Insufficiency of collagenases in establishment of primary chondrocyte culture from cartilage of elderly patients receiving total joint replacement.

Background Collagenases are frequently used in chondrocyte isolation from articular cartilage. However, the sufficiency of this enzyme in establishing primary human chondrocyte culture remains unknown. Methods Cartilage slices shaved from femoral head or tibial plateau of patients receiving total joint replacement surgery (16 hips, 8 knees) were subjected to 0.02% collagenase IA digestion for 16 h with (N = 19) or without (N = 5) the pre-treatment of 0.4% pronase E for 1.5 h. Chondrocyte yield and viability were compared between two groups. Chondrocyte phenotype was determined by the expression ratio of collagen type II to I. The morphology of cultured chondrocytes was monitored with a light microscope.Results Cartilage with pronase E pre-treatment yielded significantly higher chondrocytes than that without the pre-treatment (3,399 ± 1,637 cells/mg wet cartilage vs. 1,895 ± 688 cells/mg wet cartilage; P = 0.0067). Cell viability in the former group was also significantly higher than that in the latter (94% ± 2% vs. 86% ± 6%; P = 0.03). When cultured in monolayers, cells from cartilage with pronase E pre-treatment grew in a single plane showing rounded shape while cells from the other group grew in multi-planes and exhibited irregular shape. The mRNA expression ratio of collagen type II to I was 13.2 ± 7.5 in cells isolated from cartilage pre-treated with pronase E, indicating a typical chondrocyte phenotype. Conclusions Collagenase IA was not sufficient in establishing primary human chondrocyte culture. Cartilage must be treated with pronase E prior to collagenase IA application.

Primary culture of chondrocytes after collagenase IA or II treatment of articular cartilage from elderly patients undergoing arthroplasty.

Background: Joint replacement surgery provides articular cartilage samples for chondrocyte isolation. To our knowledge, the effect of the collagenase type on releasing of chondrocytes from the extracellular matrix of cartilage is not reported. Objectives: To determine whether cartilage digested with collagenase IA yielded more chondrocytes than that digested with collagenase II and determine whether chondrocytes isolated with collagenase IA could be cultured in vitro. Methods: Cartilage slices collected from 18 elderly patients who received joint replacement surgery (16 hips, 2 knees) were digested sequentially with 0.4% pronase E and 0.02% collagenase IA, or with 0.15% collagenase II alone, or sequentially with 0.4% pronase E and 0.02% collagenase II. We compared cell yield from each method. Cell viability by the most effective method was calculated and plotted. The morphology of cultured monolayer chondrocytes was recorded with a light microscope. Results: Sequential digestion with pronase E and collagenase IA yielded 2566 ± 873 chondrocytes per mg wet cartilage, which was more effective than the other isolation methods (P = 0.018). The average chondrocyte viability could reach 84% ± 8% (n = 11). Light microscopic images showed typical chondrocyte morphology in monolayer cultures. Conclusion: Sequential digestion of human articular cartilage with pronase E and collagenase IA was more effective than collagenase II alone or collagenase II combined with pronase E for releasing chondrocytes from extracellular matrix of cartilage. Chondrocytes isolated with this method could be maintained in monolayer cultures for at least 2 passages with unaltered morphology.

Vitamin D Status and Patient Outcomes after Knee or Hip Surgery: A Meta-Analysis.

BACKGROUND: This study evaluates the effect of vitamin D status in patient outcomes after hip or knee joint surgery. METHOD: Literature search was carried out in electronic databases, and study selection followed predetermined eligibility criteria. Data were extracted from relevant studies and meta-analyses of standardized mean differences between hypovitaminosis D (vitamin D deficiency or insufficiency) and euvitaminosis D in assessment scores of patient-reported outcomes were performed. RESULTS: A total of 12 studies (2,593 patients; age 69.89 years [95% CI 68.07-71.70]; 35.95% [29.43-42.46] males) were included in the meta-analysis. The prevalence of hypovitaminosis D (vitamin D deficiency or insufficiency) was 33.18% [25.10-41.26], but the combined prevalence of deficiency and insufficiency was 46.99 [34.02-59.96]. Hospital stay was 1.09 days [-0.39 to 2.56] longer in the hypovitaminosis D group compared to the euvitaminosis D group. Preoperatively, Harris Hip Score (HHS) and Knee Society Score were significantly lower (p = 0.001 and p = 0.00001, respectively) in the hypovitaminosis D group than in the euvitaminosis D group. Postoperatively, HHS (p = 0.004) score was significantly lower in the hypovitaminosis D group than in the euvitaminosis D group. CONCLUSION: The prevalence of hypovitaminosis D is high in osteoarthritis patients undergoing knee or hip surgery. Vitamin D deficiency may affect the outcomes of orthopedic joint surgery. However, randomized trial/s will be required to confirm these findings.

