Effects of basic fibroblast growth factor on osteoblast-related gene expression in the process of medullary bone formation induced in rat femur

 The effects of basic fibroblast growth factor (bFGF) on osteogenic differentiation in-vivo were investigated using a rat bone marrow ablation model. bFGF was infused directly into rat femora for 6 days after bone marrow ablation. The contralateral femur was infused with vehicle only and used as control. Bone formation was induced in the rat femoral cavity, and the gene expression of osteoblast markers was examined. Treatment with bFGF at 50 and 100 ng/day significantly enhanced the mRNA levels of osteopontin compared with the levels in the control leg, with increases of 25% and 24%, respectively. In contrast, bFGF infusion at 50 ng/day provoked a significant (nearly 20%) inhibition of expression for type I collagen. Infusion of bFGF at a higher dose exhibited an inhibitory tendency for bFGF action on gene expression. There were no significant changes in alkaline phosphatase and osteocalcin mRNA levels in response to any dose of bFGF. The findings presented here suggest that bFGF modulates osteogenic differentiation in-vivo and may play an important role in the process of bone remodeling. Received: February 22, 2002 / Accepted: October 7, 2002 Offprint requests to: H. Tanaka     

不同剂量重组牛碱性成纤维细胞生长因子治疗创面的比较

目的　探索外用重组牛碱性成纤维细胞生长因子（bFGF）治疗外伤创面的最佳用药浓度、给药方法与剂量,以指导临床应用。方法　设等渗盐水加灭滴灵治疗为对照组,选择bFGF的不同用药浓度（75Au/cm     

Immunohistochemical localization of fibroblast growth factor receptors in the rat mandibular condylar cartilage and tibial cartilage

The fibroblast growth factor receptors (FGFRs), members of the tyrosine-kinase receptor family, are known to play a crucial role in the growth and development of cartilaginous tissues. The mandibular condylar cartilage has been suggested to have a characteristic growth pattern compared with the tibial growth plate cartilage, e.g., cell alignment, mode of proliferation and differentiation, and response to humoral and mechanical factors. To examine the mRNA expression and localization of fibroblast growth factor receptor (FGFR)-1, -2, and -3 in the condylar and tibial growth plate cartilages, reversed transcribed polymerase chain reaction (RT-PCR) assay and immunohistochemistry were carried out using growing rats. The enzymatically isolated rat condylar and tibial chondrocytes expressed mRNA of aggrecan and type II collagen, which are together known as the major cartilaginous extracellular matrices. Both types of cells expressed mRNA of FGFR-1, -2, and -3 by RT-PCR. In the neonatal rat, immunolocalization of FGFR-1, -2, and -3 was found in the middle of the condylar cartilage, mainly in the hypertrophic zone of the tibial cartilage. At 3 weeks old, the three FGFRs were broadly observed in both cartilages. At 8 weeks old, localization of FGFR-3 was absent in the hypertrophic cell layer of the condyle, whereas it was still broadly observed in the tibial growth plate cartilage. In the same stage, FGFR-1 and FGFR-2 showed similar localization in both cartilages to that at 3 weeks of age. All these observations suggest that FGFRs play an important role in the differential growth pattern of the condylar cartilage. Received: Jan. 14, 1999 / Accepted: March 3, 1999     

不同剂量碱性成纤维细胞生长因子对神经侧枝再生的影响

目的: 探讨碱性成纤维细胞生长因子 (basicFibroblastGrowthFactor, bFGF) 能否促进神经端侧缝合后的侧枝再生, 其作用是否存在量效关系。方法: 15只Wistar大鼠随机分为A、B、C3组, 切断一侧腓总神经并将其远端与外膜开窗的胫神经做端侧缝合。A、B2组于术后 1周内每天在腹腔内注射生理盐水稀释的bFGF, A组剂量 4 000u/kg, B组剂量 2 000u/kg, C组注射等量生理盐水做对照。术后 3个月取缝合口远端的腓神经做组织学检查。结果: 应用bFGF后再生神经纤维数目、有髓纤维截面积和髓鞘截面积显著大于生理盐水对照组 (P<0. 05 ); 而A组上述三项指标均显著大于B组 (P<0. 05 )。结论: 神经端侧缝合后早期应用外源性bFGF可促进未受损神经的侧枝发芽, 其作用与剂量呈正相关。     

