Objective:Hypertrophic scars(HS)are a variety of skin tissue fibrosis disease that occurs in human skin,the effective therapeutic method of which is still inaccessible up to now.As a bioactive constituent of a well-kn...Objective:Hypertrophic scars(HS)are a variety of skin tissue fibrosis disease that occurs in human skin,the effective therapeutic method of which is still inaccessible up to now.As a bioactive constituent of a well-known medical plant,Salvia miltiorrhiza(Danshen in Chinese),tanshinone Ⅱ_(A)(TSA)is reported to inhibit cell proliferation in HS.Therefore,the aim of this study was to prepare TSA self-soluble microneedles to strengthen its dermal retention and break through the difficulty of significantly thickening epidermal connective tissue and stratum corneum at the HS site.The possible mechanism of action in suppressing HS was studied using human skin fibroblasts(HSF).Methods:Tanshinone Ⅱ_(A) self-dissolving microneedles(TSA-MN)was prepared using a negative mold casting method.The prescription process of microneedle was optimized by Box-Behnken effect surface method.Different media were selected to investigate the ability of transdermal absorption and in vitro release.Furthermore,according to Cell Counting Kit-8(CCK8)method as well as the Western blot method,the effect of TSA-MN on the biological characteristics of HSF was investigated.Results:With remarkable slow release effect and dermal retention,the release and transdermal properties of TSA-MN in vitro were better than both TSA and ordinary dosage forms.Its effect of HSF confirmed the essential decrease in cell motility during cell proliferation and cell migration in vitro,which plays a significant role in down-regulating the secretion of transforming growth factor-β1(TGF-β1)in HSF and increasing the expression level of Smad7.Conclusion:The prepared TSA self-soluble microneedles is helpful in solving the problem of hypertrophic scars,with a stable dermal retention effect after process optimization.展开更多
Background:Epidermal stem cells(ESCs)are primarily located in the basal layer of the epidermis and play a crucial role in wound healing.ESCs-derived exosomes(ESCs-Exo)are emerging as promising candidates for skin rege...Background:Epidermal stem cells(ESCs)are primarily located in the basal layer of the epidermis and play a crucial role in wound healing.ESCs-derived exosomes(ESCs-Exo)are emerging as promising candidates for skin regeneration and wound healing.However,the underlying mechanisms remain unclear.This study aims to investigate the role and mechanisms of ESCs-Exo in promoting the proliferation,migration,and collagen synthesis of human skin fibroblasts(HSFBs).Methods:This study generated,isolated,and characterized ESC-Exos.The effects of ESCs-Exo on the proliferation of human skin fibroblasts(HSFBs)were detected via Cell Counting Kit-8(CCK8),5-Ethynyl-2’-deoxyuridine(EdU),and Proliferating Cell Nuclear Antigen(PCNA)and Marker of Proliferation Ki-67(MKI67)gene expression methods.The effect of ESCs-Exo on the migration of HSFBs was detected via a transwell assay and a scratch test.The concentrations of collagen secreted by the HSFBs and the mRNAs of the two kinds of collagen expressed by the HSFBs were analyzed.We also analyzed the phosphorylation of Protein Kinase N1(PKN1)and the expression of cyclins via western blotting.Finally,the effect of ESCs-Exo on wound healing was verified by animal experiments,and the key genes and signaling pathways of ESCs-Exo were excavated by transcriptomic analysis.Results:Western blotting revealed that the exosomes of ESCs highly expressed established markers such as Alix,CD63,and CD9.ESCExos significantly promoted HSFB proliferation and migration in a dose-dependent manner,as well as HSFB collagen synthesis,and effectively increased the ratio of collagen III/I.In addition,bioinformatics analysis showed that the expression of key gene C-X-C motif chemokine ligand 9 was lower in the ESCs-Exo group,which may promote wound healing by regulating PKN1-cyclin and tumor necrosis factor signaling pathways.Animal experiments demonstrated that ESCs-Exo could reduce inflammation and accelerate wound healing.Conclusions:In this study,we found that ESCs-Exo may improve wound healing by promoting the proliferation and migration of HSFBs.展开更多
