Exploring the potential mechanism of WuFuYin against hypertrophic scar using network pharmacology and molecular docking

BACKGROUND Hypertrophic scar(HTS)is dermal fibroproliferative disorder,which may cause physiological and psychological problems.Currently,the potential mechanism of WuFuYin(WFY)in the treatment of HTS remained to be elucidated.AIM To explore the potential mechanism of WFY in treating HTS.METHODS Active components and corresponding targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform.HTSrelated genes were obtained from the GeneCards,DisGeNET,and National Center for Biotechnology Information.The function of targets was analyzed by performing Gene Ontology and Kyoto Encyclopaedia of Genes and Genome(KEGG)enrichment analysis.A protein+IBM-protein interaction(PPI)network was developed using STRING database and Cytoscape.To confirm the high affinity between compounds and targets,molecular docking was performed.RESULTS A total of 65 core genes,which were both related to compounds and HTS,were selected from multiple databases.PPI analysis showed that CKD2,ABCC1,MMP2,MMP9,glycogen synthase kinase 3 beta(GSK3B),PRARG,MMP3,and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit gamma(PIK3CG)were the hub targets and MOL004941,MOL004935,MOL004866,MOL004993,and MOL004989 were the key compounds of WFY against HTS.The results of KEGG enrichment analysis demonstrated that the function of most genes were enriched in the PI3K-Akt pathway.Moreover,by performing molecular docking,we confirmed that GSK3B and 8-prenylated eriodictyol shared the highest affinity.CONCLUSION The current findings showed that the GSK3B and cyclin dependent kinase 2 were the potential targets and MOL004941,MOL004989,and MOL004993 were the main compounds of WFY in HTS treatment.

A Case Report of Concurrent Acne-Related Occurrence Complications: Telangiectasia, Post-Inflammatory Erythema, Post-Inflammatory Hyperpigmentation, and Atrophic and Hypertrophic Scars

Prior to his initial diagnosis, a 21-year-old male had been experiencing facial acne for two years and had been treated by a doctor in private practice. The patient visited our department because the clinical manifestations of mandibular acne did not improve. At the time of initial examination, telangiectasia (TE), post-inflammatory erythema (PIE), post-inflammatory hyperpigmentation (PIH), atrophic scars (ASs), and a hypertrophic scar (HS) with induration were observed on the right neck. We diagnosed this as an acne vulgaris complication. HS lesions were topically treated by injecting triamcinolone acetonide, and the patient was prescribed 8.1 g/day of oral Saireito (Japanese herb). Adapalene benzoyl peroxide gel and topical tacrolimus hydrate ointment were used to treat PIE and TE. Both HSs and PIE improved;however, TE and AS did not improve. Currently, the patient is under observation. We consider this to be a very rare concurrent occurrence of diverse complications of acne vulgaris, and present the following case study.

Inhibition effects of a negative electret 5-FU patch on the growth of a hypertrophic scar

In this study,the hypertrophic scar（HS） model in rats was established.5-fluorouracil（5-FU）patch,-1000 V and-2000 V polypropylene（PP） electret 5-FU patches were prepared and applied onto the wound.The in vitro permeation experiment was performed using the Franz diffusion cell system to determine the permeation cumulative amount and retention amount of5-FU through/in scar skin.The inhibition effect of negative electret on growth of HS was studied by hematoxylin-eosin（HE） staining,Masson staining and the immunohistologicall methods.The permeation study indicated that a negative electret could enhance the permeation and retention of 5-FU through and in scar skin respectively.HE staining and Masson staining indicated a better effect for-1000 V and-2000 V electret 5-FU patches on HS inhibition after28 d post-wounding compared with 5-FU patch.The immunohistological study showed much more reduced expressions of collegan type I,collegan type III,TGF-β1 and HSP47 in scar tissue after application of negative electret 5-FU patches than those of 5-FU patch.A negative electret5-FU patch may be advantageous for HS treatment.

