Calculus bovis inhibits M2 tumor-associated macrophage polarization via Wnt/β-catenin pathway modulation to suppress liver cancer

BACKGROUND Calculus bovis(CB),used in traditional Chinese medicine,exhibits anti-tumor effects in various cancer models.It also constitutes an integral component of a compound formulation known as Pien Tze Huang,which is indicated for the treatment of liver cancer.However,its impact on the liver cancer tumor microenvironment,particularly on tumor-associated macrophages(TAMs),is not well understood.AIM To elucidate the anti-liver cancer effect of CB by inhibiting M2-TAM polarization via Wnt/β-catenin pathway modulation.METHODS This study identified the active components of CB using UPLC-Q-TOF-MS,evaluated its anti-neoplastic effects in a nude mouse model,and elucidated the underlying mechanisms via network pharmacology,transcriptomics,and molecular docking.In vitro assays were used to investigate the effects of CB-containing serum on HepG2 cells and M2-TAMs,and Wnt pathway modulation was validated by real-time reverse transcriptase-polymerase chain reaction and Western blot analysis.RESULTS This study identified 22 active components in CB,11 of which were detected in the bloodstream.Preclinical investigations have demonstrated the ability of CB to effectively inhibit liver tumor growth.An integrated approach employing network pharmacology,transcriptomics,and molecular docking implicated the Wnt signaling pathway as a target of the antineoplastic activity of CB by suppressing M2-TAM polarization.In vitro and in vivo experiments further confirmed that CB significantly hinders M2-TAM polarization and suppresses Wnt/β-catenin pathway activation.The inhibitory effect of CB on M2-TAMs was reversed when treated with the Wnt agonist SKL2001,confirming its pathway specificity.CONCLUSION This study demonstrated that CB mediates inhibition of M2-TAM polarization through the Wnt/β-catenin pathway,contributing to the suppression of liver cancer growth.

Th17/Treg balance and macrophage polarization ratio in lower extremity arteriosclerosis obliterans

Objective:To explore the balance of peripheral blood T helper 17 cells/regulatory T cell(Th17/Treg)ratio and the polarization ratio of M1 and M2 macrophages in lower extremity arteriosclerosis obliterans(ASO).Methods:A rat model of lower extremity ASO was established,and blood samples from patients with lower extremity ASO before and after surgery were obtained.ELISA was used to detect interleukin 6(IL-6),IL-10,and IL-17.Real-time RCR and Western blot analyses were used to detect Foxp3,IL-6,IL-10,and IL-17 expression.Moreover,flow cytometry was applied to detect the Th17/Treg ratio and M1/M2 ratio.Results:Compared with the control group,the iliac artery wall of ASO rats showed significant hyperplasia,and the concentrations of cholesterol and triglyceride were significantly increased(P<0.01),indicating the successful establishment of ASO.Moreover,the levels of IL-6 and IL-17 in ASO rats were pronouncedly increased(P<0.05),while the IL-10 level was significantly decreased(P<0.05).In addition to increased IL-6 and IL-17 levels,the mRNA and protein levels of Foxp3 and IL-10 in ASO rats were significantly decreased compared with the control group.The Th17/Treg and M1/M2 ratios in the ASO group were markedly increased(P<0.05).These alternations were also observed in ASO patients.After endovascular surgery(such as percutaneous transluminal angioplasty and arterial stenting),all these changes were significantly improved(P<0.05).Conclusions:The Th17/Treg and M1/M2 ratios were significantly increased in ASO,and surgery can effectively improve the balance of Th17/Treg,and reduce the ratio of M1/M2,and the expression of inflammatory factors.

Identification of M2 macrophage-related genes for establishing a prognostic model in pancreatic cancer: FCGR3A as key gene