The objective of this research was to assess the potential of phosphatidylcholineencapsulated resveratrol as a cosmetic ingredient.The hydrogen peroxide(H_(2)O_(2))and ultraviolet A(UVA)induced human skin fibroblasts(...The objective of this research was to assess the potential of phosphatidylcholineencapsulated resveratrol as a cosmetic ingredient.The hydrogen peroxide(H_(2)O_(2))and ultraviolet A(UVA)induced human skin fibroblasts(HSF)models of skin damage were established to compare the antioxidant and anti-wrinkle properties between phosphatidylcholine-encapsulated resveratrol and unencapsulated resveratrol.The findings reveal that encapsulating resveratrol with phosphatidylcholine not only enhances skin absorption but also significantly improves its antioxidant capabilities.In the H2O2-induced HSF injury model,phosphatidylcholine-encapsulated resveratrol demonstrates a superior ability to neutralize reactive oxygen species(ROS)generated by H2O2 compared to the resveratrol group.Further analysis indicates that this enhanced functionality is associated with increased enzymatic activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and catalase(CAT)when treated with phosphatidylcholine-encapsulated resveratrol.Additionally,in UVA-irradiated HSF cells,phosphatidylcholine-encapsulated resveratrol effectively reduces the levels of matrix metalloproteinases-1 and-3(MMP-1 and MMP-3)and increased the contents of CollagenⅠand CollagenⅢ(Col-1 and Col-3),demonstrating significant anti-wrinkle effects.These findings provide critical evaluation criteria and application references for enhancing cosmetic ingredients through phosphatidylcholine encapsulation,thereby advancing skincare formulations.展开更多
A study on the effect of the solar ultra-violet radiation on the human skin fibroblast cells revealed that the production of matrix metalloproteinase-2 was inhibited by the radiation.A CO2 incubator connected by optic...A study on the effect of the solar ultra-violet radiation on the human skin fibroblast cells revealed that the production of matrix metalloproteinase-2 was inhibited by the radiation.A CO2 incubator connected by optical fibers to a reflector telescope for collecting the solar light was built at Syowa station by the 49th Japanese Antarctica Research Expedition.The direction of the telescope was continuously controlled by a sun-tracker to follow the movement of the Sun automatically.The intensity of the collected light was monitored by a portable spectrophotometer housed inside.The human skin fibroblast cells were incubated in the CO2 chamber to investigate the effect of the solar radiation at Syowa station and were compared with those reference experiments at a laboratory in Japan.The results showed cell damage by strong UV radiation.The production of matrix metalloproteinase-2 was prompted by the moderate UV-B,but was inhibited by the strong UV-B radiation,as studied under laboratory conditions in Japan.The effect of strong solar radiation at Syowa station involving the radiation of UV-B region was estimated to be of the same extent of the radiation caused by an artificial UV-B light with the intensity more than 50 mJ/cm2.展开更多
[Objectives]To explore the effect of the extract of Growgx yeast fermentation product on ultraviolet A(UVA)-induced photo-aging of human skin fibroblasts(HFF-1)and its specific mechanisms from the level of cell biolog...[Objectives]To explore the effect of the extract of Growgx yeast fermentation product on ultraviolet A(UVA)-induced photo-aging of human skin fibroblasts(HFF-1)and its specific mechanisms from the level of cell biology,and discussed the application of the Growgx yeast fermentation product extract for skin aging.[Methods]A photo-aging model of HFF-1 cells was induced by UVA radiation meter,and it was given different concentrations of Growgx for intervention.The proliferation activity of the HFF-1 cells was detected with MTT method,and their migration and invasion ability was measured by scratch test.The SOD,HYP and MDA levels were detected with corresponding kits.After oxygen infusion apparatus assisted facial skin administration,the repair of the skin was observed through the VISIA skin tester.[Results]The survival rate of HFF-1 cells was reduced significantly by UVA with an irradiation dose of 10.8 J/cm2(P<0.05),while Growgx significantly increased their survival rate(P<0.001)and sped up the repair of damaged cells.Growgx promoted the production of SOD(P<0.05),reduced the level of MDA(P<0.05),and increased the level of HYP(P<0.05).Growgx effectively inhibited UVA-induced photo-aging of HFF-1 cells.The mechanism may be related to accelerating cell damage repair,up-regulating SOD and HYP expression levels,and reducing MDA expression level.The clinical observation showed that Growgx effectively improved skin spots and pores,making the skin smoother and more delicate.[Conclusions]Growgx can effectively fight against photo-aging caused by ultraviolet rays,and can significantly improve skin wrinkles.展开更多
