Treatment of Streptococcus mutans biofilms with a nonthermal atmospheric plasma

AIMS: A nonthermal atmospheric plasma, designed for biomedical applications, was tested for its antimicrobial activity against biofilm cultures of a key cariogenic bacterium Streptococcus mutans. METHODS AND RESULTS: The Strep. mutans biofilms were grown with and without 0.15% sucrose. A chlorhexidine digluconate rinse (0.2%) was used as a positive antimicrobial reference. The presence of sucrose and the frequency of plasma application during growth were shown to have a significant effect on the response to treatment and antibacterial activity. CONCLUSIONS: A single plasma treatment for 1 min on biofilms cultured without sucrose caused no re-growth within the observation period. However, with either single or repeated plasma treatments of 1 min, on biofilms cultured with 0.15% sucrose, growth was only reduced. SIGNIFICANCE AND IMPACT OF THE STUDY: In summary, there may be a role for nonthermal plasma therapies in dental procedures. Sucrose and associated growth conditions may be a factor in the survival of oral biofilms after treatment.     

Dental plaque as a biofilm

Dental plaque is the diverse microbial community found on the tooth surface embedded in a matrix of polymers of bacterial and salivary origin. Once a tooth surface is cleaned, a conditioning film of proteins and glycoproteins is adsorbed rapidly to the tooth surface. Plaque formation involves the interaction between early bacterial colonisers and this film (the acquired enamel pellicle). To facilitate colonisation of the tooth surface, some receptors on salivary molecules are only exposed to bacteria once the molecule is adsorbed to a surface. Subsequently, secondary colonisers adhere to the already attached early colonisers (co-aggregation) through specific molecular interactions. These can involve protein-protein or carbohydrate-protein (lectin) interactions, and this process contributes to determining the pattern of bacterial succession. As the biofilm develops, gradients in biologically significant factors develop, and these permit the co-existence of species that would be incompatible with each other in a homogeneous environment. Dental plaque develops naturally, but it is also associated with two of the most prevalent diseases affecting industrialised societies (caries and periodontal diseases). Future strategies to control dental plaque will be targeted to interfering with the formation, structure and pattern of development of this biofilm.     

Destruction of oral biofilms formed in situ on machined titanium (Ti) surfaces by cold atmospheric plasma

The decontamination of implant surfaces represents the basic procedure in the management of peri-implant diseases, but it is still a challenge. The study aimed to evaluate the degradation of oral biofilms grown in situ on machined titanium (Ti) discs by cold atmospheric plasma (CAP). ～ 200 Ti discs were exposed to the oral cavities of five healthy human volunteers for 72?h. The resulting biofilms were divided randomly between the following treatments: CAP (which varied in mean power, treatment duration, and/or the gas mixture), and untreated and treated controls (diode laser, air-abrasion, chlorhexidine). The viability, quantity, and morphology of the biofilms were determined by live/dead staining, inoculation onto blood agar, quantification of the total protein content, and scanning electron microscopy. Exposure to CAP significantly reduced the viability and quantity of biofilms compared with the positive control treatments. The efficacy of treatment with CAP correlated with the treatment duration and plasma power. No single method achieved complete biofilm removal; however, CAP may provide an effective support to established decontamination techniques for treatment of peri-implant diseases.     

等离子体医学及其在肿瘤治疗中的应用

大气压冷等离子体是近年来学术界兴起的新研究领域,由于其在大气压下产生,气体温度低、粒子活性高,在众多领域尤其是生物医学方面的应用引起了人们广泛的关注.等离子体医学是一个革新的、新兴的交叉学科研究领域,结合了等离子体物理学、化学、生命科学和临床医学等.本文首先介绍了大气压冷等离子体的产生及其粒子成分,与液体和生物组织的相互作用,并介绍了大气压冷等离子体在生物医学领域的一些主要应用,如杀菌消毒、凝血、牙科应用、伤口愈合及皮肤病治疗等方面.同时,重点介绍大气压冷等离子体在肿瘤治疗方面的研究进展.大气压冷等离子体可有效诱导肿瘤细胞死亡、抑制增殖及迁徙、诱导肿瘤细胞分化并抑制干细胞潜能,同时能提高化疗药物敏感性,在肿瘤治疗领域具有很好的应用前景.     

