激光造型及RGD表面修饰仿生基质对成骨化ADSCs的作用研究

目的 评估激光造型及RGD联合修饰支架后的细胞生物学特征,验证优化表面改性的方法.方法 Nd:YAG激光处理nβTCP/Cs/PCL仿生基质,RGD表面修饰,如下接种羊骨化脂肪基质细胞:A组(Nd:YAG激光+RGD表面改性基质+ADSCs),B组(RGD表面改性基质+ADSCs),C组(Nd:YAG激光改性基质+ADSCs),D组(无表面改性基质+ADSCs);第1、4、8、12、16、20、24、28天,倒置、荧光镜及共聚焦观察细胞生长与存活,检测碱性磷酸酶及Ⅰ型胶原含量,分析细胞的活性和功能水平.结果 A组细胞生长旺盛、细胞外基质丰富,存活率高于其他组,增殖活力0.55±0.13、碱性磷酸酶61.49±7.18及Ⅰ型胶原水平73.49±9.21均高于其他组(B组0.42±0.06、50.32±3.18、48.91±6.74;C组0.31±0.09、38.19±5.12、36.91±8.12;D组0.12±0.03、18.13±1.21、17.91±2.16),细胞骨架发育良好.结论 Nd:YAG激光及RGD联合修饰支架能促进细胞粘附与增殖,为理想的表面改性方法.     

人重组骨形态发生蛋白9基因修饰的兔骨髓间充质干细胞异位成骨实验研究

目的 探讨人重组骨形态发生蛋白9(human recombinant bone morphogenetic 9,rh-BMP-9)基因治疗与骨组织工程技术相结合的可行性.方法 采用阳离子脂质体介导rhBMP-9基因转染兔骨髓间充质干细胞(mesenchymal stem cells,MSCs),荧光显微镜观察及流式细胞学检测,碱性磷酸酶(ALP)活性定量测定及Vbn Kossa's钙结节染色.MSCs与聚乳酸-羟基乙酸(polylactide-co-glycolide,PLGA)支架共培养,荧光显微镜及扫描电镜观察MSCs在PLGA上的粘附、生长情况.将转染与未转染rhBMP-9的MSCs分别与PLGA支架复合培养,构建组织工程化骨并回植兔肌肉内,组织植入4周后进行HE染色观察.结果 菜流式细胞仪测定质粒转染率为34.15%.转染组MSCs的ALP活性较未转染组明显增高(P<0.01),Von Kossa's钙结节染色比较转染细胞形成的钙结节明显大于未转染组.荧光显微镜及扫描电镜观察见MSCS在PLGA支架上的粘附、生长良好.异位回植术后4周组织HE染色见肌肉内有新生骨基质分泌,成骨面积定量分析转染组与对照组相比差异有统计学意义(P<0.05).结论 阳离子脂质体介导的质粒能稳定转染Mscs,rhBMP-9能刺激MsCs的ALP活性增强,诱导钙结节形成,rhBMP-9基因转染的MSCs作为一种新型的种子细胞,其与PUGA支架复合用于骨缺损的修复具有较强的可行性.     

骨髓间充质干细胞的旁分泌作用研究进展

随着近些年来对骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)研究的不断深入,其在多种组织损伤中的修复作用也逐步被揭示。而近几年来发现,BMSCs 除了能通过直接分化为靶细胞而发挥修复作用,其旁分泌作用也在对组织的修复过程中发挥重要作用,BMSCs 分泌的多种细胞因子对损伤组织的修复有重要意义。     

Bone malformations in Proteus syndrome: an analysis of bone structural changes and their evolution during growth

The radiographic follow-up of a patient with Proteus syndrome is presented. Review of radiographs obtained at 3 years 10 months, 10 years, and 17 years 8 months indicated that the rate of growth in length of the oversized tubular bones of the hands was similar to that of the normal bones of the same hand. This observation supports the view that the primary lesion occurs in the early embryonic period, when the limb bud mesenchyme cells condense and cartilage differentiates producing oversized cartilage anlages, rather than being a defect of bone cell-mediated apposition and modelling processes of bone. Additional radiographs of the pelvis and spine were obtained at age 4 years 10 months and head CT at 8 years 10 months. This pathogenetic mechanism fits well with the hypothesis of somatic mosaicism, which is at present the most credible explanation for the aetiology of Proteus syndrome. Other skeletal malformations recognized as typical of the syndrome can be interpreted as secondary adaptations to the altered mechanical conditions induced by overgrowth of bones.     

