To study the expression of the bFGF and its receptor in the mouse bone marrow by treatment with acute radioactive injury and Ligustrazine, 56 mice were divided into 3 groups: normal group, radiation-injured group and Ligustrazine group. After irradiation by 6.0 Gy 60Co gamma-ray, each mouse was orally given 0.1 ml Ligustrazine twice a day for 13 days in Ligustrazine group, and each mouse in radiation injured group was orally given equal amount of saline. On the 3rd, 7th, 14th day after irradiation, bone marrow mono-nuclear cells (BMMNC) were counted, and the expression levels of bPGF and bFGFR in bone marrow were evaluated by immunohistochemistry and flow cytometry analysis respectively. On the 3rd, 7th, 14th day after irradiation, expression of bFGF in bone marrow were significantly lower than in normal group (P<0.05 or P<0.01). Expressions of bFGF and bFGFR were much higher in Ligustrazine treated group than that in the control group (P<0.05 or P<0.01). Ligustrazine potentiate the expression of bFGF and bFGFR in bone marrow MNC to recover the bone marrow hematopoiesis inductive microenvironment, which is one of the mechanisms by which Ligustrazine rebuild the bone marrow hematopoiesis after acute radioactive injury.
Bone Marrow Cells/metabolism ; Fibroblast Growth Factor 2/*biosynthesis ; Hematopoiesis/drug effects ; Pyrazines/*pharmacology ; Radiation Injuries, Experimental/*metabolism ; Radiation-Protective Agents/pharmacology ; Receptors, Fibroblast Growth Factor/*biosynthesis

OBJECTIVETo investigate the effect of recombinant human basic fibroblast growth factor (rhbFGF) on angiogenesis during mandible fracture healing in rabbit.

METHODSFifty adult white rabbits were used for animal model and randomly divided into a control group (25 rabbits) and an experimental group (25 rabbits). The membranous complex of rhbFGF and bovine type I collagen was prepared and implanted into the rabbit mandible fracture site under periosteum. The animals were sacrificed on 7, 14, 28, 56 and 84 days respectively after operation and the whole mandibles were harvested. The expression of factor VIII related antigen (F8-RA) in callus was examined with immunohistochemical staining.

RESULTSThe amounts of microvascular formation in calluses in the rhbFGF-treating group on days 7, 14, 28 and 56 were more than those of the control group (P<0.01).

CONCLUSIONSThe results indicated that rhbFGF could stimulate microvascular formation during mandible fracture healing in rabbits.

Animals ; Fibroblast Growth Factor 2 ; pharmacology ; Fracture Healing ; physiology ; Mandibular Fractures ; physiopathology ; Neovascularization, Physiologic ; drug effects ; Rabbits ; Recombinant Proteins ; pharmacology

OBJECTIVETo evaluate the effect of basic fibroblast growth factor (bFGF) treated collagen composite sponge on vascularization of HA orbital implants.

METHODSNew Zealand rabbits received three different orbital implants:naked implants, implants wrapped with collagen composite sponge and implants wrapped with bFGF treated collagen composite sponge.Implants were harvested 2, 4, 6, 8 and 12 weeks after surgery. The vascularization of implants was then assessed by light and electron microscopy.

RESULTSAt post-surgery weeks of 2, 4 and 6, bFGF treated collagen composite sponge induced the highest degree of vascularization of orbital implants. Collagen composite sponge alone resulted in higher extent of vascularization than naked implants. Complete vascularization of implants was observed at post-surgery 6 weeks by bFGF treated collagen composite sponge, which was not observed in the other two groups until post-surgery 8 weeks. There were significant differences in the average length of fibrovasculature and in the degree of vascularization among each group at post-surgery 2, 4 and 6 weeks (P<0.05), while no statistical difference was observed at post-surgery 8 and 12 weeks (P>0.05).

CONCLUSIONSbFGF treated collagen composite sponge facilitates fibrovascularization of orbital implants, and shortens the time required for complete vascularization. Collagen composite sponge alone promotes early-stage fibrovascularization, but fails to facilitate complete vascularization of orbital implants.

Animals ; Collagen ; pharmacology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Male ; Neovascularization, Physiologic ; drug effects ; Orbital Implants ; Rabbits

OBJECTIVETo investigate the mechanism and the accelerating effect of rhEGF and rhbFGF on wound healing.

METHODSTwelve New Zealand rabbits with 72 incised wounds on ventral side of 24 ears were randomly divided into two therapeutic groups (rhEGF of 10 ug/cm(2) and rhbFGF of 100 AU/cm(2)) and a control group (1% silver sulfadiazine cream, SD-Ag). The general conditions of the wound healing was observed grossly. Biopsies were harvested at different time points for the pathomorphological examination, the electron microscopic examination, and for assessment of integrin beta1 mRNA expression by in situ hybridization.

RESULTSThe expressions of integrin beta 1 mRNA in two therapeutic groups were significantly higher than that of control group. The quality of the wound healing was improved in therapeutic group with its healing time shortened when compared with that in control group (P < 0.05). There was an obvious difference in the number of fibroblasts and capillary gemmules between the therapeutic and control groups (P < 0.05).

