Time course of de novo adipogenesis in matrigel by gelatin microspheres incorporating basic fibroblast growth factor

Controlled release of basic fibroblast growth factor (bFGF) from gelatin microspheres achieved de novo adipogenesis at the implanted site of a basement membrane extract (Matrigel). Following subcutaneous co-implantation of Matrigel and gelatin microspheres incorporating 0.1 microg of bFGF into the back of mice, adipose tissue was formed at the implanted site after 4 weeks postoperatively although the extent increased with implantation time. Formation of adipose tissue was significantly faster than the co-implantation of Matrigel, and 0.1 microg of free bFGF while a larger volume of the adipose tissue formed was retained 15 weeks later. When measured in Matrigel co-implanted with the gelatin microspheres incorporating bFGF, the number of cells infiltrated into Matrigel increased to a significantly high extent compared with the bFGF co-implantation. Matrigel alone was much less effective in inducing formation of adipose tissue. We conclude that gelatin microspheres incorporating bFGF enable Matrigel to efficiently induce de novo adipogenesis at the implanted site in respect to the formation rate and volume of adipose tissue.     

Neovascularization effect of biodegradable gelatin microspheres incorporating basic fibroblast growth factor

Biodegradable microspheres were prepared through glutaraldehyde cross-linking of gelatin without using any surfactants as a carrier matrix of basic fibroblast growth factor (bFGF). In the in vitro system, bFGF was sorbed to microspheres of acidic gelatin with an isoelectric point (IEP) of 5.0, but not to those of basic gelatin with an IEP of 9.0. The rate of bFGF sorption to the acidic gelatin microsphere in phosphate-buffered saline solution (pH 7.4) was smaller than that in water. Following incorporation of bFGF into the microspheres at 4 degrees C for 12 h, bFGF release from the bFGF-incorporating microspheres was studied. Approximately 30% of incorporated bFGF was released from the acidic gelatin microsphere within the initial 3 h, followed by no substantial release, whereas the basic gelatin microsphere released almost completely the incorporated bFGF within 1 day. It is likely that when basic bFGF molecules were immobilized to the acidic gelatin constituting microspheres through polyion complexation, they were not readily released under the in vitro nondegradation condition of gelatin. Incorporation of anionic carboxylmethyl cellulose (CMC) into the acidic gelatin microspheres reduced the amount of bFGF desorbed initially. This indicates that the initial burst is ascribed to free bFGF which is not ionically interacted with the acidic gelatin. CMC will function as a bFGF sorbent to suppress the initial leakage from the microspheres. When injected subcutaneously into the mouse back, bFGF-incorporating acidic gelatin microspheres were degraded over time and induced neovascularization around the injection site, in marked contrast to bFGF in the solution form. CMC incorporation slowed down the biodegradation and vascularization effect of bFGF-incorporating gelatin microspheres. It was concluded that the gelatin microsphere was a promising carrier matrix of bFGF to enhance the vascularization effect.     

