The development of bone regenerative substitutes capable of orchestrating osteogenesis within inflammatory immune microenvironments remains a critical challenge. This study investigates the dual-functional immunomodulatory peptide DP7-C as a microRNA (miRNA) co-delivery system to regulate osteogenic differentiation and macrophage polarization synchronously. Through systematic screening of DP7-C/miRNA nanocomplexes (miR-21, -26a, -29a, -34a, -124, -125a) in bone marrow mesenchymal stem cells (BMSCs) and RAW264.7 macrophages, we identified DP7-C/miR-124 as the optimal nanocomplex, demonstrating synergistic osteoimmunomodulatory effects. Results demonstrated that the DP7-C/miR-124 combination raised the expression of anti-inflammatory factors in inflammatory macrophages and decreased the expression of pro-inflammatory factors. It also stimulated the production of osteogenesis-related proteins BMP2 and Runx2 to promote BMSC osteogenesis. Mechanistic studies revealed bidirectional cellular crosstalk, where DP7-C/miR-124 enhanced IL-10-mediated anti-inflammatory macrophage polarization while reciprocally promoting BMSC differentiation through paracrine modulation. These findings establish DP7-C/miRNA nanocomplexes as next-generation osteoimmunomodulatory biomaterials that concurrently resolve inflammation and amplify bone regeneration through epigenetic-immune circuit regulation, offering a promising strategy for functionalized bone defect repair in inflammatory microenvironments.

BACKGROUND:Oral mucositis is the most common and troublesome complication for cancer patients receiving radiotherapy or chemotherapy. Recent research has shown that Lycium barbarum, an important economic crop widely grown in China, has epithelial protective effects in several other organs. However, it is unknown whether or not Lycium barbarum can exert a beneficial effect on oral mucositis. Network pharmacology has been suggested to be applied in "multi-component-multi-target" functional food studies. The purpose of this study is to evaluate the effect of Lycium barbarum on oral mucositis through network pharmacology, molecular docking and experimental validation. AIMS:To explore the biological effects and molecular mechanisms of Lycium barbarum in the treatment of oral mucositis through network pharmacology and molecular docking combined with experimental validation. METHODS:Based on network pharmacology methods, we collected the active components and related targets of Lycium barbarum from public databases, as well as the targets related to oral mucositis. We mapped protein- protein interaction (PPI) networks, performed gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment, and constructed a 'components-disease-targets' network and 'components-pathways-targets' network using Cytoscape to further analyse the intrinsic molecular mechanisms of Lycium barbarum against oral mucositis. The affinity and stability predictions were performed using molecular docking strategies, and experiments were conducted to demonstrate the biological effects and possible mechanisms of Lycium barbarum against oral mucositis. RESULTS:A network was established between 49 components and 61 OM targets. The main active compounds were quercetin, beta-carotene, palmatine, and cyanin. The predicted core targets were IL-6, RELA, TP53, TNF, IL10, CTNNB1, AKT1, CDKN1A, HIF1A and MYC. The enrichment analysis predicted that the therapeutic effect was mainly through the regulation of inflammation, apoptosis, and hypoxia response with the involvement of TNF and HIF pathways. Molecular docking results showed that key components bind well to the core targets. In both chemically and radiation-induced OM models, Lycium barbarum significantly promoted healing and reduced inflammation. The experimental verification showed Lycium barbarum targeted the key genes (IL-6, RELA, TP53, TNF, IL10, CTNNB1, AKT1, CDKN1A, HIF1A, and MYC) through regulating the HIF and TNF signaling pathways, which were validated using the RT-qPCR, immunofluorescence staining and western blotting assays. CONCLUSION:In conclusion, the present study systematically demonstrated the possible therapeutic effects and mechanisms of Lycium barbarum on oral mucositis through network pharmacology analysis and experimental validation. The results showed that Lycium barbarum could promote healing and reduce the inflammatory response through TNF and HIF signaling pathways.

