In this study,different molecular weight(MW)carboxymethyl chitosans(CM-chitosan)nanoparticles were prepared by ionic gelification.The particle size of nanoparticles was around 180–250 nm by dynamic light scattering(D...In this study,different molecular weight(MW)carboxymethyl chitosans(CM-chitosan)nanoparticles were prepared by ionic gelification.The particle size of nanoparticles was around 180–250 nm by dynamic light scattering(DLS)and transmission electron microscope(TEM).With the increase of CM-chitosan nanoparticles concentration from 2 to 200μg/mL,the growth inhibition effects on the keloid fibroblast increased.At the concentration of 100μg/mL,CM-chitosan nanoparticles withMW6.3 kDa had a significant inhibitory effect(inhibition ratio 48.79%)of the proliferation of keloid fibroblast.Compared with CM-chitosan solution,the inhibition of CM-chitosan nanoparticles were lower in prior period and similar in later period.By analyzing the different effects of chitosan,CM-chitosan solution and CM-chitosan nanoparticles on proliferation of keloid fibroblast,we have found that the carboxylmethyl groups of CM-chitosan play an important role in inhibition of proliferation of keloid fibroblast.展开更多
AIM: To study the expression of SMAD3 and SMAD7 of transforming growth factor β1 (TGF β1)on keloid derived fibroblasts. METHODS: The expression of SMAD3 (at 1,2,4,24,48 h)and SMAD7(at 0.5 1,1.5, 2,4,24 h) were detec...AIM: To study the expression of SMAD3 and SMAD7 of transforming growth factor β1 (TGF β1)on keloid derived fibroblasts. METHODS: The expression of SMAD3 (at 1,2,4,24,48 h)and SMAD7(at 0.5 1,1.5, 2,4,24 h) were detected by using methods of Western blot after 500 pmol/L TGF β1 were added to the monolayer culture system.RESULTS: The level of SMAD3 were down regulated at 24 h and the SMAD7 were maximum up regulated at 4 h when TGF β1 were used.CONCLUSION: TGF β1 may down regulate the expression of SMAD3,but up regulate that of SMAD7 .展开更多
Background Transforming growth factor-β1 （TGF-β1） is known to have a role in keloid formation through the activation of fibroblasts and the acceleration of collagen deposition. The objective of this current study ...Background Transforming growth factor-β1 （TGF-β1） is known to have a role in keloid formation through the activation of fibroblasts and the acceleration of collagen deposition. The objective of this current study was to isolate TGF-β1 phage model peptides from a phage display 7-mer peptide library to evaluate their therapeutic effect on inhibiting the activity of keloid fibroblasts.Methods A phage display 7-mer peptide library was screened using monoclonal anti-human TGF-β1 as the target to obtain specific phages containing ectogenous model peptides similar to TGF-β1. Enzyme-linked immunosorbent assay （ELISA） was performed to select monoclonal phages with good binding activity, which underwent DNA sequencing. MTT assay and apoptosis assessment were used to evaluate the biological effects of the phage model peptides on keloid fibroblasts. Immunofluorescence assay was employed to show the binding affinity of the model peptides on phages causing keloid fibroblasts. Quantitative real-time PCR analysis was carried out to detect the expressions of nuclear factor κB （NF-κB） mRNA, connective tissue growth factor （CTGF） mRNA and TGF-β receptor Ⅱ （TβRII） mRNA in keloid fibroblasts.Results Specific phages with good results of ELISA were beneficiated. Four phage model peptides were obtained. The data of MTT showed that TGF-β1 and one phage model peptide （No. 4） could promote keloid fibroblasts proliferation,however, three phage model peptides （No. 1-3） could inhibit keloid fibroblasts proliferation. The results of apoptosis assessment showed that the three phage model peptides could slightly induce the apoptosis in keloid fibroblasts. The data of immunofluorescence assay revealed that the model peptides on phages rather than phages could bind to keloid fibroblasts. The findings of quantitative real-time PCR analysis suggested that the expressions of NF-κB mRNA and CTGF mRNA in the three phage model peptide groups decreased, while the expression of TβRII mRNA slightly increased.Conclusions Three phage model peptides isolated from a phage display 7-mer peptide library can inhibit keloid fibroblasts proliferation and induce the apoptosis in keloid fibroblasts. They can inhibit the activity of keloid fibroblasts by blocking TGF-β1 binding to its receptor and then regulating the expressions of NF-κB, CTGF and TβRII.展开更多
