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BACKGROUND:Studies have demonstrated that the success rate of isolation of umbilical cord blood mesenchymal stem cells (UCB-MSCs)is low,which also lacks of unified identification method.OBJECTIVE:To modify the traditional in vitro isolation and culture method of UC8-MSCs to obtain a higher success rate and to observe its biological characteristics.DESIGN,TIME AND SETTING:In vitro observation of cytology.The study was performed at the Ninth People's Hospital of Shanghai Jiao Tong University Medical College between April 2006 and January 2007.MATERIALS:A totaI of 28 UCB samples were obtained from full-term normal delivery.Department of Matemity,Shanghai Red House Hospital.The written informed consent was obtained from the puerpera and their families.METHODS:NeonataI umbilical cord blood was collected under sterile condition.Mononuclear cells (MNCs) were separated from using lymphocyte isolation medium by centrifugation and suspended in α-minimum essential medium(α-MEM)containing 10% fetal bovine serum.The medium was half exchanged after 5-7 days of primary culture and then was totally exchanged every 3-4 days.The confluent cells were divided into 2 groups.In group 1.when the round megakaryocytes were confluent and fusiform fibroblast-like cells were fell off.the cell suspension was transferred to new culture dish;in group 2.when the round megakaryocytes dominated the majodty,the culture medium was replaced by α-MEM containing 15% calf serum,followed by culture in α-MEM containing 10% fetal bovine serum when the round megakaryocytes fell off.The fifth passage of UCB-MSCs was harvested for subsequent osteoinduction in vitro.MAIN OUTCOME MEASURES:The cell morphology was observed by microscopy;Flow cytometry was used to examine the surface antigen phenotype;alkaline phosphatase and oil red staining was performed to detect cell differentiation capacity.RESULTS:Of 28 samples of UCB,attaching cells were obtained from 20 samples(6/10 in group 1,14/18 in group 2),fibroblast-like cells that could passage steadily(4 samples in group 1,9 in group 2)were cultured from 13 0f 20 samples with success rate of 46.4%,among which MSCs were passaged steadily up to P22.UCB-MSCs were all positive for MSC-related antigens such as CD29 and CD105,but negative for CD34,CD45 and CD106.Incubation of UCB-MSCs under special condition resulted in a differentiation of osteoblast and adipocyte.CONCLUSIONMSCs exist in UCB,which have multi-differentiation capacity,and passage steadily.The modified in vitro culture method improves culture success rale of UCB-MSCs.
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BACKGROUND: Bone and exact reduction and internal fixation should be kept to fully repair the function of patella in comminuted patellar fracture operation. Currently, comminuted patellar fracture without a unified therapeutic standard has been a difficulty in clinic.OBJECTIVE: To compare the effects of modified AO tension band, titanium cable cerclage and nickel-titanium (Ni-Ti) patellar concentrator fixation in the treatment of comminuted patellar fracture.DESIGN: Prospective comparative observation.PARTICIPANTS: Totally 60 patients with simple unilateral closed comminuted patellar fracture were enrolled at Department of Orthopaedics of Changzheng Hospital from November 2004 to November 2006, including 32 males and 28 females aged 28-59 years. Inclusive criteria contained diagnosis of having simple unilateral closed comminuted patellar fracture and injury timing of less than 7 days. The patients all signed the informed consent.METHODS: The patients were assigned into modified AO tension band group (n =22), titanium cable cerclage group (n = 21) and Ni-Ti patellar concentrator group (n =17) according to their will. Twelve patients in the modified AO tension band group received simple modified AO tension band internal fixation, and ten received modified AO tension band internal fixation and extraction. Patients in the titanium cable cerclage group received titanium cable cerclage. Ten patients in the Ni-Ti patellar concentrator group received simple Ni-Ti patellar concentrator internal fixation, and seven received internal fixation and extraction.MAIN OUTCOME MEASURES: ①Postoperative host and material reaction, intraoperative blood loss, medical cost and workday loss, and ②active range of knee motion at month 3 after operation and functional recovery and curative effect at month 6 after operation.RESULTS: Totally 60 patients were involved in the result analysis. ①Host and material reaction was not found in all patients. Blood loss was significantly more in the patients of simple Ni-Ti patellar concentrator internal fixation group than patients of simple modified AO tension band internal fixation group and titanium cable cerclage group, whereas workday loss was less than that of the simple modified AO tension band internal fixation group (P < 0.05). Medical cost was obviously more in the titanium cable cerclage group than the simple Ni-Ti patellar concentrator internal fixation group and the simple modified AO tension band internal fixation group (P < 0.05). ②Active range of knee motion was the largest, about mean 108°, and long-term functional score of injured knee was the highest, about mean 21.3 points, with the excellent and good rate of 95.2%, and no sever complication appeared in the titanium cable cerclage group. These were much better than those of the other groups (P < 0.05).CONCLUSION: Modified AO tension band fixation and patellar concentrator fixation in the treatment of comminuted patellar fracture have excellent effects in some patients. Titanium cable cerclage fits for the treatment of comminuted patellar fracture and has better recovery of knee joint function.
