Objective To evaluate the effect of composite (bFGF/PDPB) of basic fibroblast growth factor（bFGF） and partially deproteinized bone (PDPB) on the repair of femoral head defect. Methods Forty-eight femoral heads with defect derived from 24 New Zealand rabbits were divided into 3 groups at random, which were implanted with bFGF/PDPB(group A), PDPB(group B) and nothing(group C) respectively.The rabbits were sacrificed at 2,4,and8 weeks after operation, and then the femoral heads were obtained. The specimens injected with Chinese ink were created. Then X-ray examination, histopathological and morphological examination of blood vessel, and image analysis were made. Results The bone defects healed completely 8 weeks after operation in group A. The implants in the repaired tissue were not substituted completely in group B. The bone defects did not heal completely in group C. Two weeks after operation, affluent newly formed vessels were seen in repaired areas in groupA. No significant difference between group A and group B was observed 8 weeks after operation. In group C, newly formed vessels were scarce 2, 4, and 8 weeks after operation. There were 3 sides rated excellent, 2 good and 1 fair in group A; 1 excellent, 2 good, 2 fair and 1 poor in group B; and 1 fair and 5 poor in group C according to the X-ray evaluation 8 weeks after operation. Eight weeks after operation, the volume fraction of bone trabecula in repaired tissue was higher in group A than that in group B (Plt;0.05), and the fraction in group C was thelowest among the 3 groups (Plt;0.05). Conclusion The composite ofbFGF and PDPB can effectively promote the repair of femoral head defect of rabbit.
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 localize the distribution of basic fibroblast growth factor (bFGF) and transforming growth factor-beta(TGF-beta) in tissues from dermal chronic ulcer and hypertrophic scar and to explore their effects on tissue repair. METHODS: Twenty-one cases were detected to localize the distribution of bFGF and TGF-beta, among them, there were 8 cases with dermal chronic ulcers, 8 cases with hypertrophic scars, and 5 cases of normal skin. RESULTS: Positive signal of bFGF and TGF-beta could be found in normal skin, mainly in the keratinocytes. In dermal chronic ulcers, positive signal of bFGF and TGF-beta could be found in granulation tissues. bFGF was localized mainly in fibroblasts cells and endothelial cells and TGF-beta mainly in inflammatory cells. In hypertrophic scar, the localization and signal density of bFGF was similar with those in granulation tissues, but the staining of TGF-beta was negative. CONCLUSION: The different distribution of bFGF and TGF-beta in dermal chronic ulcer and hypertrophic scar may be the reason of different results of tissue repair. The pathogenesis of wound healing delay in a condition of high concentration of growth factors may come from the binding disorder of growth factors and their receptors. bFGF may be involved in all process of formation of hypertrophic scar, but TGF-beta may only play roles in the early stage.
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 influences of estradiol (E2), basic fibroblast growth factor (bFGF), and tamoxifen (TAM) on the proliferation of hemangioma vascular endothelial cell (HVEC). Methods Two strawberry hemangioma from 2 infants (case 1 and case 2) were prepared for HVEC culture. The HVEC on passage 3 were cultured in estrogenfree improved minimum essential medium (IMEM) and subjected to various treatments with 100 pg/ml 17-β-E2, 10 ng/ml bFGF, and 1×10-6 mol/L 4-OH-tamoxifen(4-OH-TAM). The experiment was divided into 5 groups: group 1(IMEM, control group), group 2(17-β-E2), group 3(bFGF), group 4(17-β-E2／bGFG) and group 5(17-β-E2／bGFG／4-OH-TAM). The cell count(CC) and DNA proliferation index (PI) were determined. Results Two cases of HVEC were successfully cultured in vitro. The HVEC showed cobblestoneslike under microscopy and factor Ⅷrelated antigen(also named as von Willebrand factor，vWF) was positive by immunochemical staining. At 9 days in case 1: CC and PI remained unchanged in the control group; CC and PI were slightly increased in group 2, being 1.4 and 1.6 times as much as those in the control group respectively (P＜0.05); CC and PI significantly increased in group 3, being2.6 and 2.3 times as much as those in the control group respectively (P＜0.01); CC and PI increased remarkably in group 4, being 3.7 and 2.9 times as much as those in thecontrol group respectively (P＜0.01); CC and PI were down to the levels of controls in group 5(P＞0.05). The results in case 2 were similar to those in case 1. Conclusion In vitro, the promoting effect of bFGF on HVEC proliferation is much ber than that of estrogen. Estrogen and bFGF enhance this proliferation in a synergistic manner, which can be inhibited by tamoxifen.
