TGF-β change in blister fluid of hypertrophic scar and its significance

Objective To investigate of the tissue TGF-β changes at early stage of hypertrophic scar formation and the value of scar blisters in hypertrophic scar.Methods The TGF-β1 content in the blister fluid and the blood were quantified with ELISA,patients(n=15)with hypertrophy scar after depth burn were included,three time point(each n=5)on early　stage(<3 months)of hypertrophy scar formation were monitored.and normal skin blister fluid and the blood(n=5)was used as control.Results The serum TGF-β1 in the both hypertrophic scar patients and normal skin group was not elevated(P>0.01),the TGF-β1 in the blister of normal skin was also not elevated(P>0.01),but TGF-β1 level in the scarblisters hypertrophic scar was elevated significantly[<60 d(158.5±69.8)pg/L,60-90 d,(181.1±40.1)pg/L,>90 d,(534.4±125.9)pg/L,P<0.01] and higher than the normal skin blister and the blood(P<15.6 pg/L.P<0.01),the increased TGF-β1 1evel in the hypertrophic scar blisters were persisted for at least three months.the TGF-β1 level of scar blister on the 3th month of hypertrophic scar formation reached a peak [(534.4±125.9)pg/L,P<0.01].Conclusions The data in this study indicates that TGF-β production at the early stage of hypertrophic scar formation is increased and may play an important role in scar formation;scar blisters is a valuable approach in hypertrophic scar study.     

增生性瘢痕组织中褪黑素受体的表达

Objective To investigate the expression and its significance of melatonin receptor in human hypertrophic scarring. Methods The expression of melatonin receptor GPR50 was detected with immunohistochemistiy and the melatonin receptors( MT1 、MT2)mRNA were assessed with RT-PCR method in 10 cases of human hypertrophic scar and normal skin. The positive production was sequenced with auto sequencing instrument. Results Positive signals of melatonin receptor could be found in the cell membrane and cytoplasm. The melatonin receptor GPR50 was located in the epithelial basal cells, sweat gland cells and hair follicle in both hypertrophic scar and normal skin. The melatonin receptor GPR50 was extensively expressed in fibroblasts of hypertrophic scar, but not in fibroblasts in normal skin. RT-PCR showed that the expression of melatonin receptor( MT1, MT2 ) mRNA in hypertrophic scar was significantly higher than that in normal skin( P <0. 05). In normal skin and hypertrophic scar group, the expression of MT1 mRNA was higher than MT2 mRNA ( P < 0. 05 ) . In normal skin and hypertrophic scar group, the expression of MT1 mRNA was 0.99081 ±0.26485 and 1.16584 ±0.21829 copy number/μl cDNA, respectively;the expression of MT2 mRNA was 0. 77083 ±0. 15927and 0. 99550 ±0. 14624 copy number/ μl cDNA, respectively. Sequencing results indicated that the positive product coincided with cDNA of human melatonin receptor in GeneBank. Conclusions Positive expression of melatonin receptor can be found in human hypertrophic scar and normal skin, but it is higher in scar. The over expression of melatonin receptor in hypertrophic scar may be related to the development of hypertrophic scar.     

增生性瘢痕组织中褪黑素受体的表达

Objective To investigate the expression and its significance of melatonin receptor in human hypertrophic scarring. Methods The expression of melatonin receptor GPR50 was detected with immunohistochemistiy and the melatonin receptors( MT1 、MT2)mRNA were assessed with RT-PCR method in 10 cases of human hypertrophic scar and normal skin. The positive production was sequenced with auto sequencing instrument. Results Positive signals of melatonin receptor could be found in the cell membrane and cytoplasm. The melatonin receptor GPR50 was located in the epithelial basal cells, sweat gland cells and hair follicle in both hypertrophic scar and normal skin. The melatonin receptor GPR50 was extensively expressed in fibroblasts of hypertrophic scar, but not in fibroblasts in normal skin. RT-PCR showed that the expression of melatonin receptor( MT1, MT2 ) mRNA in hypertrophic scar was significantly higher than that in normal skin( P <0. 05). In normal skin and hypertrophic scar group, the expression of MT1 mRNA was higher than MT2 mRNA ( P < 0. 05 ) . In normal skin and hypertrophic scar group, the expression of MT1 mRNA was 0.99081 ±0.26485 and 1.16584 ±0.21829 copy number/μl cDNA, respectively;the expression of MT2 mRNA was 0. 77083 ±0. 15927and 0. 99550 ±0. 14624 copy number/ μl cDNA, respectively. Sequencing results indicated that the positive product coincided with cDNA of human melatonin receptor in GeneBank. Conclusions Positive expression of melatonin receptor can be found in human hypertrophic scar and normal skin, but it is higher in scar. The over expression of melatonin receptor in hypertrophic scar may be related to the development of hypertrophic scar.     

