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ABSTRACT Purpose: To assess the effects of the preoperative application of artificial tears combined with recombinant bovine basic fibroblast growth factor on the ocular surface function and inflammatory factor levels after operation in cataract patients complicated with dry eyes. Methods: A total of 118 cataract patients (118 eyes) complicated with dry eyes treated from February 2019 to February2020 were assigned to control and observation groups (n=59 eyes/group) using a random number table. One week before the operation, the control group was administered 0.1% sodium hyaluronate eye drops (artificial tears), based on which the observation group received Beifushu eye drops (recombinant bovine basic fibroblast growth factor), both 6 times daily for 1 week. A comparison was made between the scores of clinical symptoms and the indices of ocular surface function, inflammatory factors in tears, and oxidative stress indices before and after the operation. The ocular surface function was evaluated by an ocular surface disease index questionnaire, tear film breakup-time assay, Schirmer's I test, and corneal fluorescein stain test. The inflammatory factors in tears were measured. Results: No significant differences were noted in the general data and clinical symptom score, ocular surface disease index, tear film breakup-time, Schirmer's I test score, fluorescein stain score, interleukin-6, tumor necrosis factor-alpha, malondialdehyde, superoxide dismutase, lipid peroxide, and total antioxidant capacity before treatment between the 2 groups (p>0.05). After treatment, the clinical symptom score, ocular surface disease index, fluorescein stain score, tumor necrosis factor-alpha, interleukin-6, malondial-dehyde and lipid peroxide declined significantly, and tear film breakup-time, Schirmer's I test score, superoxide dismutase, and total antioxidant capacity increased in both the groups. The improvements in the clinical symptom score as well as in the indices of ocular surface function, inflammatory factors, and oxidative stress were more prominent in the observation group than in the control group (p<0.05). Conclusions: Artificial tears combined with recombinant bovine basic fibroblast growth factor before operation. significantly improved the ocular surface function, reduced inflammatory factors in tears, and alleviated dry eye symptoms after operation in cataract patients.
RESUMO Objetivo: Avaliar os efeitos da aplicação pré-operatória de lágrimas artificiais combinadas com o fator de crescimento de fibroblastos básicos bovinos recombinantes na função da superfície ocular e níveis de fator inflamatório após cirurgia em pacientes com catarata complicada com olhos secos. Métodos: Um total de 118 pacientes com catarata complicada com olhos secos (118 olhos), tratados entre fevereiro de 2019 e fevereiro de 2020, foram divididos em grupos de controle e de observação (n=59, 59 olhos) usando uma tabela de números aleatórios. Uma semana antes da cirurgia, o grupo controle recebeu colírio de hialuronato de sódio a 0,1% (lágrimas artificiais), enquanto o grupo de observação recebeu colírio Beifushu (fator de crescimento de fibroblastos básicos bovinos recombinantes), ambos, seis vezes ao dia, por uma semana. Antes do tratamento e um mês após a cirurgia, os escores de sintomas clínicos, índices de função da superfície ocular, níveis de fatores inflamatórios nas lágrimas e índices de estresse oxidativo foram comparados. A função da superfície ocular foi avaliada pelo questionário do índice de doença da superfície ocular, ensaio de tempo de ruptura do filme lacrimal, teste I de Schirmer e teste de coloração por fluoresceína da córnea. Os níveis de fatores inflamatórios nas lágrimas foram medidos. Resultados: Não houve diferenças significativas nos dados gerais e no escore de sintomas clínicos, índice de doença da superfície ocular, tempo de ruptura do filme lacrimal, escore do teste I de Schirmer, pontuação do teste de coloração por fluoresceína da córnea, interleucina-6, fator de necrose tumoral alfa, malondialdeído, superóxido dismutase, peróxido lipídico e capacidade antioxidante total antes do tratamento entre os dois grupos (p>0,05). Após o tratamento, o escore de sintomas clínicos, índice de doença da superfície ocular, escore do teste de coloração por fluoresceína da córnea, fator de necrose tumoral alfa, interleucina-6, malondialdeído e peróxido lipídico diminuíram significativamente, e o tempo de ruptura do filme lacrimal, escore do teste I de Schirmer, superóxido dismutase e a capacidade antioxidante total aumentou em ambos os grupos. As melhorias no escore de sintomas clínicos, bem como os índices de função da superfície ocular, fatores inflamatórios e estresse oxidativo foram mais proeminentes no grupo de observação do que no grupo controle (p<0,05). Conclusões: Lágrimas artificiais combinadas com fator de crescimento de fibroblastos básicos recombinantes antes da cirurgia melhoram notavelmente a função da superfície ocular, diminuem os níveis de fatores inflamatórios nas lágrimas e aliviam os sintomas de olho seco após a cirurgia em pacientes com catarata complicada com olhos secos.