骨髓基质细胞组成及造血调控作用的研究进展

基质细胞是构成骨髓造血微环境的重要组成部分,在各系造血细胞增殖、分化、成熟、迁入血窦并最终进入外周循环的过程中发挥重要作用.基质细胞包括树突状细胞、巨噬细胞、成纤维细胞、内皮细胞、脂肪细胞和网状细胞等.这些细胞可以通过分泌细胞因子、胞外基质和细胞直接接触调控造血的全部过程,基质细胞造血微环境的紊乱将直接造成各种血液和免...     

体外小儿骨髓间充质干细胞对牵张应力早期应答反应的研究

目的 观察体外小儿骨髓间充质干细胞(MSCs)对周期性牵张应力的早期应答反应.方法 分离并培养小儿MSCs,利用体外细胞牵张应力装置对第三～六代细胞加载短期力学信号,检测各组细胞内Oa2+以及早期反应基因蛋白(c-fos、c-jun和c-myc)的表达情况.结果 应力干预MSCs后,细胞内Ca3+浓度在受力1min后即增高,荧光强度是对照组的164.63%,3、5min升高更明显,干预10 min后Ca2+浓度升高的幅度有下降趋势;c-fos、c-jun和c-myc基因蛋白在未受力组细胞内不表达或微量表达,加载力学信号后,表达发生了明显的变化,随时间的延长,均表现为增强-持续-回复原来水平.结论 周期性牵张应力影响MSCs内Ca2+的浓度及早期反应基因蛋白(c-fos、c-jun和c-myc)的表达,推测c-fos、c-jun和c-myc参与了MSCs响应力学的信号传导途径,可能依赖Ca2+通道的活化来调控.     

Post-natal growth of 157 children born as extremely premature neonates

Aim: With increasing survival rate of extremely premature neonates, their long‐term outcomes including growth and risk factors for later disorders need to be considered. We prospectively evaluated anthropometric parameters in children born as extremely premature neonates. Methods: Anthropometric parameters were measured at the ages of 2 and 5 years in 72 extremely premature children born between the 22nd and 25 + 6th weeks of gestation (group I) and 85 children born between the 26th and 27 + 6th weeks of gestation (group II). Results: Although catch‐up in the postnatal growth was observed in both groups of children, resulting in growth improvement, the height of the extremely premature children at the ages of 2 and 5 years remains lower (P < 0.01) compared with the control population. A decline in head growth was observed in both groups between the ages of 2 and 5 years, resulting in decrease of standard deviation score (SDS) for head circumference (HC) in comparison with that of the control population, accompanied by an increased number of children with microcephaly, defined as HC < ?2 SD. At the age of five, microcephaly was found in 18% of children from group I and 11.7% of children from group II. At the age of 5 years, the waist and hip circumferences and ten skinfolds were not different between both groups of children. Conclusion: Long‐term follow‐up of extremely premature neonates is important not only to establish their growth patterns but also for risk factors assessment including adiposity for later development of adult‐onset diseases.     

骨髓间充质干细胞与支气管肺发育不良的研究进展

支气管肺发育不良是一种发生于新生儿的慢性肺疾病,近年来发病率不断上升.骨髓间充质干细胞具有多向分化功能,可分离、纯化、扩增和运用于临床实验.该文对支气管肺发育不良的病理、发病机制、骨髓间充质干细胞的生物学特性等进行阐述,提出循环中的骨髓间充质干细胞可在肺组织定植,并对肺损伤有修复功能.     

