碱性成纤维细胞生长因子与肝素结合性质的研究进展

碱性成纤维细胞生长因子 (ｂａｓｉｃｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ ,ｂＦＧＦ)是成纤维细胞生长因子 (ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒｓ ,ＦＧＦｓ)这一大家族的原型成员 ,自 1974年首次从牛垂体中分离ｂＦＧＦ以来[1] ,ＦＧＦ家族的成员目前已增加至 19位[2 ] 。ＦＧＦｓ各成员结构相关、功能相似 ,调节多种细胞的生长、分化、迁移、凋亡 ,此外 ,该家族成员均能结合肝素或硫酸肝素[3 ] 。ｂＦＧＦ和硫酸肝素的结合是它所具有的复杂生化调节机制的基础。肝素和ｂＦＧＦ及其受体三者共同组成复杂的ｂＦＧＦ…     

急性脊髓损伤后aFGF、bFGF的表达

成纤维细胞生长因子 (ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃ ｔｏｒ ,ＦＧＦ)在体外能促进多种中枢及外周神经元的存活及突起生长 ,体内也能促进神经元的修复与再生。ＦＧＦ是 1974年Ｇｏｓｐａｄａｒｏｗｉｃｚ[1] 从牛脑的垂体中分离纯化出的一种多肽分子。根据等电点的不     

纤维母细胞生长因子的提纯及抗血清的制备

1974年Ｇｒｏｓｐｏｄａｒｏｗｉｃｚ从牛垂体中提纯出碱性成纤维母细胞生长因子 (ｂａｓｉｃＦｉｂｒｏｂｌａｓｔＧｒｏｗｔｈＦａｃ ｔｏｒ ,ｂＦＧＦ)后 ,其临床意义得到广泛的重视。目前 ,ｂＦＧＦ被认为是某些恶性肿瘤诊断的一项敏感性指标 ,具有操作简便、特异性高等优点。为建立ｂＦＧＦ诊断试剂盒 ,我们将Ｇｒｏｓｐｏｄａｒｏｗｉｃｚ的方法稍加改进 ,从牛脑提取了ｂＦＧＦ ,并制备了抗ｂＦＧＦ血清。1　材料与方法1 1　材料　牛脑购自长春市屠宰厂 ;肝素 Ｓｅｐｈａ ｒｏｓｅ、ＣＭ ＳｅｐｈａｄｅｘＣ 50购自Ｐｈａｒｍａｃｉ…     

bFGF及KGFmRNA在口腔粘膜中的表达分析

成纤维细胞生长因子超家族（ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒｓ，ＦＧＦ）是包括９个成员的肽类生长因子，参与了胚胎发育、损伤修复、血管生成等多种重要的生理和病理生理过程。ｂＦＧＦ（ｂａｓｉｃＦＧＦ）及ＫＧＦ（ｋｅｒａｔｉｎｏｃｙｔｅｇｒｏｗ...     

血管生成因子及其受体表达与Keloid瘤样增生的关系

碱性成纤维细胞生长因子（ｂａｓｉｃｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃ－ｔｏｒ，ｂＦＧＦ）和血管内皮生长因子（ｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＶＥＧＦ）或血管通透因子（ｖａｓｃｕｌａｒｐｅｒｍｅａｂｉｌ－ｉｔｙｆａｃ...     

抗人aFGF单克抗抗体抗原表位的筛选

成纤维细胞生长因子ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒＦＧＦ是一类多功能的分子,现已发现其有多种同源结构形式,形成了一个生长因子的家族。酸性ＦＧＦａｃｉｄｉｃＦＧＦａＦＧＦ,也称为ＦＧＦ－１,是ＦＧＦ家族中的主要成员,其结构与功能方面的研究受到广泛的关注１２。本室曾制备了抗人ａＦＧＦｈａＦＧＦ的单克隆抗体ｍＡｂ,并建立了检测ｈａＦＧＦ的双抗体夹心测定方法３４。在此基础上,我们采用噬菌体多肽展示技术,筛选出抗ｈａＦＧＦｍＡｂ的抗原表位,为ｈａＦＧＦ的免疫分析…     