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AIM: To investigate the clinical value of serum vitamin A(Vit A)and basic fibroblast growth factor(bFGF)levels predicting retinopathy of prematurity(ROP).METHODS: Prospective cohort studies. A total of 411 premature or low birth weight infants with gestational age less than 37 wk or birth weight less than 2 500 g who were delivered in Hainan Branch, Shanghai Children's Medical Center Affiliated to Shanghai Jiao Tong University School of Medicine from January 2020 to December 2022 were selected as subjects. The Vit A and bFGF levels in peripheral blood were detected at 7 d and 35 d after birth, respectively.RESULTS: A total of 392 premature infants or low birth weight infants completed clinical study, including 51 cases in stage 1-2 ROP group, 23 cases in stage 3-5 ROP group and 318 cases in the group without ROP. At 7 d postnatal, the serum Vit A(0.44±0.17 μmol/L)and bFGF(0.53±0.16 ng/L)levels in stage 1-2 ROP group were lower than those in the group without ROP(0.50±0.12 μmol/L and 0.63±0.15 ng/L; all P&#x003C;0.05). The serum Vit A(0.34±0.18 μmol/L)and bFGF(0.44±0.18 ng/L)levels in stage 3-5 ROP group were lower than those in the group without ROP(P&#x003C;0.05). The serum Vit A and bFGF levels in stage 3-5 ROP group were lower than those in stage 1-2 ROP group(P&#x003C;0.05). At 35d postnatal, the serum Vit A(0.33±0.19 μmol/L)and bFGF(0.39±0.19 ng/L)levels in stage 3-5 ROP group were lower than those in stage 1-2 ROP group(0.43±0.16 μmol/L and 0.48±0.17 ng/L; all P&#x003C;0.05). According to the ROC curve drawn by serum Vit A, the AUC value was 0.853, the maximum Youden index was 0.68, the best sensitivity was 73%, and the best specificity was 95%. According to the ROC curve drawn by serum bFGF, the AUC value was 0.828, the maximum Youden index was 0.58, the best sensitivity was 90%, and the best specificity was 68%. According to the ROC curve drawn by serum Vit A combined with bFGF, the AUC value was 0.917, the maximum Youden index was 0.70, the best sensitivity was 70%, and the best specificity was 100%.CONCLUSION: Serum Vit A and bFGF levels are sensitive and effective indicators for predicting ROP. If the serum Vit A or bFGF levels are lower in premature infants or low birth weight infants, it may indicate the higher probability of ROP and its pathological stages. In addition, the clinica value of serum Vit A combined with bFGF in the diagnosis of ROP is higher than that of Vit A or bFGF alone, and the misdiagnosis rate is reduced.
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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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@#Basic fibroblast growth factor (bFGF) exhibits superior biological functions by improving periodontal inflammation, promoting the migration and proliferation of periodontal-related stem cells, promoting the formation of blood vessels and periodontal ligament-like tissue, and regulating the formation of bone/cementum. It plays an important role in tooth development, repair and regeneration. bFGF can be combined with seed cells and scaffold materials for periodontal tissue regeneration, which has been verified in a number of experimental studies. However, the application of bFGF alone as a drug in clinical treatment requires further research.
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Objective To study the influence of recombinant bovine basic fibroblast growth factor as an adjuvant therapy on scar alleviation and inflammatory cytokines in patients with atrophic acne scar. Methods The random number table was employed to randomly assign 120 patients with atrophic acne scar into a test group and a control group.Both groups of patients were treated with CO2 lattice laser.After the operation,the control group was routinely smeared with erythromycin ointment and the test group was coated with recombinant bovine basic fibroblast growth factor gel.The clinical efficacy,clinical indicators,scar alleviation,and inflammatory cytokine levels before and after treatment were compared,and adverse reactions were counted. Results The test group had higher total effective rate(P=0.040) and lower total incidence of adverse reactions(P=0.028) than the control group.Compared with the control group,the test group showcased short erythema duration after treatment(P=0.025),early scab forming(P=0.002),and early edema regression(P<0.001).After treatment,the proportion of grade 1 scars graded by Goodman and Baron's acne scar grading system in the test group and control group increased(P=0.001,P=0.027),and the proportion of grade 4 scars decreased(P<0.001,P=0.034).Moreover,the proportion of grade 1 scars in the test group was higher than that in the control group(P=0.031) after treatment,and the proportion of grade 4 scars presented an opposite trend(P=0.031).After treatment,the levels of tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) in both groups declined(all P<0.001),and the test group had lower TNF-α and IL-1β levels than the control group(all P<0.001). Conclusion The recombinant bovine basic fibroblast growth factor gel as an adjuvant therapy of CO2 lattice laser can effectively alleviate the atrophic acne scar,relieve local inflammatory reaction,and has good curative effect and less adverse reactions.
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Animals , Cattle , Humans , Acne Vulgaris/drug therapy , Atrophy/complications , Carbon Dioxide , Cicatrix/pathology , Fibroblast Growth Factor 2/therapeutic use , Treatment Outcome , Tumor Necrosis Factor-alphaABSTRACT
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.