白藜芦醇上调碱性成纤维细胞生长因子、胰岛素样生长因子1表达治疗骨骼肌损伤

文题释义： 白藜芦醇：非黄酮类的多酚化合物,分子式为C₁₄H₁₂O₃,相对分子质量228.25,为白色针状晶体,易溶于乙醚、氯仿、甲醇、乙醇、丙酮、乙酸、乙酯等有机溶剂。别名：3,4',5-三羟基芪、虎杖甙元,是相关植物在受到病菌侵染或环境恶化时产生的植物抗毒素,主要存在于葡萄、虎杖、决明、花生、桑葚等植物中。 骨骼肌急性钝挫伤：钝挫伤指在钝器作用下,造成以皮内或皮下及软组织出血为主要改变的闭合性损伤。骨骼肌急性钝挫伤指钝器在短时间内伤到肌肉层造成皮下及软组织出血,肌肉没有断裂或者撕裂的闭合性损伤。临床上90%的骨骼肌损伤属于钝挫伤与扭伤。 背景：近年来国内外对白藜芦醇抑制机体组织纤维化方面做了大量研究,但其在肌组织损伤康复方面的作用却鲜有报道。 目的：观察骨骼肌急性钝挫伤修复过程中碱性成纤维细胞生长因子、胰岛素样生长因子1蛋白表达规律,探讨白藜芦醇促进受损骨骼肌结构与功能恢复的作用机制。 方法：33只新西兰兔随机分为3组：正常组(3只)、自然恢复组(15只)、白藜芦醇组(15只),除正常组外均采用钝性暴力法制造骨骼肌钝挫伤模型,损伤后自然恢复组不予处理,白藜芦醇组给予白藜芦醇灌胃治疗,分别于伤后1,3,7,14,21 d处死动物,采用苏木精-伊红染色、Masson染色观察炎症细胞浸润情况、胶原纤维形成情况,免疫组织化学、免疫印记法检测骨骼肌中碱性成纤维细胞生长因子、胰岛素样生长因子1蛋白表达。 结果与结论：①苏木精-伊红染色显示：正常组兔肌纤维多边形、形态规则、排列紧密,肌核均匀分布于肌膜下,无增生与固缩,肌膜完整;自然恢复组伤后1 d见血细胞渗出,3 d炎症细胞开始浸润,至7 d达峰值,21 d肌纤维形态基本恢复正常;白藜芦醇组在炎症细胞浸润、修复时间上整体优于自然恢复组;②Masson染色显示：正常肌细胞中胶原纤维含量极少;自然恢复组随着瘢痕组织的形成,胶原纤维逐渐增加,于14 d达高峰;白藜芦醇组胶原纤维含量低于自然恢复组;③免疫组织化学和免疫印记检测显示：碱性成纤维细胞生长因子、胰岛素样生长因子1蛋白在骨骼肌修复过程中呈现先升高后降低的变化规律,两组均于7 d达高峰,21 d时仍高于正常,且白藜芦醇组峰值高于自然恢复组;④整体来看,白藜芦醇组在炎症反应以及修复程度上均优于自然恢复组,白藜芦醇通过上调碱性成纤维细胞生长因子、胰岛素样生长因子1蛋白表达来促进骨骼肌修复,但其并不改变骨骼肌损伤修复过程中蛋白表达量的整体变化规律。 ORCID: 0000-0001-6570-2052(刘杏) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

De novo formation of adipose tissue by controlled release of basic fibroblast growth factor

De novo adipogenesis at the implanted site of a basement membrane extract (Matrigel) was induced through controlled release of basic fibroblast growth factor (bFGF). bFGF was incorporated into biodegradable gelatin microspheres for its controlled release. When the mixture of Matrigel and bFGF-incorporated gelatin microspheres was implanted subcutaneously into the back of mice, a clearly visible fat pad was formed at the implanted site 6 weeks later. Histologic examination revealed that the de novo formation of adipose tissue accompanied with angiogenesis was observed in the implanted Matrigel at bFGF doses of 0.01, 0.1, and 1 microg/site, the lower and higher doses being less effective. The de novo formation induced by the bFGF-incorporated microspheres was significantly higher than that induced by free bFGF of the same dose. The mRNA of a lipogenesis marker protein, glycerophosphate dehydrogenase, was detected in the formed adipose tissues, biochemically indicating de novo adipogenesis. Free bFGF, the bFGF-incorporated gelatin microspheres, or Marigel alone and bFGF-free gelatin microspheres with or without Matrigel did not induce formation of adipose tissue. This de novo adipogenesis by mixture of Matrigel and the bFGF-incorporated gelatin microspheres will provide a new idea for tissue engineering of adipose tissue.     

Sequential delivery of basic fibroblast growth factor and platelet-derived growth factor for angiogenesis

An externally regulated delivery model that permits temporal separation of multiple angiogenic factors was used for the delivery of basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF). While bFGF plays a significant role in the sprouting of new capillaries, PDGF plays a role in the recruitment of mural cells, which stabilize neovessels. However, these two factors have been shown to inhibit each other, when presented together. Using the externally regulated model, sequential delivery of bFGF and PDGF led to not only increased endothelial cell migration, but also endothelial cell and vascular pericyte colocalization. More importantly, this delivery strategy was able to induce red blood cell-filled neovessels, suggesting integration of angiogenesis with the existing vasculature.     

