Streptomyces-derived actinomycin D inhibits biofilm formation via downregulating ica locus and decreasing production of PIA in Staphylococcus epidermidis.

AIM: The objective of this study was to investigate the biofilm inhibitory activity of Streptomyces-derived actinomycin D against biofilm formation by Staphylococcus epidermidis. METHODS AND RESULTS: The microtitre plate method and microscopy were used to detect the biofilm formation of S. epidermidis. And an attempt was made to detect the effect of actinomycin D on important biofilm components, exopolysaccharides (EPS) in S. epidermidis using precolumn derivation HPLC. Also cell surface hydrophobicities of S. epidermidis were assessed to explore action mechanisms. The qPCR was performed to demonstrate the genetic mechanisms of biofilm formation by S. epidermidis. Unlike other antibiotics, actinomycin D (1·5 µg ml-1 ) from Streptomyces luteus significantly inhibited biofilm formation by S. epidermidis. Additionally, it effectively inhibited S. epidermidis cells from adhering to glass slides. Actinomycin D downregulated ica locus and then the reduced polysaccharide intercellular adhesin production caused S. epidermidis cells to become less hydrophobic, thus supporting its anti-biofilm effect. CONCLUSION: Streptomyces-derived actinomycin D is active in inhibiting the biofilm formation of S. epidermidis. SIGNIFICANCE AND IMPACT OF THE STUDY: Actinomycin D can be used as a promising antibiofilm agent in inhibiting S. epidermidis biofilm formation. The study is also the first insight into how actinomycin D inhibited the biofilm formation of S. epidermidis. Actinomycin D could potentially be used to reduce the risk of biofilm-associated infections. Our study also suggests that the metabolites from Actinomycete strains keep further attention as potential antibiofilm agents against biofilm formation of S. epidermidis, even biofilm infections of the other bacteria.

In vitro study on the influence of strontium-doped calcium polyphosphate on the angiogenesis-related behaviors of HUVECs.

Angiogenesis is of great importance in bone tissue engineering, and has gained large attention in the past decade. Strontium-doped calcium polyphosphate (SCPP) is a novel biodegradable material which has been proved to be able to promote in vivo angiogenesis during bone regeneration. An in vitro culture system was developed in the present work to examine its influence on angiogenesis-related behaviors of human umbilical vein endothelial cells (HUVECs), including cell adhesion, spreading, proliferation and migration. The effects of microtopography, chemical property and the ingredients in the degradation fluid (DF) on cell behaviors were discussed. The results showed that cells attached and spread better on SCPP scaffold than on calcium polyphosphate (CPP), which might partially result from the less rough surface of SCPP scaffold and the less hydrogel formed on the surface. In addition, cell proliferation was significantly improved when treated with SCPP DF compared with the treatment with CPP DF. Statistical analysis indicated that Sr(2+) in SCPP DF might be the main reason for the improved cell proliferation. Moreover, cell migration, another important step during angiogenesis, was evidently stimulated by SCPP DF. The improved in vivo angiogenesis by SCPP might be assigned to its better surface properties and strontium in the DF. This work also provides a new method for in vitro evaluation of biodegradable materials' potential effects on angiogenesis.

Effect of polymerization degree of calcium polyphosphate on its microstructure and in vitro degradation performance.

Preparation, characterization and in vitro study of a series of calcium polyphosphate (CPP) with different polymerization degree were reported. A series of CPP with different polymerization degree were prepared by controlling calcining time. Average polymerization degree was analyzed by liquid state 31P nuclear magnetic resonance (NMR). The microstructure was observed by scanning electric microscope (SEM). X-ray diffraction (XRD) analysis was used to demonstrate that polymerization degree would not affect the crystal system and space group of CPP. The results showed that polymerization degree increased with the increase of calcining time. Degradation studies were performed during 32 days in physiological saline solution (aqueous solution, 0.9 wt.%NaCl) to assess the effect of polymerization degree on the degradation velocity of the samples. It was also shown that the degradation velocity of CPP (polymerization degree=13) doubles than another two samples (polymerization degree=9,19). The results in the present study may be able to provide some fundamental data for controlling CPP degradation.

Biodegradable porous calcium polyphosphate scaffolds for the three-dimensional culture of dental pulp cells.

AIM: To develop a three-dimensional culture model of human dental pulp cells (DPCs) with biodegradable porous calcium polyphosphate (CPP) scaffolds. METHODOLOGY: Human DPCs were isolated from three donors. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity of CPP compared with hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP). Values were analysed using unpaired t-tests. Cells were seeded onto porous CPP scaffolds with pore sizes in the range of 200-300 microm. The nature of cellular adaptation in the three-dimensional culture model was then evaluated visually by scanning electronic microscopy (SEM) and confocal laser scanning microscopy (CLSM). The apoptotic property of cells on the scaffolds was also assessed by DNA staining with CLSM. RESULTS: The cytotoxicity assay indicated that there was no significant difference between CPP and HA for each donor's original cells (P>0.05). Calcium polyphosphate had no cytotoxic effect on DPCs, whilst SEMs showed that cells successfully adhered to CPP scaffolds and spread amongst pores. On the cell surface, fine processes and matrix secretory granules were found. Confocal laser scanning microscopy showed that cells took on a three-dimensional structure with signs of vitality. CONCLUSION: Porous CPP scaffolds are promising for the establishment of a three-dimensional culture model of DPCs.

[Research progresses on degradation mechanism in vivo and medical applications of polylactic acid].

Polylactic acid (PLA), a kind of aliphatic polyester, is a biodegradable polymer with outstanding biocompatibility. The degradation of PLA is a complex process involving four main phenomena, namely water absorption, ester cleavage, diffusion of soluble oligomers, and solubilization of fragments. This paper emphasizes on the following aspects: 1) present understanding of the hydrolytic degradation and enzymatic degradation of PLA and autocatalystic degradation mechanism in the bulk PLA; 2) main factors influencing PLA degradation; 3) biological results; 4) the development in application of PLA.