Inflammation and cutaneous nervous system involvement in hypertrophic scarring

This study aimed to use a mouse model of hypertrophic scarring by mechanical loading on the dorsum of mice to determine whether the nervous system of the skin and inflammation participates in hypertrophic scarring. Results of hematoxylin-eosin and immunohistochemical staining demonstrated that inflammation contributed to the formation of a hypertrophic scar and increased the nerve density in scar tissue.Western blot assay verified that interleukin-13 expression was increased in scar tissue. These findings suggest that inflammation and the cutaneous nervous system play a role in hypertrophic scar formation.

NON-LINEAR SPECTRAL IMAGING MICROSCOPY STUDIES OF HUMAN HYPERTROPHIC SCAR

Skin scar is unique to humans,the major significant negative outcome sustained after thermal injuries,traumatic injuries,and surgical procedures.Hypertrophic scar in human skin is investigated using non-linear spectral imaging microscopy.The high contrast images and spectroscopic intensities of collagen and elastic fibers extracted from the spectral imaging of normal skin tissue,and the normal skin near and far away from the hypertrophic scar tissues in a 10-year-old patient case are obtained.The results show that there are apparent differences in the morphological structure and spectral characteristics of collagen and elastic fibers when comparing the normal skin with the hypertrophic scar tissue.These differences can be good indicators to differentiate the normal skin and hypertrophic scar tissue and demonstrate that non-linear spectral imaging microscopy has potential to noninvasively investigate the pathophysiology of human hypertrophic scar.

Dynamic changes of autophagy during hypertrophic scar formation and the role of autophagy intervention

Background:The role of autophagy in the formation of hypertrophic scars(HS)remains unclear.This study aimed to explore the role and potential mechanism of autophagy during the development of HS.Methods:RNA and protein expression levels of Beclin-1,p62,and LC3II in normal skin tissues and HS specimens from different patients were examined.Autophagy inducers and inhibitors were used to cure established HS in rabbit ears,and the expression of Beclin-1,p62,and LC3II at the RNA and protein level was determined.Lastly,the effects of autophagy inducers and inhibitors on HS development were analyzed.Results:Compared to normal skin tissues,the expression of LC3II and Beclin-1 was higher(P<0.05),while that of p62 was lower(P<0.05)in HS tissues.In addition,the LC3II/LC3I ratio was increased during HS formation,and the altered expression of the three proteins stabilized after one year.Administration of autophagy inducers enhanced the formation of HS as well as the expression levels of LC3II and Beclin-1 but decreased p62 expression.Meanwhile,administration of autophagy inhibitors increased the expression of LC3II,Beclin-1,and p62,along with reduced HS formation.Conclusion:Autophagic activity increased during HS initiation and subsequent stabilization.In addition,autophagy inhibitors were able to inhibit HS formation by suppressing autophagy,whereas autophagy inducers promoted scar hyperplasia by enhancing autophagy。

The expression of Cyclin A and p21^(cip1)in fibroblast of hypertrophic scar

Objective To study the relation of the mRNA and protein expression of CyclinA and p21cip1 in different stages hypertrophic scar fibroblast (FB) with its cell cycle,so as to provide theoretical evidence for intervention therapy of

A six-herb Chinese medicine composition ointment as a promising candidate for treatment of hypertrophic scars

Objective:To study the anti-hypertrophic scar effect of the six-herb Chinese medicine composition(SCMC) ointment on the rabbit ear hypertrophic scar models.Methods:The optimal formulation of SCMC ointment matrix was screened by the orthogonal designs and a series of evaluation tests.The SCMC ointment was prepared through emulsifying method.The rabbit ear hypertrophic scar models were established and used to investigate the anti-hypertrophic scar effect of SCMC ointment.Results:Our results demonstrated that all the quality control indications of the SCMC ointment met the requirements.Anti-hypertrophic scar activity results showed that all the rabbit ear scar tissues appeared different degrees of shrink and fading,and took an unobvious but palpable shift from hard to soft texture with the low,middle and high concentration SCMC ointments treatments in vivo.Additionally,on 21 st day the scar area and thickness in different concentrations of SCMC ointment groups were significantly reduced than control group,in a concentration-dependent manner.The immunohistochemical results also indicated that the SCMC ointment had good anti-hypertrophic scar properties and could inhibit hypertrophic scar formation.Conclusion:The SCMC ointment could improve the blood circulation condition of hypertrophic scar tissues.Our research has demonstrated the Chinese medicine composition ointment with good antihypertrophic scar properties that could be used to treat hypertrophic scars.Meanwhile,it provides a theoretical basis for further clinical application.

