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BACKGROUND:Abnormal extracellular matrix accumulation and excessive proliferation of fibroblasts are the main manifestations of pathological scars.Excessive proliferation of fibroblasts leads to the production of large amounts of collagen-based extracellular matrix.Therefore,to investigate the role of fibroblast fibrosis in the formation of pathological scar will provide a new idea for revealing the mechanism of pathological scar and biological therapy. OBJECTIVE:To investigate the effect of RAS-selective lethal small molecule 3(RSL3)on the fibrosis of human pathological scar fibroblasts. METHODS:Then cases of pathological scar tissue and normal skin tissue samples from the same individuals,provided by the Department of Burn Plastic Surgery,General Hospital of Ningxia Medical University,were collected.Fibroblasts of human pathological scar and human normal skin were extracted and used in the following experiments.The general condition of the pathological scar tissue and the normal skin tissue was detected by hematoxylin-eosin staining.The appearance of fibroblasts from pathological scar and normal skin were observed by inverted microscope.The fibroblasts were verified by immunofluorescence assay.The cells were treated with different concentrations of RSL3(1,3,5,7,9,11,13 μmol/L).The inhibitory concentration of RSL3 on fibroblasts was detected by cell counting kit-8.Control group(without treatment)and RSL3 intervention group(treated with 7 μmol/L RSL3 for 24 hours)were set up.The mRNA and protein expressions of glutathione peroxidase 4,type Ⅰ collagen,type Ⅲ collagen and α-smooth muscle actin were detected by Qrt-PCR and western blot,respectively.Level of malondialdehyde in cells was detected.The residual scratch area was measured by cell scratch test after 24 hours to calculate the percentage of residual scratch area. RESULTS AND CONCLUSION:The expression of glutathione peroxidase 4 in the pathological scar group was higher than that in the normal skin group(Mrna:t=3.252,P<0.01;protein:t=5.075,P<0.01).The expression of glutathione peroxidase 4 in the pathological scar fibroblast group was higher than that in the normal skin fibroblast group(Mrna:t=10.32,P<0.01;protein:t=26.22,P<0.01).Compared with the control group,the expression of glutathione peroxidase 4 was decreased(Mrna:t=2.798,P<0.05;protein:t=4.643,P<0.01),the content of malondialdehyde was increased(t=2.917,P<0.05),the expression of type Ⅰ collagen(Mrna:t=15.84,P<0.01;protein:t=4.610,P<0.01),type Ⅲ collagen(Mrna:t=28.86,P<0.01;protein:t=7.713,P<0.01)and α-smooth muscle actin(Mrna:t=2.671,P<0.05;protein:t=7.417,P<0.01)were decreased in the RSL3 intervention group.Compared with the control group,the migration ability was weakened in the RSL3 intervention group(t=14.06,P<0.01).To conclude,RSL3 can inhibit the expression of glutathione peroxidase 4 and then inhibit the ability of fibrosis and migration of pathological scar fibroblasts.