Long-term follow-up of tracheal cartilage growth promotion by intratracheal injection of basic fibroblast growth factor

Background

Intratracheal injection of basic fibroblast growth factor (b-FGF) has been shown to enlarge the tracheal lumen 4?weeks after treatment. The objective of this study was to investigate the long-term effect of tracheal cartilage growth promotion by intratracheal injection of b-FGF.

不同浓度碱性成纤维细胞生长因子对体外培养兔肌腱细胞增殖的影响

目的探讨不同浓度的碱性成纤维细胞生长因子(bFGF)对体外培养兔肌腱细胞增殖的影响,并确定促进肌腱细胞增殖的最佳浓度。方法在体外培养第2代兔肌腱细胞的培养液中分别加入不同浓度(0.5、1、2、5、10、20、30、40、50μg/L)的bFGF,继续培养48 h,噻唑蓝(MTT)法染色,在酶联免疫检测仪上测出不同浓度bFGF组光密度(OD)值。结果与对照组OD均值相比:5、10、20、30、40、50μg/L bFGF组差异均有统计学意义(P<0.05);各组OD均值随bFGF浓度的增加,先逐渐增大(5～20μg/L),达到峰值后又逐渐降低(20～50μg/L)。结论 bFGF有明显促肌腱细胞增殖的作用,且与浓度有一定相关性,即促肌腱细胞增殖的起始bFGF浓度为5μg/L,而20μg/L时可达到促肌腱细胞增殖的最佳浓度。     

Promotion of tracheal cartilage growth by intra-tracheal injection of basic fibroblast growth factor (b-FGF)

Purpose

Basic fibroblast growth factor (b-FGF) is a very effective growth factor that induces the proliferation of chondrocytes. This study aimed to investigate whether intra-tracheally-injected b-FGF solution promotes the growth of tracheal cartilage.

Methods

Group 1: 500 μl of distilled water was injected at the posterior wall of the cervical trachea of New Zealand white rabbits by using a tracheoscope (n = 5). Group 2: 100 μg/500 μl of b-FGF solution was injected at the posterior wall of the cervical trachea (n = 5). Group 3: Biodegradable gelatin hydrogel microspheres incorporating 100 μg/500 μl of b-FGF solution were injected at the posterior wall of the cervical trachea (n = 5). All animals were sacrificed 4 weeks later, and the outer diameter and luminal area of the cervical trachea at the site of b-FGF injection were measured.

Results

The cervical tracheas in the two b-FGF injection groups were spindle-shaped and had a maximum diameter at the injection site. The median outer diameter of the cervical trachea in Groups 1, 2, and 3 was 7.3, 8.0, and 8.0 mm, respectively, showing a significant difference among Groups 1, 2, and 3 (P = 0.04). The median luminal area in Groups 1, 2, and 3 was 27.4, 29.4, and 32.1 mm², respectively. The ad hoc test showed a marginally significant difference only between groups 1 and 3 (p = 0.056).

Conclusion

Intra-tracheal injection of slowly released b-FGF enlarged the tracheal lumen.     

