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AIM: To investigate the relationship between the levels of chemokine receptor 2(CXCR2)and basic fibroblast growth factor(bFGF)in aqueous humor and the prognosis of trabeculectomy in patients with acute primary angle-closure glaucoma(APACG).METHODS: A total of 80 cases(80 eyes)APACG patients who underwent trabeculectomy in our hospital from June 2020 to January 2022 were collected in the case group. According to the postoperative efficacy, they were grouped into a success group of 60 cases(60 eyes)and a failure group of 20 cases(20 eyes). Another 86 cataract patients(86 eyes)who underwent phacoemulsification with normal intraocular pressure in our hospital during the same period were included in the control group. Enzyme linked immunosorbent assay was applied to detect the levels of CXCR2 and bFGF in aqueous humor. ROC curve was applied to analyze the value of predicting trabeculectomy failure in APACG patients by the levels of CXCR2 and bFGF in aqueous humor. Furthermore, multivariate Logistic regression was applied to analyze the influencing factors of trabeculectomy failure in APACG patients.RESULTS: The levels of CXCR2 and bFGF in the aqueous humor of the case group were significantly higher than those of the control group(P<0.05). The levels of CXCR2 and bFGF in the aqueous humor of the failed group and the proportion of patients with postoperative shallow anterior chamber were significantly higher than those of the successful group(P<0.05). The AUC for predicting trabeculectomy failure in APACG patients using CXCR2 and bFGF levels alone and in combination was 0.885, 0.883 and 0.953, respectively. CXCR2 and bFGF were independent risk factors for trabeculectomy failure in APACG patients(P<0.05).CONCLUSION: The levels of CXCR2 and bFGF in the aqueous humor of APACG patients are obviously elevated, and both are risk factors for trabeculectomy failure.
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Abstract Introduction Postoperative dysphonia is mostly caused by vocal fold scarring, and careful management of vocal fold surgery has been reported to reduce the risk of scar formation. However, depending on the vocal fold injury, treatment of postoperative dysphonia can be challenging. Objective The goal of the current study was to develop a novel prophylactic regenerative approach for the treatment of injured vocal folds after surgery, using biodegradable gelatin hydrogel microspheres as a drug delivery system for basic fibroblast growth factor. Methods Videoendoscopic laryngeal surgery was performed to create vocal fold injury in 14 rabbits. Immediately following this procedure, biodegradable gelatin hydrogel microspheres with basic fibroblast growth factor were injected in the vocal fold. Two weeks after injection, larynges were excised for evaluation of vocal fold histology and mucosal movement. Results The presence of poor vibratory function was confirmed in the injured vocal folds. Histology and digital image analysis demonstrated that the injured vocal folds injected with gelatin hydrogel microspheres with basic fibroblast growth factor showed less scar formation, compared to the injured vocal folds injected with gelatin hydrogel microspheres only, or those without any injection. Conclusion A prophylactic injection of basic fibroblast growth factor -containing biodegradable gelatin hydrogel microspheres demonstrates a regenerative potential for injured vocal folds in a rabbit model.
Resumo Introdução A disfonia pós-operatória é causada principalmente por cicatrizes nas pregas vocais. Tem sido relatado que o manejo cuidadoso da cirurgia das pregas vocais reduz o risco de formação de cicatriz. No entanto, a depender da lesão da prega vocal, o tratamento da disfonia pós-operatória pode ser desafiador. Objetivo Desenvolver uma nova abordagem regenerativa profilática para o tratamento de pregas vocais lesionadas após a cirurgia, com microesferas biodegradáveis de hidrogel de gelatina como sistema de administração de medicamentos para o Fator Básico de Crescimento de Fibroblastos (bFGF). Método A cirurgia laríngea videoendoscópica foi feita para criar lesão nas pregas vocais em 14 coelhos. Imediatamente após esse procedimento, microesferas biodegradáveis de hidrogel de gelatina com bFGF foram injetadas na prega vocal. Duas semanas após a injeção, as laringes foram excisadas para avaliação da histologia das pregas vocais e do movimento da mucosa. Resultados A presença de função vibratória deficiente foi confirmada nas pregas vocais lesionadas. A histologia e a análise de imagem digital demonstraram que as pregas vocais lesionadas injetadas com microesferas de hidrogel de gelatina com bFGF apresentaram menor formação de cicatriz, em comparação com as pregas vocais lesionadas injetadas apenas com microesferas de hidrogel de gelatina ou aquelas sem injeção. Conclusão Uma injeção profilática de microesferas biodegradáveis de hidrogel de gelatina com bFGF demonstra um potencial regenerativo para pregas vocais lesionadas em um modelo de coelho.