目的 探讨枸杞糖肽(Lycium barbanun glycopeptide,LbGP)对放射诱导人角质形成细胞HaCaT损伤的保护作用及其机制.方法 采用直线加速器(速率6 Gy/min)分别按4、8、12、16、20、24、28 Gy剂量对HaCaT细胞进行照射,CCK-8法检测细胞活性.以0、0.05、0.1、0.5、0.8、1.0、1.5、3mg/mL的 LbGP作用 HaCaT细胞4h,进行 12 Gy剂量放射,CCK-8法检测细胞活性.将HaCaT细胞分为空白对照组(不加LbGP不放射)、放射组(12 Gy剂量照射)、LbGP+放射组(0.8 mg/mLLbGP作用24 h,12 Gy剂量照射),照射1 h后,流式细胞术检测活性氧(reactive oxygen species,ROS)产生量,WST-8法检测超氧化物歧化酶(superoxide dismutase,SOD)活性,Western blot法检测细胞中核因子E2相关因子2(nuclear factor-E2 related factor 2,Nrf2)、p-Nrf2、NADPH 氧化还原酶 1(NADPH quinone oxidoreductase 1,NQO1)、血红素加氧酶-1(heme oxygenase-1,HO-1)蛋白表达水平;分别于放射1、3、5h后,qRT-PCR法检测Nrf2、HO-1、NQO1基因mRNA的转录水平.结果 12 Gy放射辐射量可诱导约50％细胞死亡,0.8 mg/mL LbGP对放射细胞活性保护作用最佳.放射1h后,与空白对照组比较,放射组HaCaT细胞内ROS含量显著增加(F=2.55,P＜0.001),SOD活力显著降低(F=1.23,P＜0.01),NQO1、Nrf2蛋白含量差异无统计学意义(F分别为1.78和1.00,P均＞0.05),HO-1蛋白含量明显增加(F=1.37,P＜0.05),p-Nrf2蛋白含量显著下降(F=2.75,P＜0.01);与放射组比较,LbGP+放射组HaCaT细胞内 ROS含量明显降低(F=3.61,P＜0.001),SOD活性明显升高(F=1.23,P＜0.05),Nrf2,p-Nrf2、HO-1 及NQO1蛋白含量均显著上升(F分别为4.00、2.25、6.25及1.27,P均＜0.05).放射1、3、5 h,与放射组比较,LbGP+放射组Nrf2、HO-I、NQO1基因mRNA转录水平均明显升高(F=0.20～36.00,P均＜0.05).结论 LbGP可减轻放射对HaCaT细胞的氧化应激损伤,保护细胞活性,该作用可能是通过激活Nrf2及提高其下游抗氧化酶SOD、HO-1、NQO1水平而发挥作用的.

Radiation-induced oral mucositis (RIOM) is considered to be the most common acute side effect of radiation therapy and occurs during intentional or accidental radiation exposure. Antioxidant synthesis agents have been reported to protect against or alleviate the development of mucositis, but the resulting side effects of chemical synthesis agents limit their use in clinical practice. Lycium barbarum polysaccharide-glycoprotein (LBP), a polysaccharide extract of the Lycium barbarum fruit, has superior antioxidant capacity and biosafety and is a potential option for radiation prevention and treatment. Here, we aimed to investigate whether LBP conferred radioprotection against ionizing radiation-induced oral mucosal damage. We found that LBP exerted radioprotective effects in irradiated HaCaT cells, improving cell viability, stabilizing mitochondrial membrane potential, and decreasing cell death. LBP pretreatment reduced oxidative stress and ferroptosis in radioactivity-damaged cells by activating the transcription factor Nrf2 and promoting its downstream targets, such as HO-1, NQO1, SLC7A11, and FTH1. Knockdown of Nrf2 eliminated the protective effects of LBP, implying the essential role of Nrf2 in LBP activity. Additionally, the topical application of LBP thermosensitive hydrogel on rat mucosa resulted in a significant decrease in ulcer size in the irradiated group, suggesting that LBP oral mucoadhesive gel may be a potential tool for the treatment of irradiation. In conclusion, we demonstrated that LBP attenuates ionizing radiation-induced oral mucosa injury by reducing oxidative stress and inhibiting ferroptosis via the Nrf2 signaling pathway. LBP may be a promising medical countermeasure against RIOM.