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BACKGROUND:Stem cell therapy has become an emerging method of treating pathological scars.miRNAs are involved in scarring mechanisms,and the targeted action of some stem cell sources of miRNA is mediated by exosomes. OBJECTIVE:To review the biological properties of miRNA derived from mesenchymal stem cells and its derivatives,the mechanism of treatment of scarring through anti-inflammation,suppression of excessive tissue reconstruction and antioxidation. METHODS:The first author used a computer in September 2023 to retrieve the relevant literature published from January 2000 to September 2023,searching for"stem cell,exosome,miRNA,keloid"in English,and"stem cells,exosome,keloid"in Chinese,eventually incorporating 74 papers for analysis. RESULTS AND CONCLUSION:(1)miRNAs with high expression of exosomes derived from mesenchymal stem cells increase the proportion of M2-type macrophages by promoting the polarization of macrophages,target the regulation of transforming growth factor β,transforming growth factor β receptors or related signaling pathways,inhibit the expression of pro-inflammatory factors,promote the expression of anti-inflammatory factors and other mechanisms to inhibit inflammation and thus suppress scar lesions.(2)miRNAs with high expression of exosomes derived from mesenchymal stem cells can reduce the secretion of matrix metalloproteinases,regulate the balance between matrix metalloproteinase inhibitors and matrix metalloproteinases,inhibit the proliferation and migration of fibroblasts and myofibroblasts,directly reduce the production of collagen and other mechanisms,and ultimately lead to the normal degradation of extracellular matrix,thereby inhibiting excessive tissue remodeling and cicatricial lesions.(3)miRNAs with high expression of exosomes from mesenchymal stem cells can improve the resistance of scar fibroblasts to oxidative stress by regulating reactive oxygen species and hypoxia-inducing factors,and then regulate the proliferation and apoptosis of scar fibroblasts to inhibit scar lesions.(4)Exosomes derived from mesenchymal stem cells have good prospects for scar treatment.Studies on this aspect can find mirnas that regulate inflammatory cells,inflammatory factors,signaling pathways,matrix metalloproteinases,fibroblasts,reactive oxygen species,hypoxia-inducing factors and other key factors from the three aspects of inflammation,tissue remodeling and oxidative stress.Then,by inducing mesenchymal stem cells with high expression of the above miRNA,exosomes were extracted,and finally verified and clinical trials were carried out.
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Objective @#To analyze the effect of miR⁃148b⁃3p on the proliferation of keloid derived fibroblasts. @*Methods @#The expression levels of miR⁃148b⁃3p and SPARC in human keloid derived fibroblasts (HKF) and normal human fibroblasts (NFS) were analyzed by real time PCR. The expression level of SPARC protein was detected by Western blot. The effects of miR⁃148b⁃3p and SPARC on HKF proliferation were analyzed by CCK⁃8 method the luciferase reporter plasmid was constructed. The targeted binding site of miR⁃148b⁃3p and the target gene was analyzed by luciferase reporter gene method.@*Results @#miR⁃148b⁃3p was low expressed in HKF and SPARC was high expressed in HKF. Transfection of miR⁃148b⁃3p in HKF cells could down regulate the expression of SPARC and inhibit cell proliferation. Online analysis software predicted that miR⁃148b⁃3p could target the 3 ′⁃ UTR binding SPARC ; The results of dual luciferase reporter gene further confirmed that miR⁃148b⁃3p could target the 3 ′⁃ UTR of SPARC. Transfection of SPARC eukaryotic expression plasmid into HKF transfected with miR⁃148b⁃3p could counteract the effect of miR⁃148b⁃3p and restore cell proliferation. @*Conclusion @# miR⁃148b⁃3p can inhibit the proliferation of HKF by targeting the 3 ′⁃ UTR of SPARC and inhibiting its expression.
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Objective:To study the application of B-mode ultrasound in objective evaluation and dynamic monitoring of scar.Methods:The subjects were patients with scar in the outpatient and inpatient department of plastic surgery in the First Affiliated Hospital of Zhengzhou University from March 2018 to June 2020. According to the type of scar, they were divided into 3 groups: 21 patients in the normal scar group, 23 patients in the hypertrophic scar group, and 15 patients in the keloid group. All 59 patients were regularly scanned by B-mode ultrasound for scar images and the scar thickness was measured.Results:B-mode ultrasonic images of scars were analyzed in each group: The echo intensity of dermis of normal scar was uneven, hypertrophic scar and keloid dermis showed obvious hypoecho, and some cases of keloid presented a small amount of blood flow. There was no significant change in the ultrasonic images of scar in each group within 1 year. Changes in scar thickness were observed in each group. The normal scar thickness did not change significantly within 1 year. The thickness of hypertrophic scar showed a trend of first increasing and then stabilized within 1 year.Conclusions:B-mode ultrasound can assist to identify different scar types according to ultrasonic performance. B-mode ultrasound can be used to measure scar thickness objectively and accurately and monitor the dynamic changes of scars.