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Animals , Vocal Cords/surgery , Gelatin , Rabbits , Fibroblast Growth Factor 2 , Hydrogels , MicrospheresABSTRACT
@#AIM: To investigate the repairment of wound by using soft contact lenses and rb-bFGF eye drops after deep foreign body removal.<p>METHODS: Patients with deep corneal foreign body(72 cases, 72 eyes)were randomly separated into three groups and received surgery to remove the foreign bodys. Patients in group C only accepted levofloxacin eye drops and ofloxacin eye ointment while patients in group A wore soft contact lenses and group B received rb-bFGF eye drops as an extra after operation. The corneal irritation and pain(1, 3, 5d), wound healing(1wk)and relevant factors in visual acuity impairment(1mo)were observed after deep foreign body removal.<p>RESULTS: Corneal irritation and pain scores in group A were significantly lower than that in the other two groups at 1, 3 and 5d after operation. Patients felt less painful in group B than group C at 3 and 5d. Corneal wound healing in groups A and B was significantly higher than group C at 1wk after surgery(<i>P</i>>0.05). The closer corneal foreign body was to the pupil area, the more vision was affected after 1mo operation(<i>r</i>s=0.635, <i>P</i><0.05).<p>CONCLUSION: Soft contact lenses can effectively alleviate eye irritation after deep corneal foreign body removal in early time. Both SCL and rb-bFGF eye drops can accelerate the recovery of corneal wounds. Visual acuity impairment was closely related to the location of foreign body in deep corneal.
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Objective To investigate the mechanisms of basic fibroblast growth factor(bFGF) promoting blood spi-nal cord barrier(BSCB) recovery in rats after spinal cord injury(SCI).Methods Rats were randomly divided into sham group,SCI model group, bFGF intervention group(80 μg/kg). The nerve function of hind limb motor was evaluated by Basso, Beattie, and Bresnahan (BBB) scores during postoperative 14 days. Neuron loss of injured spinal cords was observed by haematoxylin and eosin(HE) staining and NeuN staining.The integrity of BSCB was investigated with Evan's Blue staining and fluorescein isothiocyanate (FITC)-dextran extravasation. The expression protein of adhesive connection protein (p120-catenin,β-catenin) and tight junction protein expression(occludin, claudin-5) were analyzed by Western blot.Results Compared with SCI model group, bFGF intervention group neuron loss decreased significantly at 3 d after injury(P<0.05);the permeability of blood spinal cord barrier obvi-ously reduced at 1d after injury; the expressions of p120-catenin, beta-catenin, occludin, claudin-5 protein of bFGF intervention group were increased dramatically at 1 d after injury(P<0.05). Conclusions bFGF improves the recovery of BSCB in an SCI model by reducing the loss of neurons and increasing the expressions of adhesion junction proteins and tight juction proteins.
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Objective: To detect the expression of basic fibroblast growth factor(bFGF) in oral mucosa with ulcer in rabbits. Methods: 72 New Zealand rabbits(with the weight of 3 000-3 500 g) were randomly divided into control group,model group,and treatment group(n = 24). 1,3,5 and 7 d after treatment buccal mucous membrane tissues of the rabbits were respectively taken from the 3 groups. The models of oral ulcer were examined by HE staining. The expression of bFGF mRNA was detected by RTPCR. The expression of bFGF protein was detected by immunohistochemistry. Results: The oral ulcer model of the rabbits was successfully established. Both RT-PCR and immunohistochemistry analyses showed that 1-7 d after treatment the expression levels of bFGF mRNA and protein were higher in treatment group than in model group(P ＜ 0. 05) and control group(P ＜ 0. 05),3-7 d after treatment were higher than in model group(P＞ 0. 05). Conclusion: bFGF may be a new therapeutic target for oral ulcer.