环核苷酸对心肌细胞bFGF与TGFβ_1基因表达的调控初探

目的 :探讨环核苷酸 (ｃＡＭＰ/ｃＧＭＰ)对心肌细胞碱性成纤维细胞生长因子 (ｂＦＧＦ)与转化生长因子 β1基因表达调节中的作用 ,为心肌肥大的防治提供一定的理论依据。方法 :将Ｗｉｓｔａｒ乳鼠无血清培养 2 4ｈ的心肌细胞分为对照组、ｃＡＭＰ组 (8 ＢｒｏｍｏｃＡＭＰ 0 .1ｍｍｏｌ/Ｌ)、ｃＧＭＰ组 [加入一氧化氮 (ＮＯ)的供体硝普钠 1ｍｍｏｌ/Ｌ ,借此升高心肌细胞内ｃＧＭＰ浓度 ],并在 30ｍｉｎ、3ｈ、2 4ｈ、48ｈ、72ｈ等 5个时相点检测如下指标 :(1)细胞生长代谢活性 (ＭＴＴ法 ) ;(2 )ｂＦＧＦ和ＴＧＦβ1的ｍＲＮＡ表达 (原位…     

环核苷酸对心肌细胞bFGF与TGFβ_1基因表达的调控初探

目的 :探讨环核苷酸 (ｃＡＭＰ/ｃＧＭＰ)对心肌细胞碱性成纤维细胞生长因子 (ｂＦＧＦ)与转化生长因子 β1基因表达调节中的作用 ,为心肌肥大的防治提供一定的理论依据。方法 :将Ｗｉｓｔａｒ乳鼠无血清培养 2 4ｈ的细胞分为对照组、ｃＡＭＰ组 ( 8-Ｂｒｏｍｏ -ｃＡＭＰ 0 .1ｍｍｏｌ/Ｌ)、ｃＧＭＰ组 [加入一氧化氮 (ＮＯ)的供体 -硝普钠 1ｍｍｏｌ/Ｌ ,借此升高心肌细胞内ｃＧＭＰ浓度 ],并在 30ｍｉｎ、3ｈ、2 4ｈ、48ｈ、72ｈ等 5个时相点检测如下指标 :( 1 )细胞生长代谢活性 (ＭＴＴ法 ) ;( 2 )ｂＦＧＦ和ＴＧＦβ1 的ｍＲＮＡ表达 …     

血管内皮生长因子与糖尿病肾病

糖 尿病肾病 (ｄｉａｂｅｔｉｃｎｅｐｈｒｏｐａｔｈｙ ,ＤＮ)是糖尿病常见的微血管并发症之一 ,发病机制尚不完全清楚。研究表明 ,血管内皮生长因子 (ＶａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｇｒｏｗｔｈｆａｃｔｏｒＶＥＧＦ)在ＤＮ的发生发展中可能起着重要作用。本文就目前国内外在这一领域中的研究进展介绍如下。1　ＶＥＧＦ及其受体ＶＥＧＦ是一种由几种类型细胞产生的同源二聚体糖蛋白生长因子 ,目前至少发现 5种亚型[1 ] ,根据分子量的不同 ,可分为ＶＥＧＦ1 2 1 ,ＶＥＧＦ1 38,ＶＥＧＦ1 65等。ＶＥＧＦ即一种能影响血管通透性的…     

转化生长因子β胞内信号转导与Smads蛋白

转化生长因子 β(ｔｒａｎｓｆｏｒｍｉｎｇｇｒｏｗｔｈｆａｃｔｏｒ -β,ＴＧＦ -β)是ＴＧＦ -β超家族成员之一 ,分布于多种细胞组织中 ,在调节细胞增殖与分化、机体生长与发育、细胞外基质形成、免疫功能等方面均有重要作用。由于它具有广泛而重要的生物学效应 ,其信号转导通路成为诸多领域研究热点之一。最近发现 ,Ｓｍａｄｓ蛋白是ＴＧＦ -β受体 (ＴβＲ)复合物的下游信号调节蛋白 ,可将信号从胞膜直接转至胞核[1,2 ] ,从而对于ＴＧＦ -β胞内信号转导机制的认识有了进一步发展。1　ＴＧＦ -β的胞外信号转导及跨膜　　ＴＧＦ -β在…     