Effect of insulin-like growth factor 1 and basic fibroblast growth factor on DNA synthesis and collagen production in cultured anterior cruciate ligament cells

This study was undertaken to investigate the effects of insulin-like growth factor 1 (IGF-1) and basic fibroblast growth factor (bFGF) on the DNA and matrix synthesis of cells out-grown from the anterior cruciate ligament (ACL). Five batches of ACL cells from five 8-week-old Japanese white rabbits were isolated and maintained in culture until the fifth passage. We analyzed the effects of various concentrations of IGF-1 (1–1000 ng/ml) on [³H]-thymidine uptake in the cells at the first and fifth passages, and collagen content in the cell layer at the third passage, in the presence or absence of bFGF (10ng/ml). In the absence of bFGF, IGF-1 caused a significant increase in the synthesis of DNA and collagen in the ACL cells. IGF-1 and bFGF, in combination, synergistically increased the DNA synthesis of ACL cells, whereas such synergistic enhancement was not observed in their, collagen production. The amounts of [³H]-thymidine incorporated into the cells incubated with IGF-1 (500ng/ml) and bFGF (10ng/ml) combined were 1.3–1.5 times greater at first passage and 1.3–1.9 times greater at fifth passage than the sum of these with the growth factors used individually. Based on this in vitro finding, we consider it clinically relevant that IGF-1 and bFGF, when used together, have the capability to enhance the primary healing of ruptured ACL. Presented at the 11th Annual Meeting for Orthopaedic Research of the Japanese Orthopaedic Association, Kagoshima, Japan, October 17, 1996.     

磁刺激和碱性成纤维细胞生长因子对脊髓损伤早期的保护作用

目的探讨脊髓损伤后,应用磁刺激和外源性碱性成纤维细胞生长因子(bFGF)对损伤脊髓组织早期的保护作用。方法实验利用Allen WD(weight drop)技术,以致伤力(砝码质量10g,下落高度2．5cm)制成wistar大鼠T8脊髓损伤模型,治疗组分别于术后即刻、1h、2h、4h、24h分别予0．5HZ、70％输出强度的磁刺激,同时经蛛网膜下腔导管注入20ul bFGF,对照组不做处理。术后2h、6h、24h取治疗组、对照组动物损伤区脊髓组织作以下检测：用干湿法测水含量;用原子吸收光谱法测钙、镁离子含量;伤后5d取损伤脊髓组织,电镜观察脊髓结构。结果损伤区脊髓组织水含量增多,钙离子水平升高,镁离子水平下降,白质内髓鞘结构紊乱,囊性变严重;而应用磁刺激和bFGF可改善上述变化。结论脊髓损伤后应用磁刺激和bFGF可减轻脊髓损伤后的离子失衡,从而对继发性脊髓损伤具有保护作用。     

羊膜移植联合碱性成纤维细胞生长因子滴眼液治疗兔角膜碱烧伤

目的 观察羊膜移植与碱性成纤维细胞生长因子(bFGF)滴眼液联合应用对兔角膜碱烧伤的治疗效果。方法 将60只新西兰白兔随机分为4组,分别为羊膜移植联合bFGF滴眼液点眼组(A组),羊膜移植组(B组),bFGF滴眼液点眼组(C组)及对照组(D组)。用1 mol/L NaOH烧伤兔右眼,术后2 ～28 d用裂隙灯显微镜观察角膜透明度,新生血管生长,角膜上皮修复及羊膜植片的变化。结果 所有烧伤角膜都能完全愈合,但在抑制炎症反应、抗新生血管的生长及角膜上皮修复等方面,羊膜移植联合bFGF滴眼液点眼组明显优于对照组,也好于其他两实验组,差异有统计学意义(P＜0.05)。结论 羊膜移植联合bFGF滴眼液点眼对治疗角膜碱烧伤有良好效果。     