Subject(s)
Animals , Vocal Cords/surgery , Gelatin , Rabbits , Fibroblast Growth Factor 2 , Hydrogels , MicrospheresABSTRACT
BACKGROUND: In recent years, resveratrol has been studied a lot on the inhibition of tissue fibrosis, but the effect of resveratrol on the rehabilitation of muscle injury has been rarely reported. OBJECTIVE: To observe the expression of basic fibroblast growth factor (bFGF) and insulin-like growth factor 1 (IGF-1) protein in the repair of acute blunt trauma of the skeletal muscle, and to explore the mechanism by which resveratrol promotes the structural and functional recovery of damaged skeletal muscle. METHODS: Thirty-three New Zealand rabbits were randomly divided into three groups: Normal group (n=3), natural recovery group (n=15) and resveratrol group (n=15). The skeletal muscle contusion model was established by blunt violence except for the normal group. The natural recovery group was not treated and the resveratrol group was intragastrically given resveratrol after injury. The animals were euthanized at 1, 3, 7,14, and 21 days after injury. Infiltration of inflammatory cells and formation of collagen fibers were observed using hematoxylin-eosin staining and Masson staining. The expression of bFGF and IGF-1 protein in the skeletal muscle was detected by immunohistochemistry and immunoblotting. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining results: In the normal group, the muscle fibers were presented with polygons, regular shape, tight arrangement, muscle nucleus evenly distributed under the sarcolemma, no hyperplasia and pyknosis, and sarcolemma intact. In the injury groups, blood cells were exuded at 1 day, and inflammatory cells infiltrated at 3 days, which reached the maximum at 7 days. The morphology of muscle fibers returned to normal at 21 days after injury. The resveratrol group was better than the natural recovery group in terms of inflammatory cell infiltration and repair time. (2) Masson staining results: There were few collagen fibers in normal muscle cells. After injury, the number of collagen fibers increased with the formation of scar tissue, and reached a peak at 14 days. The content of collagen fibers in the resveratrol group was lower than that in the natural recovery group. (3) Immunohistochemistry and immunoblotting results: The expression of bFGF and IGF-1 protein first increased and then decreased after injury. In both groups, the expression of bFGF and IGF-1 protein reached the peak at 7 days and was still at a high level at 21 days. The resveratrol group had significantly higher bFGF and IGF-1 levels than the natural recovery group (P<0.05). Overall, resveratrol can effectively accelerate the histological healing process and improve the healing quality of rabbit skeletal muscle after blunt trauma. Resveratrol significantly promotes the repair of damaged skeletal muscle by up-regulating bFGF and IGF-1 expression, but not altering the overall change of protein expression during skeletal muscle injury repair.
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BACKGROUND: At present, cell patch technology has been widely used in the repair of various tissues and organs such as periodontal ligament, cornea, heart, cartilage, and esophagus. However, the effect and mechanism of basic fibroblast growth factor (bFGF) on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cell patches are still unknown. It is of great significance for understanding how to integrate growth factor with tissueengineered cell patch technology for the final use in tissue engineering repairing. OBJECTIVE: To investigate the expression of angiogenesis-related factors in rat bone marrow mesenchymal stem cell patch during bFGF-induced osteogenic differentiation. METHODS: After isolation, purification and identification, rat bone marrow mesenchymal stem cell patches were constructed by continuous induction of vitamin C and divided into two groups: bFGF group (20 µg/L bFGF+osteoinductive medium) and control group (osteoinductive medium). The expression of angiogenesis-related genes was detected by alizarin red staining at 7 and 14 days of osteogenic induction and by fluorescence quantitative PCR method at 10 days of osteogenic induction. RESULTS AND CONCLUSION: The results of alizarin red staining showed that the number of calcified nodules in the bFGF group was significantly higher than that in the control group. The expression of transforming growth factor Β1 mRNA in the bFGF group was significantly higher than that in the control group, while the expression of vascular endothelial growth factor, angiopoietin 1 and platelet-derived factor BB was lower than that in the control group. Together, these results demonstrate that bFGF can induce the osteogenic differentiation of rat bone marrow mesenchymal stem cell patches, and increase the expression of transforming growth factor Β1 in the late osteogenic stage.
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@#AIM: To explore the effect of artificial tears combined with recombinant bovine basic fibroblast growth factor(rbFGF)on ocular surface function and inflammatory factors in patients with cataract complicated with xerophthalmia.<p>METHODS: A total of 118 cataract patients(118 eyes)with dry eyes treated in our hospital from February 2019 to February 2020 were randomly divided into control group(<i>n</i>=59)and observation group(<i>n</i>=59). The control group was treated with artificial tears before operation and the observation group was treated with artificial tears combined with rbFGF before operation. The clinical symptom score, ocular surface function index \〖ocular surface disease index(OSDI)questionnaire score, break up time(BUT), schirmer Ⅰ test(SⅠt), fluorescein stain test(FL)\〗, inflammatory factors in tears \〖interleukin-6(IL-6), tumor necrosis factor α(TNF-α)\〗 and oxidative stress indexes \〖malondialdehyde(MDA), lipid peroxide(LPO), superoxide dismutase(SOD), total antioxidant capacity(TAC)\〗 were compared between the two groups before and after treatment. The random walking model was used to evaluate the ocular surface function and the level of inflammatory factors in tears of the two groups.<p>RESULTS: Before treatment, there was no significant difference in clinical symptom score, OSDI, BUT, SⅠt, FL, IL-6, TNF-α, MDA, SOD, LPO and TAC between the two groups. Thirty days after treatment, the clinical symptom score, OSDI, FL, TNF-α, IL-6, MDA and LPO levels in two groups were significantly decreased, while the levels of BUT, SⅠt, SOD and TAC were significantly increased. The improvement of clinical symptom score, ocular surface function, inflammatory factors and oxidative stress in the observation group were significantly better than that in the control group.<p>CONCLUSION: Preoperative intervention with artificial tears combined with rbFGF can significantly improve the ocular surface function of cataract patients with xerophthalmia, reduce the level of inflammatory factors in tears, improve the symptoms of xerophthalmia, and provide reference for the clinical treatment of cataract with xerophthalmia.
