Effects of basic fibroblast growth factor,transforming growth factor-β1, insulin-like growth factor-1, and insulin on human osteoarthritic articular cartilage explants

This study evaluated the effects of basic fibroblast growth factor, transforming growth factor-β1, insulin-like growth factor-1, and insulin on the incorporation of thymidine and sulfate in human osteoarthritic articular cartilage. Tissue explants were obtained from 11 patients undergoing total knee arthroplasty and were categorized as nonfibrillated or fibrillated cartilage. The explants were cultured for 22 days, with changes of medium and growth factor every 72 hours, and labeled with [³H]thymidine and [³⁵S]sulfate. Growth factors were used in the following concentrations: basic fibroblast growth factor at 1, 10, and 100 ng/ml; transforming growth factor-β1 at 0.5, 5, and 50 ng/ml; insulin-like growth factor-1 at 0.15, 1.5, and 15 ng/ml; and insulin at 0.05, 0.5, and 5 μg/ml. Basic fibroblast growth factor decreased thymidine incorporation to 70% and sulfate incorporation to less than 20% that of the growth factor-free controls. Transforming growth factor-β1 had no significant effect on thymidine incorporation, whereas the concentrations studied inhibited sulfate incorporation to approximately 40% that of the controls. At the concentrations tested, insulin-like growth factor-1 had no significant effect on incorporation of either thymidine or sulfate. In contrast, insulin significantly stimulated the incorporation of both. Compared with growth factor-free controls, insulin maximally increased thymidine incorporation by a factor (± EM) of 2.36 ± 0.47 and 1.69 ± 0.19 in nonfibrillated and fibrillated explants, respectively; sulfate incorporation was maximally increased 1.60 ± 0.24 and 1.92 ± 0.29-fold for nonfibrillated and fibrillated explants, respectively. Of the factors tested, insulin demonstrated the greatest promise for promoting a synthetic response that may contribute to the regeneration of osteoarthritic cartilage.     

Expression of basic fibroblast growth factor and its receptor-1 in cardiac myxoma

BACKGROUND: To clarify the significance of basic fibroblast growth factor (bFGF) in angiogenesis or proliferative activity in cardiac myxoma, the expression of bFGF and its receptor (FGFR-1) were immunohistochemically examined. METHODS: Formalin-embedded tissues of cardiac myxomas were obtained by surgical resection from 15 patients and analyzed by immunostaining of bFGF and FGFR-1. The microvessel density was measured in the 15 myxomas using platelet derived endothelial cell adhesion molecule-1. For evaluation of proliferative activity of the cardiac myxomas, proliferating cell nuclear antigen (PCNA) immunostaining was performed, and the PCNA labeling index was measured in each section. RESULTS: bFGF and FGFR-1 were observed in 73.3% and 67.7% of the myxomas, respectively. There was a close correlation between the expression of bFGF and FGFR-1. This co-expression was frequently observed in the myxoma cells around the microvessels appearing as a ring structure. Regarding possible relationships between the expression of bFGF or FGFR-1 and the clinicopathologic features, there were no parameters excluding the macroscopic type of myxoma. The microvessel density in the myxomas with bFGF or FGFR-1 expression was higher than that in myxomas without it. The PCNA labeling index in myxomas with bFGF expression was higher than that in myxomas without it, and the PCNA labeling index tended to be higher in myxomas with FGFR-1 expression than that in myxomas without it. CONCLUSIONS: bFGF and/or FGFR-1 was expressed in some of cardiac myxoma, and may be an important role for tumor angiogenesis and proliferative activity.     

Expression of basic fibroblast growth factor in thyroid disorders

Morphologic and biologic studies were undertaken to clarify the biologic significance of basic fibroblast growth factor (bFGF) in human thyroid neoplasms. A total of 71 malignant tumors (50 papillary carcinomas, 14 follicular carcinomas, 7 anaplastic carcinomas), 11 follicular adenomas, 6 adenomatous goiters, and 6 Graves' disease tissues were examined employing immunohistochemical methods (avidin-biotin-peroxidase complex technique). An affinity-purified polyclonal rabbit antiserum to human bFGF was used as a primary antibody. The eluate of malignant thyroid tumor tissues from the heparin-Sepharose column was examined by Western blot analysis to elucidate the molecular weight form. With immunohistochemical staining, bFGF was frequently detected in the cytoplasm of malignant thyroid tumors compared to tissues of the benign diseases and normal controls. With anaplastic carcinoma, immunoreactivity of the tumor cells was particularly strong. In the correlative analyses between UICC TNM classification and bFGF staining in papillary carcinoma, there were significant differences when relating positive staining to the grade of nodal metastases. By Western blot analysis, the bFGF immunoreactivity was specifically detected in the two forms, with molecular weights of 18 and 33 kDa. The high-molecular-weight form was detected in only anaplastic carcinoma. The present investigations demonstrated a close correlation between the expression of bFGF and the degree of malignancy. bFGF might play an important role in promoting lymph node metastases. Moreover, the high-molecular-weight form of bFGF might have an intense influence on tumor growth.