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BACKGROUND:It has been recently indicated that nervous factors are able to adjust and dominate bone fracture healing. Type Ⅰ collagen is a major factor to promote the differentiation of osteoblasts and enhance the adhesion of osteoblasts; while, it is also a matrix protein for composing bone framework. Type Ⅱ collagen is derived from chondrocytes. OBJECTIVE: To study changing law of type Ⅰ and Ⅱ collagen expression during denervated bone fracture healing. DESIGN, TIME AND SETTING: Randomized controlled anima study was performed at the Animal Laboratory and Cell Biology Laboratory, the Second Military Medical University of Chinese PLA between May and December 2005. MATERIALS: Forty 3-month-year healthy male SD rats were randomly divided into fracture group (tibial fracture alone) and combination group (spinal cord injury combined with tibial fracture), with 20 rats at each group. METHODS: A φ 0.8 mm Kirschner wire was inserted into anterior border of left tibial plateau to establish tibial fracture models in the fracture group. A 0.3-cm spinal cord transection was cut at T10 segment to establish tibial fracture models with entire spinal cord injury. MAIN OUTCOME MEASRUES: Type Ⅰ and Ⅱ collagen protein expressions of callus were detected using Western blot technique in week 1, 2, 4, and 5 post-injury. RESULTS: One week after injury, type Ⅰ and Ⅱ collagen was represented in callus in the two groups, while the expressions in the combination group were significantly higher than fracture group (P<0.05); two weeks after injury, type Ⅱ collagen expression reached at the peak in the combination group, and the expression was significantly higher than the fracture group (P<0.05); four weeks after injury, type Ⅰ collagen expression reached at the peak in the fracture group, and the expression was significantly higher than the combination group (P<0.05), while type Ⅱ collagen still highly expressed in the combination group; five weeks after injury, type Ⅰ and Ⅱ collagen expressions were decreased in the two groups. CONCLUSION: Secretory law of type Ⅰ and Ⅱ collagen during denervated bone fracture healing is similar to normal bone fracture healing; however, the differences at time points, in particular expression at peak, are remarkable between them.
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Objective To observe the changes of the peripheral blood T lymphocyte subsets following acute spinal cord injury and investigate the possible mechanism of these changes. Methods The SCI models of rats were made by Allen's method. Forty SD rats were divided into four groups, ie,normal control group, sham operation group, 100 g·cm group and 200 g·cm group. The expressions of CD4 and CD8 subsets of the peripheral blood T lymphocyte of the injured rats were determined by immunofluorescence labelling and flow cytometry at different times after injury. Results It was found that the expression of CD4 was significantly reduced to (30.40±4.76)% in 100 g·cm group and to (26.54± 9.34) % in 200 g·cm group, which were significantly lower than that of normal control group ( P ＜0.01 ). At 36 hours after injury, the ratio of CD4/CD8 was significantly reduced to 1.81 ± 0.55 in 100 g·cm group and and 1.29 ± 0.50 in 200 g·cm group, with statistical difference (P ＜ 0.05).Conclusions The immunoreaction is significantly depressed at the early stage of acute spinal cord injury. The severer injury results in more significant decrease of CD4 and ratio of CD4/CD8. The changes of CD4 and CD4/CD8 ratio can be used to indicate the severity of spinal cord injury.
