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.

Effect of ALA-PDT on the expressions of MMP-9, MMP-13 and TIMP-1 of hypertrophic scar model in rabbit ears

Objective: To investigate the effect of 5-aminolevulinic(ALA)-photodynamic therapy(PDT) on the expressions of MMP-9, MMP-13 and TIMP-1 of hypertrophic scar model in rabbit ears, and analyze the possible therapeutic mechanisms of ALA-PDT treatment to hypertrophic scars of rabbit ears. Methods: The experimental animals were randomly divided into normal control, negative control, high concentration of ALA-PDT, low concentration of ALA-PDT and PDT groups. The latter three groups received ALA-PDT treatment or PDT treatment once a week for 3 weeks. The specimens of the rabbits were collected respectively 1, 2 and 3 months after treatment to be used for RT-PCR and Western-blot test. Results: 1, 2 and 3 months after PDT treatment, the expressions of MMP-9 and MMP-13(including mRNA and protein) in hypertrophic scar tissues of three treatment groups were significantly higher than those of the negative control group(P<0.01), and the expression of TIMP-1 mRNA and protein of three treatment groups were significantly lower than that of the negative control group(P<0.01). There were also significant differences between high-concentration ALA-PDT treatment group and the low one(P<0.05). Conclusion: ALA-PDT is effective in treating hypertrophic scars of rabbit ears, and its possible therapeutic mechanisms are that ALA-PDT treatment generates oxidation activation effect to activate the activity of MMPs and induces the photoaging of fibroblasts of hypertrophic scar tissues of rabbit ears to inhibit the activity of TIMPs, which causes the up-regulation of MMPs and the down-regulation of TIMPs. Because of this, the degradation of collagen and ECM is accelerated and the formation of scars is suppressed.

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.

STUDY ON FUNCTION OF FOCAL ADHENSIVE KINASE AND INTEGRIN α_1 IN HYPERTROPHIC SCAR FIBROBLASTS

Objective To study the function of focal adhesion kinase （FAK） in the formation of hypertrophic scar and its interrelationship with integrin α1. Methods Original fibroblasts from human hypertrophic scar and human normal dermis were cultured, and immunocytochemistry was applied to detect localization of expres- sion of FAK and integrin α1 in hypertrophic scar and human normal skin fibroblasts. The expression of integrin α1 was detected before and after FAK antibody blocking hypertrophic scar fibroblasts （HSFB） 48 h later. Meanwhile the collagen synthesis was evaluated by [^3 H]-proline incorporation and HSFB cell proliferation was measured by MTT method. Results The expression of FAK and integrin aI of hypertrophic scar fibroblasts was higher than that of the normal skin fibroblasts significantly （ P 〈 0.01 ）. The expression of integrin α1 was reduced after FAK being blocked （ P 〈 0.01 ）. Meanwhile the collagen synthesis of human scar-derived fibroblasts by [^3H] -proline incor- poration was depressed respectively （ P 〈 0. 01 ）. The cell proliferation was inhibited by using 1：100 and 1：200 FAK antibody with MTI＂ method （ P 〈 0. 01 ）. Conclusion FAK is the key point of signal transmission pathway mediated by integrin α1 , which regulates protein synthesis of integrin α1 , it may play an important role in the proliferation and constriction of hypertrophic scar. FAK antibody can inhibit the collagen synthesis and cell proliferation of hypertrophic scar fibroblasts.

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

The Effects of Laser Therapy in Treating Hypertrophic Scars and Keloids after Median Sternotomy:A Scoping Review

Background:Hypertrophic scars and keloids,common complications following median sternotomy for cardiac surgery,significantly impact patient quality of life due to their aesthetic and symptomatic burden.Recent advancements in laser therapy have made it a prominent option for managing these complex scars,yet a com-prehensive understanding of its efficacy is lacking.The aim of this scoping review is to explore the effects of laser therapy in managing hypertrophic scars and keloids after median sternotomy.Methods:This scoping review ana-lyzed studies up to February 2024 from databases including PubMed,EMBASE,CINAHL,Scopus,Web of Science,and the Cochrane Library.We included any study that assessed laser therapy’s effects on hypertrophic scars and keloids following median sternotomy.Studies were selected based on predefined inclusion criteria with-out publication year,design,or origin restrictions.Results:Six studies met the inclusion criteria,involving a total offive RCTs and one review.These studies primarily tested 585 and 595-nm pulsed dye laser(PDL)treatments,focusing on scar appearance,patient symptoms,and treatment satisfaction.Most studies reported significant improvements in scar height reduction and patient symptom relief after treatment,with mixed results for scar erythema and elasticity.Adverse events were generally mild and transient.Conclusions:Laser therapy offers a beneficial approach for improving the appearance and symptoms of hypertrophic scars and keloids post-median sternotomy.However,further research is necessary to optimize treatment parameters and explore the long-term psychosocial impacts of this therapy.This review highlights the need for more comprehensive studies to establish standardized treatment protocols and evaluate their effectiveness.