基因治疗对下颌骨牵引过程中碱性成纤维细胞生长因子表达的影响

目的 探讨电穿孔介导的基因治疗对兔下颌骨牵引成骨过程中碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)表达的影响.方法 新西兰大白兔双侧下颌骨截骨,术后3d开始以0.8 mm/d速度行下颌骨牵引,连续牵引7d,将实验动物分为:A、B、C、D、E5组.分别在牵引区注射2μg重组质粒pIRES-hVEGF165-hBMP2、pIRES-hBMP2、pIRES-hVEGF165、空质粒pIRES及生理盐水.各组实验动物均施加电穿孔刺激.各组分别于固定期第7、14、28天处死动物取材行免疫组织化学检查bFGF的表达,并利用病理图像分析系统进行分析.结果 bFGF在肉芽中的成纤维细胞、单核巨噬细胞、多核巨噬细胞、间质细胞、成骨细胞和骨细胞中表达:1周时以C组表达较强,2、4周时以A、B、C组表达仍较强,A、B、C组与D、E组相比差异有统计学意义(P＜0.01).结论 电穿孔介导的基因治疗能使bFGF在牵引区的表达增强和表达时限延长,发挥其生理作用,促进细胞的分裂增殖与分化及牵引区细胞基质的形成和新骨生成.这可能是基因治疗促进牵引区新骨生成的机制之一.     

Expression of basic fibroblast growth factor mRNA in benign prostatic hyperplasia and prostatic carcinoma

In our previous work we demonstrated that prostate-derived growth factor (PrGF) is homologous to basic fibroblast growth factor (bFGF), not acidic fibroblast growth factor (aFGF). Using Northern blot analysis we now show that the messenger RNA for bFGF but not aFGF is expressed in benign prostatic hyperplastic (BPH) tissue as well as in carcinoma of the prostate (CAP). This not only corroborates our previous results, but suggests that PrGF is produced locally and not merely stored in the prostate. The demonstration of local production of bFGF by prostate tissue may indicate that this growth factor plays a role, either alone or in conjunction with other factors, in the etiology of benign hyperplasia or prostatic cancer.     

bFGF 、TGFβ-1在膀胱出口梗阻患者逼尿肌细胞中的表达

目的 观察膀胱出口梗阻（BOO）后膀胱平滑肌细胞碱性成纤维细胞生长因子（bFGF）、转化生长因子（TGFβ-1）表达的变化,探讨生长因子在BOO后膀胱平滑肌细胞继发改变中的作用。方法 应用逆转录－聚合酶链反应（RT－PCR）和免疫组织化学SP法在mRNA和蛋白水平检测bFGF、TGFβ-1在30例BOO及4例非BOO膀胱平滑肌组织中的表达。结果 BOO组与非BOO组膀胱平滑肌组织均有bFGF mRNA表达,BO组表达水平高于非BOO组,P＜0．01;TGFβ-1 mRNA在两组未见表达。结论 BOO 后磅胱平滑肌的继发改变与bFGF mRNA的表达有关。     

bFGF缓释微胶囊诱导缺血心肌血管新生的实验研究

目的　探讨局部应用bFGF(碱性成纤维细胞生长因子 )缓释微胶囊对诱导缺血心肌血管新生的作用。方法　 2 4只新西兰大白兔随机分为对照组 (I组 ) ,空白胶囊组 (II组 ) ,bFGF缓释微胶囊组(III组 ,每只胶囊含bFGF 1μg) ,每组 8只。开胸结扎冠状动脉前降支根部 ,II、III组于左旋支、前降支交界区心外膜下埋藏空白微胶囊 ,或bFGF缓释微胶囊各 5只。术后 5周 ,EVAN蓝染色测定心肌梗死区与左心室重量之比 ,免疫组化测定心肌梗死边缘区微血管数。结果　与I、II组相比 ,III组心肌梗死区与左心室重量之比明显缩小 (I组 16 8%± 0 4% ,II组 16 7%± 0 5 % ,III组 7 0 %± 0 2 % ,P <0 0 0 1) ,微血管数目明显增多 (I组 37 75± 4 5 0 ,II组 38 37± 4 98,III组 135 5 0± 4 81,P <0 0 0 1)。结论　bFGF缓释微胶囊给药方便、剂量恒定 ,可以减小心肌梗死面积 ,诱导缺血心肌血管新生     