Background:Senescent human skin primary fibroblast(FB)models have been established for studying aging-related,proliferative,and inflammatory skin diseases.The aim of this study was to compare the transcriptome charact...Background:Senescent human skin primary fibroblast(FB)models have been established for studying aging-related,proliferative,and inflammatory skin diseases.The aim of this study was to compare the transcriptome characteristics of human primary dermal FBs from children and the elderly with four senescence models.Methods:Human skin primary FBs were obtained from healthy children(FB-C)and elderly donors(FB-E).Senescence models were generated by ultraviolet B irradiation(FB-UVB),D-galactose stimulation(FB-D-gal),atazanavir treatment(FB-ATV),and replication exhaustion induction(FB-P30).Flow cytometry,immunofluorescence staining,real-time quantitative polymerase chain reaction,co-culturing with immune cells,and bulk RNA sequencing were used for systematic comparisons of the models.Results:In comparison with FB-C,FB-E showed elevated expression of senescence-related genes related to the skin barrier and extracellular matrix,proinflammatory factors,chemokines,oxidative stress,and complement factors.In comparison with FB-E,FB-UVB and FB-ATV showed higher levels of senescence and expression of the genes related to the senescence-associated secretory phenotype(SASP),and their shaped immune microenvironment highly facilitated the activation of downstream immune cells,including T cells,macrophages,and natural killer cells.FB-P30 was most similar to FB-E in terms of general transcriptome features,such as FB migration and proliferation,and aging-related characteristics.FB-D-gal showed the lowest expression levels of senescence-related genes.In comparisons with the single-cell RNA sequencing results,FB-E showed almost complete simulation of the transcriptional spectrum of FBs in elderly patients with atopic dermatitis,followed by FB-P30 and FB-UVB.FB-E and FB-P30 showed higher similarity with the FBs in keloids.Conclusions:Each senescent FB model exhibited different characteristics.In addition to showing upregulated expression of natural senescence features,FB-UVB and FB-ATV showed high expression levels of senescence-related genes,including those involved in the SASP,and FB-P30 showed the greatest similarity with FB-E.However,D-galactose-stimulated FBs did not clearly present aging characteristics.展开更多
Skin tissue engineering with considerable skin regeneration capability is an urgent need for the wound site.The current challenge for researchers is to develop a bionic scaffold that imitates the extracellular matrix ...Skin tissue engineering with considerable skin regeneration capability is an urgent need for the wound site.The current challenge for researchers is to develop a bionic scaffold that imitates the extracellular matrix for the regeneration of the damaged regions.In our study,poly(L-lactide-co-caprolactone)(PLCL)was blended with polyurethane(PU)to obtain nanofibrous scaffolds via electrospinning.The electrospun fibers with 50%PLCL content had a certain number of intersections and jointing points,and exhibited significantly enhanced mechani-cal properties combined with suitable porosity.Moreover,cell activities demonstrated that PU/PLCL membranes had significantly biological advantages in enhanced growth of human skin fibroblasts with spreading morphology compared with PU membranes,indicating good cytocompatibility of composite scaffolds.These findings proved that PU/PLCL electrospun membranes have great potential in applications of skin tissue engineering.展开更多
基金supported by the National Natural Science Foundation of China(No.82173982,2021)the Guangdong Natural Science Project(No.2018A0303130234)Excellent Young Teachers Training Project in Higher Education of Guangdong(No.YQ2015099).