当归注射液对环磷酰胺损伤的骨髓细胞凋亡率的影响

目的研究当归注射液对环磷酰胺损伤的小鼠骨髓细胞凋亡率的影响。方法用8~12周龄Balb/c小鼠30只,随机分为6组:正常对照组;实验对照组;预防大小剂量组:治疗大小剂量组。环磷酰胺一次性腹腔注射,剂量为200mg/kg。制模完成后断颈处死,取右侧股骨,冲出骨髓细胞,制备成细胞悬液。以Annexin V-FITC试剂染色,用流式细胞仪来检测骨髓细胞的凋亡率。结果通过流式细胞仪检测骨髓细胞凋亡率发现预防组、治疗组及实验对照组与正常对照组间有显著差异(P<0.05):预防组与实验对照组间无差异(P>0.05),而治疗组与实验对照组间有显著性差异(P<0.05),且大小剂量组间也有差异性(P<0.05)。结论当归注射液在预防环磷酰胺对骨髓损伤时,对细胞凋亡率作用不明显;而在治疗时作用显著,且大剂量作用更明显。     

Growth hormone treatment enhances bone mineralisation in children with chronic renal failure

Dual energy X-ray absorptiometry of the whole body and the lumbar spine was performed to study bone mineralisation before and after 1 year of recombinant human growth hormone (rhGH) treatment in ten children with chronic renal failure. At the start, median age was 7.3 years (range 2.0–8.8 years) and median glomerular filtration rate 15 ml/min per 1.73 m² (range 7–41 ml/min per 1.73 m²). Total body mineral content (TBMC), lumbar spine mineral content (LBMC), total body bone mineral density (TBMD) and lumbar spine mineral density (LBMD) improved significantly (P < 0.05) after 1 year of treatment. Bone mineral data before and after treatment were compared with two groups of controls, i.e. ten healthy children matched for age and ten healthy children matched for height. Patients' TBMC, LBMC, TBMD and LBMD data before treatment were no different from those of height-matched controls; the same was true after 1 year of treatment except for the patients' significantly better LBMD (P < 0.05). When compared with age-matched controls, patients had significantly lower baseline TBMC and LBMC levels before treatment; after treatment LBMC was no longer different. However, there were no differences in TBMD or LBMD between patients and age-matched controls at baseline or after rhGH. Conclusion Recombinant human growth hormone treatment for 1 year results in a significant increase in both growth velocity and bone mineralisation. Comparison with height-matched controls shows a similar bone mineralisation at baseline and a better bone mineral density after treatment. Received: 10 August 2000 and in revised form 10 November 2000 and 5 January 2001 /  Accepted: 8 January 2001     

纳米相羟基磷灰石胶原复合材料与人骨髓基质干细胞体外相容性的研究

目的探讨纳米相羟基磷灰石胶原复合材料(nano-HAP／Collagen，NHAC)与人骨髓基质干细胞(hBMSC)相容性，为应用组织工程方法修复骨缺损提供依据。方法将人骨髓基质干细胞与纳米相羟基磷灰石胶原复合材料体外复合培养，进行形态学和功能测定。结果骨髓基质干细胞能在纳米相羟基磷灰石胶原复合材料上良好地粘附、增殖、生长。细胞的活性和碱性磷酸酶活性未受到纳米相羟基磷灰石胶原复合材料的影响。结论纳米相羟基磷灰石胶原复合材料具有良好的细胞相容性，可作为骨组织工程理想的骨载体材料。     