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Resumen Se emprendió un estudio morfológico y biológico, con el propósito de clarificar la significación biológica del factor básico de crecimiento de fibroblastos (FBCF) en los neoplasmas de la glándula tiroides humana.Setenta y un tumores malignos (50 carcinomas papilares, 14 carcinomas foliculares y 7 carcinomas anaplásico), 11 adenomas foliculares, 6 bocios adenomatosos y 6 tejidos de glándula con enfermedad de Graves fueron examinados mediante métodos inmunohistoquímicos (técnica del complejo avidina-biotinaperoxidasa); se utilizó un antisuero policlonal purificado contra el FBCF humano, como anticuerpo primario. Además, se utilizó el análisis de Western blot para elucidar la forma de peso molecular.Con la coloración inmunohistoquímica, el FBCF fue detectado con frecuencia en el citoplasma de los tumores malignos de la tiroides, en comparación con lo observado en la enfermedad benigna y en los controles normales. La inmunorreactividad tumoral fue particulamente fuerte en el carcinoma anaplásico. En el análisis correlativo entre la clasificiación TNM UICC y la coloración del FBCF en el carcinoma papilar, se hallaron diferencias significativas relativas a la coloración positiva y al grado de la metástasis ganglionares.En el análisis Western blot, la inmunorreactividad FBCF fue específicamente detectada en las dos formas diferentes con pesos moleculares de 18 K y 33 K. La forma de alto peso molecular fue detectada sólamante en el carcinoma anaplásico.La presente investigación demuestra una estrecha correlación entre la expresión de FBCF y el grado de malignidad. El FBCF puede jugar un papel importante en promover las metástasis ganglionares. Además, la forma de alto peso molecular del FBCF puede tener una influencia más intensa sobre el crecimiento del tumor.

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Résumé Une étude morphologique et biologique a été effectuée pour clarifier la signification du facteur de croissance des fibroblastes de base (bFGF) dans les tumeurs de la thyroïde chez l'homme. On a étudié 71 tumeurs de la thyroïde (50 cancers papillaires, 14 cancers folliculaires, 6 adénomes solitaires, et 7 cancers anaplasiques, 11 adénomes folliculaires, 6 goitres adénomateux et 6 Maladies de Basedow) en utilisant des méthodes immunohistochimiques et notamment la technique du complexe avidinebiotine-peroxydase. On a employé un anticorps primaire fabriqué à partir d'un antisérum polyclonal de lapin purifié dirigé contre le bFGF humain. L'éluat des tissus thyroïdiens malins provenant de la colonne héparine-sepharose a été examiné selon la technique Western Blot pour déterminer son poids moléculaire. Le bFGF a été détecté plus fréquemment dans le cytoplasme des tumeurs thyroïdiennes malignes que dans les maladies bénignes et les contrôles. Dans le cancer anaplasique, l'immunoréactivité des cellules cancéreuses a été particulièrement forte. En ce qui concerne la corrélation entre la classification TNM de l'Union Internationale contre le Cancer et la coloration bFGF des cancers papillaires, il y avait des différences significatives correspondant au degré d'envahissement des ganglions lymphatiques. Dans l'analyse selon la technique du Western blot, l'immunoréactivité bFGF a été détectée spécifiquement sous les deux formes de bFGF ayant des poids moléculaires de 18 et de 33 K, respectivement. Le poids moléculaire de 33K n'a été détecté que dans le cancer anaplasique. Cette étude démontre la corrélation étroite entre l'expression bFGF et le degré de malignité. Le bFGF peut probablement jouer un rôle important dans la survenue de métastase lymphatique. Le type à poids moléculaire élevé de bFGF pourrait influencer d'advantage la croissance tumorale.

     

Skull bone regeneration in primates in response to basic fibroblast growth factor.