Promoted growth of murine hair follicles through controlled release of basic fibroblast growth factor

This study is an investigation to evaluate how the controlled release of basic fibroblast growth factor (bFGF) affects the hair follicle growth of mice in different hair cycle stages: second anagen and second telogen. bFGF was incorporated into biodegradable gelatin hydrogels for its controlled release. After subcutaneous implantation of gelatin hydrogels incorporating 0, 0.7, 7, and 70 microg of bFGF or injection of 0 and 70 microg of free bFGF into the backs of mice, hair follicle growth was evaluated photometrically and histologically on the basis of three parameters: skin color of the reverse side of the implanted or injected site, skin thickness, and area occupied by hair follicle tissue. For mice in second anagen, the darkness of the reverse side of skin implanted with gelatin hydrogel incorporating 7 microg of bFGF was significantly higher than that of skin injected with 70 microg of bFGF 10 days after bFGF application. Implantation of gelatin hydrogel incorporating bFGF enabled the hair follicles to increase the area occupied in skin tissue to a significantly greater extent than in other groups, whereas no effect on skin thickness was observed. bFGF-free, empty gelatin hydrogels did not affect hair follicle growth. Moreover, hair shaft length was significantly elongated by gelatin hydrogel incorporating 7 microg of bFGF, in marked contrast to other agents. The skin of telogen mice receiving gelatin hydrogel incorporating 7 microg of bFGF did not show any change in darkness of reverse skin side or skin thickness, but a significant increase in the size of hair follicles 10 days later. These results indicate that the controlled release of bFGF positively affects the hair growth cycle of mice.     

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.     

肠道缺血再灌流对肾内源性碱性成纤维细胞生长因子和 …

目的和方法：研究肠道缺血再灌流过程对肾脏内源性碱性成纤维细胞生长因子和转化生长因子β基因表达的影响，采用肠系膜上动脉夹闭４５ｍｉｎ与再灌流６ｈ和２４ｈ动物模型，用原位杂交与逆转录ＰＣＲ（ＲＴ－ＰＣＲ）技术研究两种生长因子基因在正常，缺血以及再灌流肾组织中的表达。     