母孕前和孕期高脂饮食对新生仔鼠骨骼生长的影响及机制探讨

目的 观察母鼠孕前和孕期高脂饮食对新生仔鼠骨骼生长的影响,并探讨影响骨骼生长的机制.方法 40 只雌性Sprague-Dawley 大鼠随机分为高脂组和对照组(n=20),分别喂养35% 高脂饲料和普通饲料.喂养8 周后,高脂组和对照组各取8 只雌鼠观察其肝脏组织病理;其余与普通饲料喂养的雄性大鼠交配,孕期分别继续给予高脂饲料或普通饲料喂养.待娩出新生仔鼠后24 h 内,测量两组仔鼠体长(鼻尾长度);酶联免疫吸附试验测定血清胰岛素样生长因子-I(IGF-I)水平;光镜下观察肝脏组织病理;免疫组织化学法检测长骨(胫骨、股骨)中胰岛素受体底物1(IRS-1)和磷酸化IRS-1(Phospho-IRS-1)的表达;蛋白质印迹技术检测长骨软骨细胞中促分裂原活化蛋白激酶(MAPK)和磷酸化MAPK(Phospho-MAPK)、磷酯酰肌醇3-激酶(PI3K)和磷酸化PI3K(Phospho-PI3K)、蛋白激酶B(AKT1)和磷酸化AKT1(Phospho-AKT1)的蛋白表达.结果 高脂组仔鼠的出生体长较对照组显著降低(P<0.05).高脂组仔鼠血清IGF-I 水平较对照组下降,但差异无统计学意义(P>0.05).高脂组母鼠及仔鼠肝组织可见脂肪样变,而对照组肝脏病理形态正常.两组仔鼠长骨软骨细胞中IRS-1 的表达差异无统计学意义(P>0.05).高脂组仔鼠长骨软骨细胞中MAPK 的表达水平高于对照组(P<0.05),而PI3K 及AKT1/Phospho-AKT1 的表达水平在两组间差异无统计学意义(P>0.05).结论 母鼠孕前和孕期高脂饮食会影响胎鼠在宫内的骨骼发育,可能与IGF-I 的下降有关,但未发现IGF-I 对骨骼影响的确切发病机制.     

先心病婴幼儿骨髓间充质干细胞向心肌样细胞诱导分化的实验研究

目的探讨先心病婴幼儿骨髓间充质干细胞（bone marrow stromal cells，BMSCs）向心肌样细胞诱导分化的可行性，为心肌组织工程提供新的种子细胞的来源。方法无菌条件下抽取先天性心脏病婴幼儿胸骨骨髓3～5ml，经密度梯度离心（Percoll分离液，T为1．073），接种获得BM～SCs，常规培养。流式细胞仪检测细胞表面抗原。将已传2代的细胞应用5-氮杂胞苷（5-Azacytidine，5Aza）诱导24h，更换为完全培养基（LG-DMEM）后继续培养，2周后行免疫细胞化学及透射电镜行超微结构检查。结果先心病婴幼儿BMSCs基本上为梭型成纤维细胞样形态，表面标记为CD29、CD44、CD71、CD90表达阳性，CD3、CD14、CD34、CD45、HLA-DR不表达。BMSCs经5-Aza诱导2周后α-actin，Desmin，cTnI，Cx-43免疫细胞化学染色阳性，透射电镜示诱导后的细胞有明显的肌丝，单个细胞核呈椭圆形，位于细胞的中央。结论先心病婴幼儿骨髓阃充质干细胞可在体外经5-Aza诱导分化为心肌样细胞，有望成为应用自体细胞体外构建工程化心肌重要的种子细胞。     

自体骨髓注射治疗单纯性骨囊肿的影像学变化与临床意义

目的观测自体骨髓注射治疗单纯性骨囊肿的影像学改变，分析其临床意义。方法自体骨髓注射治疗单纯性骨囊肿患儿27例，获随访25例，随访平均3．3年。以CR摄影判定影像学上的疗效；观测术后不同时期X线骨囊肿面积、皮质厚度的变化情况；分术前合并病理骨折组及无病理骨折组，测算术前囊肿纵径／最小皮质厚度，行组间比较。通过三维CT结合X线观察骨囊肿的基本愈合形式。结果1次注射后愈合10例，缺损愈合5例，持续存在7例（含2例无反应），复发3例。除外无反应者，术后3个月病灶内均有不同程度的密度增高；X线骨囊肿面积基本稳定，主要发生在术后649个月（21／23）；X线骨囊肿皮质厚度基本稳定，主要发生在术后9个月～1年（19／23）。除外下肢持重管状骨病变，术前合并病理骨折组及无病理骨折组囊肿纵径与最小皮质厚度比值差异显著。结论自体骨髓注射治疗单纯性骨囊肿术后随访至少1年；该法通过骨皮质增厚，囊腔内骨嵴及骨小梁向心性增生，分割包绕填塞囊腔而逐步愈合。     

Effects of growth during infancy and childhood on bone mineralization and turnover in preterm children aged 8-12 years

