铜代谢与口腔疾病的诊断及治疗

背景:铜是人体的必需微量元素,在机体各项生理活动中起着重要作用,其代谢异常与多种疾病密切相关。铜代谢主要涉及铜离子的吸收、转运、储存和排泄等过程,这些过程共同调节铜稳态。近年来,多项研究证实铜稳态失调会严重影响人体的代谢活动,引起各个系统的疾病,而其中铜在口腔疾病中的作用也一直备受关注。目的:探讨铜在口腔疾病发生发展和治疗中的作用,并对这一领域的研究进展进行综合概述。方法:由第一作者应用计算机在PubMed、Web of Science、中国知网检索铜在口腔疾病中的相关研究,以“Cu,Copper,Copper metabolism,Oraldiseases,Oral squamous cell carcinoma,Periodontitis,Oral submucous fibrosis,Oral lichen planus,Recurrent oral ulceration,Pulpitis”等为英文检索词,以“铜,铜代谢,口腔疾病,口腔鳞状细胞癌,牙周炎,口腔黏膜纤维下变性,口腔扁平苔藓,口腔溃疡,牙髓炎”等为中文检索词,经过筛选后纳入78篇文献进行综述分析。结果与结论:(1)口腔鳞状细胞癌患者血清及唾液内的铜浓度升高,升高的铜通过氧化应激和促进血管生成,促进癌症进展。而通过靶向过度升高或降低肿瘤细胞内铜浓度均可抑制肿瘤细胞生长,铜与抗癌药物联合应用,能显著提高药物疗效。(2)牙周炎患者血清中铜浓度升高,铜过量可促进氧化应激,加重牙周炎。而铜与药物制剂联合作用,可促进牙周骨再生和牙周组织愈合。(3)体内铜水平与口腔黏膜纤维化程度呈正相关关系,升高的铜进入口腔黏膜内,上调赖氨酰氧化酶的活性,从而增加胶原蛋白的产生,促进口腔黏膜纤维化。(4)口腔扁平苔藓患者体内铜含量升高,升高的铜可能通过调控免疫细胞功能促进扁平苔藓的进展。(5)复发性口腔溃疡患者血清铜水平显著增高,铜利用发生障碍,可导致含铜酶活性降低,从而影响溃疡的愈合。由此可见:铜与多种口腔疾病密切相关,针对铜的相关治疗可显著提高药物疗效,但还需进一步研究其作用机制,为口腔疾病提供更完善的治疗策略奠定基础。

Pre-assembled nanospheres in mucoadhesive microneedle patch for sustained release of triamcinolone in the treatment of oral submucous fibrosis

Objective:Drug-loaded mucoadhesive silk fibroin(SF)microneedle patch can overcome the limitations of low bioavailability and significant pain associated with traditional treatment methods,such as topical application or injection of triamcinolone for oral submucous fibrosis(OSF).However,these systems release the drug too quickly,failing to meet the clinical requirements.This study aims to construct a mucoadhesive SF microneedle patch pre-assembled with silk fibroin nanospheres(SFN)and explore its ability to sustain the release of triamcinolone in the treatment of OSF.Methods:SFN was pre-assembled via precipitation reaction and characterized by scanning electron microscope(SEM)for the morphology.The particle size andζ-potential were measured by dynamic light scattering(DLS).Triamcinolone was loaded onto SFN using a diffusional post-loading method.The effective loading of triamcinolone was confirmed using Fourier-transform infrared spectroscopy(FTIR).The concentration of unloaded triamcinolone was quantified by high-performance liquid chromatography.Drug encapsulation efficiency and loading capacity of SFN were then calculated to determine the optimal amount of drug loading.The SFN suspension was pre-mixed with SF solution to prepare the microneedle under-layer.The microneedle morphology was observed by SEM.Compression mechanical tests were performed to evaluate the fracture force of microneedles at different nanosphere contents(5%,10%,and 20%),determining the optimal pre-mixing ratio.Ex-vivo mouse oral mucosa permeation studies were performed to ascertain the insertion depth of the microneedles via histological sections.The adhesive top layer was synthesized using SF and tannic acid,with FTIR confirming its successful synthesis.Its viscoelasticity was characterized by a rheometer,and differential scanning calorimetry analyzed thermal properties.Tensile tests evaluated the interfacial bonding strength between the adhesive layer and microneedle base to ensure no detachment during use.Adhesion to wet oral mucosal tissues was tested and compared to commercial oral patches.Under the optimized conditions,the double-layered mucoadhesive microneedle patch with pre-assembled nanospheres was prepared.Its cell compatibility was evaluated by cell counting kit-8(CCK-8),live/dead staining,and phalloidin staining after co culturing with fibroblasts.The drug release experiment was conducted to demonstrate its sustained release efficacy.Results:SFN(mean diameter 46.25 nm)was successfully prepared.The maximum drug encapsulation efficiency was(63.88±1.09)%(corresponding loading capacity of SFN was(27.41±3.06)%when the weight ratio of triamcinolone/SFN was 0.5.The corporation of SFN did not affect microneedle morphology.The mechanical properties of microneedles decreased with increasing nanosphere amount.Only the fracture force of the group with 5%SFN[(0.07±0.01)N/needle]exceeded the minimum force required for mucosal penetration,thus selected as the optimal pre-mixing ratio.Histological sections confirmed that the SFN microneedles could penetrate the epithelial layer and deliver drugs to OSF affected areas.Adhesion strength between the microneedle base and top layer was(94.8±6.89)kPa,confirming strong bonding with no detachment during use.The wet adhesive strength of the double-layered mucoadhesive microneedle patch[(41.28±7.43)kPa]was significantly enhanced compared to commercial oral patches(4.5 kPa,P<0.01).CCK-8 and live/dead staining results confirmed no significant cytotoxicity.Drug release experiment showed the double-layered mucoadhesive microneedle patch with pre-assembled SFN enabled sustained release time of triamcinolone from 4 days to 14 days.Conclusion:Pre-assembling nanospheres in mucoadhesive SF microneedle patches can extend triamcinolone release time,meeting clinical requirements for sustained drug delivery.