Background:Pancreatic ductal adenocarcinoma(PDAC)has a rich and complex tumor immune microenvironment(TIME).M2 macrophages are among the most extensively infiltrated immune cells in the TIME and are necessary for the growth and migration of cancers.However,the mechanisms and targets mediating M2 macrophage infiltration in pancreatic cancer remain elusive.Methods:The M2 macrophage infiltration score of patients was assessed using the xCell algorithm.Using weighted gene co-expression network analysis(WGCNA),module genes associated with M2 macrophages were identified,and a predictive model was designed.The variations in immunological cell patterns,cancer mutations,and enrichment pathways between the cohorts with the high-and low-risk were examined.Additionally,the expression of FCGR3A and RNASE2,as well as their association with M2 macrophages were evaluated using the HPA,TNMplot,and GEPIA2 databases and verified by tissue immunofluorescence staining.Moreover,in vitro cell experiments were conducted,where FCGR3A was knocked down in pancreatic cancer cells using siRNA to analyze its effects on M2 macrophage infiltration,tumor proliferation,and metastasis.Results:The prognosis of patients in high-risk and low-risk groups was successfully distinguished using a prognostic risk score model of M2 macrophage-related genes(p=0.024).Between the high-and low-risk cohorts,there have been notable variations in immune cell infiltration patterns,tumor mutations,and biological functions.The risk score was linked to the manifestation of prevalent immunological checkpoints,immunological scores,and stroma values(all p<0.05).In vitro experiments and tissue immunofluorescence staining revealed that FCGR3A can promote the infiltration or polarization of M2 macrophages and enhance tumor proliferation and migration.Conclusions:In this study,an M2 macrophage-related pancreatic cancer risk score model was established,and found that FCGR3A was correlated with tumor formation,metastasis,and M2 macrophage infiltration.

Abnormally activated wingless/integrated signaling modulates tumor-associated macrophage polarization and potentially promotes hepatocarcinoma cell growth

In this article,we comment on the article by Huang et al.The urgent development of new therapeutic strategies targeting macrophage polarization is critical in the fight against liver cancer.Tumor-associated macrophages(TAMs),primarily of the M2 subtype,are instrumental in cellular communication within the tumor microenvironment and are influenced by various signaling pathways,including the wingless/integrated(Wnt)pathway.Activation of the Wnt signaling pathway is pivotal in promoting M2 TAMs polarization,which in turn can exacerbate hepatocarcinoma cell proliferation and migration.This manuscript emphasizes the burgeoning significance of the Wnt signaling pathway and M2 TAMs polarization in the pathogenesis and progression of liver cancer,highlighting the potential therapeutic benefits of inhibiting the Wnt pathway.Lastly,we point out areas in Huang et al’s study that require further research,providing guidance and new directions for similar studies.

From traditional Chinese medicine formulations to effective anticancer agents:Insights from Calculus bovis

This editorial examines the therapeutic potential of traditional Chinese medicine(TCM)for aggressive cancers,particularly liver cancer.It highlights the study by Huang et al,which shows how Calculus bovis,a component of the TCM Pien Tze Huang,suppresses liver cancer by inhibiting M2 macrophage polarization via the Wnt/β-catenin pathway.This research emphasizes the importance of transitioning from effective TCM formulations to isolating active components and understanding their mechanisms.While the study provides valuable insights,it primarily focuses on the Wnt/β-catenin pathway and does not delve deeply into the mechanisms of individual components.Future research should aim to comprehensively study these components,explore their interactions,and validate findings through clinical trials.This approach will integrate traditional wisdom with modern scientific validation,advancing the development of innovative cancer treatments based on TCM formulations.

Anti-tumor efficacy of Calculus bovis: Suppressing liver cancer by targeting tumor-associated macrophages

Despite significant advances in our understanding of the molecular pathogenesis of liver cancer and the availability of novel pharmacotherapies,liver cancer remains the fourth leading cause of cancer-related mortality worldwide.Tumor relapse,resistance to current anti-cancer drugs,metastasis,and organ toxicity are the major challenges that prevent considerable improvements in patient survival and quality of life.Calculus bovis(CB),an ancient Chinese medicinal drug,has been used to treat various pathologies,including stroke,convulsion,epilepsy,pain,and cancer.In this editorial,we discuss the research findings recently published by Huang et al on the therapeutic effects of CB in inhibiting the development of liver cancer.Utilizing the comprehensive transcriptomic analyses,in vitro experiments,and in vivo studies,the authors demonstrated that CB treatment inhibits the tumor-promoting M2 phenotype of tumor-associated macrophages via downregulating Wnt pathway.While multiple studies have been performed to explore the molecular mechanisms regulated by CB,this study uniquely shows its role in modulating the M2 phenotype of macrophages present within the tumor microenvironment.This study opens new avenues of future investigations aimed at investigating this drug’s efficacy in various mouse models including the effects of combination therapy,and against drug-resistant tumors.