To investigate the effect of growth on later bone mass and turnover, bone mineral content (BMC) and density (BMD: dual X-ray absorptiometry (QDR 1000W) and single photon absorptiometry (Lunar SP2)) and bone turnover (plasma osteocalcin, urine deoxypyridinoline) were measured at 8-12 y in 244 preterm children who had weight and height measured at 18 mo and 7.5-8 y corrected age. Weight and length at birth, 18 mo, 7.5-8 y and current follow-up showed increasingly strong, positive correlations with bone area, BMC and BMD. After adjusting for current size, there were significant negative associations between earlier size measurements and later whole body and lumbar spine bone mass which were stronger for length than for weight, and a negative relationship between birthweight for gestation and later radial bone mass; but no relationship with bone turnover. Current calcium intake and activity level had no independent effect on bone mass. Bone mass at 8-12 y is related to current bone and body size, which tracks throughout childhood. However, amongst children of the same current size, those who have shown the greatest increase in size, particularly in height, have the highest bone mass. These findings raise the hypothesis that improving linear growth in vulnerable children may be important in maximizing bone mass.     

评价儿童青少年骨体重负荷的意义及指标选择(英文)

目的　探讨儿童青少年骨体重负荷对腰椎和髋部骨矿含量 (BMC)、骨密度 (BMD)的影响 ,并比较两指标的优次。方法　应用DXAQDR - 4 5 0 0A型扇形束骨密度仪测量长沙地区 5 4 7例 6～ 15岁儿童青少年腰椎前后位 ,仰卧侧位及髋部股骨近端的骨量。结果　不论男女 ,儿童青少年体重、体块指数 (BMI)、腰椎及髋部BMC和BMD随年龄增加而增加 (P <0 .0 5或 0 .0 1) ;体重与BMC的相关性较体重与BMD的相关性更密切 ;髋部及腰椎各部位体重标准化BMC随年龄增加而增大 ,而髋部和腰椎各部位体重标准化BMD随年龄增加反而减小。结论　 6～ 15岁儿童青少年腰椎及髋部BMC指标判断骨强度优于BMD ,尤以髋部及腰椎侧位BMC为佳。     

Relationship between chronological and bone ages and pubertal stage of breasts with bone biomarkers and bone mineral density in adolescents

ObjectiveTo study bone mineral density (BMD) in adolescent females according to five groups of chronological age (CA), bone age (BA), and breast development stage (B), and to correlate these parameters with plasma bone biomarkers (BB).MethodsThis was a cross-sectional study performed in 101 healthy adolescent females between 10 and 20 years old. The study variables were: weight, height, body mass index (BMI), CA, B, BA, calcium intake, BMD, and BB. Osteocalcin (OC), bone alkaline phosphatase (BAP), and C-terminal telopeptide (S-CTx) were evaluated for BB. BMD was measured using dual energy X-ray absorptiometry (DXA).ResultsBMD in lumbar spine, proximal femur, and total body increased with age, and the respective observed averages were: in CA1 (10 years old), 0.631, 0.692, 0.798 g/cm²; in CA2 (11 to 12 years old), 0.698, 0.763, 0.840 g/cm²; in CA3 (13 to 14 years old), 0.865, 0.889, 0.972 g/cm²; in CA4 (15 to 16 years old), 0.902, 0.922, 1.013 g/cm²; and in CA5 (17 to 19 years old), 0.944, 0.929, 1.35 g/cm². These results showed significant differences between 13 and 14 years of age (CA3) or when girls reached the B3 stage (0.709, 0.832, 0.867 g/cm²). The highest median concentrations of BB were between 10 and 12 years of age when adolescents were in the B2–B3 (p < 0.001). Median BB concentrations decreased in advanced BA and B.ConclusionsBB concentrations were positively correlated with the peak height velocity and negatively correlated with BMD in the study sites. Increased BMD and BB concentrations were observed in B3.     