文摘Objective:Hypertrophic scars(HS)are a variety of skin tissue fibrosis disease that occurs in human skin,the effective therapeutic method of which is still inaccessible up to now.As a bioactive constituent of a well-known medical plant,Salvia miltiorrhiza(Danshen in Chinese),tanshinone Ⅱ_(A)(TSA)is reported to inhibit cell proliferation in HS.Therefore,the aim of this study was to prepare TSA self-soluble microneedles to strengthen its dermal retention and break through the difficulty of significantly thickening epidermal connective tissue and stratum corneum at the HS site.The possible mechanism of action in suppressing HS was studied using human skin fibroblasts(HSF).Methods:Tanshinone Ⅱ_(A) self-dissolving microneedles(TSA-MN)was prepared using a negative mold casting method.The prescription process of microneedle was optimized by Box-Behnken effect surface method.Different media were selected to investigate the ability of transdermal absorption and in vitro release.Furthermore,according to Cell Counting Kit-8(CCK8)method as well as the Western blot method,the effect of TSA-MN on the biological characteristics of HSF was investigated.Results:With remarkable slow release effect and dermal retention,the release and transdermal properties of TSA-MN in vitro were better than both TSA and ordinary dosage forms.Its effect of HSF confirmed the essential decrease in cell motility during cell proliferation and cell migration in vitro,which plays a significant role in down-regulating the secretion of transforming growth factor-β1(TGF-β1)in HSF and increasing the expression level of Smad7.Conclusion:The prepared TSA self-soluble microneedles is helpful in solving the problem of hypertrophic scars,with a stable dermal retention effect after process optimization.
基金supported by National Natural Science Foundation of China(82272276)GuangDong Basic and Applied Basic Research Foundation(2022A1515012160,2024A1515010477)Chongqing traditional Chinese Medicine inheritance and Innovation team project(2023090006KJZX2022WJW008).
文摘Background:Epidermal stem cells(ESCs)are primarily located in the basal layer of the epidermis and play a crucial role in wound healing.ESCs-derived exosomes(ESCs-Exo)are emerging as promising candidates for skin regeneration and wound healing.However,the underlying mechanisms remain unclear.This study aims to investigate the role and mechanisms of ESCs-Exo in promoting the proliferation,migration,and collagen synthesis of human skin fibroblasts(HSFBs).Methods:This study generated,isolated,and characterized ESC-Exos.The effects of ESCs-Exo on the proliferation of human skin fibroblasts(HSFBs)were detected via Cell Counting Kit-8(CCK8),5-Ethynyl-2’-deoxyuridine(EdU),and Proliferating Cell Nuclear Antigen(PCNA)and Marker of Proliferation Ki-67(MKI67)gene expression methods.The effect of ESCs-Exo on the migration of HSFBs was detected via a transwell assay and a scratch test.The concentrations of collagen secreted by the HSFBs and the mRNAs of the two kinds of collagen expressed by the HSFBs were analyzed.We also analyzed the phosphorylation of Protein Kinase N1(PKN1)and the expression of cyclins via western blotting.Finally,the effect of ESCs-Exo on wound healing was verified by animal experiments,and the key genes and signaling pathways of ESCs-Exo were excavated by transcriptomic analysis.Results:Western blotting revealed that the exosomes of ESCs highly expressed established markers such as Alix,CD63,and CD9.ESCExos significantly promoted HSFB proliferation and migration in a dose-dependent manner,as well as HSFB collagen synthesis,and effectively increased the ratio of collagen III/I.In addition,bioinformatics analysis showed that the expression of key gene C-X-C motif chemokine ligand 9 was lower in the ESCs-Exo group,which may promote wound healing by regulating PKN1-cyclin and tumor necrosis factor signaling pathways.Animal experiments demonstrated that ESCs-Exo could reduce inflammation and accelerate wound healing.Conclusions:In this study,we found that ESCs-Exo may improve wound healing by promoting the proliferation and migration of HSFBs.