Diagnosis and Treatment of Keloids and Hypertrophic Scars—Japan Scar Workshop Consensus Document 2018

There has been a long-standing need for guidelines on the diagnosis and treatment of keloids and hypertrophic scars that are based on an understanding of the pathomechanisms that underlie these skin fibrotic diseases.This is particularly true for clinicians who deal with Asian and African patients because these ethnicities are highly prone to these diseases.By contrast,Caucasians are less likely to develop keloids and hypertrophic scars,and if they do,the scars tend not to be severe.This ethnic disparity also means that countries vary in terms of their differential diagnostic algorithms.The lack of clear treatment guidelines also means that primary care physicians are currently applying a hotchpotch of treatments,with uneven outcomes.To overcome these issues,the Japan Scar Workshop(JSW)has created a tool that allows clinicians to objectively diagnose and distinguish between keloids,hypertrophic scars,and mature scars.This tool is called the JSW Scar Scale(JSS)and it involves scoring the risk factors of the individual patients and the affected areas.The tool is simple and easy to use.As a result,even physicians who are not accustomed to keloids and hypertrophic scars can easily diagnose them and judge their severity.The JSW has also established a committee that,in cooperation with outside experts in various fields,has prepared a Consensus Document on keloid and hypertrophic scar treatment guidelines.These guidelines are simple and will allow even inexperienced clinicians to choose the most appropriate treatment strategy.The Consensus Document is provided in this article.It describes(1)the diagnostic algorithm for pathological scars and how to differentiate them from clinically similar benign and malignant tumors,(2)the general treatment algorithms for keloids and hypertrophic scars at different medical facilities,(3)the rationale behind each treatment for keloids and hypertrophic scars,and(4)the body site-specific treatment protocols for these scars.We believe that this Consensus Document will be helpful for physicians from all over the world who treat keloids and hypertrophic scars.

Pressure therapy upregulates matrix metalloproteinase expression and downregulates collagen expression in hypertrophic scar tissue

Background Pressure therapy improves hypertrophic scar healing, but the mechanisms for this process are not well understood. We sought to investigate the differential expression of matrix metalloproteinases （Mmps） and collagen in post- traumatic hypertrophic scar tissue with mechanical pressure and delineate the molecular mechanisms of pressure therapy for hypertrophic scars. Methods Fibroblast lines of normal skin and scar tissue were established and a mechanical pressure system was devised to simulate pressure therapy. Reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting assays were used to compare differences in the mRNA and protein expression of Mmps and collagen in scar fibroblasts before and after pressure therapy. Results The expression differed between the hypertrophic scar cell line and the normal cell line. RT-PCR assays showed that Collagen I, highly expressed in the hypertrophic scar cell line, decreased significantly after pressure therapy. Mmp2, Mmp9, and Mmp12 expression in the hypertrophic scar tissue increased significantly after pressure therapy （P 〈0.05）. Western blotting assays further revealed that Mmp9 and Mmp12 expression increased significantly in the hypertrophic scar tissue after pressure therapy （P 〈0.05） but not Mmp2 expression （P 〉0.05）. Conclusion Mechanical pressure induces degradation of Collagen I in hypertrophic scar tissue by affecting the expression of Mmp9 and Mmp12.

Strategies to prevent hypertrophic scar formation:a review of therapeutic interventions based on molecular evidence

Once scar tissues mature,it is impossible for the surrounding tissue to regenerate normal dermal tissue.Therefore,it is essential to understand the fundamental mechanisms and establish effective strategies to inhibit aberrant scar formation.Hypertrophic scar formation is considered a result of the imbalance between extracellular matrix synthesis and degradation during wound healing.However,the underlying mechanisms of hypertrophic scar development are poorly understood.The purpose of this review was to outline the management in the early stage after wound healing to prevent hypertrophic scar formation,focusing on strategies excluding therapeutic agents of internal use.Treatment aimed at molecular targets,including cytokines,will be future options to prevent and treat hypertrophic scars.More basic studies and clinical trials,including combination therapy,are required to investigate the mechanisms and prevent hypertrophic scar formation.