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Context: Osteosarcoma (OS) is a progressive primary bone tumor that originates from immature stromal spindle cells. After chemotherapy, the serum-related indexes which are related to the prognosis. Aims: The aim of this study is to investigate the correlation between changes in serum cyclooxygenase-2 (COX-2), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), transforming growth factor-beta (TGF-β), and tumor-specific growth factor (TSGF) levels and prognosis of patients with osteosarcoma (OS) before and after treatment. Settings and Design: Data of 75 patients with OS (observation group) and 55 healthy controls (control group) were retrospectively analyzed. Materials and Methods: Chemotherapy was administered to the observation group. Serum lactate dehydrogenase, alkaline phosphatase, and TSGF levels were measured before and after treatment. The observation group patients were classified as normal or abnormal according to the changes in serum COX-2, bFGF, VEGF, TGF-β, and TSGF levels after chemotherapy. Patients were followed up for 7.5 years, and the survival rate was determined. Statistical Analysis Used: Single-factor influencing prognosis was included in the Cox model, and independent factors influencing prognosis were analyzed. Results: After chemotherapy, the mean serum COX-2, bFGF, VEGF, and TSGF levels decreased significantly in the observation group but were still higher than those in the control group. Furthermore, serum TGF-β levels increased in the observation group but were still lower than those in the control group. The 5-year survival rate of patients with normal serum COX-2, bFGF, VEGF, and TSGF levels was significantly higher in the normal subgroup than in the abnormal subgroup. Cox analysis showed that the Enneking stage and COX-2 level after chemotherapy were independent prognostic factors. Conclusions: The serum COX-2, bFGF, VEGF, and TSGF levels of patients with OS significantly changed after chemotherapy, and the short-term survival rate of patients with normal levels of these biomarkers after chemotherapy was high
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BACKGROUND: As a necessary regulatory factor of in vitro cell culture, and in vivo cell growth and proliferation, growth factors have attracted much attention. OBJECTIVE: To investigate the effects of basic fibroblast growth factor (bFGF) combined with insulin-like growth factor-1 (IGF-1) on the proliferation and apoptosis of spermatogonial stem cells in mice. METHODS: Spermatogonial stem cells were isolated and cultured from the testis of 6-8 days old male Kunming mice and were identified. Spermatogonial stem cells were inoculated into the feeder layer of embryonic fibroblasts treated with mitomycin C, and were then divided into four groups. Control group was cultured with normal DMEM medium; bFGF and IGF-1 groups were cultured with DMEM medium containing 20 μg/L bFGF and 20 μg/L IGF-1, respectively; bFGF+IGF-1 group was cultured with DMEM medium containing 20 μg/L bFGF and 20 μg/L IGF-1. The proliferation activity of spermatogonial stem cells was detected by cell counting kit-8 assay and EDU staining. The growth cycle and apoptosis of spermatogonial stem cells were detected by flow cytometry. The expression levels of PCNA, Bax and Bcl-2 proteins were detected by western blot assay. RESULTS AND CONCLUSION: Compared with the control group, the absorbance values in the bFGF, IGF-1 and bFGF+IGF-1 groups were significantly increased. Compared with the bFGF and IGF-1 groups, the absorbance values in the bFGF+IGF-1 group were further increased (P < 0.05). EDU staining results showed the same conclusion as cell counting kit-8 assay results. The proportion of S+G2/M phase cells in the bFGF+IGF-1 group was significantly higher than that in the other three groups (P < 0.05). The proportion in the IGF-1 and bFGF groups was significantly higher than that in control group (P < 0.05). Compared with the control group, the number of apoptotic cells in the bFGF, IGF-1 and bFGF+IGF-1 groups was decreased. Compared with the bFGF and IGF-1 groups, the number of apoptotic cells in the bFGF+IGF-1 group was further decreased. Compared with the control group, the relative expression levels of Bax protein in the bFGF, IGF-1 and bFGF+IGF-1 groups were significantly decreased (P < 0.01), and the expression levels of Bcl-2 and PCNA proteins were significantly increased (P < 0.05). Compared with the bFGF and IGF-1 groups, the relative expression level of Bax protein in the bFGF + IGF-1 group was decreased further (P < 0.01), and the relative expression levels of Bcl-2 and PCNA proteins were increased further (P < 0.05). These results indicate that bFGF and IGF-1 can promote cell proliferation and inhibit cell apoptosis by up-regulating the expression of PCNA and Bcl-2 proteins and down-regulating the expression of Bax protein. The combination of bFGF and IGF-1 can achieve favorable effects.
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BACKGROUND: Nowadays, there are many studies addressing basic fibroblast growth factor (bFGF) that promotes chondrocyte differentiation and inflammatory factors that damage chondrocyte structure, such as interleukin-1, but there are no reports on the combined effect of the two factors on chondrocytes. OBJECTIVE: To observe the effect of bFGF and inflammatory cytokines on growth characteristics and differentiation potential of chondrocytes. METHODS: Passage 3 chondrocytes from Sprague-Dawley rats cultured in vitro were divided into four groups: a blank control group in which chondrocytes were cultured alone, a negative control group in which chondrocytes were cultured with serum-free DMEM/F12 containing tumor necrosis factor-α, interleukin-1 and interleukin-6, a positive control group in which chondrocytes were cultured with serum-free DMEM/F12 containing bFGF, and an experimental group in which chondrocytes were cultured with serum-free DMEM/F12 containing bFGF, tumor necrosis factor-α, interleukin-1 and interleukin-6. At 3, 5, and 7 days after culture, the proliferative activity of chondrocytes was detected by cell counting kit-8 method; the levels of inflammatory cytokines were determined by ELISA method; and mRNA expression of collagen type II and proteoglycan in chondrocytes was measured by real-time PCR. Protein levels of collagen type II and proteoglycan were assessed by immunofluorescence assay at 7 days after culture. RESULTS AND CONCLUSION: At 3, 5, 7 days after culture, the proliferation of chondrocytes ranked as follows: positive control group > experimental group > blank control group > negative control group. Compared with the blank control group, the expressions of interleukin-1, interleukin-6 and tumor necrosis factor-α were significantly increased in the negative control group. However, compared with the negative control group, the expressions of the above three inflammatory cytokines in the experimental group showed a significant decline (P < 0.05). There was positive expression of collagen type II and proteoglycan in the blank control, positive control and experimental groups, especially in the negative control group. Compared with the blank control group, the expressions of collagen type II and proteoglycan were up-regulated significantly in the positive control and experimental groups. To conclude, the rationale use of bFGF can maintain the phenotype of chondrocytes, inhibit dedifferentiation and promote cell proliferation.
