Pectin coating of titanium and polystyrene surfaces modulates the macrophage inflammatory response

Titanium has been used with success for bone anchoring of dental implants. However, when implant surfaces are exposed to the oral environment, the progression of peri-implantitis triggered by specific oral bacteria has been reported. Bacterial colonization of implants leads to prolonged immune cell activation and bone resorption. A new strategy to improve implant biocompatibility and prevent peri-implantitis is to develop pectin surface nanocoatings. These plant-derived polysaccharides are promising candidates for surface nanocoatings of titanium implants due to their osteogenic and anti-inflammatory properties. Therefore, the aim of the study was to evaluate the in vitro effect of nanocoating with plant-derived rhamnogalacturonan-I (RG-I) on pro- and anti-inflammatory responses of primary human monocyte-derived macrophages (HMDMs) induced by Escherichia coli LPS and Porphyromonas gingivalis bacteria. In the present study, two different types of surface materials, tissue culture polystyrene (TCPS) plates and titanium (Ti) discs, coated with pectic polysaccharides, potato unmodified RG-I (PU) and potato dearabinanated RG-I (PA), have been examined. The inflammatory responses of HMDMs after E. coli LPS/P. gingivalis stimulation were investigated through gene expression measurements of pro- and anti-inflammatory cytokines. The results showed that PU and PA decreased expression of the proinflammatory genes tumour necrosis factor-alpha (TNFA), interleukin-1 beta (IL1B) and interleukin-8 (IL8) in activated HMDMs cultured on TCPS/Ti surfaces. In contrast, the effects on anti-inflammatory interleukin-10 (IL10) gene expression were not significant. The results indicate that RG-Is should be considered as a candidate for organic nanocoatings of titanium implant surfaces in order to limit host proinflammatory responses and improve bone healing. DOI: http://dx.doi.org/10.5281/zenodo.125054

Depletion of Mcpip1 in murine myeloid cells results in intestinal dysbiosis followed by allergic inflammation

MCPIP1 (called also Regnase-1) is a negative regulator of inflammation. Knockout of the Zc3h12a gene, encoding Mcpip1 in cells of myeloid origin ($Mcpip1^{MKO}$), has a pathological effect on many organs. The aim of this study was to comprehensively analyze pathological changes in the skin caused by Mcpip1 deficiency in phagocytes with an emphasis on its molecular mechanism associated with microbiome dysbiosis. $Mcpip1^{MKO}$ mice exhibited spontaneous wound formation on the skin. On a molecular level, the Th2-type immune response was predominantly characterized by an increase in Il5 and Il13 transcript levels, as well as eosinophil and mast cell infiltration. Irritation by DNFB led to a more severe skin contact allergy in $Mcpip1^{MKO}$ mice. Allergic reactions on the skin were strongly influenced by gut dysbiosis and enhanced systemic dissemination of bacteria. This process was followed by activation of the C/EBP pathway in peripheral macrophages, leading to local changes in the cytokine microenvironment that promoted the Th2 response. A reduced bacterial load inhibited allergic inflammation, indicating the role of intestinal dysbiosis in the development of skin diseases. Our results clearly show that MCPIP1 in phagocytes is an essential negative regulator that controls the gut-skin axis

Plant-derived pectin nanocoatings to prevent inflammatory cellular response of osteoblasts following Porphyromonas gingivalis infection

Background: Bioengineered plant-derived Rhamnogalacturonan-Is (RG-Is) from pectins are potential candidates for surface nanocoating of medical devices. It has recently been reported that RG-I nanocoatings may prevent bacterial infection and improve the biocompatibility of implants. The aim of the study was to evaluate in vitro impact of bioengineered RG-I nanocoatings on osteogenic capacity and proinflammatory cytokine response of murine osteoblasts following Porphyromonas gingivalis infection. Methods: Murine MC3T3-E1 osteoblasts and isolated primary calvarial osteoblasts from C57BL/6J (B6J osteoblasts) mice were infected with P. gingivalis and incubated on tissue culture polystyrene plates with or without nanocoatings of unmodified RG-Is isolated from potato pulps (PU) or dearabinanated RG-Is (PA). To investigate a behavior of infected osteoblasts cultured on RG-Is cell morphology, proliferation, metabolic activity, mineralization and osteogenic and pro-inflammatory gene expression were examined. Results: Following P. gingivalis infection, PA, but not PU, significantly promoted MC3T3-E1 and BJ6 osteoblasts proliferation, metabolic activity, and calcium deposition. Moreover, Il-1b, Il-6, TNF-α, and Rankl gene expressions were downregulated in cells cultured on PU and to a higher extent on PA as compared to the corresponding control, whereas Runx, Alpl, Col1a1, and Bglap gene expressions were upregulated vice versa. Conclusion: Our data clearly showed that pectin RG-Is nanocoating with high content of galactan (PA) reduces the osteoblastic response to P. gingivalis infection in vitro and may, therefore, reduce a risk of inflammation especially in immunocompromised patients with rheumatoid or periodontal disorders