Growth and bone mineralization in patients with juvenile idiopathic arthritis

Objective  To investigate growth, development and bone mineralization of children with juvenile idiopathic arthritis (JIA). Methods  Thirty patients between 4–17 years of age (mean 11.34 ± 3.88) resistant to therapy were studied. Enrollment began in November 1999 and continued through November 2004 and children with chronic disease were excluded. Data like height, weight, medications and acute phase reactants were obtained from medical records. On study-visit, puberty was assessed by physical examination and bone mineral density (BMD) was measured. Serum Ca, P, ALP, insulin-like growth factor-1 (IGF-1) and urinary Ca/Cr and hydroxyproline /Cr levels were measured. Results were compared with the control group that consisted of 30 cases of similar age and gender. Results  Patients with JIA had decreased height standard deviation score (SDS) and growth retardation. BMD of the cases in the study group was lower than the control group (p<0.05). Patients who were at younger age at the onset of the disease had lower BMD. Among the drugs, only steroids had a negative effect on growth. Serum IGF-1 levels of the study group were significantly lower than the control group (p<0.0001). Conclusion  Early diagnosis and suppression of disease activity is important in prevention of osteoporosis and growth retardation in children with JIA. BMD has to be measured yearly in patients for accurate diagnosis of osteoporosis. Vitamin D and Ca-rich nutrition with promotion of physical activity and controlled use of steroids may protect the children against bone loss.     

MDR1基因转染的骨髓单个核细胞对放疗后造血功能重建的研究

目的 研究BALB/c小鼠放射治疗后经尾静脉回输携带多药耐药MDR1基因的骨髓单个核细胞(bone marrow mononuclear cells,BM-MNCs)对造血功能重建的影响.方法 BAB/c近交系小鼠32只,随机分为4组:正常对照组、空白对照组、阴性对照组和实验组.正常对照组不做任何处理,其他3组首先接受1.5Gy60Co-γ射线全身照射,空白对照组尾静脉回输等量生理盐水,阴性对照组经尾静脉回输未转染MDR1基因BM-MNCs,实验组回输已转染MDR1基因BM-MNCs.动态观察各组外周血变化.结果 同种异体回输后第七天,阴性对照组和实验组与空白对照组比较均表现为白细胞恢复提前.实验组7、10、14 d白细胞计数分别为(2.9±0.3)×109/L、(3.2±0.2)×109/L、(4.2±0.3)×109/L,阴性对照组为(2.7±0.2)×109/L、(2.8±0.2)×109/L、(3.5±0.3)×109/L,实验组与阴性对照组比较,白细胞数量增加,差异有统计学意义(t=2.21、3.53、4.73,P＜0.05),14 d后差异无统计学意义(t=0.79,P＞0.05).结论 同种异体携MDR1基因的骨髓单个核细胞移植,能在早期显著提高外周血白细胞数量,利于骨髓造血微环境恢复,加快辐射损伤后骨髓早期造血功能重建.     

新生适于胎龄儿骨转换标志物与骨声波传导速度的关系

目的：研究新生适于胎龄儿Ⅰ型前胶原羧基端前肽（PICP）、尿脱氧吡啶啉（DPD）及骨声波的传导速度（SOS）随胎龄变化的规律,探讨骨转换标志物和骨SOS的关系。方法：共选取65例新生适于胎龄儿为研究对象,根据胎龄分为早产儿组（≤34周,14例）,晚期早产儿组（>34周至<37周,13例）,足月儿组（≥37周,38例）。所有研究对象均测量出生体重、身长,采用Ponderal指数（PI）估测新生儿营养状态;生后7?d内采集静脉血测定血PICP水平;收集尿液测定尿DPD、尿肌酐（Cr）水平;生后7 d内超声定量技术测量左侧胫骨声波的传导速度（SOS）。结果：胎龄、出生体重、身长及PI值在3组间的差异均有统计学意义（分别F=140.199、47.042、46.877、11.898,均P<0.01）,且出生体重、身长和PI值随胎龄增加而增加。3组之间PICP、DPD/Cr、SOS的差异有统计学意义（分别F=30.384、21.761、20.052,均P<0.01）,且胎龄越大,PICP及DPD/Cr水平越低,骨SOS越高。PICP、DPD/Cr水平与胎龄、出生体重及骨SOS呈负相关（P<0.01）;而骨SOS与胎龄和出生体重呈正相关（P<0.01）,在校正了胎龄、出生体重后依然存在相关关系。结论：新生适于胎龄儿骨转换生化指标和胎龄、出生体重及骨SOS呈负相关;新生适于胎龄儿的高骨转换状态对骨健康不利。