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AIM:To investigate the effect of recombinant bovine basic fibroblast growth factor (rb-bFGF) eye drops and hydroxyl indican eye drops on tear film stability and dry eye symptoms after age-related cataract surgery.METHODS:Totally 80 patients with 80 affected eyes undergoing age-related cataract surgery in our hospital from January 2015 to October 2016 were selected as study subjects,and they were randomly divided into control group and experimental group with 40 patients (40 affected eyes) in each group.The two groups were treated with hydroxyl indican eye drops and rb-bFGF eye drops,respectively.The clinical curative effect,inflammation related factors [interleukin 6 (IL-6),tumor necrosis factor α (TNF-α)],Schirmer test (S Ⅰ t),break-up time (BUT) of tear film,corneal sodium fluorescein staining (FL) and scores of dry eye symptoms in the two groups were observed.RESULTS:The total treatment effective rate of experimental group after treatment was significantly higher than that of the control group (90.0％ vs 72.5％;x2 =4.021,P＜ 0.05).Before treatment,there was no significant difference in IL-6,TNF-α,S Ⅰ t,BUT,FL score and scores of dry eye symptoms between the two groups (P＞0.05).After treatment,IL-6,TNF-α,S Ⅰ t,FL score and scores of dry eye symptoms in two groups significantly decreased while BUT significantly increased (P＜ 0.05),and changes of the indexes were more significant in the experimental group than the control group (P＜0.05).CONCLUSION:In the treatment of patients after age-related cataract after surgery,rb-bFGF eye drops has more advantages over hydroxyl indican eye drops in regulating the expression of inflammatory factors,improving the tear film stability and relieving dry eye symptoms.
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Objective To observe the effect of external recombinant human basic fibroblast growth factor (rh-bFGF) on the skin and muscles.Methods 40 BALB/c mice of 6 weeks were randomly divided into skin and muscle groups.The skin group was randomly divided into group A and group B.The skin group was injected with 100U rh-bFGF and the same dose of saline in the chin.The muscle groups were randomly divided into C,D,E,and F,and the muscle group was injected into the left calf muscle in order of 200,400,800U rh-bFGF and the same dose of normal saline.The materal was taken at week 8.HE staining was used to observe the submental tissue and muscle fiber morphology.Two groups of muscle fibers,collagen and fibrous tissue were detected by trichrome stain.Immunohistochemistry was used to determine the blood vessel density of the chin and gastrocnemius.Results In week 8,10 mice in group A had a mass in the lower jaw,and the gastrocnemius of the groups C and D showed hypertrophy.Group A showed the thickness of epidermis,dermal papillary layer and mucosa,hair follicle and blood vessel quantity,glandular cavity,collagen and fiber content were sigificantly greater than that of group B (P＜0.05).In the groups C,D and E,the gastrocnemius muscle was significantly thicker than that of group F,and the thickening of group D was the most obvious (P＜0.05),and the contents of collagen and fiber in the groups C,D and E were significantly higher than that in group F.Conclusions Subcutaneous injection of rh-bFGF leads to submental skin thickening,vascular hyperplasia,increased diameter of hair follicle and higher collagen and fiber contents;intramuscular injection of it may induce the muscular hypertrophy and vascular proliferation.
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AIM To investigate the effect and mechanism of methanolic extract of Eupatorium (MEOE) to model rats with chronic soft tissue injury.METHODS The model rats were established by mechanical injury and a subsequent two-week normal feeding for respective administration of high,medium and small dosage of MEOE once a day successively for 14 days.An array of indices,the level of superoxide dismutase (SOD),malondialdehyde (MDA),prostaglandin E2 (PGE2),nitric oxide (NO),interleukin-6 (IL-6) and histamine,the expression of tumor necrosis factor-alpha (TNF-α),nitric oxide (NO) and Collagen-Ⅰ/Ⅲ,and the activity of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were measured to analyze the effect of MEOE to model rats with chronic muscle injury.RESULTS MEOE resulted in apparent reduction of contents of MDA,PGE2 and NO,and the levels of TNF-α and IL-6 in muscular tissue (P ＜ 0.05),significantly increased of the SOD in muscular tissue (P ＜ 0.01),a remarkably inhibited expression of the tissue Collagen-Ⅰ/Ⅲ protein (P ＜ 0.01),and significantly improved activity of tissue VEGF and bFGF (P ＜ 0.01).CONCLUSION The certain therapeutic effects of MEOE to rats with chronic muscle injury may correlate with its influence to the levels of inflammatory factors inhibition,the oxidative stress relief,the overexpression of collagen-Ⅰ/Ⅲ inhibition,the VEGF and bFGF activity improvement,and the time spare from the repairing.