Dextranase from Arthrobacter oxydans KQ11-1 inhibits biofilm formation by polysaccharide hydrolysis

Dental plaque is a biofilm of water-soluble and water-insoluble polysaccharides, produced primarily by Streptococcus mutans. Dextranase can inhibit biofilm formation. Here, a dextranase gene from the marine microorganism Arthrobacter oxydans KQ11-1 is described, and cloned and expressed using E. coli DH5α competent cells. The recombinant enzyme was then purified and its properties were characterized. The optimal temperature and pH were determined to be 60°C and 6.5, respectively. High-performance liquid chromatography data show that the final hydrolysis products were glucose, maltose, maltotriose, and maltotetraose. Thus, dextranase can inhibit the adhesive ability of S. mutans. The minimum biofilm inhibition and reduction concentrations (MBIC₅₀ and MBRC₅₀) of dextranase were 2 U ml^?1 and 5 U ml^?1, respectively. Scanning electron microscopy and confocal laser scanning microscope (CLSM) observations confirmed that dextranase inhibited biofilm formation and removed previously formed biofilms.     

Novel mutation breeding method for Streptomyces avermitilis using an atmospheric pressure glow discharge plasma

Aims: Avermectins are major antiparasitic agents used commercially in animal health, agriculture and human infections. To improve the fermentation efficiency of avermectins, for the first time a plasma jet generated by a novel atmospheric pressure glow discharge (APGD) was employed to generate mutations in Streptomyces avermitilis. Methods and Results: The APGD plasma jet, driven by a radio frequency (RF) power supply with water‐cooled and bare‐metallic electrodes, was used as a new mutation method to treat the spores of S. avermitilis. The plasma jet yielded high total (over 30%) and positive (about 21%) mutation rates on S. avermitilis, and a mutated strain, designated as G1‐1 with high productivity of avermectin B1a and genetic stability, was obtained. Conclusions: Because of the low jet temperature, the high concentrations of the chemically reactive species and the flexibility of its operation, the RF APGD plasma jet has a strong mutagenic effect on S. avermitilis. Significance and Impact of the Study: This is a proof‐of‐concept study for the use of an RF APGD plasma jet for inducing mutations in microbes. We have shown that the RF APGD plasma jet could be developed as a promising and convenient mutation tool for the fermentation industry and for use in biotechnology research.     

L-异亮氨酸高产菌选育及其培养基优化

旨在诱变选育L-异亮氨酸高产菌,并探索突变株最佳发酵条件。利用传统化学诱变结合常压室温等离子体生物诱变体系对实验室保藏的Brevibacterium flavum I-12进行逐级诱变,选育2-噻唑丙氨酸(2-TA)和磺胺胍(SG)高抗性和在琥珀酸平板上能快速生长的突变菌株。随后,在单因素实验的基础上,利用响应面设计优化出目的突变株摇瓶发酵培养基组分的最佳参数水平。结果显示,经过一系列诱变和筛选,成功选育出一株在40 g/L的2-TA和5 g/L的SG,且以琥珀酸为唯一碳源的培养基上快速生长突变株,命名为B. flavum TA-6,该菌株产酸达26.2±0.5 g/L,比出发菌株提高了44.75%,而副产物L-缬氨酸和L-亮氨酸积累量明显降低。经响应面法优化发酵条件后,突变株产酸可达27.8±0.5 g/L,比优化前提高了6.1%。通过传统化学诱变结合ARTP生物诱变体系,成功选育出一株杂酸降低的L-异亮氨酸高产菌TA-6,该菌株具有潜在生产应用价值。     

低龄婴幼儿龋相关致龋微生物研究进展

低龄婴幼儿龋（Earlychildhoodcaries,ECC）发生、发展是由多种因素共同作用,多种微生物共同参与的复杂过程。因此、了解儿童口腔内致龋微生物的种类及其与龋病的相关关系,对预防和治疗儿童龋病有着重要意义。本研究就ECC相关致龋菌及菌斑生物膜研究进展作一综述。     

Lipoprotein(a): its inheritance and molecular basis of its atherothrombotic role

Lipoprotein(a) or Lp(a), is a member of the plasma lipoproteins with general properties of LDL but with a protein moiety represented by apoB100 disulfide linked to apolipoprotein(a) or apo(a). Apo(a) is polymorphic in size; at present a total of 11 isoforms have been reported, but more are likely to be identified in view of the fact that at least 19 alleles of the apo(a) gene have recently been reported. There are remarkable variations in the plasma Lp(a) levels; but uncertainties still exist about the factors responsible for this variability. High plasma Lp(a) levels have been associated with an increased incidence of cardiovascular disease, mainly based on epidemiological evidence. Both atherogenic and thrombogenic potentials have been suggested; the first attributable to the LDL-like properties of Lp(a) and the other to the plasminogen-like characteristics of apo(a). From the mechanistic viewpoint in vitro studies suggest that the thrombogenic action may occur at the level of the endothelium whereas Lp(a) that localizes in the sub-endothelial intima is expected to undergo complexation with matrix components and favor the formation of the atherosclerotique plaque. How Lp(a) polymorphism relates to the postulated cardiovascular pathogenicity of this lipoprotein remains to be established.     