背景:前期研究显示,微渠多孔羟基磷灰石具有良好骨诱导性、生物相容性、骨传导性,为临床修复颌骨缺损扩大种植适应证提供了新思路.目的:观察微渠多孔羟基磷灰石修复犬下颌骨大面积缺损后与牙种植体骨结合的情况.方法:制作微渠多孔羟基磷灰石支架.将8只比格犬随机分为实验组和对照组,每组4只,拔除犬双侧下颌前磨牙,拔牙3个月后,在双侧前磨牙处各制造2个近远端为8 mm、颊舌侧为10 mm、冠根方向为8 mm的圆柱型骨缺损,实验组植入微渠多孔羟基磷灰石支架,对照组植入自体骨;3个月后于骨缺损修复区植入直径4 mm、高度10 mm的Dentium种植体.种植体植入4,12周后取材,进行锥形束CT、Micro-CT、组织学与种植体共振频率分析.结果 与结论:①锥形束CT:植入4周后,两组种植体与周围骨界面处有部分低密度影;12周后,两组种植体与骨结合的面积增加,种植体与植入材料结合紧密,无明显暗影;②Micro-CT检测:实验组植入4周后的植体表面被覆骨量小于对照组(P<0.05),植入12周后的植体表面被覆骨量比较差异无显著性意义(P>0.05);③组织学形态:苏木精-伊红、Masson与亚甲基蓝-酸性品红染色显示,两组植入4周后术区均可见新骨形成,实验组种植体-骨结合率低于对照组(P<0.05);12周后两组植体周围骨量明显增加,两组种植体-骨结合率比较差异无显著性意义(P>0.05);④种植体共振频率分析:实验组植入4,12周后的种植体稳定系数均低于对照组(P<0.05),但两组种植体在各时间段均有良好的稳定性;⑤结果表明:微渠多孔羟基磷灰石支架能修复犬颌骨大面积缺损,诱导新生骨形成,与种植体形成良好的骨结合.

"骨免疫学"的提出为修复骨缺损提供了新的方向.通过加载生物因子赋予生物材料骨免疫调节能力是常用的方法之一.microRNAs作为内源性的非编码小RNA,调节人体至少30％的蛋白基因编码,并且具有低免疫原性,作为改性生物材料的生物因子具有巨大潜力.但鲜有调节骨免疫微环境的microRNAs报道.本综述阐述了生物材料植入体内后的宿主反应、巨噬细胞在骨愈合中的作用、miR-21、miR-26a、miR-29等靶向调节成骨细胞及免疫细胞的microRNAs.试图探寻一种新的生物材料改性策略,将具备骨免疫微环境调节能力的microRNA作为生物活性因子加载到生物材料上,未来应用这一策略使生物材料植入体内后靶向调节骨免疫微环境,获得理想的成骨效果.

AIMS:Accumulating evidence suggests that Porphyromonas gingivalis is closely associated with the development of various chronic inflammatory diseases, particularly periodontitis. This study investigated the antibacterial activity and action mechanism of a novel antimicrobial peptide (AMP), DP7, against P. gingivalis.METHODS AND RESULTS:The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) for DP7 were determined via a broth microdilution method, revealing an MIC of 8 μg ml-1 and MBC of 32 μg ml-1 . Growth inhibition and killing assays confirmed the bactericidal effect of DP7, and treatment with DP7 at MBC eliminated P. gingivalis within 8 h. DP7 had a low cytotoxic effect against human cells. Transmission electron microscopy revealed that DP7 destroyed the bacterial membrane, and confocal laser scanning microscopy revealed its inhibitory effect on P. gingivalis biofilms. Quantitative reverse transcription-polymerase chain reaction revealed DP7-mediated inhibition of several virulence factor genes, partially explaining its antibacterial mechanism.CONCLUSIONS:DP7, a novel AMP with low mammalian cytotoxicity, inhibits both planktonic and biofilm forms of P. gingivalis by destroying the bacterial membrane and reducing virulence factor gene expression.SIGNIFICANCE AND IMPACT OF THE STUDY:DP7 has potential clinical application in the prevention and treatment of P. gingivalis-associated diseases.

目的 探讨儿童龋齿发病趋势、相关风险因素以及预防对策.方法 采用统一制定的调查方案对南充市107名儿童进行追踪调查,采用问访的方式对儿童的饮食习惯和口腔保健行为进行调查.结果 2013年107名儿童患龋人数为36(33.6％),龋均0.570.2014年儿童患龋人数58(54.2％),龋均1.215.2015年患龋70(65.4％)人,龋均1.831.2013年儿童患龋前牙所占比例较高,后期所占龋齿比例下降.结论 南充市学龄前儿童患龋率较高,口腔保健行为较差,应该加强相关教育,提高儿童和家长的口腔保健意识.

目的 了解成都市大邑县居民对古镇文化保护的情况,为古镇保护提出可参考意见方法 自制问卷,随机抽取356名居民进行问卷调查结果 90.2%的居民对古镇文化有一定的保护意识,但仅41.0%的居民有较强的主动保护行为结论 建议通过培养居民地方依恋,加强居民古镇文化保护意识,并同时加强古镇文化保护的宣传力度.