Exploring a new chapter in traditional Chinese medicine:The potential of Calculus bovis in liver cancer treatment

In the ongoing quest for new treatments in medicine,traditional Chinese medicine offers unique insights and potential.Recently,studies on the ability of Calculus bovis to inhibit M2-type tumour-associated macrophage polarisation by modulating the Wnt/β-catenin signalling pathway to suppress liver cancer have undoubtedly revealed new benefits and hope for this field of research.The purpose of this article is to comment on this study and explore its strengths and weaknesses,thereby providing ideas for the future treatment of liver cancer.

ScRNA-seq reveals the correlation between M2 phenotype of tumorassociated macrophages and lymph node metastasis of breast cancer

The process of lymphatic metastasis was proved to be associated with podoplanin-expressing macrophages in breast cancer(BC).This study aimed to investigate the role of the M2 phenotype of tumor-associated macrophages and mine the key M2 macrophages-related genes for lymph node metastasis in BC.We downloaded the GSE158399 dataset from the Gene Expression Omnibus(GEO)database,which includes transcriptomic profiles of individual cells from primary tumors,negative lymph nodes(NLNs),and positive lymph nodes(PLNs)of breast cancer patients.The cell subsets were identified by clustering analysis after quality control of the scRNA-seq using Seurat.The activation and migration capability of M2 macrophages were evaluated with R package“GSVA”.The key M2 macrophages-related genes were screened from the differential expressed genes(DEGs)and M2 macrophages activation and migration gene sets collected from MSigDB database.Our analysis identified three main cell types in primary tumors,NLNs,and PLNs:basal cells,luminal cells,and immune cell subsets.The further cell type classification of immune cell subsets indicated M2 macrophages accumulation in NLs and PLs.The GSVA enrichment scores for activation and migration capability were increased significantly in M2 macrophages from primary tumors than NLNs and PLNs(pvalue<0.001).Seven M2 macrophages activation-related and 15 M2 macrophages migration-related genes were significantly up-regulated in primary tumors than NLNs and PLNs.The proportion and GSVA enrichment scores for activation and migration of M2 macrophages may be potential markers for lymph node metastasis in breast cancer.Our study demonstrated that twenty-two up-regulated mRNA may be possible therapeutic targets for lymph node metastasis in breast cancer.

Growth differentiation factor 11 promotes macrophage polarization towards M2 to attenuate myocardial infarction via inhibiting Notch1 signaling pathway

Background:Myocardial infarctions(MI)is a major threat to human health especially in people exposed to cold environment.The polarization of macrophages towards different functional phenotypes(M1 macrophages and M2 macrophages)is closely related to MI repairment.The growth differentiation factor 11(GDF11)has been reported to play a momentous role in inflammatory associated diseases.In this study,we examined the regulatory role of GDF11 in macrophage polarization and elucidated the underlying mechanisms in MI.Methods:In vivo,the mice model of MI was induced by permanent ligation of the left anterior descending coronary artery(LAD),and mice were randomly divided into the sham group,MI group,and MI+GDF11 group.The protective effect of GDF11 on myocardial infarction and its effect on macrophage polarization were verified by echocardiography,triphenyl tetrazolium chloride staining and immunofluorescence staining of heart tissue.In vitro,based on the RAW264.7 cell line,the effect of GDF11 in promoting macrophage polarization toward the M2 type by inhibiting the Notch1 Signaling pathway was validated by qRT-PCR,Western blot,and flow cytometry.Results:We found that GDF11 was significantly downregulated in the cardiac tissue of MI mice.And GDF11 supplementation can improve the cardiac function.Moreover,GDF11 could reduce the proportion of M1 macrophages and increase the accumulation of M2 macrophages in the heart tissue of MI mice.Furthermore,the cardioprotective effect of GDF11 on MI mice was weakened after macrophage clearance.At the cellular level,application of GDF11 could inhibit the expression of M1 macrophage(classically activated macrophage)markers iNOS,interleukin(IL)-1β,and IL-6 in a dose-dependent manner.In contrast,GDF11 significantly increased the level of M2 macrophage markers including IL-10,CD206,arginase 1(Arg1),and vascular endothelial growth factor(VEGF).Interestingly,GDF11 could promote M1 macrophages polarizing to M2 macrophages.At the molecular level,GDF11 significantly down-regulated the Notch1 signaling pathway,the activation of which has been demonstrated to promote M1 polarization in macrophages.Conclusions:GDF11 promoted macrophage polarization towards M2 to attenuate myocardial infarction via inhibiting Notch1 signaling pathway.