OBJECT: The feasibility of using a biodegradable hydrogel incorporating basic fibroblast growth factor (bFGF) to induce bone regeneration at the site of a skull defect in monkeys was investigated. METHODS: Basic fibroblast growth factor was incorporated into a bioabsorbable hydrogel, which was prepared through glutaraldehyde crosslinking of gelatin. Following treatment of monkey skull defects measuring 6 mm in diameter (six defects/experimental group) with gelatin hydrogel incorporating bFGF, skull bone regeneration was evaluated using soft x-ray studies, dual x-ray absorptometry, and histological examinations. The water content of the hydrogels varied according to the glutaraldehyde concentration in the hydrogel preparation. Gelatin hydrogels incorporating 100 microg of bFGF significantly promoted bone regeneration and the skull defect was completely closed 21 weeks after implantation. This is in marked contrast with the effect of the same dose of bFGF in solution form. Bone mineral density (BMD) measured at the sites of skull defect was enhanced by the bFGF-incorporating hydrogels. The BMD enhancement was more prominent at lower water contents of hydrogel. Empty gelatin hydrogels neither induced nor interfered with skull bone regeneration. CONCLUSIONS: The findings of this study indicate that bFGF coupled with bioabsorbable hydrogel is a very promising tool to assist in the regrowth of bone at the site of a skull defect, which clinically has been recognized as almost impossible.     

Expression of basic fibroblast growth factor during distraction osteogenesis

Distraction osteogenesis is a process of tissue regeneration under an external mechanical stimulation. In the study, the spatial and temporal expression patterns of basic fibroblast growth factor in the newly formed osseous tissue during distraction osteogenesis in goats were studied using immunohistochemistry. During the distraction period, the expression of basic fibroblast growth factor was observed in the osteoblasts on the newly formed trabecular bone and the bone formation front. The cells of osteoblastic lineage and the mesenchymal cells in the distraction callus also expressed basic fibroblast growth factor. The expression of basic fibroblast growth factor in the distraction period was stronger than that during the latency and consolidation periods. However, some osteoblasts still were expressing basic fibroblast growth factor in the consolidation periods. According to these results, basic fibroblast growth factor may have a local regulatory role during distraction osteogenesis. The tensile force may stimulate the expression of basic fibroblast growth factor in osteoblasts and other cell types.     

Healing large tympanic membrane perforations using hyaluronic acid, basic fibroblast growth factor, and epidermal growth factor.

Large tympanic membrane perforations usually require a surgical tympanoplasty for closure. Reducing surgical costs and risks has encouraged investigators to examine nonsurgical office procedures for healing these perforations. Growth accelerators are the most promising agents. We study here the closure of large acute perforations using weekly applications of 1 mg of 1% hyaluronic acid (HA), 0.4 microg basic fibroblast growth factor (bFGF), or 1.0 microg epidermal growth factor (EGF) directly to the tympanic membranes of the experimental ears. Control ears were treated with 0. 1 mL Vasocidin. Complete closure was obtained in 100% of the ears treated with HA and EGF and 85.7% of those treated with bFGF by day 21, compared with 63.6% of the controls by day 32. Moderate-to-severe ipsilateral and contralateral external canal hypertrophy was noted in 14.2% and 37.5% of the ears treated with bFGF and HA, respectively, but was not seen in ears treated with EGF or in the control group.     

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。     

Effect of prostatic growth factor,basic fibroblast growth factor,epidermal growth factor,and steroids on the proliferation of human fetal prostatic fibroblasts

To study the relationship between androgen metabolism and the pathogenesis of benign prostatic hypertrophy, we purified a growth factor from benign hyperplastic tissue of human prostates and assayed the proliferative responses of human fetal prostatic fibroblasts to the purified growth factor (hPGF), basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), dihydrotestosterone (DHT), and estradiol (E₂). Prostatic tissue extracts were fractionated using heparin-Sepharose chromatography. The fraction that eluted with 1.3–1.7 M NaCl contained the majority of mitogenic activity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) of the lyophilyzed active fraction showed a band at 17,000 daltons. Human prostatic fibroblasts were isolated from fetal prostate and tested for their proliferative responses to hPGF, bFGF, EGF, DHT, and E₂. hPGF, as well as bFGF and EGF, did increase tritiated thymidine incorporation into the cultured fibroblasts. DHT(10⁻⁷ M) had a significant stimulatory effect on cell growth in serum-free media after 6 days of culture. E₂(10⁻⁷ M) had no effect on cell proliferation. The combination of DHT and E₂ showed no synergistic effect. We conclude that our purified hPGF, bFGF, and EGF promote cell growth directly, DHT indirectly, while E₂ does not. The effect of DHT appears to be mediated via the increased production and/or secretion of growth factor(s). Possibly, the bFGF-like hPGF purified from human benign hyperplastic prostatic tissue is such a mediator. © 1996 Wiley-Liss, Inc.     