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Objective To study the effects of supplemental Dangguibeimukushen pill prescription on the expressions of MMP13 and bFGF in tumor tissues on H22 hepatoma bearing mice.Methods H22 hepatoma bearing mice was employed to carry out anti-tumor experiment in vivo in this study.A total of 60 mice were randomly divided into model group,DDP positive control group,low and high dosage of supplemental Dangguibeimukushen pill treatment group,supplemental Dangguibeimukushen pill combined with DDP group.In each gruop,gastric infusion was performed continuously 14 d.The tumor growth and the general conditions of mice were recorded.After the last administration of gavages orally treatment,all mice were anaesthetized and killed by cervical disloca-tion method to obtain completely tumor tissue for further HE staining measure and detection of MMP13 and bFGF in tumor tissue with the method of RT-qPCR and immunohistochemistry.Results The tumor volume of model group and low dosage of supple-mental Dangguibeimukushen pill treatment group were larger than that of other groups,futhermore,ulceration were found inside tumor,and basic integrity were observed of tumor capsule.However,in all the treatment groups,the tumor volume was relatively smaller,and the capsule of tumor was incomplete,ulceration of different degrees and bleeding when tumor was peeled off.In combi-nation groups,the tumor volume was much smaller,and the tumor had incomplete capsule and less ulceration inside.Pathological observation showed that connective tissue was found inside tumor,cells were sparsely spread and the number of necrosis cancer cells were increased in all treatment groups.RT-qPCR detection showed that the mRNA expression of MMP13 and bFGF in each treat-ment group were decreased comparing with the model group,and the difference was statistically significant(P <0.05).Immunohis-tochemical detection showed that the expression of MMP13 and bFGF in the combined group were less colored and weakly positive expression comparing with the positive group or the pure Chinese medicine treatment group,the difference was statistically signifi-cant(P <0.05).Conclusion Dangguibeimukushen pill could downregulate the expression of MMP13 and bFGF of tumor tissue in H22 hepatocarcinoma xenograft mice at mRNA and protein levels,and played a vital role in inhibiting cancer invasion and angiogen-esis,reducing toxics and side effects and enchancing its efficiency.
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Objective To analyze the effects of Xiaogan capsule, Xiaoyao pill combined with Shu er capsule on hyperplasia of mammary gland and its effect on vascular endothelial growth factor ( VEGF ) and basic fibroblast growth factor ( bFGF ) .Methods 106 cases of patients with breast hyperplasia treated in our hospital from April 2013 to August 2016 were randomly divided into experimental group and control group according to the order of admission, with 53 cases in each group.The control group was treated with Xiaojin Capsule and Xiaoyao Pills.The experimental group was treated with Xiaojin Capsule, Xiaoyao Pill and Shuerjing Capsule.Analyze the total effective rate of clinical treatment, incidence of adverse reactions, visual analogue pain score and the influence on the expression of VEGF and bFGF in the two groups.Results The total effective rate was 96.23%(51/53) in the experimental group and 86.79%(46/53) in the control group,the total effective rate in the experimental group was much higher than that in the control group (P<0.05).The incidence of adverse reactions was 11.32%(6/53) in the experimental group and 13.21%(7/53) in the control group,there was no significant difference in adverse reactions between the two groups.The visual analogue pain score of the experimental group was (2.85 ±0.41) points, the visual analogue pain score of the control group was (5.25 ±1.84),the visual analogue pain score of the experimental group was significantly lower than that of the control group ( P <0.05 ) .After treatment, the expression of VEGF and bFGF in the patients with hyperplasia of mammary gland decreased, and the levels of VEGF and bFGF in the patients with hyperplasia of mammary glands were significantly higher than those in the untreated group (P<0.05),the expression of VEGF and bFGF in the control group was enhanced.Conclusion Xiaojin capsule,Xiaoyao pill and Shu er jing capsule have higher application value in the patients with hyperplasia of mammary gland , and significantly lower the expression of VEGF and bFGF .