碱性成纤维细胞生长因子及胰岛素样生长因子1对精原干细胞增殖凋亡影响的机制

文题释义：碱性成纤维细胞生长因子：是细胞生长和分化的重要调节因子,具有促血管生成、细胞增殖、细胞趋化、细胞迁移等活性,在细胞分化和机体发育过程中发挥重要作用。碱性成纤维细胞生长因子通过与细胞膜表面的特异性配体结合,进而引发细胞内的一系列级联反应,从而产生各种生物学效应。 胰岛素样生长因子1：是多功能细胞增殖调控因子,主要分布在肝脏中,其与胰岛素样生长因子2、胰岛素及其受体一起构成了胰岛素样生长因子家族,其对细胞生长和代谢具有多效作用。 背景：生长因子作为体外细胞培养和体内细胞生长及增殖必需的调节因子,一直被广泛的关注。 目的：探讨碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)与胰岛素样生长因子1(insulin-like growth facter,IGF-1)联合作用对小鼠精原干细胞增殖、凋亡的影响。 方法：从6-8 d龄昆明雄性小鼠睾丸内分离培养精原干细胞并进行鉴定。将精原干细胞接种于经丝裂霉素C处理过的胚胎成纤维细胞饲养层上,分组干预：对照组加入正常DMEM培养基进行培养;bFGF、IGF-1组分别加入含20 μg/L bFGF、20 μg/L IGF-1的DMEM培养基进行培养;bFGF+IGF-1组同时加入含20 μg/L bFGF及20 μg/L IGF-1的DMEM培养基进行培养。采用CCK-8、EDU染色法分别检测精原干细胞增殖活性,流式细胞仪检测精原干细胞生长周期和细胞凋亡情况,Western blot检测增殖和凋亡相关蛋白PCNA、Bax、Bcl-2的表达。 结果与结论：①与对照组比较,bFGF组、IGF-1组、bFGF+IGF-1组吸光度值显著升高,与bFGF组、IGF-1组比较,bFGF+IGF-1组吸光度值进一步升高(P < 0.05),EDU染色得到与CCK-8实验一致的结论;②bFGF+IGF-1组S+G₂/M期细胞比例明显高于其他3组(P < 0.05),IGF-1组、bFGF组S+G₂/M期细胞比例高于对照组(P < 0.05);③与对照组比较,bFGF组、IGF-1组、bFGF+IGF-1组凋亡细胞降低;与bFGF组、IGF-1组比较,bFGF+IGF-1组凋亡细胞进一步降低;④与对照组比较,bFGF组、IGF-1组、bFGF+IGF-1组细胞中Bax蛋白相对表达水平显著下降(P < 0.01),Bcl-2和PCNA蛋白相对表达水平均显著升高(P < 0.05)。与bFGF组、IGF-1组比较,bFGF+IGF-1组细胞中Bax蛋白相对表达水平进一步下降(P < 0.01),Bcl-2和PCNA蛋白相对表达水平进一步升高(P < 0.05);⑤结果表明,bFGF、IGF-1通过上调PCNA和Bcl-2蛋白的表达,下调Bax蛋白的表达,促进细胞增殖,抑制细胞凋亡,二者联合作用效果最佳。 ORCID: 0000-0001-5693-3713(李宏) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

Importance of vascular phenotype by basic fibroblast growth factor, and influence of the angiogenic factors basic fibroblast growth factor/fibroblast growth factor receptor-1 and ephrin-A1/EphA2 on melanoma progression

The expression of several angiogenic factors and receptors was examined in a series of vertical growth phase cutaneous melanomas using high-throughput tissue microarray technology and immunohistochemistry. The results were correlated with microvessel density, clinicopathological features, and patient survival. Expression of basic fibroblast growth factor (bFGF) was significantly associated with increased microvessel density. Also, we found an independent prognostic importance of vascular phenotype by endothelial cell expression of bFGF; cases with positive vessels had the best prognosis and these tumors revealed a low frequency of vascular invasion (14%) when compared with bFGF-negative vessels (47%). This bFGF-negative phenotype was significantly increased in metastatic lesions. Strong tumor cell expression of FLT-4, ephrin-A1, and EphA2 was associated with increased melanoma thickness, and ephrin-A1 staining was related to decreased survival (P = 0.039). Expression of EphA2 in tumor cells was associated with increased tumor cell proliferation (Ki-67 positivity), indicating possible autocrine growth stimulation. Thus, our findings indicate the presence of phenotypic diversity among tumor-associated vessels, and subgroups defined by bFGF expression may be of clinical importance. bFGF was associated with microvessel density, whereas the ephrin-A1/EphA2 pathway might also be important for tumor cell proliferation and patient survival.     

碱性成纤维细胞生长因子共聚物缓释微球与兔跨区轴型皮瓣的成活

背景：研究表明使用生长因子直接或间接刺激新生血管形成可以促进皮瓣远端缺血部分的成活。 目的：观察碱性成纤维细胞生长因子-聚乳酸-聚羟基乙酸共聚物缓释微球对家兔侧腹制作跨区轴型皮瓣成活的影响。 方法：取24只健康家兔制作侧腹壁跨区轴型皮瓣,随机分组：实验组术前5 d皮内注入碱性成纤维细胞生长因子-聚乳酸-聚羟基乙酸共聚物微球,对照组术前5 d皮内注入碱性成纤维细胞生长因子+空微球悬浊液,空白对照组术前5 d皮内注入生理盐水。5 d后掀起皮瓣原位缝合。 结果与结论：①皮瓣成活率：实验组显著高于对照组和空白对照组(P < 0.01),②皮瓣组织学变化：实验组新生血管增生明显,以毛细血管为主。③CD34+免疫组织化学结果：实验组新生血管大量形成,平均血管数目高于对照组和空白对照组(P < 0.05)。表明术前5 d局部注射碱性成纤维细胞生长因子-聚乳酸-聚羟基乙酸共聚物缓释球可促进皮瓣新生血管形成,增加皮瓣血运,促进皮瓣成活。     