Macrophage colony-stimulating factor expressed in non-cancer tissues provides predictive powers for recurrence in hepatocellular carcinoma

AIM To investigate the role of macrophage colony-stimulating factor(M-CSF) in patients with hepatocellular carcinoma(HCC) after surgery. METHODS Expression of M-CSF, distribution of M2 macrophages(Mφs), and angiogenesis were assessed in the liver, including tumors and peritumoral liver tissues. The prognostic power of these factors was assessed. Mouse isolated hepatic Mφs or monocytes were cultured with media containing M-CSF. The concentration of vascular endothelial growth factor(VEGF) in media was assessed. Furthermore, the role of the M-CSF-matured hepatic Mφs on proliferation of the vascular endothelial cell(VEC) was investigated. RESULTS A strong correlation between the expressions of M-CSF and CD163 was observed in the peritumoral area. Also, groups with high density of M-CSF, CD163 or CD31 showed a significantly shorter time to recurrence(TTR) than low density groups. Multivariate analysis revealedthe expression of M-CSF or hepatic M2Mφs in the peritumoral area as the most crucial factor responsible for shorter TTR. Moreover, the expression of M-CSF and hepatic M2Mφs in the peritumoral area had better predictable power of overall survival. Values of VEGF in culture media were significantly greater in the hepatic Mφs compared with the monocytes. Proliferation of the VEC was greatest in the cells co-cultured with hepatic Mφs when M-CSF was present in media.CONCLUSION M-CSF increases hepatocarcinogenesis, most likely by enhancing an angiogenic factor derived from hepatic Mφ and could be a useful target for therapy against HCC.

Exosomal microRNA-588 from M2 polarized macrophages contributes to cisplatin resistance of gastric cancer cells

BACKGROUND Gastric cancer is a prevalent malignant cancer with a high incidence and significantly affects the health of modern people globally.Cisplatin(DDP)is one of the most common and effective chemotherapies for patients with gastric cancer,but DDP resistance remains a severe clinical challenge.AIM To explore the function of M2 polarized macrophages-derived exosomal microRNA(miR)-588 in the modulation of DDP resistance of gastric cancer cells.METHODS M2 polarized macrophages were isolated and identified by specific markers using flow cytometry analysis.The exosomes from M2 macrophages were identified by transmission electron microscopy and related markers.The uptake of the PKH67-labelled M2 macrophages-derived exosomes was detected in SGC7901 cells.The function and mechanism of exosomal miR-588 from M2 macrophages in the modulation of DDP resistance of gastric cancer cells was analyzed by CCK-8 assay,apoptosis analysis,colony formation assay,Western blot analysis,qPCR analysis,and luciferase reporter assay in SGC7901 and SGC7901/DDP cells,and by tumorigenicity analysis in nude mice.RESULTS M2 polarized macrophages were isolated from mouse bone marrow stimulated with interleukin(IL)-13 and IL-4.Co-cultivation of gastric cancer cells with M2 polarized macrophages promoted DDP resistance.M2 polarized macrophagesderived exosomes could transfer in gastric cancer cells to enhance DDP resistance.Exosomal miR-588 from M2 macrophages contributed to DDP resistance of gastric cancer cells.miR-588 promoted DDP-resistant gastric cancer cell growth in vivo.miR-588 was able to target cylindromatosis(CYLD)in gastric cancer cells.The depletion of CYLD reversed miR-588 inhibition-regulated cell proliferation and apoptosis of gastric cancer cells exposed to DDP.CONCLUSION In conclusion,we uncovered that exosomal miR-588 from M2 macrophages contributes to DDP resistance of gastric cancer cells by partly targeting CYLD.miR-588 may be applied as a potential therapeutic target for the treatment of gastric cancer.