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Objective To investigate the long-term efficacy of calcium phosphate cement combined with bFGF in repairing pulp chamber perforation and to analyze the correlation between the diameter of the perforation and the curative effect.Methods 75 patients with pulp chamber perforation (82 teeth) were enrolled and divided into the observation group and the control group according to the repair material and method.A series of subgroups were also modeled according to the diameter of the perforation,which include the control group A (≤ 1.5 mm),control group B (1.6～3 mm) and control group C (＞3 mm),as well as the observation group A (≤ 1.5 mm),observation group B (1.6～3 rmm) and observation group C (＞3 mm).The observation group was treated with calcium phosphate cement combined with bFGF,and the control group was treated with calcium phosphate cement alone.Results The total effective rate of the observation group was 97.8％,which was significant higher than 80.6％ in the control group (P＜0.05).The cure rate of the observation group A was 100％,which was significant higher than 73.3％ in the observation group B and 41.7％ in the observation group C (all P＜0.05).The total effective rates of the observation group A and B were significantly higher than 91.7％ in the observation group C (all P＜0.05).The cure rate of the control group A was 92.9％,which was significant higher than 60.0％ in the control group B and 25.0％ in the control group C (all P＜0.05).The total effective rates of the control group A (100％) and B (90.0％) were significantly higher than 91.7％ in the control group C (all P＜0.05).Conclusions Calcium phosphate cement combined with bFGF in repairing the pulp chamber perforation was significantly better than calcium phosphate cement alone.The cure rate of perforation repairing is closely related to the perforation size.The perforation with small diameter may achieve a better repairing effect.
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Objective:To investigate the effects of total flavonoids from Cycas Revolute on expression of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF),Hypoxia inducible factor-1α (HIF-1α) and nuclear factor-κB(NF-κB) in model mice of Lewis lung cancer.Methods: The expressions of VEGF,bFGF,HIF-1α and NF-κB in tumor tissues were detected by immunohistochemistry and Western blot.The expression of VEGF,VEGF and NF-κB in tumor tissues were detected by fluorescent quantitative PCR.BFGF,HIF-1α and NF-κB mRNA were detected by immunohistochemistry.Results: The results of immunohis-tochemistry,Western blot and Real-time PCR showed that the results were basically the same,compared with model group,the expression of VEGF,bFGF,HIF-1α,NF-κB mRNA and the expression of VEGF,bFGF,HIF-1α and NF-κB were decreased,the difference was highly significant (P<0.05).Conclusion: The mechanism of total flavonoids from Cycas Revolute in the treatment of lung cancer may be through inhibition of the expression of VEGF,bFGF,HIF-1α,NF-κB in invasion and metastasis,and further inhibit the expression of VEGF,bFGF,HIF-1α and NF-κB in invasion and metastasis-related proteins,thus play a role of anti-lung cancer invasion and metastasis.
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Objective:To observe the effects of Er ∶YAG(erbium:yttrium-aluminum-garnet) laser combined with diode laser in the treatment of chronic periodontitis.Methods:30 patients with chronic periodontitis patients were treated by mechanical scaling and root planning(SRP),pure Er ∶ YAG laser(ERT),diode + Er ∶ YAG + diode laser therapy(TPT) respectively.In the treatment VAS pain score was recorded.Probing deepth (PD),clinical attachment level (CAL) and bleeding on probing (BOP) were examined,IL-1 β,TNF-α and b-FGF in GCF,Porphyromonas gingivalis(Pg) in subgingiva plaqure were measured before and after treatment.Data were analyzed by SPSS 21.0 software.Results:6 weeks,12 weeks after treatment PD,BOP and CAL in TPT group were lower than those in ERT and SRP groups(P ＜0.05),TGF-β and b-FGF in TPT group were higher,IL-1β and TNF-α were lower than those in ERT and SRP groups (P ＜ 0.05).The content of Pg relative to total bacteria in TPT group were lower than that in ERT and SRP groups (P ＜ 0.05),VAS value in twinlight laser group was significantly lower than hand instruments group(P ＜ 0.05).Conclusion:The twinlight laser is effective in the treatment of periodontitis.