bFGF-PLGA微球促进随意皮瓣早期断蒂

目的 :探讨碱性成纤维细胞生长因子 -聚乳酸 /聚乙醇酸 (bFGF PLGA)微球促进随意皮瓣早期断蒂。方法 :通过复乳溶剂挥发法制备bFGF PLGA微球 ,采用60 Co辐照灭菌。将bFGF PLGA微球注入大鼠背部随意皮瓣下方 ,4d后断蒂 ,断蒂后 10d观察皮瓣成活率、新生微血管数量。并与单独使用bFGF组进行对比。结果 :皮瓣成活率A组为 (89 2± 2 6 0 ) %,B组为 (85 6± 2 9 7) %,C组为 (76 6± 3 2 2 ) %。新生血管数A组为 2 9 7,B组为 2 4 6,C组为 12。经统计学分析 ,各组间存在显著差异。结论 :bFGF PLGA微球能够更好地促进随意皮瓣的早期断蒂。     

Demonstration of fibroblast growth factor receptor-I in human prostate by polymerase chain reaction and immunohistochemistry

The expression and localization of fibroblast growth factor receptor-1 were investigated in human prostatic tissues with or without benign hyperplasia. Using a polymerase chain reaction method, we were able to demonstrate that prostatic tissues with benign hyperplasia expressed a significantly higher level of fibroblast growth factor receptor-1 mRNA than normal prostatic tissues (P < 0.01 by Anova). Western blot analysis using an antiserum against the receptor gave 2 bands with molecular weights of about 140 kDa and 80 kDa; these correspond to the expected sizes of the long and secreted forms of the fibroblast growth factor receptor-1, respectively. An immunohistochemical study using the same antiserum further demonstrated that the immunoreactive staining occurred mainly in the basal cells of the glandular epithelium and occasionally in the stromal cells. These results suggest that fibroblast growth factors may influence, at least in part, the proliferation of the epithelial cells seen in benign hyperplasia of human prostate. © 1995 Wiley-Liss, Inc.     

碱性成纤维细胞生长因子在膀胱移行细胞癌中表达及相关性研究

目的 探讨膀胱移行细胞癌(TCC)中碱性成纤维细胞生长因子(bFGF)的表达及其与膀胱癌移行细胞癌生物学行为的关系。方法 应用免疫组化方法及图象分析技术检测54例膀胱移行细胞癌标本中bFGF的表达,并进行分组比较。结果 bFGF的表达在不同的病理分级、临床分期及有否复发组之间均有显著性差异(P<0.05)。结论 bFGF的表达与膀胱移行细胞癌发生发展密切相关,其主要由TCC肿瘤组织分泌,这为临床更好地诊断和治疗膀胱肿瘤提供了新的方向和理论依据。     

前列腺增生组织中bFGF和PCNA区域表达的研究

为了探讨碱性成纤维细胞生长因子（ｂＦＧＦ）和ＰＣＮＡ在前列腺增生（ＢＰＨ）组织中区域分布及其作用,应用免疫组化结合计算机图像分析方法检测ｂＦＧＦ和ＰＣＮＡ在２４例ＢＰＨ组织的尿道周围区、中间区和包膜下区中表达。结果显示尿道周围区ｂＦＧＦ和ＰＣＮＡ在ＢＰＨ组织中存在在区域分布差异,两者在尿道周围区表达增高,提示其与ＢＰＨ发生发展有密切联系。     

bFGF基因转染促进股骨头坏死修复的实验研究

目的 :为临床治疗股骨头及其它骨缺血性坏死探索新方法。方法 :将碱性成纤维细胞生长因子 (bFGF)真核表达质粒pCD rbFGF与胶原混合植入坏死的兔股骨头内 ,术后RT PCR及免疫组化方法检测bFGF表达情况 ,组织切片及组织形态学分析股骨头内血管生长及新骨形成情况。结果 :术后 2周RT PCR及免疫组化证实转染bFGF基因的股骨头内有bFGF表达。术后 2周股骨头内血管生长与对照组相比 ,术后 8周股骨头内新骨形成与对照组相比 ,差异均有非常显著意义 (P <0 .0 1)。结论 :利用bFGF基因转染可刺激股骨头坏死内血管再生和新骨形成 ,为临床治疗骨缺血性坏死提供了新的研究方向