Enhanced angiogenesis through controlled release of basic fibroblast growth factor from peptide amphiphile for tissue regeneration

In the present study, we hypothesized that a novel approach to promote vascularization would be to create injectable three-dimensional (3-D) scaffolds with encapsulated growth factor that enhance the sustained release of growth factor and induce the angiogenesis. We demonstrate that a 3-D scaffold can be formed by mixing of peptide-amphiphile (PA) aqueous solution with basic fibroblast growth factor (bFGF) suspension. PA was synthesized by standard solid phase chemistry that ends with the alkylation of the NH(2) terminus of the peptide. A 3-D network of nanofibers was formed by mixing bFGF suspensions with dilute aqueous solutions of PA. Scanning electron microscopy (SEM) observation revealed the formation of fibrous assemblies with an extremely high aspect ratio and high surface areas. In vitro and in vivo release profile of bFGF from 3-D network of nanofibers was investigated while angiogenesis induced by the released bFGF was assessed. When aqueous solution of PA was subcutaneously injected together with bFGF suspension into the back of mice, a transparent 3-D hydrogel was formed at the injected site and induced significant angiogenesis around the injected site, in marked contrast to bFGF injection alone or PA injection alone. The combination of bFGF-induced angiogenesis is a promising procedure to improve tissue regeneration.     

Enhancing the vascularization of three-dimensional porous alginate scaffolds by incorporating controlled release basic fibroblast growth factor microspheres

Site-specific delivery of angiogenic growth factors from tissue-engineered devices should provide an efficient means of stimulating localized vessel recruitment to the cell transplants and would ensure cell survival and function. In the present article, we describe the construction of a novel porous alginate scaffold that incorporates tiny poly (lactic-co-glycolic acid) microspheres capable of controlling the release of angiogenic factors, such as basic fibroblast growth factor (bFGF). The microspheres are an integral part of the solid alginate matrix, and their incorporation does not affect the scaffold porosity or pore size. In vitro, bFGF was released from the porous composite scaffolds in a controlled manner and it was biologically active as assessed by its ability to induce the proliferation of cardiac fibroblasts. The controlled delivery of bFGF from the three-dimensional scaffolds accelerated the matrix vascularization after implantation on the mesenteric membrane in rat peritoneum. The number of penetrating capillaries into the bFGF-releasing scaffolds was nearly fourfold higher than into the control scaffolds (those incorporating microspheric BSA and heparin but not bFGF). At day 10 posttransplantation, capillary density in the composite scaffolds was 45 +/- 3/mm(2) and it increased to 70 +/- 7/mm(2) by day 21. The released bFGF induced the formation of large and matured blood vessels, as judged by the massive layer of mural cells surrounding the endothelial cells. The control over bFGF delivery and localizing its effects to areas of need, may aid in the wider application of bFGF in therapeutic angiogenesis as well as in tissue engineering.     

Coexpression of heparanase, basic fibroblast growth factor and vascular endothelial growth factor in human esophageal carcinomas

Heparan sulfate (HS), which is degraded by heparanase, plays an important role in cell adhesion, insolubility of the extracellular matrix (ECM) and as a reservoir for various growth factors such as basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF). In the present study, we examined the immunohistochemical expression of heparanase, bFGF and VEGF, and evaluated the correlation between their expression and microvessel density (MVD) in human esophageal carcinomas. Heparanase, bFGF and VEGF were immunolocalized predominantly to the carcinoma cells, but they were also localized to the endothelial cells of microvessels near the carcinoma cell nests. In carcinomas with invasion of the muscular layer or adventitia, heparanase staining was stronger at the invasive areas of carcinomas than the intraepithelial spread. Expression of heparanase and bFGF and the degree of MVD were associated with tumor invasion, lymph node metastasis and pathological stages. Cases with positive staining for heparanase, bFGF or VEGF tended to have a higher MVD than those without staining, and carcinomas with concomitant expression of heparanase, bFGF and VEGF showed the highest MVD. The level of heparanase mRNA expression was directly correlated with the MVD. In addition, heparanase-positive cases had a higher positive ratio of bFGF and VEGF compared with the heparanase-negative cases. These data suggest the possibility that heparanase may contribute to not only cancer cell invasion but also angiogenesis probably through degradation of HS in the ECM and release of bFGF and VEGF from the HS-containing ECM.     