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Pathological scar is a kind of skin fibrotic disease caused by abnormal wound healing, including hypertrophic scar and keloid. Pathological scar may lead to aesthetic flaws, limb dysfunction and local discomfort in patients. Due to the complexity of the wound healing process, the formation of scar is affected by many factors. In addition to traditional surgical, laser, cryostatic and hormone injection methods for the treatment of pathological scar, there are new therapies, such as mesenchymal stem cell therapy, fat transplantation, interferon, and botulinum toxin. They are widely used in clinical practice, but with such problems as high prices and many side effect. Traditional Chinese medicine (TCM) has a long history in treating pathological scar. In recent years, in vivo and in vitro studies have shown that TCM has effect IN reducing inflammation, inhibiting fibroblast proliferation, regulating fibroblast activation and migration, inducing fibroblast apoptosis and autophagy, promoting the degradation of extracellular matrix (ECM) and reducing angiogenesis in general. Besides, TCM has also a certain regulatory role in the signaling pathways, such as transforming growth factor-β1 (TGF-β1)/Smads, phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) and sonic hedgehog (Shh). There are still some contradictions in relevant studies, and specific mechanisms remain to be further improved. This paper summarizes the study content, findings and relevant mechanisms of different TCM based on in vivo and in vitro experiments, analyzes the advantages and disadvantages of TCM in the prevention and treatment of pathological scar, and its prospects in clinical application, so as to provide basis and ideas for future scar studies.
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Objective To explore the role of macrophage migration inhibitory factor (MIF) in the pathogenesis of pathological scar and the effect of ISO-1 on the behavior of scar fibroblasts.Methods Samples of normal skin,normal scar,and pathological scar were collected and detected by hematoxylin-eosin staining and immumohistochemical staining.Human fibroblasts were isolated from the samples and then divided into different groups with the intervention with ISO-1 (0 ～ 100 μ mol/mL).Fibroblast proliferation was detected by Alamber dyeing and cell apoptosis was detected by TUNEL staining.Expressions of fibroblast specific proteins and PI3K/Akt/mTOR signaling pathways wcre detected by Western Blot and RT-PCR.Results The positive rates of MIF for hyperplastic scar and keloid were greater than those for normal scar and normal skin (P ＜ 0.01).Apoptotic cells occurred less in the group without intervention.The apoptotic rate increased gradually as the concentration of ISO-1 increased.There were significant statistical differences in the migration rate among all the groups (P ＜ 0.05).As concentration of ISO-1 increased,the protein and gene expressions of type I collagen,FN and CTGF were decreased.Expressions of activated PI3K and Akt decreased as ISO-1 concentration increased.Conclusions The expression of MIF is different in different types of scar tissue.ISO-1 inhibits the biological behavior of fibroblasts derived from pathological scar through PI3K/Akt/mTOR pathways.
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Objective To explore the effect of hypoxia-inducible factor 1α(HIF-1α)in pathological scar and its specific mechanism,and the therapy target of pathological scar.Methods Real time PCR and Western blot was used to test the expression of HIF-1αin normal tissue and pathological scar,meanwhile to detect the effect of hypoxic environment on the expression of HIF-1α.Fibroblasts activity in pathological scar and normal tissue under different oxygen concentration(20%,10%,5%,2% and 1%)was determined by MTT method.Determine the different expression of HIF-1αmRNA and protein in normal environment(20% oxygen) and hypoxic environment(5%oxygen).The changes of the number of fibroblasts after the silence of HIF-1αby HIF-1αshRNA virus using flow cytometer.Results The Real time PCR and Western blot results showed that the expressions of HIF-1αmRNA and protein in fibroblasts of pathological scar was significantly higher than that in normal tissue(P<0.05),and the activity of the keloid fibroblasts in hypoxia environment(5%,2% and 1%)was also higher than that in normal tissue(P<0.05).The expressions of HIF-1αmRNA and protein in keloid fiber cells were higher in hypoxia environment(5% oxygen)than those in normal environment(20% oxygen),but when silence the expression of HIF-1αin keloid fibroblasts,the apoptosis of fibroblast in hypoxia environment(5%oxygen)were significantly higher than control group[(0.021 ±0.001)%vs.(3.739 ± 0.039)%,P<0.05].Conclusion HIF-1αincreases significantly in the pathological keloid fibroblasts and could promote the proliferation of fibroblasts and its vitality,which could accelerate the formation of pathological scar.