Objective: To define the mechanisms underlying the keloid formation. Methods: The gap junctional intercellu- lar commumication (GJIC) of fibroblasts derived from pathological scars and noed skin was investigated. Six ...Objective: To define the mechanisms underlying the keloid formation. Methods: The gap junctional intercellu- lar commumication (GJIC) of fibroblasts derived from pathological scars and noed skin was investigated. Six Samples of hyperplastic scars, keloids and normal skin were obtained. Fibroblasts from these samples were cultured in ho and vended by type Ⅰ collagen, and the GJIC among the fibroblasts was investigated by adherent cell analysis with soning interactive laser coytometer(ACAS 570) for fibroblasts culturing. Results: The fibroblasts from normal skin showed nounal GJIC, which are depressed between fibroblasts from hyperplastic scare and was completely blocked in fibroblasts from the keloids. Conclusion: The blockade of interoellular communication may play an important role in the pathogenesis of excessive and invasive growth of fibroblasts derived from the keloids.展开更多
The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many s...The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many similar characteristics. Antineoplastic drugs can downregulate fibroblast activity and cytokine growth factors. This study evaluates the effect of six antineoplastic drugs on keloid and Dupuytren’s disease fibroblasts. Keloid, normal scar, Dupuytren’s affected palmar fascia, and normal palmar fascia fibroblasts were grown and seeded into Fibroblast Populated Collagen Lattices (FPCLs). The FPCLs were treated with one of six antineoplastic drugs or left untreated as controls. At 7 days, supernatants were extracted from all FPCLs and assayed for expression of Transforming Growth Factor beta (TGF)-β<sub>1</sub> and TGF-β<sub>2</sub>. All six antineoplastic drugs significantly inhibited FPCL contraction in both fibroproliferative conditions compared with the untreated controls (p β<sub>1</sub> and TGF-β<sub>2</sub> expression was downregulated in the supernatants of all FPCLs by the drug exposure. Cytotoxicity did not occur in these studies and was not the reason for the results. Although antineoplastic drugs can have significant side effects when given systemically, these results may be minimized when given to small areas involved in fibroproliferative scarring or when given topically or intralesionally. These in vitro results suggest that antineoplastic drugs may have a utility for treating various fibroproliferative disorders and warrant further investigation.展开更多
基金This work was supported by the National Natural Science Foundation of China(Grant No.81071274).
文摘In this study,different molecular weight(MW)carboxymethyl chitosans(CM-chitosan)nanoparticles were prepared by ionic gelification.The particle size of nanoparticles was around 180–250 nm by dynamic light scattering(DLS)and transmission electron microscope(TEM).With the increase of CM-chitosan nanoparticles concentration from 2 to 200μg/mL,the growth inhibition effects on the keloid fibroblast increased.At the concentration of 100μg/mL,CM-chitosan nanoparticles withMW6.3 kDa had a significant inhibitory effect(inhibition ratio 48.79%)of the proliferation of keloid fibroblast.Compared with CM-chitosan solution,the inhibition of CM-chitosan nanoparticles were lower in prior period and similar in later period.By analyzing the different effects of chitosan,CM-chitosan solution and CM-chitosan nanoparticles on proliferation of keloid fibroblast,we have found that the carboxylmethyl groups of CM-chitosan play an important role in inhibition of proliferation of keloid fibroblast.
文摘AIM: To study the expression of SMAD3 and SMAD7 of transforming growth factor β1 (TGF β1)on keloid derived fibroblasts. METHODS: The expression of SMAD3 (at 1,2,4,24,48 h)and SMAD7(at 0.5 1,1.5, 2,4,24 h) were detected by using methods of Western blot after 500 pmol/L TGF β1 were added to the monolayer culture system.RESULTS: The level of SMAD3 were down regulated at 24 h and the SMAD7 were maximum up regulated at 4 h when TGF β1 were used.CONCLUSION: TGF β1 may down regulate the expression of SMAD3,but up regulate that of SMAD7 .
基金This study was supported by grants from the Nationa1 Natura1 Science Foundation of China （No. 30670571, 30772258 and 81071560）, Science and Technology Research Program of Shandong Province （No. 2009GG10002078）, Scientific Research Development Plan of the Department of Education of Shandong Province （No. J07WD03） and National Basic Research Program of China （973 Program） （No. 2005CB522603）.