重组牛碱性成纤维细胞生长因子与胰岛素联合应用治疗压疮创面：随机对照

背景：长期卧床可导致压疮,促进压疮创面的愈合,缩短压疮的治愈时间是压疮治疗的关键。 目的：观察胰岛素对重组牛碱性成纤维细胞生长因子治疗压疮创面愈合的影响。 方法：选择2009-01/2010-05四川省南充市中心医院收治的压疮病例34例共45处压疮,采用单盲随机分组原则分为实验组和对照组。TDP-L-1-2特定电磁波理疗后,实验组创面加入重组牛碱性成纤维细胞生长因子外用溶液1~3 mL(轻度1 mL,中度2 mL,重度3 mL)+生理盐水3~5 mL(轻度3 mL,中度4 mL,重度5 mL)+胰岛素;对照组不加胰岛素。记录两组治疗两周显效情况和创面愈合时间,并进行对比分析。 结果与结论：实验组治疗有效率、压疮创面治愈时间明显高于对照组(P < 0.05),但两组患者治疗前血糖浓度差异无显著性意义(P > 0.05)。提示胰岛素的加入可以促进压疮的愈合,而不引起患者血糖浓度的改变。     

Controlled and modulated release of basic fibroblast growth factor.

Basic fibroblast growth factor has multivariate effects in stimulating cell growth and the processes that surround tissue repair. Pathophysiologic studies have been hampered by the stability of the compound. Though very potent, basic fibroblast growth factor is rapidly degraded when injected or ingested. Controlled release of basic fibroblast growth factor would allow for examination of the chronic effects of this compound. Conventional matrix polymer-based release devices were fabricated and basic fibroblast growth factor released in a sustained fashion, but 99% of basic fibroblast growth factor mitogenic activity was lost. The source of these losses was identified and preventative measures examined. Preservation and stabilization of basic fibroblast growth factor was accomplished by binding the factor to heparin-Sepharose beads. This permitted prolonged storage, repeated handling, and the encapsulation of basic fibroblast growth factor within a microspherical controlled-release device using a naturally occurring polymer material, alginate. Encapsulation was accomplished with 77% efficiency and 87.5 +/- 12% of the basic fibroblast growth factor was released in a biologically active form. Release activation and regulation was achieved when cleavage of the basic fibroblast growth factor-heparin bonds was enhanced (e.g. by enzymatic bond cleavage with heparinase). Kinetic profiles were identified for a variety of experimental conditions and the effects of the controlled release of basic fibroblast growth factor on BALBc/3T3 fibroblasts examined.     

碱性成纤维细胞生长因子及胰岛素样生长因子影响骨髓间充质干细胞增殖及胶原合成

BACKGROUND: How to control the orderly formation of collage in skin repair and scarring process is worthy of attention. OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF) combined with insulin-like growth factor 1 (IGF-1) on the proliferation and collagen synthesis of rat bone marrow mesenchymal stem cells in vitro. METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured to induce adipogenic differentiation assessed by oil red O staining and osteogenic differentiation identified by alizarin red staining in vitro. Passage 3 cells were cultured in the medium containing bFGF, IGF-1, combination of them or the control fluid, respectively. MTT assay was used to detect cell proliferation at 12, 24, 48, 72 and 96 hours of culture. The expression of type I collagen and type III collagen were detected by RT-PCR and western blot after 10 days of incubation. RESULTS AND CONCLUSION: Compared with the control group, bFGF or IGF-1 alone significantly promoted the proliferation of bone marrow mesenchymal stem cells, and inhibited the expression of type I collagen and type III collagen. After combined use of bFGF and IGF-1, the proliferation of bone marrow mesenchymal stem cells was improved more significantly, and the expression of type I collagen and type III collagen returned to normal levels. These findings indicate that the combination of IGF-1 and bFGF can promote proliferation of bone marrow mesenchymal stem cells and restrain the expression of type I collagen and type III collagen, which may be helpful for control and repair of scar formation during wound healing.      