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BACKGROUND: Prostaglandin E1 and basic fibroblast growth factor can promote the proliferation of human dental pulp stem cells, but the effects of their combinations on the proliferation of human dental pulp stem cells and angiogenesis are unknown. OBJECTIVE: To investigate the effects of prostaglandin E1 combined with basic fibroblast growth factor on the proliferation and angiogenesis of human dental pulp stem cells. METHODS: (1) Human dental pulp stem cells were isolated and cultured in vitro. After detection and identification of surface markers, prostaglandin E1 and basic fibroblast growth factor at concentrations of 5, 10, 20, 50 and 100 μg/L were used to treat human dental pulp stem cells in vitro. The untreated cells served as control group. The cell proliferation was detected by cell counting kit-8 assay, and the optimum drug concentration and time of drug action were screened. (2) The in vitro cultured human dental pulp stem cells were divided into four groups: blank control group, prostaglandin E1 group, basic fibroblast growth factor group and combination group. The cell proliferation was detected by cell counting kit-8 assay. Human dental pulp stem cell conditioned medium was extracted. The levels of vascular endothelial growth factor and endostatin in the culture medium were detected by ELISA. The in vitro tubular formation ability of human umbilical vein endothelial cells after culture in conditioned medium was tested by tubule formation experiment. RESULTS AND CONCLUSION: The optimum concentration of prostaglandin E1 and basic fibroblast growth factor was 20 μg/L, and the optimum time of action was 2 days. Compared with the blank control group, the relative proliferation rate, level of vascular endothelial growth factor and the angiogenesis ability of human umbilical vein endothelial cell in vitro in the prostaglandin E1, basic fibroblast growth factor and combination groups were significantly increased (P < 0.05), while the level of endostatin was significantly decreased (P < 0.05). All above index levels in the combination group were significantly superior to those in the prostaglandin E1 and basic fibroblast growth factor groups (P < 0.05). In summary, prostaglandin E1 combined with basic fibroblast growth factor can promote the proliferation of human dental pulp stem cells and enhance the tubular formation ability of human umbilical vein endothelial cells in vitro.
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Objective: To observe the expressions of transforming growth factor β 1 (TGF-β 1) and basic fibroblast growth factor (bFGF) induced membrane by Masquelet technique in rats treated with glycoside of short-horned epimedium Herb, and to explore the effect of glycoside of short-horned epimedium Herb on Masquelet induced membrane. Methods: Sixty 3-month-old male Wistar rats were randomly divided into 3 groups with 20 rats in each group; a tibial bone defect (6 mm in length) model was established. The blank group (group A) was not treated; the control group (group B) and the experimental group (group C) were filled with vancomycin antibiotic bone cement in the drawing stage, and the bone cement was completely solidified. Group C was given perfused flavonoids glycoside of short-horned epimedium Herb (10 μmol/L) by gavage once a day (0.3 mL) from 1 day after operation, and groups A and B were given the same amount of normal saline by gavage. After operation, the recovery and wound healing of experimental animals were observed; at 4 weeks after operation, X-ray film was taken to observe the recovery of bone defect of proximal tibia; at 6 weeks after operation, the bone defect was observed, and the morphological changes and vascularization degree of granulation tissue and induction membrane tissue were observed; the expressions of TGF-β 1 and bFGF were observed by immunohistochemistry staining and ELISA detection. Results: The bone defect models of the 3 groups were established successfully, and there was no abnormality after operation. The incisions healed by first intention after operation. At 4 weeks after operation, X-ray films of proximal tibial defect showed that there was obvious space in group A, while bone cement was filled and Kirschner wire fixation was good in groups B and C. At 6 weeks after operation, the gross observation showed that the granulation tissue was filled in the defect area in group A; transparent membrane was formed in groups B and C, and microvessels were seen in some areas, and the microvessels in group C were significantly more than those in group B. Immunohistochemical staining showed that the expressions of TGF-β 1 and bFGF were negative in group A, but they were expressed in groups B and C, and the expressions of TGF-β 1 and bFGF in group B were significantly less than those in group C. ELISA detection showed that the expressions of TGF-β 1 and bFGF in group C were significantly higher than those in groups A and B ( P0.05). Conclusion: Glycoside of short-horned epimedium Herb can significantly increase the expressions of TGF-β 1 and bFGF, accelerate the process of osteogenesis, and contribute to bone shaping and reconstruction.