肉毒毒素A对大鼠皮肤及创面组织中SP、CGRP、TGF-β1和α-SMA的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

A型肉毒毒素局部应用对兔耳增生性瘢痕创面愈合和瘢痕增生的影响

Objective To investigate the effect of botulinum toxin type A (Botox A) injection on hypertrophic scar in rabbit ear model. Methods The hypertrophic scar model was established in 16 Japanese rabbits' ears. These wounds were divided into two groups as group T(treated with Botox A, n =48) and group S (not treated, n = 48). The wounds healing times and scar hypertrophy were observed with 8 specimen of normal skin at the rabbit ears as sham group B. HE stain was used to assess the hypertrophic index(HI). The expression of collagen Ⅰ and Ⅲ was tested by western-blot. The cell cycle of fibroblasts was studied by flow cytometry. Results The [] was significantly lower in group T than in group S(P < 0.01). The expression of collagen Ⅰ and Ⅲ, as well as the ratio of Ⅰ to Ⅲ, was markedly stronger in group S than in group T(P < 0.01). Compared with group T, more fibroblasts were in G2-M in gToup S and fewer in G0-G1 (P <0.05). Conclusions Local injection of Botox A can inhibite the formation of hypertrophic scar and the activity of fibroblasts in rabbit ear model. It can significantly decrease the expression of collagen Ⅰ and Ⅲ in hypertrophic scar, as well as the ratio of collagen Ⅰ to Ⅲ. It serves as the basis for the treatment of hypertrophic scar with Botox A.     

Expression of MCP-1 and TGF-β1 and its significance in early stage of obstructive jaundice in rats

Objective To investigate the expression of MCP-1 and TGF-β1 in early stage of ob-structive jaundice and explore its relation to liver correlated injury indexes in rats. Methods Fifty male Wistar rats were randomized into the control group, sham-operated group and biliary obstruction group. On day 10, serum ALT and BIL-T levels were determined in inferior caval blood and hepatic MDA was estimated in liver homogenates. Meanwhile, serum TGF-β1 level was measured by ELISA and the MCP-1 infiltration determined with immunohistochemistry. Results Serum ALT and BIL-T values were elevated. Meanwhile, the liver MDA content was increased. The positive expression of MCP-1 was augmented and serum TGF-β1 was over-expressed in the early stage after obstructive jaun-dice in rats. The hepatic MCP-1 expression had a positive correlation with the elevated ALT, BIL-T and MDA levels. However, the serum TGF-β1 content only had a positive correlation with ALT and BIL-T. Conclution Hepatic MCP-1 and serum TGF-β1 expression in the early stage after biliary tract obstruction is related to liver injury and extrahepatic cholestasis. Meanwhile, the hepatic MCP-1 ex-pression is correlated with hepatic oxidation stress. They have an important significance in liver injury in rats during the early period of obstructive jaundice.     

Differentiation of human epidermal stem cells into fibroblasts induced by TGF-β1 in vitro

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.     

Differentiation of human epidermal stem cells into fibroblasts induced by TGF-β1 in vitro

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.     

Differentiation of human epidermal stem cells into fibroblasts induced by TGF-β1 in vitro

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.     

Differentiation of human epidermal stem cells into fibroblasts induced by TGF-β1 in vitro

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.     

Differentiation of human epidermal stem cells into fibroblasts induced by TGF-β1 in vitro

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.     

Differentiation of human epidermal stem cells into fibroblasts induced by TGF-β1 in vitro

Objective To investigate the correlation between human epidermal stem cell (hESCs) and hypertrophic scar or keloid. Methods Improved collagen Ⅳ-coated adhesion methods was used to isolate and culture the epidermal stem cells after neutral protease selectively digested the dermo-epidermal junctions. After the cells were cultured and expanded in vitro, and passage 3 hESCs were induced by different concentrations of TGF-β1 (0.1, 5.0, and 10.0 ng/ml). Morphological fea-tures and identification of these cells were meseasured by HE, Masson, immunohistochemical staining on the days 3 and 7, respectively. Results After induced by TGF-β1 for 3 and 7 days, the morpholo-gy of the epidermal stem cell (hESCs) was changed into fusiform shape, similar to fibroblasts. 70 % ofthe cell which was induced by TGF-β1 were blue stained in the cytoplasm by Masson stain, which is the distinctive method for collagen, suggesting collagen appeared or increased in the cells. The collagen concentrations in supernatants of hESCs were 0.4150±0.0014, 0.3380±0. 0020, and 0.3870±0.0020, much higher than that in control group (0.0780±0.0025) and normal skin fibro-blast group (0.15004±0.0051) (P<0.05). Immunohistochemical staining revealed that positive rates of these cells for anti-vimentin staining were more than (95.00±1.20)% in experiments and (5.70±0.20)% in control group. Conclusion The differentiantion of hESCs induced by TGF-β1 into fibro-blasts indicates that hESCs may play a role in the pathogenesis of hypetrophic scar and keloid.