cells have favourable ectopic osteogenesis and can be applied as seeded cells in bone tissue engineering.
Abstract Objective: This study aims to provide theoretical basis for treatment and prevention of osteomyelitis by investigating and analyzing clinical features and pathogen distribution among 282 patients with chronic tibial osteomyelitis. Methods: A total of 282 patients with chronic tibial osteomyelitis from January 1, 2012 to January 1, 2022 in the First Affiliated Hospital of Xinjiang Medical University were retrospectively analyzed. All data were collected from electronic medical record (EMR) system including demographics, etiology, risk factors, osteomyelitis location and clinical classification. Results: Farmers, students, unemployed and retirees accounted for a relatively large proportion of the 282 patients. There were 233 males and 49 females with a gender ratio of 4.75:1. The average age was 40.21±15.68 years and was mainly concentrated in 41-50 years, specifically, the mean age of females was slightly older than that of males. Education level was mostly primary and secondary school education, and illiteracy. Risk factors of chronic tibial osteomyelitis included history of smoking and drinking, history of multiple repeated surgeries, and impaired immunity. Frequent clinical symptoms were in the order of pain, local swelling,pus discharge and skin ulceration. Among all inflammatory markers, proportion of positive results were 30.85%, 59.93% and 53.90% for white blood cell (WBC), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), respectively. Positive rate of pathogenic microorganism culture was low and the three most common bacteria were Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa) and Escherichia coli (E. coli). The most frequent site of infection was middle tibia. According to Cierny-Mader osteomyelitis classification, the most common types were type IIIA, IVA and IIA. Conclusion: Number of visits due to chronic osteomyelitis increased year by year, with young and middle-aged male farmers and low education level as the main groups. Smoking and drinking were two considerable risk factors that should be attached to a great importance. No significant increase was found in inflammatory markers and lower positive rate of pathogenic microorganism culturewas observed. Multi-drug resistant bacteria were common and S. aureus remained the most frequent pathogen. Elevated ESR had certain diagnostic value for osteomyelitis. Type III and type IV osteomyelitis accounted for a large proportion which posed great challenges for clinical diagnosis and treatment.
To provide the seed cells for bone tissue engineering, to establish immortalized human bone marrow mesenchymal stem cells (MSCxj) and to investigate the ectopic osteogenesis of MSCxj.MSCxjs of the 35th and 128th generations were maintained and harvested when the cell density reached 2 x 10(9). Then, these cells were co-cultured with heterogeneous bone scaffold in groups A (the 35th generation, n = 12) and group B (the 128th generation, n = 12); heterogeneous bone alone was used in group C (n = 12). The cell proliferation was observed by scanning electron microscopy (SEM) after 48 hours and 18 days of osteogenic induction culture. The complex was implanted subcutaneously through a 3-mm-incision at both sides of the back in 18 nude mice. Tetracycline labeling was performed before the animals were sacrificed. Tetracycline fluorescence staining, HE staining, ponceau staining, and immunohistochemistry staining for osteocalcin were performed at 4, 8, and 12 weeks after transplantation; the morphologic quantitative analysis was made.After 48 hours, SEM showed that MSCxjs adhered to heterogeneous bone and grew well; after 18 days, a large number of new filamentous extracellular matrix and small granules were found to cover the cells. The results of tetracycline fluorescence staining, HE staining, and ponceau staining in groups A and B showed that the osteogenesis was not obvious at 4 weeks after transplantation; osteoid matrix deposition was noted around and in the heterogeneous bone at 8 weeks; and osteogenesis was increased at 12 weeks. There was no significant difference in bone formation between groups A and B. Osteogenesis was not observed in group C. The osteocalcin expressions were positive in groups A and B. The bone ingrown percentages of groups A and B were 5.64% +/- 2.68% and 4.92% +/- 2.95% at 8 weeks, and 13.94% +/- 2.21% and 14.34% +/- 3.46% at 12 weeks, showing significant differences between 8 weeks and 12 weeks at the same group (P < 0.05) and no significant difference between groups A and B at the same time (P > 0.05).MSCxj has favorable abilities of ectopic osteogenesis and can be applied as seeded cells in bone tissue engineering.
AIM: To build a method to culture bone marrow derived mesenchymal stem cells (MSCs) in vitro and to investigate the feasibility of inducing MSCs into osteoblasts. METHODS: Human MSCs were isolated from adult bone marrow and purified by percoll density gradient centrifugation and cultured in vitro . The MSCs attachment formed after 7-10 d and the MSCs of the passage 3 were chosen to induce into osteoblasts. Fifteen days later, the alkaline phosphatase assay was conducted using modified calcium cobalt staining method, type Ⅰ collagen and osteocalcin assay was conducted using immunohistochemistry and calcium node assay was done using alizarin red staining. RESULTS: Homogeneous MSCs were obtained by percoll density gradient centrifugation. The induced MSCs had typical appearance of osteoblasts. The rate of ALP expression was 81%, the expressions of collagen type Ⅰand osteocalcin were positive, and calcium nodes were seen. CONCLUSION: The MSCs can be separated from adult bone marrow and cultured in vitro , which can be induced into osteoblasts.
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