Macrophages promote the transition from myocardial ischemia reperfusion injury to cardiac fibrosis in mice through GMCSF/CCL2/CCR2 and phenotype switching.

Following acute myocardial ischemia reperfusion (MIR), macrophages infiltrate damaged cardiac tissue and alter their polarization phenotype to respond to acute inflammation and chronic fibrotic remodeling. In this study we investigated the role of macrophages in post-ischemic myocardial fibrosis and explored therapeutic targets for myocardial fibrosis. Male mice were subjected to ligation of the left coronary artery for 30 min. We first detected the levels of chemokines in heart tissue that recruited immune cells infiltrating into the heart, and found that granulocyte-macrophage colony-stimulating factor (GMCSF) released by mouse cardiac microvascular endothelial cells (MCMECs) peaked at 6 h after reperfusion, and c-c motif chemokine ligand 2 (CCL2) released by GMCSF-induced macrophages peaked at 24 h after reperfusion. In co-culture of BMDMs with MCMECs, we demonstrated that GMCSF derived from MCMECs stimulated the release of CCL2 by BMDMs and effectively promoted the migration of BMDMs. We also confirmed that GMCSF promoted M1 polarization of macrophages in vitro, while GMCSF neutralizing antibodies (NTABs) blocked CCL2/CCR2 signaling. In MIR mouse heart, we showed that GMCSF activated CCL2/CCR2 signaling to promote NLRP3/caspase-1/IL-1ß-mediated and amplified inflammatory damage. Knockdown of CC chemokine receptor 2 gene (CCR2-/-), or administration of specific CCR2 inhibitor RS102895 (5 mg/kg per 12 h, i.p., one day before MIR and continuously until the end of the experiment) effectively reduced the area of myocardial infarction, and down-regulated inflammatory mediators and NLRP3/Caspase-1/IL-1ß signaling. Mass cytometry confirmed that M2 macrophages played an important role during fibrosis, while macrophage-depleted mice exhibited significantly reduced transforming growth factor-ß (Tgf-ß) levels in heart tissue after MIR. In co-culture of macrophages with fibroblasts, treatment with recombinant mouse CCL2 stimulated macrophages to release a large amount of Tgf-ß, and promoted the release of Col1α1 by fibroblasts. This effect was diminished in BMDMs from CCR2-/- mice. After knocking out or inhibiting CCR2-gene, the levels of Tgf-ß were significantly reduced, as was the level of myocardial fibrosis, and cardiac function was protected. This study confirms that the acute injury to chronic fibrosis transition after MIR in mice is mediated by GMCSF/CCL2/CCR2 signaling in macrophages through NLRP3 inflammatory cascade and the phenotype switching.

SVF-GEL Cryopreserved for Different Times Exhibits Varied Preservation and Regeneration Potential After Transplantation in a Mouse Model.

BACKGROUND: Matrix vascular component (SVF) gels derived from fat preserve tissue integrity and cell viability under cryopreserved conditions, making them easy to inject again for later use. Here, we compared the preservation power and regeneration potential of SVF-gel under different cryopreservation times. METHODS: The SVF-gel stored under - 20 °C, without cryoprotectant cryopreservation for 5, 15, and 45 days, with fresh SVF-gel as control. We evaluated the rate of volume retention after thawing the SVF-gel and the apoptosis rate of adipose-derived stem cells. Next, we analyzed retention rated, adipogenesis, angiogenesis, and connective tissue hyperplasia of the grafts, one month after subcutaneously transplanting the specimen into immunodeficient mice. RESULTS: SVF-gel cryopreserved for 5 and 15 days exhibited no significant different in apoptosis rates relative to the control group. Extending the cryopreservation time to 45 days resulted in significantly increased and decreased apoptosis and volume retention rates of SVF-gel, respectively. SVF-gel grafts cryopreserved for 5 and 15 days exhibited no significant differences from those in the control group, although their weights and volumes still fluctuated. Extending the cryopreservation time to 45 days resulted in significantly decreased retention rates of the grafts. Histologically, extending freezing time resulted in a gradual decline in the graft's health adipose tissue, as well as decreased angiogenesis, and connective tissue hyperplasia. CONCLUSION: Simple freezing of SVF-gel at - 20 °C conferred them with sufficient cell viability. Notably, short-term cryopreservation did not significantly increase the apoptosis rate, and it still had a certain regeneration after transplantation. However, prolonging freezing time to 45 days resulted in increased apoptosis rate and worsened transplantation effect. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Treatment with injectable platelet-rich fibrin in a rat model of melasma.