Bilayer dissolving microneedle array containing 5-fluorouracil and triamcinolone with biphasic release profile for hypertrophic scar therapy

Hypertrophic scar(HS)is an undesirable skin abnormality following deep burns or operations.Although intralesional multi-injection with the suspension of triamcinolone acetonide(TA)and 5-fluorouracil(5-Fu)has exhibited great promise to HS treatment in clinical,the difference of metabolic behavior between TA and 5-Fu remarkably compromised the treatment efficacy.Besides,the traditional injection with great pain is highly dependent on the skill of the experts,which results in poor compliance.Herein,a bilayer dissolving microneedle(BMN)containing TA and 5-Fu(TA-5-Fu-BMN)with biphasic release profile was designed for HS therapy.Equipped with several micro-scale needle tips,the BMN could be self-pressed into the HS with uniform drug distribution and less pain.Both in vitro permeation and in vivo HS retention tests revealed that TA and 5-Fu could coexist in the scar tissue for a sufficient time period due to the well-designed biphasic release property.Subsequently,the rabbit ear HS model was established to assess therapeutic efficacy.The histological analysis showed that TA-5-Fu-BMN could significantly reduce abnormal fibroblast proliferation and collagen fiber deposition.It was also found that the value of scar elevation index was ameliorated to a basal level,together with the downregulation of mRNA and protein expression of Collagen I(Col I)and transforming growth factor-β1(TGF-β1)after application of TA-5-Fu-BMN.In conclusion,the BMN with biphasic release profiles could serve as a potential strategy for HS treatment providing both convenient administrations as well as controlled drug release behavior.

Role of HLA-DR and CD1a molecules in pathogenesis of hypertrophic scarring and keloids

Keyword: hypertrophic scars · keloids · HLA DR · CD1a

The molecular basis of hypertrophic scars

Hypertrophic scars(HTS)are caused by dermal injuries such as trauma and burns to the deep dermis,which are red,raised,itchy and painful.They can cause cosmetic disfigurement or contractures if craniofacial areas or mobile region of the skin are affected.Abnormal wound healing with more extracellular matrix deposition than degradation will result in HTS formation.This review will introduce the physiology of wound healing,dermal HTS formation,treatment and difference with keloids in the skin,and it also review the current advance of molecular basis of HTS including the involvement of cytokines,growth factors,and macrophages via chemokine pathway,to bring insights for future prevention and treatment of HTS.

Process of Hypertrophic Scar Formation： Expression of Eukaryotic Initiation Factor 6

Background： Hypertrophic scar is one of the most common complications and often causes the disfigurement or deformity in bum or trauma patients. Therapeutic methods on hypertrophic scar treatment have limitations due to the poor understanding of mechanisms of hypertrophic scar formation. To throw light on the molecular mechanism of hypertrophic scar formation will definitely improve the outcome of the treatment. This study aimed to illustrate the negative role of eukaryotic initiation factor 6 （elF6） in the process of human hypertrophic scar tbrmation, and provide a possible indicator of hypertrophic scar treatment and a potential target molecule for hypertrophic scar. Methods： In the present study, we investigated the protein expression of elF6 in the human hypertrophic scar of different periods by immunohistochemistry and Western blot analysis. Results： In the hypertrophic scar tissue, elF6 expression was significantly decreased and absent in the basal layer of epidermis in the early period, and increased slowly and began to appear in the basal layer of epidermis by the scar formation time. Conclusions： This study confirmed that elF6 expression was significantly related to the development of hypertrophic scar, and the elF6 may be a target molecule for hypertrophic scar control or could be an indicator of the outcomes for other treatment modalities.