Gas-Phase Photoluminescence and Photodissociation of Silver-Capped Hexagold Clusters.

We report on the radiative and nonradiative deactivation pathways of selected charge states of the stoichiometric hexagold phosphine-stabilized ionic clusters, [(C)(AuDppy)6Ag2·(BF4) x](4- x)+ with x = 2 and 3 (Dppy = diphenylphosphino-2-pyridine), combining gas-phase photoluminescence and photodissociation with quantum chemical computations. These clusters possess an identical isostructural core made of a hyper-coordinated carbon at their center octahedrally surrounded by six gold ions, and two silver ions at their apexes. Their luminescence and fragmentation behavior upon photoexcitation was investigated under mass and charge control in an ion trap. The experimental and computational results shed light on the electronic states involved in the optical transitions as well as on their core, ligand, or charge transfer character. Gas-phase results are discussed in relation with condensed phase measurements, as well as previous observations in solution and on metal-organic frameworks. The monocationic species ( x = 3) is found to be less stable than the dicationic one ( x = 2). In the luminescence spectrum of the monocationic species, a shoulder at short wavelength can be observed and is assigned to fragment emission. This fragment formation appears to be favored for the monocation by the existence of a low lying singlet state energetically overlapping with the triplet state manifold, which is populated quickly after photoexcitation.

[Percutaneous vertebroplasty and open vertebroplasty for metastatic spinal tumor].

OBJECTIVE: To explore the clinical application and therapeutic effect of percutaneous vertebroplasty(PVP) and open vertebroplasty for metastatic spinal tumor. METHODS: The clinical data of 126 patients with metastatic spinal tumor underwent surgery and obtained follow-up from January 2012 to March 2016 were retrospectively analyzed. These 126 cases were divided into two groups according to different operative methods. The metastatic tumor of 43 cases encroached vertebral canal oppressing spinal cord and nerve root, they were treated with open operation(open vertebroplasty group);and other 83 cases without obviously spinal cord or nerve root compression, or unfit for open operation, were treated with PVP (percutaneous vertebroplasty group) . VAS score, ECOG and Frankel grade were used to evaluate the pain and neurofunction in two groups.All out-hospital patients were followed up every 3 months for 1 time. X-ray, CT, MRI were examined in follow-up. RESULTS: A total of 112 vertebrae underwent PVP with the median surgical time of 50 min;VAS scores decreased significantly at 2 days after operation, which maintained till 1 month later; ECOG grade at 1 month decreased significantly;44 of 112 vertebrae suffered from asymptomatic bone cement leakage, no complications such as nerve injury or pulmonary embolism was found; the median survival time was 16 months. While for open vertebroplasty group, the median surgical time was 160 min and blood loss was 1 000 ml; postoperative VAS scores and ECOG grade at 1 month decreased significantly. Postoperative Frankel grade of 36 patients got improvement in 41 patients with spinal cord functional disturbance(87.8%); and 29 of 40 patients with incompleteness out of motor function were full recovery(76.3%); 12 cases (27.9%) occurred complications and the median survival time was 11 months. CONCLUSIONS: The different vertebroplasty treatments can be selected for patients with metastatic spinal tumor, which can relieve the pain, improve the nerve function, reconstruct the spinal stabilization, maintain the local control and raise the life quality.

Clinical and radiographic results of Bryan cervical total disc replacement: 4-year outcomes in a prospective study.

INTRODUCTION: Early results have indicated that the Bryan cervical total disc replacement (TDR) favorably compares to anterior cervical decompression and fusion, while it is associated with fewer complications and higher levels of satisfaction. In this study, we sought to prospectively report the midterm outcomes of the Bryan TDR. PATIENTS AND METHODS: A total of 20 patients had performed their 4-year follow-up visit and had been assessed clinically and radiologically. Clinical outcomes (JOA, VAS, NDI, SF-36) and ROM measurements were investigated preoperatively and at 1 and 6 months, and 1, 2 and 4 years after operation. Complications were also investigated. Occurrences of heterotopic ossifications (HOs) and adjacent-level degeneration (ALD) radiographic changes were detected from 4-year follow-up X-rays. RESULTS: The mean JOA score, VAS score for arm and neck, NDI score and SF-36 score for PCS and MCS were reduced significantly at each postoperative time point when compared with the preoperative condition. The range of movement of the cervical spine, functional spinal unit, treated segment and the adjacent segment temporarily decreased at the early assessment, but all recovered to preoperative levels over a 6-month to 4-year time period. HO was evident in 6 of the 23 operated segments, which did not restrict the movement of the prosthesis. No obvious ALD was found on MRI. There were no cases of prosthesis migration, subsidence, loosening or wear. CONCLUSION: The midterm outcomes demonstrated that the Bryan TDR maintains favorable clinical and radiological results, with preservation of movement and satisfactory clinical outcome. There were no serious complications or cases of prosthetic wear or failure. The long-term benefits are yet to be examined.

