OBJECTIVE: To determine the influence of basic fibroblast growth factor (bFGF) on endothelial cell (EC) proliferation in vitro and its possible mechanisms, and to examine the effect of both TNP-470 and dexamethasone (Dex) on the EC proliferation induced by bFGF. METHODS: Human umbilical vein endothelial cells were cultured and the proliferation of EC was quantified by a colorimetric assay using MTT reagent. The expression of nuclear factor-kappa B (NF-kappa B) and ki-67 was detected with SABC immunohistochemical method. RESULTS: bFGF stimulated the EC proliferation and enhanced the expression of NF-kappa B and ki-67 in nucleus; TNP-470 and Dex suppressed EC proliferation induced by bFGF, and reduced the expression of NF-kappa B and ki-67 in nucleus. CONCLUSION: The above results indicate that the possible mechanisms of EC proliferation stimulated by bFGF come from that bFGF can activate NF-kappa B to promote the synthesis of DNA and EC mitosis. TNP-470 and Dex inhibited EC proliferation stimulated by bFGF by inhibiting NF-kappa B.
Objective To observe the proliferation and migration of endothelial cells after 30% total burn surface area (TBSA) of deep partial thickness scald, and the effect of basic fibroblast growth factor (bFGF) on angiogenesis during wound healing.Methods A total of 133 male Wistar ratswere divided randomly into normal control (n=7), injured control group (n=42), bFGF group (n=42) andanti-c-fos group (n=42). The apoptosis expression of fibroblasts was determinedwith in situ hybridization and the changes of proliferation cell nuclear antigen(PCNA), focal adhesion rinase(FAK), c-fos and extracellular signalregulated kinase(ERK) proteins expression were detected with immunohistochemistry staining technique after 3 hours, 6 hours, 1 day, 3 days, 7 days, 14 days and 21 days of scald.Results In injured control group and bFGF group, theproliferation rate of the vascular endothelial had evident changes 7 days and14 days after scald； the expression of FAK was increased 14 days after scald. ERK proteins expression was different between injury control group and bFGF group at initial stage after scald. Stimulation of ERKs by bFGF led to up-regulation of c-fos and b expression of FAK. Conclusion Exogenous bFGF extended the influence on wound healing process by ERK signaling pathway, affecting migration cascade of vascular endothelial cell. The oncogene proteins play an important role on accelerating angiogenesis duringwound healing.
OBJECTIVE: To investigate the effects of basic fibroblast growth factor (bFGF) on the promoter activities of human alpha 1(I) procollagen gene and the interaction between bFGF and transforming growth factor-beta 1 (TGF-beta 1). METHODS: Fibroblasts of the hypertrophic scar and normal skin from a 3-year-old patient were primarily cultured and subcultured in vitro. Both of the fibroblasts were transient transfected with phCOL 2.5, containing -2.5 kb of 5’f lank sequence of human alpha 1(I) procollagen gene and CAT reporter gene by FuGENE transfection reagent; and treated thereafter by 16 ng/ml bFGF, 2 ng/ml TGF-beta 1 and 16 ng/ml bFGF + 2 ng/ml TGF beta 1 for 24 hours. The relative CAT expression values were determined by CAT-ELISA. RESULTS: TGF-beta 1 bly induced the CAT expression level, however, bFGF not only inhibited the basal CAT expression but also reduced the CAT expression up-regulated by TGF-beta 1 in normal skin and hypertrophic scar fibroblasts (P lt; 0.05). CONCLUSION: bFGF can reduce the promoter activities of human alpha 1(I) procollagen gene and antagonize the role of TGF-beta 1 in up-regulating the promoter activities of human alpha 1(I) procollagen gene in normal skin and hyertrophic scar fibroblasts.
Objective To study the adhesion-preventing effect of basic fibroblast growth factor(bFGF) combined slow-releasing degradable membrane.Methods The bFGF combined slow-releasing degradable membrane was made from bFGF and the reagent which could promote fibrinogen synthesize. Sixty-six SD rats were divided into groups A,B,C randomly (22 rats each group). In group A, sutured achilles tendon were encapsulated with bFGF combined slow-releasing degradable membrane;in group B, sutured achilles tendon were encapsulated with degradable membrane without any drug; in group C, achilles tendon were only sutured. Ninety days later, light-microscope, electronmicroscopoe, figureanalysing, hydroxyproline content, extent of peritendon adhesion and biomechanic test were evaluated.Results ①The amount of fibroblast and fibrinogen inside the sutured tendon in group A was larger than that inits peripheral connective tissue and in groups B and C (P＜0.05). Thecontent of hydroxyproline and the ultimate tensile strength in group A was higher than those in groups B and C(P＜0.01).② The peripheral tissue in group A almostremains the formal loose connective tissue, but it became dense connective tissue in groups B and C and grew into the tendon. Moreover, the extent of adhesion in group A was lesser than that in groups B, C according to the mensuration of peritendon adhesion.Conclusion The bFGF combined slow-releasing degradable membrane can make the intrinsic healing of tendon faster than peripheral
OBJECTIVE: To explore the expression of basic fibroblast growth factor(bFGF) during the wound healing of human fetal and adult skin and its significance. METHODS: We established the animal model of fetal scarless healing by transplanting full-thickness skin grafts from human fetus to a subcutaneous location on the athymic mouse recipient, and then making the linear incisions. The expression of bFGF was observed in the normal adult skin, normal fetal skin and during wound healing by immunohistochemical method. The positive staining cells were counted under selected high-power focus randomly. RESULTS: bFGF staining was not observed in the normal fetal skin and the wounded one. However, bly positive staining was shown around the vessels in normal adult skin. Moreover, the positive straining became ber in the wounded skin, especially in dermal fibroblasts and endotheliocytes. The number of positive staining cell was 2.1 +/- 0.1 in normal fetal skin, and 2.2 +/- 0.1, 2.1 +/- 0.3, 2.1 +/- 0.3 and 2.0 +/- 0.1 in the fetal skins after 12 hours, 1 day, 3 days and 7 days of wound respectively. The number of positive staining cell were 23.2 +/- 4.2 in normal adult skin and 40.5 +/- 3.6 in the wound adult skin. There was significant difference between the fetal skin and adult skin (P lt; 0.01). CONCLUSION: The negative expression of bFGF in the fetal skin may be one of the important reasons for fetal scarless healing.