Effect of Abnormal Savda Munziq on Hypertrophic Scar Formation in A Rabbit Ear Model

Objective： To investigate whether administrating Abnormal Savda Munziq （ASMq）, a traditional Uighur herbal preparation used for the prevention or treatment of diseases, affects hypertrophic scar （HTS） formation by using an established rabbit ear model. Methods： The HTS rabbit model was created by circular full- thickness skin excisions on both ears of rabbits. Twenty rabbits were randomized into four groups, with 5 rabbits and 60 wounds in each group. Group A was the control group, treated with normal saline daily. Groups B, C, and D were the treatment groups at three different doses of ASMq （400, 800, and 1200 mg/kg body weight, respectively, daily, by gastrogavage）. Twenty wounds were randomly chosen from each group on the 40th day after treatment and specimen were examined. Scar elevation index （SEI） was analyzed with histological assessment, and ultrastructure analysis was analyzed with a transmission electron microscopy. Results： Groups B, C, and D demonstrated significant reductions in SEI as compared with the control group at 35.9% （P=0.0212）, 48.2% （P=0.0108）, and 52.7% （P=0.0103）, respectively in a dose-response manner. SEI was lowered in Group D compared with Group B with a significant difference （P=0.015）. However, there were no significant differences between Groups B and C, or between Groups C and D. Histological analysis showed that high- dose ASMq （1200 mg/kg） could enhance the softening of HTS of rabbit ears and increase the compliance as shown in general. Ultrastructure analysis showed that with increased ASMq dose, the fibroblasts, pro-collagen, collagen, endoplasmic reticulum and ribosomes were reduced gradually. Conclusions： Orally administered ASMq significantly reduces the severity of HTS in the rabbit ear model. The findings of this study may have clinical implications on the management of human HTS.

Overexpression of miR-101 suppresses collagen synthesis by targeting EZH2 in hypertrophic scar fibroblasts

Background:MicroRNA-101(miR-101)is a tumor suppressor microRNA(miRNA)and its loss is associated with the occurrence and progression of various diseases.However,the biological function and target of miR-101 in the pathogenesis of hypertrophic scars(HS)remains unknown.Methods:We harvested HS and paired normal skin(NS)tissue samples from patients and cultured their fibroblasts(HSF and NSF,respectively).We used quantitative reverse transcriptase polymerase chain reaction(qRT-PCR),fluorescence in situ hybridization(FISH),enzyme-linked immunosorbent assays(ELISA)and Western blot analyses to measure mRNA levels and protein expression of miR-101,enhancer of zeste homolog 2(EZH2),collagen 1 and 3(Col1 and Col3)andα-smooth muscle actin(α-SMA)in different in vitro conditions.We also used RNA sequencing to evaluate the relevant signaling pathways and bioinformatics analysis and dual-luciferase reporter assays to predict miR-101 targets.We utilized a bleomycin-induced fibrosis mouse model in which we injected miR-101 mimics to evaluate collagen deposition in vivo.Results:We found low expression of miR-101 in HS and HSF compared to NS and NSF.Overexpressing miR-101 decreased Col1,Col3 andα-SMA expression in HSF.We detected high expression of EZH2 in HS and HSF.Knockdown of EZH2 decreased Col1,Col3 andα-SMA in HSF.Mechanistically,miR-101 targeted the 3-untranslated region(3UTR)of EZH2,as indicated by the decreased expression of EZH2.Overexpressing EZH2 rescued miR-101-induced collagen repression.MiR-101 mimics effectively suppressed collagen deposition in the bleomycin-induced fibrosis mouse model.Conclusions:Our data reveal that miR-101 targets EZH2 in HS collagen production,providing new insight into the pathological mechanisms underlying HS formation.

Control of fibrosis and hypertrophic scar formation via glycolysis regulation with IR780

Background:Hypertrophic scars(HS)represent one of the most common clinical challenges due to unsatisfactory therapeutic results.HS formation is associated with the abnormal activation of fibroblasts and their excessive fibrotic behavior.Glycolysis dysregulation has been shown to participate in the incidence and progression of various fibrotic diseases and shows potential as a means of controlling HS formation.This work aimed to discuss the impact of augmented glycolysis on HS and to propose a method for controlling HS formation through glycolysis regulation.Methods:Here,augmented glycolysis was confirmed together with enhanced fibrotic activity in both HS fibroblasts(HFs)and HS tissues,and the suppression of glycolysis also attenuated fibroblast activation.We also introduced IR780,a heptamethine cyanine dye,to regulate glycolysis for the control of HS formation.Results:In vitro,cell studies indicated that IR780 significantly down-regulated glycolysis and suppressed the fibrotic activity of HFs.In vivo,the intralesional injection of IR780 into rabbit HS models led to the downregulation of glycolysis and the control of HS formation.Furthermore,IR780 accumulated preferentially in activated fibroblasts in both in vitro and in vivo studies,and thus specifically downregulated glycolysis and efficiently controlled fibrosis by targeting activated fibroblasts.Conclusions:This work identified a strategy for controlling fibrosis and HS formation from the perspective of glycolysis regulation with IR780 targeting of activated fibroblasts.