Plasma concentrations of vascular endothelial growth factor and basic fibroblast growth factor in lymphoproliferative disorders

Angiogenesis plays a major role in the development and progression of haematological malignancies. In our study we measured plasma concentrations of key angiogenic activators vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) using comercially available sandwich enzyme-linked immunosorbent assay (ELISA) in 37 patients with lymphoid malignancies and 20 healthy donors. We found a statistically significant increase in bFGF concentrations in patients with B-cell chronic lymphocytic leukemia (B-CLL, n=18) compared to the control group (median 118.8 vs. 9.3 pg/ml, p<0.001). However, we didn't find any significant difference in VEGF concentrations between B-CLL patients and the control group. There was also no significant increase in bFGF or VEGF in patients with multiple myeloma (n=7) and non-Hodgkin's lymphoma (n=12). Our pilot study shows that measurement of angiogenic activators in plasma is a feasible and reproducible method of angiogenesis assessment. Larger studies are needed for correlation between serum and plasma concentrations and detailed statistical evaluation including the impact on patients' survival.     

Expression of vascular endothelial growth factor and basic fibroblast growth factor in extramammary Paget disease

Extramammary Paget’s disease (EMPD) is a special type of cancers. The etiology of the disease is still unclear. We aimed to study the expression differences of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in EMPD tissues and corresponding adjacent normal tissues. The mRNA expression was detected by RT-PCR and the protein expression was explored by immunohistochemistry. Higher immunostaining signal scores of bFGF and VEGF in EMPD tissues had been found (z = -3.827, P < 0.001, z = -3.729, P < 0.001, respectively). In addition, the mRNA expression of bFGF and VEGF was higher in EMPD tissues, which had been validated by RT-PCR (t = 5.771, P < 0.001, t = 3.304, P = 0.004, respectively). The VEGF and bFGF might be the key signaling proteins in angiogenesis of EMPD. How to block the VEGF and bFGF in EMPD and to destroy the blood supply of the tumor cells becomes the focus of our future research.     

Injectable glycosaminoglycan hydrogels for controlled release of human basic fibroblast growth factor

Synthetic hydrogel mimics of the extracellular matrix (ECM) were created by crosslinking a thiol-modified analog of heparin with thiol-modified hyaluronan (HA) or chondroitin sulfate (CS) with poly(ethylene glycol) diacrylate (PEGDA). The covalently bound heparin provided a crosslinkable analog of a heparan sulfate proteoglycan, thus providing a multivalent biomaterial capable of controlled release of basic fibroblast growth factor (bFGF). Hydrogels contained >97% water and formed rapidly in <10min. With as little as 1% (w/w) covalently bound heparin (relative to total glycosaminoglycan content), the rate of release of bFGF in vitro was substantially reduced. Total bFGF released increased with lower percentages of heparin; essentially quantitative release of bFGF was observed from heparin-free hydrogels. Moreover, the hydrogel-released bFGF retained 55% of its biological activity for up to 28 days as determined by a cell proliferation assay. Finally, when these hydrogels were implanted into subcutaneous pockets in Balb/c mice, neovascularization increased dramatically with HA and CS hydrogels that contained both bFGF and crosslinked heparin. In contrast, hydrogels lacking bFGF or crosslinked heparin showed little increase in neovascularization. Thus, covalently linked, heparin-containing glycosaminoglycan hydrogels that can be injected and crosslinked in situ constitute highly promising new materials for controlled release of heparin-binding growth factors in vivo.