Expression of oncoproteins c-fos and c jun in hypertrophic scars and chronic dermal ulcers and their regulation of basic fibroblast growth factor

Objective To explore the characteristics of oncoprotein expression of c-fos and c-jun in hypertrophic scars and chronic dermal ulcers and their regulation of basic fibroblast growth factor (bFGF). Methods Tissues of hypertrophic scars (n=8), chronic dermal ulcers (n=8) and normal skin (n=5) were taken from 21 patients with burns and chronic dermal ulcers in operation. The ABC immunohistochemical method was used to characterize the gene product expression of c-fos, c-jun and bFGF in the above tissues. Results In normal skin, both c-fos and c-jun protein expression and bFGF protein expression were observed. The signals of both oncoproteins were localized mainly in subcutaneous fibroblasts, but, positive expression of the bFGF protein was mainly in keratinocytes. In hypertrophic scars, positive expression of both oncoproteins could be found mainly in fibroblasts, but bFGF was mainly in fibroblasts and endothelial cells. In chronic dermal ulcers, endothelial cells, some of inflammatory cells and fibroblasts were positive for both of oncoproteins, but the expression of bFGF was only seen in fibroblasts and endothelial cells. Conclusions The results indicate that the interaction between both oncoproteins and bFGF exists, and the regulating action between protooncogenes and bFGF is a major course in wound healing. The different expressions of c-fos and c-jun gene products play an important role in regulate bFGF action, thus affecting wound healing.

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.

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.

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.

Endostatin inhibits hypertrophic scarring in a rabbit ear model

Objective： The present study was designed to use an in vivo rabbit ear scar model to investigate the efficacy of systemic administration of endostatin in inhibiting scar formation. Methods： Eight male New Zealand white rabbits were randomly assigned to two groups. Scar model was established by making six full skin defect wounds in each ear. For the intervention group, intraperitoneal injection of endostatin was performed each day after the wound healed （about 15 d post wounding）. For the control group, equal volume of saline was injected. Thickness of scars in each group was measured by sliding caliper and the scar microcirculatory perfusion was assessed by laser Doppler flowmetry on Days 15, 21, 28, and 35 post wounding. Rabbits were euthanatized and their scars were harvested for histological and proteomic analyses on Day 35 post wounding. Results： Macroscopically, scars of the control group were thicker than those of the intervention group. Significant differences between the two groups were observed on Days 21 and 35 （p〈0.05）. Scar thickness, measured by scar elevation index （SEI） at Day 35 post wounding, was significantly reduced in the intervention group （1.09±0.19） compared with the controls （1.36±0.28）. Microvessel density （MVD） observed in the intervention group （1.73±0.94） was significantly lower than that of the control group （5.63±1.78） on Day 35. The distribution of collagen fibers in scars treated with endostatin was relatively regular, while collagen fibers in untreated controls were thicker and showed disordered alignment. Western blot analysis showed that the expressions of type I collagen and Bcl-2 were depressed by injection of endostatin. Conclusions： Our results from the rabbit ear hypertrophic scar model indicate that systemic application of endostatin could inhibit local hypertrophic scar formation, possibly through reducing scar vascularization and angiogenesis. Our results indicated that endostatin may promote the apoptosis of endothelial cells and block their release of platelet-dedved growth factor （PDGF） and fibroblast growth factor （FGF）, thereby controlling collagen production by fibroblasts. Blood vessel-targeted treatment may be a promising strategy for scar therapy.

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.

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.

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.

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

Keyword: hypertrophic scars · keloids · HLA DR · CD1a