增殖性肾小球肾炎中碱性成纤维细胞生长因子的表达及其作用

目的探讨碱性成纤维细胞生长因子（ｂＦＧＦ）在人类增殖性肾小球肾炎（ＰＧＮ）中的表达及作用。方法采用原位杂交、免疫组化和放免测定方法检测肾活检组织中的ｂＦＧＦｍＲＮＡ及蛋白质表达、肾小球内细胞外基质（ＥＣＭ）面积和有核细胞数、层粘蛋白（ＬＮ）和前胶原Ⅲ（ＰＣⅢ）含量。结果在以细胞增殖和ＥＣＭ积聚为特征的ＰＧＮ组中ｂＦＧＦ表达水平、ＬＮ和ＰＣⅢ含量均显著高于非增殖性肾小球肾炎（ＮＰＧＮ）组和正常组。结论过度表达的ｂＦＧＦ参与肾小球内细胞增殖和ＥＣＭ积聚，从而影响肾小球肾炎和肾小球硬化进程。     

椎间盘退变过程中碱性成纤维细胞生长因子和转移生长因子-β1的表达及其意义

目的：研究椎间盘退变过程中碱性成纤维细胞生长因子(bFGF)和转移生长因子-β1(TGF-β1)的表达及其意义。方法：收集腰椎后路手术切除的15例椎间盘源性下腰痛患者的21个通过腰椎间盘造影术证实的疼痛椎间盘，同时收集16个在MRIT2加权像上信号强度明显减弱的无腰痛症状的生理老化椎间盘和10个正常对照椎间盘，行组织学检查并用免疫组化方法检测bFGF、TGF-β1及其受体在不同椎间盘组织中的表达，观察增殖细胞核抗原在不同椎间盘的表达。结果：免疫组化染色显示bFGF、TGF-β1及其受体在疼痛椎间盘大量表达，生理老化椎间盘有少量表达，正常对照椎间盘没有表达。增殖细胞核抗原在疼痛椎间盘的肉芽组织区大量表达，在非肉芽组织区有少量表达；生理老化椎间盘有少量表达，正常对照椎间盘组织没有表达。结论：椎间盘退变和椎间盘源性下腰痛起源于椎间盘纤维环的损伤修复过程，bFGF、TGF-β1在纤维环外层的损伤修复和随之的椎间盘退变过程中可能起关键作用。     

Expression of epidermal growth factor and epidermal growth factor receptor genes in healing colonic anastomoses in rats.

This study uses complementary DNA probes to find out if epidermal growth factor (EGF) and EGF receptor genes are activated in healing colonic anastomoses in rats. 35 rats had a colocolonic anastomosis which was removed for study 1, 2, 3, 5, 7, or 14 days after operation. Six animals without surgery served as controls. The specimens were examined by northern blotting, in situ hybridisation, and conventional microscopy. Microscopic healing progressed normally. The expression of the EGF gene was minimal and surgery did not activate it. However, surgical trauma did increase the expression of the EGF receptor gene by 2.2 times when compared to day one. In situ hybridisations localised a strong EGF receptor expression in mucosal epithelial cells in all specimens, and a moderate reaction in fibroblasts in the repair tissue in the anastomotic line. The enhanced EGF receptor gene expression suggests that anastomotic healing is associated with the presence of EGF or an EGF-like substance and its activity increases during the first postoperative week.     