The management of melasma is challenging. Platelet-rich plasma (PRP) therapy has been shown to be beneficial, however, the use of anticoagulants for PRP is dangerous. To evaluate the efficacy of recently developed blood-derived biomaterials (injectable platelet-rich fibrin [I-PRF]) in a rat model of melasma. Sprague Dawley (SD) rats were used to replicate an experimental animal model of melasma. SD rats exhibiting melasma were randomly divided into experimental and control groups. The experimental group was administered weekly intradermal injections of I-PRF, whereas the control group received an equivalent amount of saline. After four weeks, back skin was removed and evaluated based on (1) gross observation, (2) pathological examination and imaging analysis, and (3) biochemical detection. Data were analysed using SAS9.4 software. I-PRF, a safe blood product without anticoagulants, inhibited melanin production in the epidermis and reduced oxidative stress damage in the cortex, improving melasma. I-PRF is a safe and cost-effective blood-derived biomaterial which is useful for the treatment of melasma.

Application of Modified Triangular Perforator Flap in Repair of Small and Medium-Sized Facial Defect.

Background: We investigated the application of local perforator flap or island flap with a modified triangular to repair small and medium facial defects. Methods: (1) Before the operation, a Doppler flowmeter was used to investigate the superficial exit point of the perforator artery. The length to breadth ratio of the flap was more than 3-4 times, and it contained 1-2 perforator vessels. (2) The lesion was excised, and the skin was cut along the design line of the flap. The flap was separated and trimmed based on the defect degree. (3) The blood supply was confirmed, and the defect was then covered with the flap by "rotation and advancement" approach without causing tension. The incision was finally sutured in layers. (4) Postoperative routine care was performed according to the situation. Results: The functional morphology and appearance of all 23 cases of skin flaps successfully recovered during follow-up. There was no major aesthetic and malformation recorded. Conclusion: In summary, the modified triangular perforator flap can improve the functional appearance at the repaired small and medium facial defects.

The Modified Suction-curettage Technique for Minimally Invasive Treatment of Axillary Osmidrosis.

BACKGROUND: Axillary osmidrosis (AO) is a common and nonnegligible disease, the treatment of which is currently lacking a consensus. AIMS: The aim of this study was to introduce a modified suction-assisted technique as a safer and more efficient surgical procedure. METHODS: This retrospective clinical study included 80 patients who recieved a modified suction-curettage procedure (group A) or a subcutaneous gland excision procedure (group B). Intraoperative assessment (endoscopy and pathological biopsy) and postoperative assessment (complications, therapeutic effect, and satisfaction) were performed for both groups. RESULTS: The endoscopy and pathological biopsy results demonstrated that the modified suction-curettage technique could remove the apocrine gland efficiently. Compared with group B, a lower complication rate (long-term, 5.00%; P=0.014, and short-term, 11.10%; P=0.001) and higher patient satisfaction (98.00%, P=0.012) were observed in group A. CONCLUSION: The modified suction-curettage procedure is an effective and safe treatment for axillary osmidrosis.

MicroRNA-145 inhibits hepatic stellate cell activation and proliferation by targeting ZEB2 through Wnt/ß-catenin pathway.

The activation of hepatic stellates cells (HSCs) is well believed to play a pivotal role in the development of liver fibrosis. MicroRNA-145 (miR-145) is known to suppress the progression of hepatocellular carcinoma, and is previously reported to be associated with Wnt/ß-catenin pathway, but its role in the progression of hepatic fibrosis and activation of HSCs remains unknown and is warranted for investigation. In the present study, we found that the expression of miR-145 is significantly down-regulated in vivo in CCl4-induced mice liver fibrosis as well as in transforming growth factor-ß1 (TGF-ß1) induced HSC-T6 cell lines and human hepatic stellate cell line LX-2 in vitro. Furthermore, over-expression of miR-145 inhibited TGF-ß1-induced the activation and proliferation of HSC-T6 cells in vitro. Mechanistically, we identified that zinc finger E-box-binding homeobox 2 (ZEB2), a key mediator of epithelial-to-mesenchymal transition, acted as a functional downstream target for miR-145. Interestingly, ZEB2 was shown to be involved in the TGF-ß1-induced HSCs activation by regulating Wnt/ß-catenin signaling pathway. Taken together, our results revealed the critical regulatory role of miR-145 in HSCs activation and implied miR-145 as a potential candidate for therapy of hepatic fibrosis by regulation of Wnt/ß-catenin through targeting ZEB2.