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OBJECTIVE@#To build the rabbit model of VX2 hepatic carcinoma, examine the tumor body using the ultrasonic contrast and study the correlation between the blood circulation grading and angiogenesis.@*METHODS@#The VX2 tumor strain was prepared in the lateral muscle of the hind legs of 40 male New Zealand rabbits (which were purchased from Nanjing Senbao Biotech Co., Ltd.). The tumor block was embedded in the center of left liver lobe directly to build the rabbit model of VX2 hepatic carcinoma. The ultrasonic contrast was performed 14 d after implanting the tumor body. The semi-quantitative classification (0-IV level) was taken according to the blood flow of tumor vessel. Animals were executed using the air embolism method. The liver was separated to extract RNA and total protein respectively. The real-time PCR and western blotting method were employed to detect the expression of angiogenesis-related factors of VEGF, bFGF and TNF-α, while the ultrasonic contrast to detect the correlation with blood circulation grading. The Pearson product moment correlation coefficient was used to measure the linear relationship between these two variables and analyze the correlation between the blood circulation grading and angiogenesis using the ultrasonic contrast.@*RESULTS@#Thirty-three rabbits had the successful model of VX2 hepatic carcinoma. The blood circulation grading by ultrasonic contrast was: 2 cases at level 0 (6.60%), 5 cases at level I (16.7%), 12 cases at level II (40.0%), 6 cases at level III (20.0%) (local dense or clustered blood flow) and 5 cases at level IV (16.7%). The results showed that there was positive correlation between three angiogenesis-related factors and the blood circulation grading. The correlation coefficient between three angiogenesis-related factors and the blood circulation grading was over 0.9, which indicated the relatively high correlation.@*CONCLUSIONS@#The ultrasound blood circulation grading for the hepatic carcinoma can clearly reflect the changes of blood vessel, which will be of critical significance for the early diagnosis of hepatic carcinoma and clinical evaluation of angiogenesis indicators.
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Objective: To build the rabbit model of VX2 hepatic carcinoma, examine the tumor body using the ultrasonic contrast and study the correlation between the blood circulation grading and angiogenesis. Methods: The VX2 tumor strain was prepared in the lateral muscle of the hind legs of 40 male New Zealand rabbits (which were purchased from Nanjing Senbao Biotech Co., Ltd.). The tumor block was embedded in the center of left liver lobe directly to build the rabbit model of VX2 hepatic carcinoma. The ultrasonic contrast was performed 14 d after implanting the tumor body. The semi-quantitative classification (0-IV level) was taken according to the blood flow of tumor vessel. Animals were executed using the air embolism method. The liver was separated to extract RNA and total protein respectively. The real-time PCR and western blotting method were employed to detect the expression of angiogenesis-related factors of VEGF, bFGF and TNF-α, while the ultrasonic contrast to detect the correlation with blood circulation grading. The Pearson product moment correlation coefficient was used to measure the linear relationship between these two variables and analyze the correlation between the blood circulation grading and angiogenesis using the ultrasonic contrast. Results: Thirty-three rabbits had the successful model of VX2 hepatic carcinoma. The blood circulation grading by ultrasonic contrast was: 2 cases at level 0 (6.60%), 5 cases at level I (16.7%), 12 cases at level II (40.0%), 6 cases at level III (20.0%) (local dense or clustered blood flow) and 5 cases at level IV (16.7%). The results showed that there was positive correlation between three angiogenesis-related factors and the blood circulation grading. The correlation coefficient between three angiogenesis-related factors and the blood circulation grading was over 0.9, which indicated the relatively high correlation. Conclusions: The ultrasound blood circulation grading for the hepatic carcinoma can clearly reflect the changes of blood vessel, which will be of critical significance for the early diagnosis of hepatic carcinoma and clinical evaluation of angiogenesis indicators.