Accelerated tissue regeneration through incorporation of basic fibroblast growth factor-impregnated gelatin microspheres into artificial dermis

The objective of this study was to evaluate the effect of incorporation of basic fibroblast growth factor (bFGF)-impregnated gelatin microspheres into an artificial dermis on the regeneration of dermis-like tissues. When used in the free form in vivo, bFGF cannot induce sufficient wound healing activity, because of its short half-life. Therefore, sustained release of bFGF was achieved by impregnation into biodegradable gelatin microspheres. A radioisotope study revealed that incorporation of bFGF-impregnated gelatin microspheres significantly prolonged in vivo retention of bFGF in the artificial dermis. Artificial dermis with incorporated bFGF-impregnated gelatin microspheres or bFGF in solution was implanted into full-thickness skin defects on the back of guinea pigs (1.5 cm x 1.5 cm) (n = 4). Incorporation of bFGF into the artificial dermis accelerated fibroblast proliferation and capillary formation in a dose-dependent manner. However, the accelerated effects were more significant with the incorporation of bFGF-impregnated gelatin microspheres than with free bFGF at doses of 50 microg or higher. We conclude that the gelatin microsphere is a promising tool to accelerate bFGF-induced tissue regeneration in artificial dermis.     

Dramatic promotion of wound healing using a recombinant human-like collagen and bFGF cross-linked hydrogel by transglutaminase

Abstract

The basic fibroblast growth factor (bFGF) plays an important role in the wound repair process. However, lacking of better biomaterials to carry bFGF still is a challenge in skin repair and regeneration. In this study, the human-like collagen (HLC) cross-linked with transglutaminase (TG) to fabricate a HLC/TG hydrogel to load bFGF. The physical properties of hydrogel, such as interior structure, mechanical property, were characterized in vitro using scanning electron microscopy (SEM), rheometer. Then, the effects of the HLC/TG hydrogel on the bFGF and cell attachmentwere evaluated, and the results showed that the HLC/TG hydrogel has good biocompatibility towards bFGF and cells. Finally, skin wound healing test was performed for the evaluation of HLC/TG hydrogels with bFGF in a mouse model. All results of macroscopic and microscopic analysis indicated that not only our HLC/TG hydrogel provide a delivery of growth factors, but also the HLC/TG hydrogel with bFGF achieving better skin regeneration in the structure and function.     

Alveolar bone regeneration using poly-(lactic acid-co-glycolic acid-co-{varepsilon}-caprolactone) porous membrane with collagen sponge containing basic fibroblast growth factor: An experimental study in the dog

The aim of this study was to evaluate the effects of combining porous poly-lactic acid-co-glycolic acid-co-ε-caprolactone (PLGC) as a barrier membrane and collagen sponge containing basic fibroblast growth factor (bFGF) to promote bone regeneration in the canine mandible. In six beagle dogs, two lateral bone defects per side were created in the mandible. The lateral bone defects on the left side were treated with a PLGC membrane plus a collagen sponge containing bFGF. In half of these, the collagen sponge contained 50?μg of bFGF. In the other half, it contained 250?μg of bFGF. As a control, we treated the right-side bone defects in each animal with the same PLGC membrane but with a collagen sponge containing phosphate buffered saline. Computed tomography (CT) images were recorded at 3 and 6 months post-op to evaluate regeneration of the bone defects. After a healing period of 6 months, whole mandibles were removed for micro-CT and histological analyses. The post-op CT images showed that more bone had formed at all experimental sites than at control sites. At 3 months post-op, the volume of bone at defect sites covered with PLGC membrane plus 250?μg of bFGF was significantly greater than it was at defect sites covered with PLGC membrane plus 50?μg of bFGF. At 6 months post-op, however, this difference was smaller and not statistically significant. Micro-CT measurement showed that the volume of new bone regenerated at bone-defect sites, covered with PLGC membrane plus bFGF, was significantly greater than that of control sites. However, the presence or absence of bFGF in the collagen sponge did not significantly affect the bone density of new bone. These results suggest that the macroporous bioresorbable PLGC membrane plus collagen sponge containing bFGF effectively facilitates healing in GBR procedures.     