Objective To observe the expression of adenovirus vector coding for mouse endostatin gene（Ad-mES） in lung cancer cells and its antiangiogenic activity in human umbilical vein endothelial cells（ECV304） in vitro.Methods Lewis lung cancer（LLC） cells were transfected with Ad-mES at different multiplicity of infection（MOI）.The expression of mES in LLC cells and supernatant after 48 hours was detected by immunohistochemical staining and Western blot respectively.The inhibitory effect of supernatant at different MOI on ECV304 non-stamulated and stimulated by basic fibroblast growth factor（bFGF） was measured by methyl thiazolyl tetrazolium（MTT） assay.Results After transfected for 48 hours，endostatin was identified in the cell plasma of infected LLC and negative result was founded in non-infected LLC.Western blot revealed band of endostatin in 20 kDa in culture supernatant of infected LLC and negative results in non-infected LLC.The inhibitory effects on ECV304 cell proliferation were ber at higher MOI，and the difference was significant between stimulated and non-stamulated cells by bFGF（Plt;0.05）.Conclusion Ad-mES can transfect and express endostatin effectively in LLC with biological activity
OBJECTIVE: To evaluate the effect of vascular endothelial growth factor(VEGF) 165 or basic fibroblast growth factor (bFGF), which was slowly-released in fibrin glue patch, on expanded prefabricated flaps in rabbits to facilitate the neoangiogenesis process. METHODS: A total of 53 rabbits were divided randomly into 6 groups. The central auricular vascular bundle of the ear was implanted into the expanded prefabricated flap as the pedicle. Fibrin glue, sandwiched between the expander and the implanted vessels, was adopted for topical delivering and slow-releasing of VEGF(625 ng) or bFGF(2880U). After 14 days, the island flap with the implanted vascular bundles as the pedicle was elevated, sutured back to its original position and then harvested more 3 days later. Neoangiogenesis was measured by digital recording of survival area, laser Doppler flowmetry, PCNA immunohistochemistry, TUNEL, ink and PbO infusions. RESULTS: When compared with the other groups, flap survival improved; neoangiogenesis of flaps increased, together with the blood flow enhanced in the groups applied growth factors. The reduced cellular apoptosis and the increased proliferation were also observed. CONCLUSION: VEGF or bFGF slowly-released by fibrin glue shows the potential to facilitate neoangiogenesis and accelerate maturation of the expanded prefabricated flap.
ObjectiveTo investigate the effects of liver X receptor agonist, T0901317, on the proliferation, migration and hydroxyproline production of human embryonic lung fibroblasts (HELF). MethodsHELF cells were devided into a control group, a growth factor group, a T0901317 group and three growth factor+T0901317 groups. The cells of the control group were treated with Dulbecco's modified Eagle medium. The cells of T0901317 group were treated with 1.00 μmol/L T0901317. The growth factor+T0901317 groups were incubated with different doses of T0901317 (0.25 μmol/L, 0.50 μmol/L and 1.00 μmol/L) for 2 h. Then the cells of the growth factor+T0901317 groups and the growth factor group were incubated with basic fibroblast growth factor and transforming growth factor-β1 for 24 h. The proliferation, migration and collagen production of HELF were determined by cell counting kit-8 method, transwell chamber, and hydroxyproline method. ResultsCompared with the control group, T0901317 had no effect on the proliferation, migration and hydroxyproline production of HELF. Growth factors could promote the proliferation, migration and hydroxyproline production of HELF significantly. T0901317 could inhibit those effects of growth factors with a dosage-dependent manner. ConclusionT0901317 may inhibit the proliferation, migration and hydroxyproline production of HELF induced by growth factors.
OBJECTIVE: To investigate the influence of basic fibroblast growth factor (bFGF) on adhesion characteristics of osteoblasts, aimed at the important problem in bone tissue engineering of how to promote the adherence of osteoblasts to extracellular matrix materials. METHODS: 5 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml bFGF were used to induce bone marrow stromal-derived osteoblasts of rabbit for 24 hours before incubation, and the common culture medium as the control. The attached cells were calculated with stereology method at 0.5 hour, 1st hour, 2nd hour, 4th hour, 8th hour after seeding. RESULTS: The number of attached cells was significant higher in the experimental group when induced by 10 ng/ml bFGF than that in the control group (P lt; 0.01); the number did not increase with the increase of bFGF concentration and there was no significant difference between the experimental group induced by 100 ng/ml bFGF and control group, and the number was even obviously lower in the experimental group when induced by 200 ng/ml than the control group (P lt; 0.01). CONCLUSION: bFGF can influence the adhesion characteristics of osteoblasts, 10 ng/ml bFGF can promote the adherence of osteoblasts to matrix materials, but 200 ng/ml bFGF may inhibit cell adhesion.