Objective To explore the effect of basic fibroblast growth factor（bFGF）and epidermal growth factor（EGF）on the growth of muscle derived stem cells(MDSCs). Methods MDSCs were isolated from hindlimb muscle of 15 new born Kunming mice through serial preplates. 2% fetal bovine serum-containing DMEM was used to induce MDSCs to differentiate into skeletal muscle lineage. The expressions of stem cell marker Sca-1 and skeletal musclecell marker αSarcomeric actin were examined by immunocytochemistry. The effect of bFGF and EGF on the proliferation of MDSCs was determined by MTT colorimetric microassay. The solo effect of bFGF or EGF at different concentrations (6.25,12.50, 25.00, 50.00, and 100.00 ng/ml) was examined at 96 h and the combined effect (100.00 ng/ml) was examined at 24,48,72 and 96 h.Results MDSCs were successfully isolated from the hindlimb of neonatal mice. Over 90% of MDSCs showed Sca-1 positive immunoreactivity. MDSCs could give rise to α Sarcomeric actin positive myotubes in differentiation cultures. The proliferative effect of bFGF and EGF on MDSCs increased with the elevated concentration.bFGF began to show significant proliferative effect at 12.50 ng/ml (P＜0.05). The effect increased significantly when the concentration reached 25.00 ng/ml from 12.50 ng/ml (P＜0.01) and reached a saturation point. The effect at 50.00 ng/ml or 100.00 ng/ml showed no significant increase when compared with thatat 25.00 ng/ml. EGF had a similar effect to bFGF except that the saturation concentration was 50.00 ng/ml. EGF showed significant effect at 72 h and bFGF at 96 h (Plt;0.01). When they were applied together, significant effect was shownat 24 h (Plt;0.01) and much higher effect was observed at 48, 72 and 96 h (Plt;0.05). Conclusion Both bFGF and EGF can promote the proliferation of MDSCs. The combined application reacts faster and ber.
Objective To select the appropriate media to culture the epidermal stem cells in vitro,and to observe the biological characteristics of the epidermal stem cells. Methods The epidermal stem cells were cultured in five different media, including FAD, FAD+1 ng/ml bFGF, FAD+5 ng/ml bFGF, FAD+10 ng/ml bFGF and K-SFM, and the same fetous fibroblasts were used as the nutrient cells. The proliferation ability was investigated by cell growth curve and MTT detection. Then the biological characteristics of epidermal stem cells were observed through phasecontrast microscope, cell growth curve, BrdU detection and FBM analysis. Results The epidermal stem cells grew best in FAD with bFGF and nutrient cells. And the epidermal stem cells retained proliferative capacity, and formed larger and more expandable clones in vitro. And 80.2% of the cells show a G0/G1 cycle, and the cells had long cell proliferation cycle. Conclusion The above results demonstrate that the media with bFGF and the use of nutrient layer were appropriate to culture epidermal stem cell in vitro. And the epidermal stem cells have a slow cell cycle, characteristics of immaturity.
OBJECTIVE The biological effects of recombinant human epidermal growth factor (rhEGF) and recombinant human fibroblast growth factor (rhFGF) were evaluated on the model of incised wounds in mini pigs. METHODS Total of 160 incised wounds in 16 mini pigs were divided into two groups (rhEGF group and rhFGF group), each containing 80 wounds. In rhEGF group, 60 incised wounds were treated with different dosages of rhEGF (50, 10 and 0.5 micrograms/wound), and another 20 wounds were treated with solvent as control group. In rhFGF group, all wounds were treated in the same way as described in rhEGF group, the dosages of rhFGF were 150, 90 and 30 U/cm2 respectively. The measurements of cavity volume and area in wound, histological examination were used to evaluate the results of wound healing. RESULTS The results showed that wound healing was accelerated in all wounds treated with rhEGF and rhFGF. In rhEGF group, the velocity of re-epithelialization was faster than that of rhFGF group, however, new granulation tissue in rhFGF was more than that of rhEGF group. CONCLUSION The results indicate that rhEGF and rhFGF can stimulate wound healing, however, the mechanisms and the biological effects involved in these processes are quite different. It suggests that it is better to use rhFGF in those wounds which need more granulation tissue formation and use rhEGF in the wounds which mainly need re-epithelialization.