Clinical trial of allogeneic cultured dermal substitutes for intractable skin ulcers

The effect of allogeneic cultured dermal substitute (CDS) on wound healing was evaluated in 9 intractable skin ulcers in 5 patients who had failed to improve despite conventional topical treatment with basic fibroblast growth factor (bFGF) for more than 2 months. In general, the topical application of bFGF is effective in facilitating wound healing. However, skin regeneration was very slow in the present 9 cases. In this study, to improve the condition of these wounds, allogeneic CDS was applied once a week for 2 months. The wound healing process was evaluated, focusing on the reduction ratio of wound size through the granulation tissue formation associated with epithelialization. In all 9 cases, the wound size was successfully decreased after the application of CDS, and ulcers were completely resurfaced in 2 cases. In all cases, except the 2 cases showing complete wound closure, the mean wound size decreased to 33.3% of the original size, i.e., a mean reduction ratio of 33.3%. The present results indicate that allogeneic CDS can promote wound healing of intractable skin ulcers that fail to improve despite treatment with bFGF.     

Basic Fibroblast Growth Factor Promotes Extension of Regenerating Axons of Peripheral Nerve. In Vivo Experiments Using a Schwann Cell Basal Lamina Tube Model

Schwann cell basal lamina tubes serve as attractive conduits for regeneration of peripheral nerve axons. In the present study, by using basal lamina tubes prepared by in situ freeze-treatment of rat saphenous nerve, the effects of exogenously applied basic fibroblast growth (bFGF) on peripheral nerve regeneration was examined 2 and 5 days after bFGF administration. Regenerating axons were observed by light and electron microscopy using PG9.5-immunohistochemistry for specific staining of axons. In addition, the localizations of bFGF and its receptor (FGF receptor-1) were examined by immunohistochemistry using anti-bFGF antibody and anti-FGF receptor-1 antibody, respectively. Regenerating axons extended further in the bFGF-administered segment than the bFGF-untreated control segment. Electron microscopy showed that regenerating axons grew out unaccompanied by Schwann cells. Findings concerning angiogenesis and Schwann cell migration were very similar between the bFGF treated and control nerve segment. bFGF-immunoreactivity was not detected in the control nerve segment. In contrast, bFGF-immunoreactivity was detected on the basal lamina tubes as well as on the plasmalemma of regenerating axons facing the basal lamina in the bFGF treated nerve segment up to 5 days after administration, suggesting that exogenous bFGF can be retained in the basal lamina for several days after administration. FGF receptor was detected on the plasma membrane of regenerating axons where they abutted the basal lamina. These results indicate that bFGF could promote the extension of early regenerating axons by directly influencing the axons, but not via Schwann cells or angiogenesis.     

脑出血对大鼠前脑碱性成纤维细胞生长因子影响的实验研究

用免疫组织化学方法 ,对胶原酶注入尾壳核诱导的脑出血模型大鼠前脑中碱性成纤维细胞生长因子的分布进行了观察。结果显示 :脑出血后大鼠脑内此因子明显上调 ,除了病灶周围、海马及额叶、扣带皮质的神经元和神经胶质细胞表达碱性成纤维细胞生长因子外 ,还见室管膜细胞、伸展细胞以及脑室周围器官 (穹窿下器、连合下器、正中隆起 )等处的细胞表达增强。另外 ,软脑膜细胞、脑蛛网膜细胞、软脑膜下胶质界膜星形胶质细胞也表达碱性成纤维细胞生长因子。脑出血后脑内神经元和非神经元碱性成纤维细胞生长因子的上调提示它可拮抗神经元的减少与溃变、保持神经元的存活和损伤组织的修复。根据实验结果对碱性成纤维细胞生长因子的分泌和转运途径进行了讨论     