生长因子与老年性骨质疏松症相关性的临床研究

目的观察老年性骨质疏松患者转化生长因子β1(TGF-β1)、胰岛素样生长因子-1(IGF-1),表皮生长因子(EGF)血清水平以及与骨质疏松的相关性.方法采用双能X线骨密度仪测量了86例在本院老年病门诊和住院的老年人腰椎正位以及股骨颈、ward三角区、大粗隆部位的骨密度(BMD),受试者均为老年男性,平均年龄为69.97±5.73岁,按照WHO推荐的诊断标准,参考中国人骨质疏松症建议诊断标准(第二稿),将其分为非骨质疏松组(NOP)、腰椎骨质疏松组(OP1)和股骨上段骨质疏松组(OP2),测定所有受试者血清TGF-β1,IGF-1,EGF水平.结果腰椎骨质疏松组和股骨上段骨质疏松组血清IGF-1浓度值均低于非骨质疏松组((P＜0.01)).腰椎骨质疏松组血清TGF-β1浓度值与非骨质疏松组无明显差异((P>0.05)),股骨上段骨质疏松组血清TGF-β1浓度值低于非骨质疏松组(0.01<P<0.05),骨质疏松组血清EGF值与非骨质疏松组无差异(P>0.5).腰椎骨密度值与血清IGF-1值呈正相关(相关系数分别为0.212、0.325、0.228),腰椎骨密度值与血清TGF-β1值呈显著正相关(相关系数分别为0.373、0.328、0.341、P<0.05).股骨颈、Ward三角、大粗隆的骨密度值与IGF-1和TGF-β1无显著相关性.骨密度值与表皮生长因子无显著相关性.结论老年骨质疏松患者血清TGF-1含量低而血清TGF-β1浓度值差异较大,但从相关分析结果看,骨密度值与IGF-1和TGF-β1有较密切关系,为进一步探讨骨质疏松的发病机理,需做生长因子的骨组织表达.血清表皮生长因子(EGF)可能与骨重建无明显相关性.     

Expression of transforming growth factor beta1 gene, basic fibroblast growth factor gene and hydroxyproline in diabetes-induced bladder dysfunction in a rat model

AIMS: To investigate the content of hydroxyproline (Hyp) and the expression of transforming growth factor beta1 (TGF beta1) and basic fibroblast growth factor (bFGF) in the bladder 8 weeks after diabetes induction. METHODS: Thirty wistar rats were divided into three groups: control (n = 10), streptozotocin-induced diabetic group (n = 10), TAD group (n = 10; diabetic rats were fed with Tadenan 100 mg kg(-1) day(-1)). Eight weeks later, the bladders were dissected. RT-PCR, immunohistochemistry, and ELISA were used to detect the expression of TGF beta1 and bFGF in the bladder. Also hydroxyproline (Hyp) was measured using a method based on alkaline hydrolysis. RESULTS: The content of hydroxyproline in the diabetic group was greater than that of control group (P < 0.05); we found significantly increased expression of TGF beta1 mRNA and bFGF mRNA in the bladder from the diabetic group compared with the control group; immunohistochemical and ELISA studies showed a statistically significant increased expression of TGF beta1 protein and bFGF protein in the bladder from the diabetic group compared with the control group (P < 0.05). The content of hydroxyproline in TAD group was less than that of diabetic group (P < 0.05); mRNA expression of TGF beta1 and bFGF greatly decreased in TAD group compared with that of the diabetic group; immunohistochemical and ELISA studies showed decreased levels of TGF beta1 protein and bFGF protein in the bladder from TAD group compared with the diabetic group (P < 0.05). CONCLUSIONS: Rats with streptozoticin-induced diabetes mellitus showed significant increase in hydroxyproline, TGF beta1 and bFGF levels in their bladders, which may be an important mechanism inducing diabetic cystopathy. Tadenan could effectively reduce hydroxyproline, TGF beta1, and bFGF levels.     

FGFR1在增生性瘢痕和正常皮肤中的表达

目的　研究FGFR1在增生性瘢痕和正常皮肤中表达的差异。方法　用免疫组织化学、Western -blot、流式细胞仪分析等方法检测FGFR1在增生性瘢痕和正常皮肤组织及相关细胞中的表达差异以及在成纤维细胞中的亚细胞分布状况。结果　FGFR1阳性细胞主要存在于角质形成细胞、汗腺、皮脂腺、血管内皮细胞及成纤维细胞等 ,细胞爬片观察到阳性颗粒主要位于细胞核 ,细胞浆中不如细胞核中明显。FGFR1在单个细胞中的表达量无明显差异。结论　FGFR1在增生性瘢痕和正常皮肤中表达无差异     

FGFR1在增生性瘢痕和正常皮肤中的表达

目的研究FGFR1在增生性瘢痕和正常皮肤中表达的差异.方法用免疫组织化学、Western-blot、流式细胞仪分析等方法检测FGFR1在增生性瘢痕和正常皮肤组织及相关细胞中的表达差异以及在成纤维细胞中的亚细胞分布状况.结果 FGFR1阳性细胞主要存在于角质形成细胞、汗腺、皮脂腺、血管内皮细胞及成纤维细胞等,细胞爬片观察到阳性颗粒主要位于细胞核,细胞浆中不如细胞核中明显.FGFR1在单个细胞中的表达量无明显差异.结论 FGFR1在增生性瘢痕和正常皮肤中表达无差异.