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Epithelial mesenchymal transition (EMT) of lens epithelial cells (LECs) may contribute to the development of posterior capsular opacification (PCO), which leads to visual impairment. Andrographolide has been shown to have therapeutic potential against various cancers. However, its effect on human LECs is still unknown. The purpose of this study is to evaluate the effect of andrographolide on EMT induced by growth factors in the fetal human lens epithelial cell line (FHL 124). Initially the LECs were treated with growth factors (TGF-β2 and bFGF) to induce EMT. Subsequently these EMT-induced cells were treated with andrographolide at 100 and 500 nM concentrations for 24 h. Our results showed that FHL 124 cells treated with growth factors had a significant decrease in protein and m-RNA levels of epithelial markers pax6 and E-Cadherin. After administering andrographolide, these levels significantly increased. It was noticed that EMT markers α-SMA, fibronectin and collagen IV significantly decreased after treatment with andrographolide when compared to the other group. Treatment with andrographolide significantly inhibited phosphorylation of ERK and JNK. Cell cycle analysis showed that andrographolide did not arrest cells at G0/G1 or G2/M at tested concentrations. Our findings suggest that andrographolide helps sustain epithelial characteristics by modulating EMT markers and inhibiting the mitogen-activated protein kinase (MAPK) signalling pathway in LECs. Hence it can prove to be useful in curbing EMT-mediated PCO.
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Background: Immortalized human endothelial cells are widely used as in vitro models for debilitating conditions such as cancer, cardiovascular and ocular diseases. Human microvascular endothelial cell (HMEC-1) is immortalized via stable transfection with a gene encoding SV40 large antigen whilst telomerase-immortalized human microvascular endothelial (TIME) cells is immortalized by engineering the human telomerase catalytic protein (hTERT) into primary microvascular endothelial cells. Here, we established a three-dimensional (3D) spheroid invasion assay with HMEC-1 and TIME and compared the difference in their ability to invade through the collagen matrix in response to exogenous growth factors, namely vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Methods: TIME and HMEC-1 spheroids were embedded in a collagen matrix. The spheroids were stimulated with exogenous growth factors, namely VEGF (50ng/mL) and bFGF (200ng/mL). Twelve points of invasion length from a spheroid was measured using image analysis software, Image J. Three independent experiments were conducted and data was analysis by GraphPad Instat software, version 3.05. Results: TIME spheroid invasion was 16.5 fold higher with exogenous VEGF (50ng/mL) and bFGF (200ng/mL) treatment as compared to those cultured in complete growth medium only. In contrast, no significant difference was observed between HMEC-1 spheroids stimulated with and without exogenous growth factors, VEGF and bFGF. Conclusions: This is the first report on the establishment of a 3D-spheroid invasion assay with TIME cells. The requirement of VEGF and bFGF for TIME spheroids invasion is a novel finding. In addition, this assay offers an advantage over HMEC-1 for testing novel angiogenic agents since it is not affected by endogenously secreted growth factors.
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OBJECTIVE@#To investigate the synergistic effect and mechanism of the combined application of recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF).@*METHODS@#24 KM male mice were randomly divided into 6 groups with 4 mice in each group, namely, Group A (control group), Group B (only treated with collagen), Group C (treated with 2 ng bFGF+collagen), Group D (treated with 4 μ g rhBMP-2+collagen), Group E (treated with 4 μ g rhBMP-2+2 ng bFGF+collagen) and Group F (treated with 4 μ g rhBMP-2+4 ng bFGF+collagen). The composites were implanted into the intermuscular septum of hind legs mice; whereas in control group, intermuscular septum of mice was separated and no implantation was performed. General observation, detection of concentration of calcium content, micro computed tomography (Micro-CT), three-dimensional reconstruction scan, measurement of bone mineral density (BMD), bone volume fraction (BVF) and trabecular thickness (Tb.Th), as well as histological observation with HE staining and ALP and CD34 immumohistochemical staining were performed.@*RESULTS@#Ectopic osteogenesis was found in Groups D, E and F mice. The difference in concentration of calcium contents was statistically significant between Groups D and E (P0.05). Micro-CT and three-dimensional reconstruction revealed continuous newborn bone substance in external surface of ectopic bone formation, and the center of bone formation did not show obvious substantial filling by bone substance. The differences in BMD, BVF and Tb.Th were statistically significant between Groups D and E or F (P<0.01 or <0.05). HE staining showed that in Groups D, E and F, newborn bone substance was mainly located at the edge of ectopic bone formation, and the bone formation in Groups E and F was better than that in Group D. ALP and CD34 immumohistochemical staining revealed the positive expression mainly at the edge of ectopic bone formation, and area of positive expression in Groups E and F was larger than that in Groups D.@*CONCLUSIONS@#rhBMP-2 possesses the capacity to induce ectopic osteogenesis independently, but bFGF does not have this ability; the combined application of rhBMP-2 and bFGF can enhance the synergetic effect on inducing ectopic osteogenesis.