Transdermal delivery of Chinese herbal medicine extract using dissolvable microneedles for hypertrophic scar treatment

Hypertrophic scars are unfavorable skin diseases characterized by excessive collagen deposition.Although systemic treatments exist in clinic to manage hypertrophic scars,they pose significant side effects and tend to lose efficacy over prolonged applications.Traditional Chinese medicine(TCM)offers as a promising candidate to treat pathological scars.A large number of TCMs have been studied to show anti-scarring effect,however,the natural barrier of the skin impedes their penetration,lowering its therapeutic efficacy.Herein,we reported the use of dissolvable hyaluronic acid(HA)microneedles(MNs)as a vehicle to aid the transdermal delivery of therapeutic agent,a model TCM called shikonin for the treatment of hypertrophic scars.Here,shikonin was mixed with HA to make MNs with adequate mechanical strength for skin penetration,making its dosage controllable during the fabrication process.The therapeutic effect of the shikonin MNs was studied in vira using HSFs and then further verified with quantitative reverse transcriptase polymerase chain reaction.Our data suggest that the shikonin HA MNs significantly reduce the viability and proliferation of the HSFs and downregulate the fibrotic-related genes(i.e.,TGFβ1,FAP-αand COL1 A1).Furthermore,we observed a localized therapeutic effect of the shikonin HA MNs that is beneficial for site-specific treatment.

Combination of ablative fractional carbon dioxide laser and platelet-rich plasma treatment to improve hypertrophic scars:a retrospective clinical observational study

Background:Hypertrophic scars are one of the main complications that affect the quality of life of patients after burns.Many methods have been shown to be effective in the treatment of hypertrophic scars,such as ablative fractional CO_(2) laser(AFCL)and platelet-rich plasma(PRP).However,there are few studies on the effect of the combined application of these measures.The purpose of this study was to explore the therapeutic effect of AFCL combined with PRP on hypertrophic burn scars.Methods:A retrospective clinical observation study was conducted on 50 patients with hypertrophic burn scars.The AFCL+PRP group included 31 patients who received AFCL combined with PRP treatment;the AFCL group included 19 patients who received AFCL treatment only.The University of North Carolina 4P Scar Scale(UNC4P)and the Vancouver Scar Scale(VSS)scores that were collected before each treatment were used as indicators of the effectiveness of the previous treatment.The scores recorded at the second,fourth and seventh months were analysed.Results:The demographic data of the 2 groups were not significantly different.Before treatment,therewas no difference in the UNC4P and VSS scores between the 2 groups.Therewas a significant decline in the UNC4P and VSS total scores over 6 months in both groups(p<0.05)and scores in the 2 groups were comparable after 3 and 6 months(p<0.05).UNC4P scores in the AFCL+PRP group decreased from a mean of 8.26 to 2.61(p<0.05)with a concomitant drop in VSS scores from a mean of 11.74 to 6.06(p<0.01).In the AFCL group UNC4P and VSS scores decreased from 7.68 to 4.63(p<0.05)and from 10.89 to 8.16(p<0.05),respectively.The sub-items of these 2 assessments were analysed and the results suggest that AFCL combined with PRP can comprehensively improve scarring.Conclusions:This study shows that PRP is an effective adjunct for AFCL in the treatment of hypertrophic burn scars and that the combination of PRP and AFCL proved to be more useful than AFCL alone.This combination may be a new and effective clinical practice for the treatment of scars.However,larger and higher-level clinical studies are still needed to determine its efficacy and possible mechanisms.