Macrophage-derived exosomal miR-342-3p promotes the progression of renal cell carcinoma through the NEDD4L/CEP55 axis

Due to its difficulty in early diagnosis and lack of sensitivity to chemotherapy and radiotherapy,renal cell carcinoma(RCC)remains to be a frequent cause of cancer-related death.Here,we probed into new targets for its early diagnosis and treatment for RCC.microRNA(miRNA)data of M2-EVs and RCC were searched on the Gene Expression Omnibus database,followed by the prediction of the potential downstream target.Expression of target genes was measured via RT-qPCR and Western blot,respectively.M2 macrophage was obtained viaflow cytometry with M2-EVs extracted.The binding ability of miR-342-3p to NEDD4L and to CEP55 ubiquitination was studied with their roles in the physical abilities of RCC cells assayed.Subcutaneous tumor-bearing mouse models and lung metastasis models were prepared to observe in vivo role of target genes.M2-EVs induced RCC growth and metastasis.miR-342-3p showed high expression in both M2-EVs and RCC cells.M2-EVs carrying miR-342-3p promoted RCC cell abilities to proliferate,invade and migrate.In RCC cells,M2-EV-derived miR-342-3p could specifically bind to NEDD4L and consequently elevate CEP55 protein expression via suppressing NEDD4L,thereby exerting tumor-promoting effects.CEP55 could be degraded by ubiquitination under the function of NEDD4L,and miR-342-3p delivered by M2-EVs facilitated the RCC occurrence and development by activating the PI3K/AKT/mTOR signaling pathway.In conclusion,M2-EVs promote RCC growth and metastasis by delivering miR-342-3p to suppress NEDD4L and subsequently inhibit CEP55 ubiquitination and degradation via activation of the PI3K/AKT/mTOR signaling pathway,strongly driving the proliferative,migratory and invasive of RCC cells.

Effect of Pingchuan granule on typeⅡinnate lymphocytes and M2 polarization of macrophages in asthmatic mice

Objective:To observe the effect of Pingchuan granule on the number of typeⅡinnate lymphocytes(ILC2)and M2 polarization of macrophages in the lung tissue of asthmatic mice;Methods:Ovalbumin sensitized and challenged asthmatic mouse models were established,and then Pingchuan granules or IL-33 neutralizing antibody were given to intervene.The pathological morphology of lung tissue was observed by HE,PAS and Masson staining,and the expressions of IL-4,IL-5,IL-13 and IL-33 in BALF and lung tissue were detected by ELISA and qRT-PCR,Flow cytometry was used to detect the number of type II innate lymphocytes and type M2 macrophages in lung tissue.Western blot was used to detect the protein expression levels of ST-2,FIZZ1 and Arg-1 in lung tissue;Results:Compared with the control group,the inflammation score,PAS score and collagen staining area of the model group were significantly increased,the expressions of IL-4,IL-5,IL-13 and IL-33 in BALF and lung tissue were significantly increased,the number of ILC2 and M2 macrophages,the expression of ST-2,FIZZ1 and Arg-1 protein in lung tissue were significantly increased,and the differences were statistically significant(P<0.05);Compared with the model group,Pingchuan granule could significantly reduce the inflammation score,PAS score and collagen staining area of asthmatic mice,down-regulate the expression of IL-4,IL-5,IL-13 and IL-33 in BALF and lung tissue,reduce the number of ILC2 and M2 macrophages,and the expression of ST-2,FIZZ1 and Arg-1 protein in lung tissue(P<0.05);Conclusion:Pingchuan granule improve the airway remodeling of asthma by inhibiting the polarization of M2 macrophages mediated by ILC2.

Morphofunctional Characteristics of Pulmonary Surfactant System and Its Effect on Immune Cells in Influenza A (H1N1) Pathogenesis

There is an annual increase of influenza-related SARI cases in winter months. Despite the high relevance of this problem, influenza pathogenesis and the role of surfactant system and its SP-A (surfactant protein A) enzyme in antiviral defense remain poorly understood. SP-A activates macrophage M1 polarization and triggers an antiviral response due to the activation of T-cells and dendritic cells. Therefore, surfactant system is an important element of infection protection and a promising therapeutic target.

Thylakoid engineered M2 macrophage for sonodynamic effect promoted cell therapy of early atherosclerosis

Atherosclerosis is the most common cause of cardiovascular diseases that contribute to the major morbidity worldwide,but still lacking of effective treatment strategy.Here,a hybrid cell is constructed for the sonodynamic effect promoted cell therapy of early atherosclerosis by fusing M2 macrophages with thylakoid(TK)membranes.After systemic administration,the obtained TK-M2 actively accumulates in the early atherosclerotic plaques,wherein M2 macrophages relieve the cholesterol accumulation and the inflammation in the foam cells.Meanwhile,the TK membranes decorated on the M2 macrophages exhibit both type I and type II sonodynamic effects under ultrasound(US)activation,inducing the direct apoptosis of foam cells.The cooperation of M2 and TK leads to significant outcome in eliminating atherosclerotic plaques without obvious side-effects,providing a new avenue for atherosclerosis treatment.