文摘Background Transforming growth factor-β1 （TGF-β1） is known to have a role in keloid formation through the activation of fibroblasts and the acceleration of collagen deposition. The objective of this current study was to isolate TGF-β1 phage model peptides from a phage display 7-mer peptide library to evaluate their therapeutic effect on inhibiting the activity of keloid fibroblasts.Methods A phage display 7-mer peptide library was screened using monoclonal anti-human TGF-β1 as the target to obtain specific phages containing ectogenous model peptides similar to TGF-β1. Enzyme-linked immunosorbent assay （ELISA） was performed to select monoclonal phages with good binding activity, which underwent DNA sequencing. MTT assay and apoptosis assessment were used to evaluate the biological effects of the phage model peptides on keloid fibroblasts. Immunofluorescence assay was employed to show the binding affinity of the model peptides on phages causing keloid fibroblasts. Quantitative real-time PCR analysis was carried out to detect the expressions of nuclear factor κB （NF-κB） mRNA, connective tissue growth factor （CTGF） mRNA and TGF-β receptor Ⅱ （TβRII） mRNA in keloid fibroblasts.Results Specific phages with good results of ELISA were beneficiated. Four phage model peptides were obtained. The data of MTT showed that TGF-β1 and one phage model peptide （No. 4） could promote keloid fibroblasts proliferation,however, three phage model peptides （No. 1-3） could inhibit keloid fibroblasts proliferation. The results of apoptosis assessment showed that the three phage model peptides could slightly induce the apoptosis in keloid fibroblasts. The data of immunofluorescence assay revealed that the model peptides on phages rather than phages could bind to keloid fibroblasts. The findings of quantitative real-time PCR analysis suggested that the expressions of NF-κB mRNA and CTGF mRNA in the three phage model peptide groups decreased, while the expression of TβRII mRNA slightly increased.Conclusions Three phage model peptides isolated from a phage display 7-mer peptide library can inhibit keloid fibroblasts proliferation and induce the apoptosis in keloid fibroblasts. They can inhibit the activity of keloid fibroblasts by blocking TGF-β1 binding to its receptor and then regulating the expressions of NF-κB, CTGF and TβRII.
文摘Objective: To define the mechanisms underlying the keloid formation. Methods: The gap junctional intercellu- lar commumication (GJIC) of fibroblasts derived from pathological scars and noed skin was investigated. Six Samples of hyperplastic scars, keloids and normal skin were obtained. Fibroblasts from these samples were cultured in ho and vended by type Ⅰ collagen, and the GJIC among the fibroblasts was investigated by adherent cell analysis with soning interactive laser coytometer(ACAS 570) for fibroblasts culturing. Results: The fibroblasts from normal skin showed nounal GJIC, which are depressed between fibroblasts from hyperplastic scare and was completely blocked in fibroblasts from the keloids. Conclusion: The blockade of interoellular communication may play an important role in the pathogenesis of excessive and invasive growth of fibroblasts derived from the keloids.
文摘The various fibroproliferative disorders affecting humans have in common excess fibroblast activity and persistent overexpression or dysregulated activity of transforming growth factor beta (TGF-β). Cancer has many similar characteristics. Antineoplastic drugs can downregulate fibroblast activity and cytokine growth factors. This study evaluates the effect of six antineoplastic drugs on keloid and Dupuytren’s disease fibroblasts. Keloid, normal scar, Dupuytren’s affected palmar fascia, and normal palmar fascia fibroblasts were grown and seeded into Fibroblast Populated Collagen Lattices (FPCLs). The FPCLs were treated with one of six antineoplastic drugs or left untreated as controls. At 7 days, supernatants were extracted from all FPCLs and assayed for expression of Transforming Growth Factor beta (TGF)-β<sub>1</sub> and TGF-β<sub>2</sub>. All six antineoplastic drugs significantly inhibited FPCL contraction in both fibroproliferative conditions compared with the untreated controls (p β<sub>1</sub> and TGF-β<sub>2</sub> expression was downregulated in the supernatants of all FPCLs by the drug exposure. Cytotoxicity did not occur in these studies and was not the reason for the results. Although antineoplastic drugs can have significant side effects when given systemically, these results may be minimized when given to small areas involved in fibroproliferative scarring or when given topically or intralesionally. These in vitro results suggest that antineoplastic drugs may have a utility for treating various fibroproliferative disorders and warrant further investigation.