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OBJECTIVE: To discuss the possibility and prospect of umbilical cord blood mesenchymal stem cells as seed cells of bone tissue engineering. DATA SOURCES: References came from Medline and Embase between January 1980 and December 2005 with the keywords of "umbilical cord blood,mesenchymal stem cell,bone tissue engineering" in English. Besides, CBM and CBMdisc databases were retrieved by computer for the articles of umbilical cord blood mesenchymal stem cells in bone tissue engineering published between January 1990 and April 2004 with the same key words in Chinese. STUDY SELECTION: Related articles of culture and identification of umbilical cord blood mesenchymal stem cells and the progress of bone tissue engineering. Inclusive criteria: ①basic research of culture and identification of umbilical cord blood mesenchymal stem cells, and ②the progress of bone tissue engineering. Exclusive criteria: reduplicative studies. DATA EXTRACTION: There were 47 relevant articles of umbilical cord blood mesenchymal stem cells and bone tissue engineering, and 30 were accorded with the inclusive criteria. The excluded 17 were due to the reduplicative contents. DATA SYNTHESIS: Under special conditioned medium, umbilical cord blood mesenchymal stem cells not only could differentiate into osteogenic, chondrogenic and adipogenic cells, but also transform into nerve cells, liver cells, somatic muscle cells and so on. Umbilical cord blood mesenchymal stem cells were safe and easy to obtain from umbilical cord blood, safe to transplant, easy to expand in vitro, stable in biological characteristics, which met the requirements of bone tissue engineering. CONCLUSION: Umbilical cord blood mesenchymal stem cells are a promising alternative of ideal seed cells of future bone tissue engineering on the basis of its characteristics.
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[Objective] To explore the effect of low intensity pulsed ultrasound(LIPUS)on the ability of repairing rabbit radius defect by cell-scaffold complex composed with bone marrow stromal cells(BMCs)and ?-tricalcium phosphate(?-TCP).[Method]Bone marrow of a rabbit was drawn and cultured by differentiation culture medium to harvest BMCs and the third generation BMCs were cultivated on ?-TCP for one week in vitro.Bilateral distal radial defects were made using a 1-mm saw in 20 mature New Zealand white rabbits.The cell-scaffold complexs stimulated by LIPUS were implanted in rabbit radial defects(cell-scaffold complexs without stimulation as control).All the rabbits were killed at 4 or 8 weeks after operation.The defect healing state was observed with X-ray and histological technic.The gray density of CA was analyzed by computer image analysis system.[Result]It was found that the callus in the experimental group grew more quickly and apparently than that in the control group.At 4 weeks after operation,there was significantly statistical difference(P0.05).Histological slices in the experimental group showed that the absorption,organization of hematoma,the formation of trabeculae and matrix preceded those in the control group.There were more fibroblast,osteoblasts and callus in the experimental group.[Conclusion]Cell-scaffold complex composed with BMCs and ?-TCP stimulated by LIPUS can repair bone defect more effectively than that without LIPUS stimulation at the early stage of healing,but the effect tend to decrease at the late stage.
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[Objective]To evaluate the outcome of tibiotalocalcaneal fusion in traumatic arthritis patients using the locked retrograde intramedullary nail with graft.[Method]Eighteen ankles underwent this procedure.There were 8 males and 10 females,9 with valgus and 2 with talipes equinovarus,all unilateral.The mean age of patients was 57.5 years(ranged,42~69) and the mean time between the procedure from primary trauma was 7.6 years(5~13 years).Bone graft were picked from distal fibula.The nail was inserted through a plantar approach with locking screws placed both proximally and distally.Patients were evaluated by a standardized follow-up examination using the AOFAS and the fusion outcome were observed radiographically.[Result]All patients were available for an average duration of follow-up of 13.5 months(ranged,6~22 months).All wound healed normally.The mean time of full bearing from operation was 13.6 weeks(ranged,9-16 weeks).One patient was found cerebral thrombosis after operation.One patient had moderate pain while walking on even surface,2 mild to moderate pain on uneven surface,1mild pain while standing and 14 relieved from pain.The average AOFAS score improved from 43.5(ranged,40~52) to 69(ranged,56~84) points.Solid fusion was achieved in 13 ankles(72%) after a mean follow-up of 16.8 months(ranged,12~24 months).Partial union was observed in 3 patients(17%) and non-union in 2 patients(11%).[Conclusion]Locked retrograde intramedullary nail fusion with bone graft is effective for traumatic tibiotalocalcaneal arthritis.