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Objective: To investigate the effects of Purendan Superfine Powder (PRD) on the retinal hemodynamics and the expressions of cytokines in the diabetic rats, and to elucidate its protective effect on diabetic retinopathy (DR). Methods: Thirty-six male SD rats were randomly divided into control group, diabetes group, and PRD group ( n= 12). The diabetes rat models were established by intraperitoneal injecting streptozotocin (STZ) in diabetes group and PRD group. After successful modeling, the diabetic rats in PRD group were given PRD (1. 8 g • kg-1 ) by gavage for 3 months. The hemodynamics parameters of central retinal artery (CRA) of the rats were measured with color Doppler ultrasound. The expression levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA in retina tissue were detected by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemical staining and Western blotting methods were used to detect the expression levels of VEGF and bFGF proteins in retina tissue of the rats. Results: Compared with control group, the blood glucose, urine volume, liver index and kidney index of the rats in diabetes group were significantly increased (P<0. 05), the peak systolic velocity (PSV), end diastolic velocity (EDV) and mean velocity (MV) of CRA were significantly decreased ( P<0. 05), the resistant index (RI) and pulsatility index (PI) of CRA were significantly increased ( P
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Objective: To prepare the vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)-loaded poloxamer thermosensitive gels and investigate the in vitro drug release property as well as the therapeutic effect of the drug-loaded thermosensitive gels on the full-thickness skin defected wound healing in rats. Meth- ods: The release of VEGF and bFGF from the drug-loaded gels in vitro was determined by the membrane release meth- od. The full-thickness skin defected model of rats was established and divided into four groups: the normal saline(Con- trol)group, poloxamer thermosensitive gel(Gel)group, freeze-dried VEGF and bFGF powder(VEGF/bFGF powder) group, and the VEGF/bFGF-loaded poloxamer thermosensitive gel(VEGF/bFGF-loaded gel)group. The wound healing was observed on the 3rd, 7th and 14th day of injury. The wound tissue was stained with hematoxylin-eosin. The wound healing and inflammation were observed under the microscope. The expression of CD31 was measured by the immunohis- tochemical staining and the number of new blood vessels was counted. The content of inflammatory factors in the wound was detected by ELISA. Results: The VEGF/bFGF-loaded poloxamer thermosensitive gels were successfully prepared. The in vitro drug release test showed that only about 5% of VEGF and bFGF in the drug-loaded gels was released from the gels within 1 h of the test, however, the subsequent VEGF and bFGF release could reach 60% on the fifth day of the re- lease test, suggesting the well sustained drug-release behavior of the VEGF/bFGF-loaded gels. In the wound healing test using the full-thickness skin defected rat model, the wound healing rate reached(90.3±2.4)% on the 14th day of injury, in the VEGF/bFGF-loaded gel group, which was significantly higher than that in the other groups(P<0.05). On the 3rd day of injury, compared with the Control and Gel groups, the level of IL-6 and TNF-α in the wound was significantly de- creased in the VEGF/bFGF-loaded gel group(P<0.05). The tissue HE staining showed that the skin wound healed well in the VEGF/bFGF-loaded gel and the VEGF/bFGF powder groups, and a lager number of neovascularization and the epi- dermis appeared earlier in both the VEGF/bFGF-loaded gel and the VEGF/bFGF powder groups than in the Control and the Gel groups. The immunohistochemical results further confirmed that the number of neovascularization was significant- ly higher in the VEGF/bFGF-loaded gel and the VEGF/bFGF powder groups than in the Control and Gel groups(P< 0.05). Conclusion: The prepared VEGF/bFGF-loaded poloxamer thermosensitive gels in the present study could be used to repair the full-thickness skin defected wound in rats, which could reduce the early inflammatory reaction, pro- mote the angiogenesis, and thus accelerate the healing.
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Objective@#To observe the effects of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor C (VEGF-C) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into lymphatic endothelial cells (LECs).@*Methods@#The third to the fifth passage of BMSCs of rats were collected for the following experiments. (1) BMSCs of rats were collected and divided into negative control group, CD90 group, CD44 group, and CD34 group according to the random number table (the same grouping method below), with 3 samples in each group. Phosphate buffer of 5 μL was added to cells in negative control group, and cells in the other 3 groups were added with 5 μL corresponding antibodies respectively. The positive expression of cell surface antigen was detected by flow cytometer. (2) BMSCs of rats in 3 batches were collected and divided into blank control group, VEGF-C group, HGF group, bFGF group, VEGF-C+ HGF group, VEGF-C+ bFGF group, HGF+ bFGF group, and VEGF-C+ HGF+ bFGF group, with 3 samples in each group. Cells in