Dual-functional microneedle with programmatic regulation of macrophage for autoimmune psoriasis treatment

Modulating the immune microenvironment to establish sustained positive feedback within immune pathways represents a promising avenue for the treatment of autoimmunity.However,the precise and efficient delivery of therapeutic systems to the subcutaneous basal layer to modulate immune disorders is a major challenge in the treatment of autoimmune psoriasis.In this project,we introduce a dual-functional microneedle(DF-MN)designed to combine MNs with multiple release kinetics and immunotherapy,the programmed treatment is achieved through segmented design of the MN structure,realizing the unification of rapid and long-lasting treatment of autoimmune psoriasis.In vivo imaging results showed that GelMA@M-CSF showed fluorescent signals after 5 days of delivery to subcutaneous tissues,whereas HA@IL-13 showed minimal fluorescent signals after 2 days.The multistage release behavior of MNs and the diffusion mechanism of drugs were explained at the molecular level,in combination with coarse-grained molecular dynamics.Additionally,DF-MN can successfully induce macrophage reprogramming in vitro and ameliorate overall symptoms in a psoriasis mice model,suggesting that it has the potential to be an effective strategy for the treatment of psoriasis and portends to be a transformative platform for the treatment of other autoimmune diseases.

Effect of Platelet-rich Plasma in Stimulating Macrophage Transformation into M2 Type under AMPK Signaling Pathway

Objective:To explore the effect of platelet-rich plasma(RPR)in stimulating the transformation of pro-inflammatory(M1)macrophages into antiinflammatory(M2)under the adenosine-monophosphate-dependent protein kinase(AMPK)signaling pathway.Methods:Rat peritoneal macrophages(RAW264.7)were cultured and randomly divided into 8 groups:blank control group,LPS group,RPR group A,RPR group B,LPS+RPR(12 h)group,LPS+RPR(24 h)group A,LPS+RPR(24 h)group B,LPS+RPR(24 h)group C.RPR was prepared based on blood donors.The expressions of AMPK signaling pathway-related proteins(AMPK,ULK1,m TOR)and macrophage markers(i NOS,Arg-1)in the blank control group,LPS group,LPS+RPR(12 h)group and LPS+RPR(24 h)group were observed and compared.The expressions of macrophage markers in LPS+RPR(24 h)B and C groups were compared,and the expressions of AMPK and TGF-βin RPR A and B groups were compared.Results:The gray values of AMPK and ULK1 in LPS cells decreased significantly,while those in LPS+RPR(12 h)and LPS+RPR(24 h)A cells increased significantly.The gray values of AMPK and ULK1 in LPS+RPR(24 h)A cells were higher than those in LPS+RPR(12 h)cells(P<0.05).The m TOR gray value of LPS cells was significantly higher than that of LPS+RPR(24 h)A cells,and the m TOR gray value of LPS+RPR(24 h)A cells was significantly lower than that of LPS+RPR(12 h)cells(P<0.05).The expression of i NOS in LPS cells was significantly decreased,the expression of i NOS in LPS+RPR(12 h)and LPS+RPR(24 h)cells was significantly increased,and the expression of i NOS in LPS+RPR(24 h)cells was higher than that in LPS+RPR(12 h)cells(P<0.05).The expression of Arg-1 in LPS cells was significantly decreased,the expression of Arg-1 in LPS+RPR(12 h)and LPS+RPR(24 h)A cells was significantly increased,and the expression of Arg-1 in LPS+RPR(24 h)A cells was higher than that in LPS+RPR(12 h)cells(P<0.05).The i NOS expression level of LPS+PRP(24 h)C cells was significantly higher than that of LPS+PRP(24 h)B cells,and the Arg-1 expression level was significantly lower than that of LPS+PRP(24 h)B cells(P<0.05).The gray values of AMPK and TGF-βin PRP B cells were significantly lower than those in PRP A cells(P<0.05).Conclusion:RPR can stimulate macrophage transformation from M1 to M2 by up-regulating AMPK signaling pathway.