CONCLUSIONThe wound healing and quality could be improved by both rhEGF and rhbFGF, but rhbFGF seemed better to be employed during the early and middle stages of the wound repair for the growth of granulation tissue, while rhEGF should be applied at the late stage of wound repair to accelerate the re-epithelialization of the wound. Combined application of rhEGF with rhbFGF according to time effect could be more beneficial to the wound repair.

Animals ; Epidermal Growth Factor ; pharmacology ; Female ; Fibroblast Growth Factor 2 ; pharmacology ; Integrin beta1 ; genetics ; Male ; Microscopy, Electron ; RNA, Messenger ; analysis ; Rabbits ; Recombinant Proteins ; pharmacology ; Wound Healing ; drug effects

OBJECTIVETTo explore the method for inducing the dedifferentiation of epidermal cells into their progenitor stem cells in vitro without external gene intervention.

METHODSHEK cells obtained from Casacade were induced to reverse their differentiated process and produce immature stem-like cells, namely the dedifferentiation derived epidermal stem cells (dESCs), by induction with basic fibroblasts growth factors (bFGF) in vitro. Immunochemical staining, flow FACS analysis, RT-PCR and immunofluorescent staining were used to detect the phenotypic and functional changes of the differentiated epidermal cells, using human epidermal stem cells (ESCs) as the positive control.

RESULTSImmunohistochemical staining revealed that the expressions of β₁-integrin, CK19 and CK14 were up-regulated, while CK10 expression was down-regulated significantly after bFGF treatment. Two-color flow cytometric analysis of α₆-integrin and CD71 showed that the percentages of α₆(+)CD71(-), α₆(+)CD71(+) and CD71(+) expressing populations reached 13.24%, 58.26% and 23.12% of the total isolated cells, as compared with those of the control (0.12%, 3.06%, 51.50%) and positive control cells (37.49%, 45.13%, 5.86%). RT-PCR analysis indicated that the relative gene expressions of β₁-integrin, CK19 and CK14 increased in bFGF treatment group, whereas the expression of CK10 was significantly suppressed. Although there was no significant difference in the expression levels of β₁ integrin, CK19 and CK10 between the bFGF-treated and the positive controls, the expression of CK14 in bFGF-treated cells showed a 1.4-fold increase as compared with that in ESCs (P < 0.05). Immunofluorescent staining showed that a regional difference in the subcellular localization of telomerase between dESCs and ESCs.

CONCLUSIONbFGF can induce the epidermal cells to convert into epidermal precursor cells. Although they are more likely to be transient amplifying cells, the method for reprogramming somatic epidermal cells into their progenitors by bFGF induction other than genetic manipulation offers a new approach to generate residual healthy stem cells for wound repair and regeneration.

Cell Dedifferentiation ; Cell Proliferation ; Cells, Cultured ; Epidermis ; cytology ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Induced Pluripotent Stem Cells ; cytology

OBJECTIVETo study the preparative method of controlled release microspheres incorporating basic fibroblast growth factor (bFGF) and the bioactivities of bFGF, which were released from bFGF microspheres, on the cultured Schwann cells.

METHODSbFGF was microcapsulated with the multiple emulsion encapsulative method using polylactic-co-glycolic acid (PLGA) as coating material. Its morphology, particle size distribution, drug loading, enveloping rate and in vitro release property were studied. The cultured Schwann cells were grouped according to the different ingredients being added to the culture medium of bFGF group or bFGF-PLGA group. Then the cytometry, cytoactivity detection and mitotic cycle analysis of Schwann cells were performed.

RESULTSThe morphology and the particle size distribution of the bFGF-PLGA microspheres were even and good. The drug loading and enveloping rate of microspheres were (27.18 x 10(-3))%+/-(0.51 x 10(-3))% and 66.43%+/-1.24%. The release property of microspheres in vitro was good and the overall release rate was 72.47% in 11 days. The in vitro cellular study showed that: at the first 2 days of plate culture, the cell number and viability of the bFGF group were statistically higher than the bFGF-PLGA group; at the 3rd and 4th days of plate culture, the cell number and viability of bFGF and bFGF-PLGA groups showed no difference; at the 6th and 8th days of the plate culture, the cell number and viability of the bFGF-PLGA group were statistically higher than the bFGF group. By flow cytometry examination, at the 2nd day of plate culture, the G2/M+S percentage of bFGF group was statistically higher than the bFGF-PLGA group, at the 4th and 8th days of plate culture, the G2/M+S percentage of the bFGF-PLGA group was statistically higher than the bFGF group.

CONCLUSIONSIt is practical to prepare the bFGF-PLGA microspheres with the multiple emulsion encapsulative method. bFGF-PLGA microspheres can preserve the bioactivities of bFGF effectively and promote the proliferation of Schwann cells in a long period because of the controlled release of bFGF from the microspheres.