blank control group were added with 2 mL complete medium, cells in VEGF-C group were added with 2 mL complete medium and 10 μL VEGF-C of 10 μg/mL, cells in HGF group were added with 2 mL complete medium and 16 μL HGF of 10 μg/mL, and cells in bFGF group were added with 2 mL complete medium and 20 μL bFGF of 1 μg/mL. Cells in VEGF-C+ HGF group, VEGF-C+ bFGF group, HGF+ bFGF group, and VEGF-C+ HGF+ bFGF group were added with 2 mL complete medium and induction factors with corresponding concentration and volume as above. On 10 d of culture, the morphology of the cells was observed by the inverted phase contrast microscope, and the protein and mRNA expressions of lymphatic vessel endothelial hyaluronic acid receptor 1 (LYVE-1), VEGF receptor 3 (VEGFR3), and integrin α9 were detected by Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction respectively. (3) BMSCs of rats were collected and divided into blank control group, HGF+ VEGF-C+ bFGF group, bFGF+ VEGF-C+ HGF group, and VEGF-C+ HGF+ bFGF group, with 3 samples in each group. Cells in blank control group were added with 2 mL complete medium. Cells in HGF+ VEGF-C+ bFGF group were added with 2 mL complete medium, 16 μL HGF of 10 μg/mL, and 10 μL VEGF-C of 10 μg/mL, after 6 hours, 20 μL bFGF of 1 μg/mL was added. Cells in bFGF+ VEGF-C+ HGF group were added with 2 mL complete medium, 20 μL bFGF of 1 μg/mL, and 10 μL VEGF-C of 10 μg/mL, after 6 hours, 16 μL HGF of 10 μg/mL was added. Cells in VEGF-C+ HGF+ bFGF group were simultaneously added with 2 mL complete medium and the same concentration and volume of three inducing factors as above. In addition, BMSCs of rats in another 2 batches were collected and grouped, and they were dealt with the same methods as above except that the interval time of 6 hours in HGF+ VEGF-C+ bFGF group and bFGF+ VEGF-C+ HGF group was adjusted to 12 and 24 hours. On 10 d of culture, protein expressions of LYVE-1, VEGFR3, and integrin α9 were detected by Western blotting. Data were processed with analysis of variance of factorial design, one-way analysis of variance, and least significant difference t test, and Bonferroni correction.@*Results@#(1) The positive expression rates of surface antigen of cells in negative control group, CD90 group, CD44 group, and CD34 group were 0.39%, 99.84%, 99.90%, and 0.57%, respectively. (2) On 10 d of culture, cells in blank control group, HGF group, bFGF group, and HGF+ bFGF group presented long fusiform, while cells in the other groups presented polygonal shape. (3) On 10 d of culture, there were no protein expressions of LYVE-1, VEGFR3, and integrin α9 in cells of blank control group, HGF group, bFGF group, and HGF+ bFGF group. On 10 d of culture, protein expressions of LYVE-1, VEGFR3, and integrin α9 in cells of VEGF-C+ HGF+ bFGF group were significantly higher than those in VEGF-C group (t=24.21, 11.04, 15.43, P<0.01), VEGF-C+ HGF group (t=10.81, 9.93, 10.20, P<0.01), and VEGF-C+ bFGF group (t=11.67, 6.32, 19.00, P<0.01). Protein expressions of LYVE-1 in cells of VEGF-C+ HGF group and VEGF-C+ bFGF group were significantly higher than the protein expression in VEGF-C group (t=8.69, 15.20, P<0.01). Protein expression of VEGFR3 in cells of VEGF-C+ bFGF group was obviously higher than the protein expressions in VEGF-C group and VEGF-C+ HGF group (t=8.67, 7.21, P<0.01). Protein expression of integrin α9 in cells of VEGF-C+ HGF group was obviously higher than the protein expressions in VEGF-C group and VEGF-C+ bFGF group (t=8.80, 8.83, P<0.01). (4) On 10 d of culture, there were no mRNA expressions of LYVE-1, VEGFR3, and integrin α9 in cells of blank control group, HGF group, bFGF group, and HGF+ bFGF group. On 10 d of culture, mRNA expressions of LYVE-1 and VEGFR3 in cells of VEGF-C group were significantly lower than those in VEGF-C+ bFGF group and VEGF-C+ HGF+ bFGF group (tLYVE-1=6.22, 18.01, tVEGFR3=8.49, 15.34, P<0.01), and mRNA expression of integrin α9 were significantly lower than that in VEGF-C+ HGF group and VEGF-C+ HGF+ bFGF group (t=13.24, 9.65, P<0.01). The mRNA expressions of LYVE-1, VEGFR3, and integrin α9 in cells of VEGF-C+ HGF+ bFGF group were obviously higher than those in VEGF-C+ HGF group and VEGF-C+ bFGF group (t=13.92, 11.95, 13.72, 5.27, 5.64, 9.10, P<0.01). Compared with those of VEGF-C+ bFGF group, the mRNA expression of VEGFR3 of cells in VEGF-C+ HGF group was significantly lower (t=6.91, P<0.01), while the mRNA expression of integrin α9 of cells in VEGF-C+ HGF group was significantly higher (t=11.69, P<0.01). (5) On 10 d of culture at interval time of 6, 12, 24 h, there were no protein expressions of LYVE-1, VEGFR3, or integrin α9 in cells of blank control group. On 10 d of culture at interval time of 6, 12, 24 h, the protein expressions of LYVE-1, VEGFR3, and integrin α9 in cells of HGF+ VEGF-C+ bFGF group, bFGF+ VEGF-C+ HGF group, and VEGF-C+ HGF+ bFGF group were close (F6 h=2.25, 2.47, 2.19, F12 h=2.93, 1.47, 3.25, F24 h=0.28, 0.20, 1.01, P>0.05).@*Conclusions@#VEGF-C is a necessary factor for inducing BMSCs to differentiate into LECs. HGF and bFGF may promote the differentiation by up-regulating the expressions of integrin α9 and VEGFR3 respectively. But the induction effects of the two factors may be independent. The combination of VEGF-C, HGF, and bFGF have the best effects of promoting differentiation.