文摘The objective of this research was to assess the potential of phosphatidylcholineencapsulated resveratrol as a cosmetic ingredient.The hydrogen peroxide(H_(2)O_(2))and ultraviolet A(UVA)induced human skin fibroblasts(HSF)models of skin damage were established to compare the antioxidant and anti-wrinkle properties between phosphatidylcholine-encapsulated resveratrol and unencapsulated resveratrol.The findings reveal that encapsulating resveratrol with phosphatidylcholine not only enhances skin absorption but also significantly improves its antioxidant capabilities.In the H2O2-induced HSF injury model,phosphatidylcholine-encapsulated resveratrol demonstrates a superior ability to neutralize reactive oxygen species(ROS)generated by H2O2 compared to the resveratrol group.Further analysis indicates that this enhanced functionality is associated with increased enzymatic activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and catalase(CAT)when treated with phosphatidylcholine-encapsulated resveratrol.Additionally,in UVA-irradiated HSF cells,phosphatidylcholine-encapsulated resveratrol effectively reduces the levels of matrix metalloproteinases-1 and-3(MMP-1 and MMP-3)and increased the contents of CollagenⅠand CollagenⅢ(Col-1 and Col-3),demonstrating significant anti-wrinkle effects.These findings provide critical evaluation criteria and application references for enhancing cosmetic ingredients through phosphatidylcholine encapsulation,thereby advancing skincare formulations.
基金fnancially supported partly by the Grand-In-Aid for Scientifc Research(C)18510022 and (C)21510032 of Japan Society for the promotion of Science
文摘A study on the effect of the solar ultra-violet radiation on the human skin fibroblast cells revealed that the production of matrix metalloproteinase-2 was inhibited by the radiation.A CO2 incubator connected by optical fibers to a reflector telescope for collecting the solar light was built at Syowa station by the 49th Japanese Antarctica Research Expedition.The direction of the telescope was continuously controlled by a sun-tracker to follow the movement of the Sun automatically.The intensity of the collected light was monitored by a portable spectrophotometer housed inside.The human skin fibroblast cells were incubated in the CO2 chamber to investigate the effect of the solar radiation at Syowa station and were compared with those reference experiments at a laboratory in Japan.The results showed cell damage by strong UV radiation.The production of matrix metalloproteinase-2 was prompted by the moderate UV-B,but was inhibited by the strong UV-B radiation,as studied under laboratory conditions in Japan.The effect of strong solar radiation at Syowa station involving the radiation of UV-B region was estimated to be of the same extent of the radiation caused by an artificial UV-B light with the intensity more than 50 mJ/cm2.
基金Supported by General Program of Guangxi Natural Science Foundation(2018GXNSFAA138098).
文摘[Objectives]To explore the effect of the extract of Growgx yeast fermentation product on ultraviolet A(UVA)-induced photo-aging of human skin fibroblasts(HFF-1)and its specific mechanisms from the level of cell biology,and discussed the application of the Growgx yeast fermentation product extract for skin aging.[Methods]A photo-aging model of HFF-1 cells was induced by UVA radiation meter,and it was given different concentrations of Growgx for intervention.The proliferation activity of the HFF-1 cells was detected with MTT method,and their migration and invasion ability was measured by scratch test.The SOD,HYP and MDA levels were detected with corresponding kits.After oxygen infusion apparatus assisted facial skin administration,the repair of the skin was observed through the VISIA skin tester.[Results]The survival rate of HFF-1 cells was reduced significantly by UVA with an irradiation dose of 10.8 J/cm2(P<0.05),while Growgx significantly increased their survival rate(P<0.001)and sped up the repair of damaged cells.Growgx promoted the production of SOD(P<0.05),reduced the level of MDA(P<0.05),and increased the level of HYP(P<0.05).Growgx effectively inhibited UVA-induced photo-aging of HFF-1 cells.The mechanism may be related to accelerating cell damage repair,up-regulating SOD and HYP expression levels,and reducing MDA expression level.The clinical observation showed that Growgx effectively improved skin spots and pores,making the skin smoother and more delicate.[Conclusions]Growgx can effectively fight against photo-aging caused by ultraviolet rays,and can significantly improve skin wrinkles.