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Objective To observe the changes of MMP-1 and TIMP-1 expression in the pathological scar tissue after scar-skin replantation and to explore the mechanism of treating pathological scars with scar-skin replantation through a rabbit ear model.Methods Rabbit ears were used to establish the hypertrophic scar animal model in this study.Specimens were taken for three times:normal skin,hypertrophic scar and scar-skin replantion separately.We then performed HE staining,Masson staining and immunohistochemical staining to observe the expression of MMP-1 and TIMP-1 in these three groups of specimens.Results The expression of both MMP-1 and TIMP-1 significantly increased in the hypertrophic scar tissue after scar-skin replantation compared with the control group (P＜0.01).The expression of MMP-1 increased more significantly than that of TIMP-1 (P＜0.01).Conclusions The mechanism of scar-skin replantation's effect in the treatment for hypertrophic scar is relevant to the imbalance in the interaction between MMP-1 and TIMP-1 in the scar tissue.
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Objective:To explore the inhibitive effects of exogenous nitric oxide on the proliferation of in vitro cultured human keloid fibroblasts.Methods:Fibroblasts was isolated from fresh pathological scar tissue and cultured.Fibroblasts were distributed in five groups.Fibroblasts in control group A was cultured without SNP;in experiment group B～D,SNP was used in the concentration of 100、200、300 ?mol/L each;in experiment group E,Fibroblasts was cultured with 200 ?mol/L SNP and 10 ?mol/L methylene blue.After 24h,Griess and MTT were used to detect the concentration of NO and cell vigor.Results:The release of NO increased along with the enhancement of the concentration of SNP.The amount of living cells in experiment groups decreased compared with the controls as shown by MTT tes(tP
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Objective To study the role of sebaceous gland and its secretion in the progress of abnormal scar and to provide a new idea to prevent pathological scar. Method The sites, pathogenesis and pathological changes were compared between pathological and normal scars.Results Pathological scar often occurred at post-ear, upper lips, maxillofacial skin, and deltoid muscle skin of upper limbs; normal scar often occurred at mid-face, forehead, nose’s tip, and proximal extremity of limbs. Several rows of sebaceous gland and its secretion could be found at the edges of pathological scar, large quantity of sebaceous gland had changes of ductal dilation, and large amount of the secretion accumulated in the cavity in pathological scar; however, on the normal scar base and edges, seldomly showed the expanded sebaceous gland and accumulated secretion. Conclusions Sebaceous gland and its secretion could stimulate the growth of scar. To control the action of sebaceous gland and the formation of secretion could inhibit the formation of pathological scar.
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Objective To investigate the effects of substance P (SP) on the expression of apoptosis-associated genes in fibroblasts derived from pathological scars. Methods Fibroblasts derived from keloid, hypertrophic scar and normal skin were cultured separately in media containing SP and SP receptor antagonist. PCNA, bcl-2 and bax protein in fibroblasts were assessed by means of immunohistochemistry. Results SP enhanced PCNA and bcl-2 expression in all three kinds of fibroblasts, whereas, bax expression was inhibited significantly. SP inhibited the expression of bax in keloid scar fibroblasts (KSFB) more remarkably than that in hypertrophic scar fibroblasts (HSFB) or normal fibroblasts (NFB), and the effect was stronger on HSFB than on NFB. SP receptor antagonist could inhibit those effects of SP totally or partially. Conclusion SP may play an important role in the formation of pathological scars by modulating the expression of apoptosis-associated genes, which is mediated by SP receptor.