Improved neovascularization and wound repair by targeting human basic fibroblast growth factor (bFGF) to fibrin

Targeted therapy is a new generation of therapeutics, where two critical factors are involved. One is the particular molecular target, and the other is the specific target-binding drug. In this work, the fibrin, a main component of plasma clot at wound sites, was used as the target for human bFGF, aiming to improve therapeutic neovascularization and wound repair. To endow bFGF with fibrin-targeting ability, a fibrin-binding peptide Kringle1 (K1), derived from human plasminogen, was fused to human bFGF. The recombinant K1bFGF showed high fibrin and plasma-clot-binding ability. When applied to the wound sites with plasma clots, K1bFGF induced robust neovascularization and improved wound healing. To extend the application of K1bFGF to other cases where no plasma clots exist, we developed a fibrin-scaffold/K1bFGF system. This system could induce localized neovascularization by delivery of K1bFGF in a sustained and site-targeting manner, and provide a microenvironment promoting cell growth and tissue regeneration. In summary, we successfully used the pathologic environment fibrin clot as the target for bFGF, and based on which bFGF was designed into a targeting agent by introduction of a fibrin-binding peptide. This provides a potential approach to improve therapeutic neovascularization and wound repair.     

Promotion of skin regeneration in diabetic rats by electrospun core-sheath fibers loaded with basic fibroblast growth factor

Diabetic skin ulcer is difficult to heal due to the lack of cellular and molecular signals required for normal wound repair. Emulsion electrospinning was adopted to imbed basic fibroblast growth factor (bFGF) into ultrafine fibers with a core-sheath structure to promote the wound healing process. An initially burst release as low as 14.0 ± 2.2% was achieved, followed by gradual release for around 4 wk. In vitro investigations on mouse embryo fibroblasts indicated that bFGF-loaded fibrous mats enhanced cell adhesion, proliferation, and secretion of extracellular matrix (ECM). Skin wounds were created in the dorsal area of diabetic rats for in vivo evaluation of skin regeneration after covered with bFGF-loaded fibrous mats. Compared with fibrous mats infiltrated with free bFGF, bFGF-loaded scaffolds revealed significantly higher wound recovery rate with complete re-epithelialization and regeneration of skin appendages. Higher density and mature capillary vessels were generated during 2 wk after treatment with bFGF-loaded fibers, and there was no fiber fragment observed in the histological sections at week 4 after operation. The gradual release of bFGF from fibrous mats enhanced collagen deposition and ECM remodeling, and the arrangement and component of collagen fibers were similar to normal tissues. The above results demonstrate the potential use of bFGF-loaded electrospun fibrous mats to rapidly restore the structural and functional properties of wounded skin for patients with diabetic mellitus.     

Roles of exogenously regulated bFGF expression in angiogenesis and bone regeneration in rat calvarial defects

Regulation of transgene expression and function is important for gene therapy because it allows complex biological processes to be controlled and monitored. Basic fibroblast growth factor (bFGF) is an effective angiogenic factor and bone regeneration factor; it can induce differentiation of mesenchymal stem cells (MSCs) in?vitro and bone regeneration in?vivo. Further, exogenous regulation of controllable bFGF expression in the bone regeneration area safely allows bone formation and regeneration. In our study, we constructed a recombinant adeno-associated virus type 2 (rAAV2)-based bFGF gene delivery system, which is regulated by tetracycline or doxycycline (Dox, an analogue of tetracycline). We evaluated the regulatory effects of this system on bFGF transgenic expression in?vitro and in?vivo. We found that bFGF could increase the mRNA expression levels of osteoblast differentiation factor and the activity of alkaline phosphatase (ALP). Dox could effectively regulate bFGF expression, thus controlling MSC differentiation. After in?vivo transplantation of genetically engineered MSCs, animals not treated with Dox showed significant bone formation and angiogenesis compared with the group treated with Dox. Dox may also effectively regulate angiogenesis and bone regeneration in?vivo. Therefore, the inducible bFGF system is an effective way of regulating bone regeneration and formation.