A phosphorescent silver(I)-gold (I) cluster complex that specifically lights up the nucleolus of living cells with FLIM imaging.

The phosphorescent silver(I)-gold(I) cluster complex [CAu6Ag2(dppy)6](BF4)4 (N1) selectively stains the nucleolus, with a much lower uptake in the nucleus and cytoplasm, and exhibits excellent photostability. This Ag-Au cluster, which has a photoluminescent lifetime of microseconds, is particularly attractive as a probe in applications of time-gated microscopy. Investigation of the pathway of cellular entry indicated that N1 permeates the outer membrane and nuclear membrane of living cells through an energy-dependent and non-endocytic route within 10 min. High concentrations of N1 in the nucleolus have been quantified by inductively coupled plasma atomic emission spectroscopy (ICP-AES) and transmission electron microscopy coupled with an energy dispersive X-ray analysis (TEM-EDXA), which also helped to elucidate the mechanism of the specific staining. Intracellular selective staining may be correlated with the microenvironment of the nucleolus, which is consistent with experiments conducted at different phases of the cell cycle. These results prove that N1 is a very attractive phosphorescent staining reagent for visualizing the nucleolus of living cells.

Clinical and radiographic results after treatment of cervical degenerative disc disease with the bryan disc prosthesis: a prospective study with 2-year follow-up.

The purpose of this study was to prospectively evaluate the clinical and radiographic results obtained with the Bryan disc prosthesis (Medtronic Sofamor Danek, Memphis, TN) in 20 consecutive patients with single-level or two-level cervical degenerative disc disease. All patients were followed up for at least two years. The mean Neck Disability Index and the visual analogue scale for neck and arm pain scores were significantly reduced for all postoperative periods compared with the mean preoperative values. SF-36 outcome measures, including the physical and mental component scores, significantly improved after surgery and remained unchanged or improved at two years postoperatively. According to Odom's scale, 18 patients (90%) had good to excellent outcomes. The range of movement of the cervical spine, of the functional spinal unit, the treated segment and the adjacent segment temporarily decreased at the early assessment, but all recovered to preoperative levels over a 6-24 months time period. There was no heterotopic ossification at the implanted levels, no adjacent level degeneration and no prosthesis subsidence or excursion. Cervical disc replacement using the Bryan disc prosthesis appeared to be safe and demonstrated encouraging clinical and radiological outcomes. However, further studies are required to assess the long-term efficacy of this prosthesis and its protective influence on adjacent levels.

[Analysis of surgical treatment and effect of spinal metastatic tumors].

OBJECTIVE: To investigate clinical outcome of surgical treatment for spinal metastatic tumors. METHODS: Thirty-six patients with metastatic spine tumors treated surgically were retrospectivly reviewed from September 2005 to August 2010. There were 15 males and 21 females with an average age of 58 years old (ranged, 27 to 79 years). The site of origin of primary cancer included the breast, prostate, colon, lung, liver, esophagus, kidney, carcinoma, bladder, and 10 patients were with unidentified primary cancer. Five lesions were located in the cervical spine, 17 in the thoracic spine, 13 in the lumbar spine and 1 in the sacral vertebrae. Preoperative evaluation was conducted according to Tokuhashi system: total score 0 to 8 in 9 cases, 9 to 11 in 25 cases, 12 to 15 in 2 cases. The surgical procedures, including PVP, tumor resection and decompression, titanium nets implant, artificial vertebra replacement, bone cement stuffiness and inter fixation, were performed based on Tokuhashi score, location of lesions and neurological symptoms. RESULTS: All the patients were followed up, and the during ranged from 2 months to 60 months with a mean time of 10.8 months. Pain relief was obtained in all patients after operation, and the VAS pain scores declined after operation. Among 14 patients suffering from spinal cord and nerve compromise, 12 patients improved 1 to 2 grades after surgery according to the Frankel grading system. According to the system of the ECOG performance status: 28 patients improved 1 to 2 grades in performance status after surgery. The postoperative survival rates at 3 months, 6 months, 1 year and 2 years were 97.2%, 63.9%, 38.9% and 16.7% respectively. Six patines had bone cement leakage after PVP, and none resulted in severe complications. CONCLUSION: According to single or multiple spinal metastases, neurological symptoms, spinal stability and patients' condition, the different surgical treatments can be selected for spinal metastatic patients, which can relieve pain, maintain or improve the neurological status, improve quality of life, improve survival rate of patients.