Animals ; Delayed-Action Preparations ; Fibroblast Growth Factor 2 ; administration & dosage ; pharmacology ; In Vitro Techniques ; Microspheres ; Rabbits ; Schwann Cells

This study was aimed to investigate the influence and significance of celecoxib (specific inhibitor of cyclooxygenase-2) on mRNA expressions of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (b-FGF), transforming growth factor β (TGF-β) in acute leukemia cells. The expressions of VEGF, b-FGF, TGF-β mRNA were measured by RT-PCR in acute leukemia cells treated with celecoxib (80 µmol/L, for 48 h) or with PBS. The results showed that the obvious expressions of VEGF, b-FGF, TGF-β mRNA were observed in acute leukemia cells. By using Pearson correlation analysis, there was positive correlation between VEGF mRNA and b-FGF mRNA expressions (r = 0.559, P = 0.001), and negative correlation between VEGF and TGF-β mRNA expressions (r = -0.4, P = 0.029). Expression levels of VEGF, b-FGF, TGF-β mRNA in experimental group were lower than that in control group (P < 0.01). It is concluded that COX-2 inhibitor celecoxib can inhabit vascular endothelial growth through down-regulating the mRNA expression of VEGF, b-FGF and TGF-β in acute leukemia cells. COX-2 inhibitor may offer supplemental effect for treating acute leukemia.
Adult ; Celecoxib ; Cyclooxygenase 2 Inhibitors ; pharmacology ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Leukemia ; drug therapy ; metabolism ; Male ; Pyrazoles ; pharmacology ; RNA, Messenger ; genetics ; Signal Transduction ; Sulfonamides ; pharmacology ; Transforming Growth Factor beta ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effect of Hirudin on random skin flap survival in rats.

METHODS24 SD rats were randomly divided into control group and experimental group. The "McFarlane flap (3 cm x 9 cm)" rat models were established on the rat dorsum. 3 ml Hirudin (30 ATU) was injected into the flap in the experimental group, while 3 ml saline in the control group. The injection was performed for 7 days. The flap survival area in the two groups was measured. The tissue samples were taken from proximal (I), middle (II) and distal (III) portions of flaps for histologic study. The VEGF and bFGF expression was also detected with immunohistochemistry method.

RESULTS7 days after operation, the flap survival rate was (69.52 +/- 3.23)% in the experimental group, while (50.36 +/- 2.37)% in control group, showing a significant difference between the two groups (P < 0. 01). In the middle portion, tissue edema and infiltration of neutrophils in experimental group was markedly slighter than that in control group. The VEGF and bFGF expression and neovascularization was enhanced markedly in experimental group.

CONCLUSIONSHirudin can increase the survival of random pattern skin flaps. It may increase the VEGF, bFGF expression through a series of complex regulatory pathway. Then flap neovascularization is promoted and the flap blood supply is increased.

Animals ; Fibroblast Growth Factor 2 ; metabolism ; Graft Survival ; drug effects ; Hirudins ; pharmacology ; Male ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; Surgical Flaps ; Vascular Endothelial Growth Factor A ; metabolism

This paper sums up some studies in the last decade regarding the inhibitory effects of traditional Chinese herbs on growth factor induced proliferation of vascular smooth muscle cell (VSMC) via directly measuring the mRNA expression of its growth factors and the related receptors by electron microscope, immunohistochemistry, blot and hybridization in situ.
Cell Proliferation ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Fibroblast Growth Factor 2 ; antagonists & inhibitors ; Humans ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Platelet-Derived Growth Factor ; antagonists & inhibitors

BACKGROUNDKeratinocyte growth factor (KGF) significantly influences epithelial wound healing. The aim of this study was to isolate KGF phage model peptides from a phage display 7-mer peptide library to evaluate their effect on promoting epidermal cell proliferation.

METHODSA phage display 7-mer peptide library was screened using monoclonal anti-human KGF antibody as the target. Enzyme linked immunosorbent assay (ELISA) was performed to select monoclonal phages with good binding activity. DNA sequencing was done to find the similarities of model peptides. Three-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, immunofluorescence assay and quantitative real-time PCR analysis were employed to evaluate the effect of the phage model peptides on epidermal cells.

RESULTSThirty-three out of fifty-eight (56.9%) of the isolated monoclonal phages exhibited high binding activity by ELISA. Ten of fifteen obtained phage model peptides were similar to KGF or epidermal growth factor (EGF). MTT assay data showed that four (No. 1 - 4) of the ten phage model peptides could promote epidermal cell proliferation. The expression of keratinocyte growth factor receptor (KGFR) mRNA in the KGF control group and the two phage model peptide groups (No. 1 and No. 2) increased. Expression of c-Fos mRNA and c-Jun mRNA in the KGF control group increased, but did not increase in the four phage model peptide groups (No.1 - 4).

CONCLUSIONFour phage model peptides isolated from the phage display 7-mer peptide library can safely promote epidermal cell proliferation without tumorigenic effect.

Cell Proliferation ; drug effects ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Epidermis ; cytology ; Fibroblast Growth Factor 7 ; chemistry ; pharmacology ; Humans ; Peptide Library ; Peptides ; chemistry ; pharmacology ; Polymerase Chain Reaction ; Receptor, Fibroblast Growth Factor, Type 2 ; genetics