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Objective:To investigate the synergistic effect and mechanism of the combined application of recombinant human bone morphogenetic protein-2(rhBMP-2) and basic fibroblast growth factor (bFGF).Methods:24KM male mice were randomly divided into6 groups with4 mice in each group, namely,GroupA(control group),GroupB(only treated with collagen),GroupC(treated with 2 ng bFGF+collagen),GroupD(treated with4 μg rhBMP-2+collagen),GroupE(treated with4 μg rhBMP-2+2 ng bFGF+collagen) andGroupF(treated with4 μg rhBMP-2+4 ng bFGF+collagen). The composites were implanted into the intermuscular septum of hind legs mice; whereas in control group, intermuscular septum of mice was separated and no implantation was performed. General observation, detection of concentration of calcium content, micro computed tomography (Micro-CT), three-dimensional reconstruction scan, measurement of bone mineral density(BMD), bone volume fraction(BVF) and trabecular thickness(Tb.Th), as well as histological observation withHE staining andALP andCD34 immumohistochemical staining were performed.Results:Ectopic osteogenesis was found inGroupsD,E andF mice.The difference in concentration of calcium contentswas statistically significant betweenGroupsD andE(P0.05).Micro-CT and three-dimensional reconstruction revealed continuous newborn bone substance in external surface of ectopic bone formation, and the center of bone formation did not show obvious substantial filling by bone substance.The differences in BMD,BVF andTb.Th were statistically significant betweenGroupsD andE orF(P<0.01 or <0.05). HE staining showed that inGroupsD,E andF, newborn bone substance was mainly located at the edge of ectopic bone formation, and the bone formation inGroupsE andF was better than that in GroupD.ALP andCD34 immumohistochemical staining revealed the positive expression mainly at the edge of ectopic bone formation, and area of positiveexpression inGroupsE andF was larger than that inGroupsD.Conclusions:rhBMP-2 possesses the capacity to induce ectopic osteogenesis independently, but bFGF does not have this ability; the combined application of rhBMP-2 and bFGF can enhance the synergetic effect on inducing ectopic osteogenesis.
ABSTRACT
Objective: To investigate the synergistic effect and mechanism of the combined application of recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF). Methods: 24 KM male mice were randomly divided into 6 groups with 4 mice in each group, namely, Group A (control group), Group B (only treated with collagen), Group C (treated with 2 ng bFGF+collagen), Group D (treated with 4 μ g rhBMP-2+collagen), Group E (treated with 4 μ g rhBMP-2+2 ng bFGF+collagen) and Group F (treated with 4 μ g rhBMP-2+4 ng bFGF+collagen). The composites were implanted into the intermuscular septum of hind legs mice; whereas in control group, intermuscular septum of mice was separated and no implantation was performed. General observation, detection of concentration of calcium content, micro computed tomography (Micro-CT), three-dimensional reconstruction scan, measurement of bone mineral density (BMD), bone volume fraction (BVF) and trabecular thickness (Tb.Th), as well as histological observation with HE staining and ALP and CD34 immumohistochemical staining were performed. Results: Ectopic osteogenesis was found in Groups D, E and F mice. The difference in concentration of calcium contents was statistically significant between Groups D and E (. P0.05). Micro-CT and three-dimensional reconstruction revealed continuous newborn bone substance in external surface of ectopic bone formation, and the center of bone formation did not show obvious substantial filling by bone substance. The differences in BMD, BVF and Tb.Th were statistically significant between Groups D and E or F (. P<0.01 or <0.05). HE staining showed that in Groups D, E and F, newborn bone substance was mainly located at the edge of ectopic bone formation, and the bone formation in Groups E and F was better than that in Group D. ALP and CD34 immumohistochemical staining revealed the positive expression mainly at the edge of ectopic bone formation, and area of positive expression in Groups E and F was larger than that in Groups D. Conclusions: rhBMP-2 possesses the capacity to induce ectopic osteogenesis independently, but bFGF does not have this ability; the combined application of rhBMP-2 and bFGF can enhance the synergetic effect on inducing ectopic osteogenesis.