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[Objective]To compare the result of treatment with double bands of Atlas Ti-cable and clavicular hook plate for total acromioclavicular dislocation.[Method]Fifty-two total acromioclavicular dislocation were treated with two different fixation.1)by double bands of Atlas cable(DC group,28 cases) and 2)clavicular hook plate(HP group,24 cases).There were 38 males and 14 females,39 acute and 13 chronic,all unilateral.The mean age of patients was 38.5 years(22~65).A sling of double cable was fixed through the base of the coracoid process and the clavicle at 1.5 cm upper medially above coracoid process.All hook plate were 6 holes.Coracoclavicular ligament was reconstructed routinely.[Result]All patients were available for an average duration of follow-up of 18.8 months(range,6~42 months).The results were evaluated by radiographic measurement and clinical recovery(muscle strength in upper arms,pain in shoulder,range of joint motion) by Lazcano standard.In HP group,14 patients were assessed as good,6 as fair and 4 as poor(good and fair 83%).In DC group,22 patients were assessed as good,5 as fair and 1 as poor(good and fair 96%).There was no statistical difference between the two groups.[Conclusion]In the treatment of acromioclavicular dislocation,fixation of double bands of Atlas cable is believed to be as good as hook plate fixation.Meanwhile,Atlas cable is featured with less pain followed operation,which in favor of early exercise.For it's good biocompatibility,titanium cable can be retained in body and does not need to be removed.
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[Objective]To discuss the factors affecting the healing of trochanteric fracture in the eldly and the way to avoid the medical misplay which may occur.[Method]1.The outcome of 100 trochanteric fracture in the eldly were reviewed in order to analyze the flaw in their treatment and the reasons for non-union.Case history before injuries were inspected and active measures were taken.Some patients with severe condition in cardiovascular or respiratory system were treated by special department.2.Early and concrete surgical intervention were taken.3.Early exercise were discouraged to avoid fracture diseases(eg.Osteoporosis,articular degeneration,muscular flappy).[Result]Totally 100 patients were followed up for 1～5 years(average,3.8 years).Two patients(2%) died and 78(78%) regained normal living activities,12(12%) partially regained normal activities and 6(6%) lost their ability to normal life.Totally 98 patients receiving complete follow up,68 healed within 8～10 months,22 patients' healing time were longer than 10 months(included 3 mal-union with chronic infection).Internal fixation complication occurred in 12 patients(8 looses,4 ruptures).[Conclusion]For trochanteric fracture in the eldly people,early operation in the context of effective control over the systemic disorders could decrease the incidence of post-traumatic complication.Active exercise after operation could effectively improve the fracture healing and patient's life quality.
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Objective To explore rational operative methods to treat hyperextension injuries of cervical spine. Methods 47 cases of hyperextension injury of cervical spine were treated in our department from September, 2000 to October, 2004. They were followed up and analyzed retrospectively. According to the operative methods, there were 3 groups: anterior approach, posterior approach, combined approach. Their therapeutic effects were compared respectively. Results After follow- ups for 6 to 12 months, preoperative and postoperative Frankel classifications and ASIA (American Spinal Injury Association) grades were compared between the 3 groups. It was found that neural function improved significantly in the 3 groups after operation. The anterior approach and combined approach groups had significantly better outcome in neural function than the posterior approach group ( P0.05) . Conclusions Anterior decompression- graft- internal fixation is the preferred consideration to treat hyperextension injury of cervical spine, and the posterior approach is only the secondary choice. The combined approach should be carefully selected under appropriate conditions
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Many basic and clinical studies have showed that through a variety of biological and biophysical mechanisms, low-intensity pulsed ultrasound improves the fracture healing and provides an ideal repairing environment.Thus,it can not only accelerate the healing of fractures,but also enhance the healing rates;not only accelerate the healing of fresh fractures, but also the healing of delayed unions and nonunions.This review elaborates on the effects of low-intensity pulsed ultrasound on the healing of fractures,its mechanisms and its prospective applications. [