基金supported by grants from the National Key R&D Program of China(No.2022YFC3601800)National Natural Science Foundation of China(Nos.82103735,82373489,and 82273542)CAMS Innovation Fund for Medical Sciences(Nos.CIFMS,2021-I2M-1-059,2022-I2M-C&T-B-096).
文摘Background:Senescent human skin primary fibroblast(FB)models have been established for studying aging-related,proliferative,and inflammatory skin diseases.The aim of this study was to compare the transcriptome characteristics of human primary dermal FBs from children and the elderly with four senescence models.Methods:Human skin primary FBs were obtained from healthy children(FB-C)and elderly donors(FB-E).Senescence models were generated by ultraviolet B irradiation(FB-UVB),D-galactose stimulation(FB-D-gal),atazanavir treatment(FB-ATV),and replication exhaustion induction(FB-P30).Flow cytometry,immunofluorescence staining,real-time quantitative polymerase chain reaction,co-culturing with immune cells,and bulk RNA sequencing were used for systematic comparisons of the models.Results:In comparison with FB-C,FB-E showed elevated expression of senescence-related genes related to the skin barrier and extracellular matrix,proinflammatory factors,chemokines,oxidative stress,and complement factors.In comparison with FB-E,FB-UVB and FB-ATV showed higher levels of senescence and expression of the genes related to the senescence-associated secretory phenotype(SASP),and their shaped immune microenvironment highly facilitated the activation of downstream immune cells,including T cells,macrophages,and natural killer cells.FB-P30 was most similar to FB-E in terms of general transcriptome features,such as FB migration and proliferation,and aging-related characteristics.FB-D-gal showed the lowest expression levels of senescence-related genes.In comparisons with the single-cell RNA sequencing results,FB-E showed almost complete simulation of the transcriptional spectrum of FBs in elderly patients with atopic dermatitis,followed by FB-P30 and FB-UVB.FB-E and FB-P30 showed higher similarity with the FBs in keloids.Conclusions:Each senescent FB model exhibited different characteristics.In addition to showing upregulated expression of natural senescence features,FB-UVB and FB-ATV showed high expression levels of senescence-related genes,including those involved in the SASP,and FB-P30 showed the greatest similarity with FB-E.However,D-galactose-stimulated FBs did not clearly present aging characteristics.
基金funded by National Natural Science Foundation of China(No.31870934).
文摘Skin tissue engineering with considerable skin regeneration capability is an urgent need for the wound site.The current challenge for researchers is to develop a bionic scaffold that imitates the extracellular matrix for the regeneration of the damaged regions.In our study,poly(L-lactide-co-caprolactone)(PLCL)was blended with polyurethane(PU)to obtain nanofibrous scaffolds via electrospinning.The electrospun fibers with 50%PLCL content had a certain number of intersections and jointing points,and exhibited significantly enhanced mechani-cal properties combined with suitable porosity.Moreover,cell activities demonstrated that PU/PLCL membranes had significantly biological advantages in enhanced growth of human skin fibroblasts with spreading morphology compared with PU membranes,indicating good cytocompatibility of composite scaffolds.These findings proved that PU/PLCL electrospun membranes have great potential in applications of skin tissue engineering.