Engineered M2 macrophage-derived extracellular vesicles with platelet membrane fusion for targeted therapy of atherosclerosis

Atherosclerosis is featured as chronic low-grade inflammation in the arteries,which leads to the formation of plaques rich in lipids.M_(2)macrophage-derived extracellular vesicles(M_(2)EV)have significant potential for anti-atherosclerotic therapy.However,their therapeutic effectiveness has been hindered by their limited targeting capability in vivo.The objective of this study was to create the P-M_(2)EV(platelet membrane-modified M_(2)EV)using the membrane fusion technique in order to imitate the interaction between platelets and macrophages.P-M_(2)EV exhibited excellent physicochemical properties,and microRNA(miRNA)-sequencing revealed that the extrusion process had no detrimental effects on miRNAs carried by the nanocarriers.Remarkably,miR-99a-5p was identified as the miRNA with the highest expression level,which targeted the mRNA of Homeobox A1(HOXA1)and effectively suppressed the formation of foam cells in vitro.In an atherosclerotic low-density lipoprotein receptor-deficient(Ldlr􀀀^(-/-)􀀀)mouse model,the intravenous injection of P-M_(2)EV showed enhanced targeting and greater infiltration into atherosclerotic plaques compared to regular extracellular vesicles.Crucially,P-M_(2)EV successfully suppressed the progression of atherosclerosis without causing systemic toxicity.The findings demonstrated a biomimetic platelet-mimic system that holds great promise for the treatment of atherosclerosis in clinical settings.

Genetically engineered M2-like macrophage-derived exosomes for P.gingivalis-suppressed cementum regeneration:From mechanism to therapy

Cementum,a thin layer of mineralized tissue covering tooth root surface,is recognized as the golden standard in periodontal regeneration.However,current efforts mainly focus on alveolar bone regeneration rather than cementum regeneration,and rarely take Porphyromonas gingivalis(Pg),the keystone pathogen responsible for periodontal tissue destruction,into consideration.Though M2 macrophage-derived exosomes(M2-EXO)show promise in tissue regeneration,the exosome-producing M2 macrophages are induced by exogenous cytokines with transitory and unstable effects,restricting the regeneration potential of M2-EXO.Here,exosomes derived from genetically engineered M2-like macrophages are constructed by silencing of casein kinase 2 interacting protein-1(Ckip-1),a versatile player involved in various biological processes.Ckip-1 silencing is proved to be an effective gene regulation strategy to obtain permanent M2-like macrophages with mineralization-promoting effect.Further,exosomes derived from Ckip-1-silenced macrophages(sh-Ckip-1-EXO)rescue Pg-suppressed cementoblast mineralization and cementogenesis.Mechanismly,sh-Ckip-1-EXO delivers Let-7f-5p targeting and silencing Ckip-1,a negative regulator also for cementum formation and cementoblast mineralization.More deeply,downregulation of Ckip-1 in cementoblasts by exosomal Let-7f-5p activates PGC-1α-dependent mitochondrial biogenesis.In all,this study provides a new strategy of genetically engineered M2-like macrophage-derived exosomes for cementum regeneration under Pg-dominated inflammation.

Multi-walled carbon nanotubes reversing the bone formation of bone marrow stromal cells by activating M2 macrophage polarization

Multi-walled carbon nanotubes(MWCNTs)are an excellent bone tissue repair material both in vitro and in vivo.The interactions between MWCNTs and single type of cells of bone tissue,including osteoblasts,bone marrow stromal cells(BMSCs)or osteoclasts,have been extensively studied.However,the interactions between MWCNTs with different types of cells in the bone microenvironment remain elusive.Bone microenvironment is a complex system composed of different types of cells,which have interactions between each other.In this work,the effects of MWCNTs on bone microenvironment were firstly studied by culture of MWCNTs with BMSCs,osteoblasts,osteoclasts,macrophages and vascular endothelial cells,respectively.Then,co-culture systems of macrophages-BMSCs,macrophages-calvaria and macrophages-BMSCs-vascular endothelial cells were treated with MWCNTs,respectively.The osteogenic differentiation of BMSCs and osteoblasts was inhibited when these two types of cells were cultured with MWCNTs,respectively.Strikingly,when co-culture MWCNTs with BMSCs and macrophages,the osteogenesis of BMSCs was promoted by inducing the M2 polymerization of macrophages.Meanwhile,MWCNTs promoted the bone formation in the osteolysis model of calvaria ex vivo.In addition,the formation of osteoclasts was inhibited,and angiogenesis was increased when treated with MWCNTs.This study revealed the inconsistent effects of MWCNTs on single type of bone cells and on the bone microenvironment.The results provided basic research data for the application of MWCNTs in bone tissue repair.