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AIM:To study the effects of basic fibroblast growth factor (bFGF) on brain edema, nerve function damage and autophagy related proteins in rats with head injury.METHODS:The rat model of craniocerebral injury (CI) was constructed.The rats were divided into control group, CI group, and low-, middle-and high-dose bFGF groups (n=10).The CI model was established in CI group, while the rats in control group were not given epidural impact.The rats in low-dose, middle-dose and high-dose bFGF groups were given bFGF at 2, 4 and 6μg, respectively, by intraperitoneal injection after 30 min.The neurological function in the rats was evaluated by improved neurological function scoring.The rat brain tissues were taken, and the water content was detected.The levels of tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6) and IL-1βin the brain tissue were measured by ELISA.The malondialdehyde (MDA) content was analyzed by thiobarbituric acid method.The activity of superoxide dismutase (SOD) was examined by WST-8 assay.The glutathine peroxidase (GSH-Px) activity was detected by colorimetric method.The protein levels of autophagy related proteins LC3-II and beclin-1 in the brain tissues were determined by Western blot.RESULTS:The neurological function score was increased significantly of the rats in CI group.The rat model of craniocerebral injury was successfully constructed.Neurological function scores in the rats in low-dose, middle-dose and high-dose bFGF groups were reduced, the water content of the brain tissue was also reduced (P<0.05).The levels of TNF-α, IL-6 and IL-1βwere decreased in the brain tissues (P<0.05) , the content of MDA was declined (P<0.05) , the activities of SOD and GSH-Px were increased (P<0.05) , the protein levels of LC3-II and beclin-1 were decreased, compared with the untreated rats in CI group (P<0.05).CON-CLUSION:bFGF improves the nerve function of the rats with craniocerebral injury, reduces the water content of the brain tissue, reduces the expression of autophagic protein LC3-II and beclin-1.The mechanism is related to the inhibition of inflammatory reaction and oxidative damage.
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Objective: To observe the effect of vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor (VEGF/PELA/bFGF) mixed microcapsules in promoting the angiogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: The BMSCs were isolated by the method of whole bone marrow adherent, and sub-cultured. The passage 3 BMSCs were identified by Wright-Giemsa staining and flow cytometry, and used for subsequent experiments. VEGF/PELA/bFGF (group A), PELA/bFGF (group B), VEGF/PELA (group C), and PELA (group D) microcapsules were prepared. The biodegradable ability and cytotoxicity of PELA microcapsule were determined，and the slow-released ability of VEGF/PELA/bFGF mixed microcapsules was measured. The passage 3 BMSCs were co-cultured with the extracts of groups A, B, C, and D, separately. At 1, 3, 7, 14, and 20 days after being cultured, the morphological changes of induced BMSCs were recorded. At 21 days, the induced BMSCs were tested for DiI-labeled acetylated low density lipoprotein (Dil-ac-LDL) and FITC-labeled ulex europaeus agglutinin I (FITC-UEA-I) uptake ability. The tube-forming ability of the induced cells on Matrigel was also verified. The differences of the vascularize indexes in nodes, master junctions, master segments, and tot.master segments length in 4 groups were summarized and analyzed. Results: The isolated and cultured cells were identified as BMSCs. The degradation time of PELA was more than 20 days. There was no significant effect on cell viability under co-culture conditions. At 20 days, the cumulative release of VEGF in the mixed microcapsules exceeded 95%, and the quantity of bFGF exceeded 80%. The morphology of cells in groups A, B, and C were changed. The cells in groups A and B showed the typical change of cobble-stone morphology. The numbers of double fluorescent labeled cells observed by fluorescence microscope were the most in group A, and decreases from group B and group C, with the lowest in group D. The cells in groups A and B formed a grid-like structure on Matrigel. Quantitative analysis showed that the differences in the number of nodes, master junctions, master segments, and tot.master segments length between groups A, B and groups C, D were significant ( P0.05). Conclusion: VEGF/PELA/bFGF mixed microcapsules have significantly ability to promote the angiogenic differentiation of rat BMSCs in vitro.
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Objective To investigate the effect of acupuncture and moxibustion plus Zushima on serum vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b FGF) in patients with knee osteoarthritis.Method Two hundred and thirty-six patients with knee osteoarthritis were allocated, using a random number table, to two groups, 118 cases each. The control group was treated with Celecoxib and the observation group, with acupuncture and moxibustion plus Zushima. The therapeutic effects, and pre-treatment and post-treatment VAS scores and WOMAC scores and serum VEGF and bFGF levels were compared between the two groups of patients. The adverse reaction incidences were compared between the two groups of patients. Result The total efficacy rate was 87.3％ in the control group and 94.9％ in the observation group and was significantly higher in the observation group than in the control group (P<0.05). There were no statistically significant pre-treatment differences in the VAS score and WOMAC score between the two groups (P>0.05). The VAS score and WOMAC score decreased significantly in both groups after treatment (P<0.05). After treatment, there was no statistically significant difference in the VAS score between the two groups (P>0.05) but the WOMAC score was significantly lower in the observation group than in the control group (P<0.05). There were no statistically significant pre-treatment differences in VEGF and bFGF between the two groups (P>0.05). After treatment, VEGF and bFGF decreased significantly in both groups (P<0.05) and were significantly lower in the observation group than in the control group (P<0.05). The adverse reaction incidence was3.4％ in the control group and 1.7％ in the observation group with no statistically significant difference between the two groups (P> 0.05). Conclusion Acupuncture and moxibustion plus Zushima can reduce inflammatory reactions, inhibit synovial angiogenesis, and effectively relive the pain, and improve articular functions in the treatment of knee osteoarthritis.