Activity of Nidus Vespae extract and chemical fractions against Streptococcus mutans biofilms

AIMS: To evaluate the effect of Nidus Vespae extract and chemical fractions on the viability and architecture of Streptococcus mutans biofilms. METHODS AND RESULTS: The raw material was first extracted using 95% ethanol/water. Subsequent fractions were prepared from this extract using cyclohexane/ethyl acetate, petroleum ether/ethyl acetate and chloroform/methanol. The biomass dry weight and total protein of samples treated with Nidus Vespae extract and chemical fractions were significantly less than those treated with the vehicle control (P < 0.05). Biofilms treated with Nidus Vespae also resulted in lower percentage of polysaccharide composition. The pH decrease in the biofilm matrix was retarded by Nidus Vespae compared with the vehicle control. Architecture of biofilms treated with Nidus Vespae was different than in the vehicle control and 0.05% chlorhexidine. CONCLUSIONS: Chloroform/methanol fraction was the most effective treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The significant antibiofilm activity demonstrated by Nidus Vespae shows it to be a promising source of novel anticariogenic agents.     

益生菌辅助治疗牙周病的研究进展

牙周病是累及牙周支持组织的慢性感染性疾病。牙菌斑生物膜中的微生物及其代谢产物是其必不可少的始动因素,可导致口腔微生态失衡和宿主免疫反应,最终引起牙周病的发生发展。目前,牙周病的基础治疗主要是机械清除牙菌斑生物膜和牙石,但治疗效果具有局限性。益生菌通过产生抑菌物质、刺激局部免疫反应、与致病菌争夺黏膜受体和营养物质,从而改善口腔微生态平衡,促进牙周病的治疗。本文就近年来益生菌在牙周病治疗上的实验和临床研究、作用机制、安全性等进行综述,为将来益生菌辅助治疗牙周病的应用提供参考。     

牙菌斑生物膜的形成与控制

牙菌斑生物膜是附着于牙釉质表面,由复杂的微生物群落构成的一种聚集体。牙菌斑生物膜的形成与生长对口腔健康有着直接或间接的影响,许多研究证实口腔疾病如龋齿和牙周病都与细菌的积累及牙菌斑的形成有关。在牙菌斑生物膜形态建成过程中,牙齿表面最初的定殖菌对生物膜的微生物组成和结构至关重要,这些初级定殖菌决定了后续与之结合形成共生体的微生物种类和数量。不同的微生物组成可能在与生物膜形成相关的口腔病理状况中发挥不同的作用。因此,本文就牙菌斑生物膜的生长及控制进行综述,介绍其微生物的早期定殖和成熟过程、以及通过物理和化学方法对牙菌斑生物膜的控制,以期为了解牙菌斑生物膜的形成机制及相关口腔疾病的预防和治疗提供有价值的参考。     

Environmental and centrifugal factors influencing the visco-elastic properties of oral biofilms in vitro

Centrifugal compaction causes changes in the surface properties of bacterial cells. It has been shown previously that the surface properties of planktonic cells change with increasing centrifugal compaction. This study aimed to analyze the influences of centrifugal compaction and environmental conditions on the visco-elastic properties of oral biofilms. Biofilms were grown out of a layer of initially adhering streptococci, actinomyces or a combination of these. Different uni-axial deformations were induced on the biofilms and the load relaxations were measured over time. Linear-Regression-Analysis demonstrated that both the centrifugation coefficient for streptococci and induced deformation influenced the percentage relaxation. Centrifugal compaction significantly influenced relaxation only upon compression of the outermost 20% of the biofilm (p < 0.05), whereas biofilm composition became influential when 50% deformation was induced, invoking re-arrangement of the bacteria in deeper biofilm structures. In summary, the effects of centrifugal compaction of initially adhering, centrifuged bacteria extend to the visco-elastic properties of biofilms, indicating that the initial bacterial layer influences the structure of the entire biofilm.