Porównanie trójwymiarowej i dwuwymiarowej analizy ilościowej tętnic wieńcowych w odniesieniu do pomiaru długości segmentów wieńcowych u pacjentów poddawanych diagnostycznej koronarografii

Cel: Określenie, czy trójwymiarowa, ilościowa analiza angiograficzna naczyń wieńcowych (3D QCA) ma przewagę nad standardową oceną dwuwymiarową (2D QCA) polegającą na zwiększeniu dokładności i precyzji pomiarów długości oraz wyeliminowaniu skrócenia typowego dla analiz 2D. Materiał i metody: Wykonano ogółem 800 pomiarów długości odcinka pomiędzy markerami na prowadnikach wieńcowych wprowadzonych do tętnicy wieńcowej (8 długości na prowadniku w zakresie 5–40 mm) z zastosowaniem oprogramowania 3D i standardowego oprogramowania 2D w porównaniu ze znanymi odległościami znaczników na prowadniku wieńcowym u 21 pacjentów poddanych koronarografii. Wyniki: W analizie 2D QCA pomiary były z reguły zaniżone w stosunku do rzeczywistej długości mierzonego odcinka, przy czym wielkość błędu wzrastała wraz z bezwzględną długością mierzonego odcinka. Analizy za pomocą 3D QCA wykazały minimalne różnice pomiarów w stosunku do rzeczywistych odległości pomiędzy markerami w całym zakresie badanych długości. Wnioski: Technika 3D QCA minimalizuje błędy pomiarów długości związane z artefaktem skrócenia obrazowego obiektu (foreshortening) typowego dla 2D QCA wykazuje minimalne odstępstwa od rzeczywistych długości w stosunku do analiz 2D QCA. 3D QCA ma szczególną przewagę w przypadku większych długości.Aim: To determine if three-dimensional quantitative coronary angiographic analysis (3D QCA) provides an advantage over twodimensional (2D QCA) by increasing accuracy and precision of length measurements through compensating for foreshortening inherent to 2D images. Material and methods: A total of 800 inter-marker length measurements were performed (8 per wire, 5-40 mm) with novel 3D software and standard 2D software and compared with the true lengths of the inter-marker distances on the wire in coronary vessels of 21 patients recruited. Results: 2D QCA generally underestimated true length in comparison to 3D, and the discrepancy increased with absolute length. In contrast, 3D QCA showed a minimal difference from true length over the examined range of lengths. Conclusions: 3D QCA minimizes errors in length measurements associated with foreshortening, shows minimal difference from true length, and performs significantly better in comparison to 2D QCA. The advantage of 3D QCA is more pronounced at longer lengths

Detection of antibiotic resistance genes in wastewater treatment plant – molecular and classical approach

Antibiotics are a group of substances potentially harmful to the environment. They can play a role in bacterial resistance transfer among pathogenic and non-pathogenic bacteria. In this experiment three representatives of medically important chemotherapeutics, confirmed to be present in high concentrations in wastewater treatment plants with HPLC analysis were used: erythromycin, sulfamethoxazole and trimethoprim. Erythromycin concentration in activated sludge was not higher than 20 ng L−1. N-acetylo-sulfamethoxazole concentration was 3349 ± 719 in winter and 2933 ± 429 ng L−1 in summer. Trimethoprim was present in wastewater at concentrations 400 ± 22 and 364 ± 60 ng L−1, respectively in winter and summer. Due to a wide variety of PCR-detectable resistance mechanisms towards these substances, the most common found in literature was chosen. For erythromycin: erm and mef genes, for sulfamethoxazole: sul1, sul2, sul3 genes, in the case of trimethoprim resistance dhfrA1 and dhfr14 were used in this study. The presence of resistance genes were analyzed in pure strains isolated from activated sludge and in the activated sludge sample itself. The research revealed that the value of minimal inhibitory concentration (MIC) did not correspond with the expected presence of more than one resistance mechanisms. Most of the isolates possessed only one of the genes responsible for a particular chemotherapeutic resistance. It was confirmed that it is possible to monitor the presence of resistance genes directly in activated sludge using PCR. Due to the limited isolates number used in the experiment these results should be regarded as preliminary

Isomalto/Malto-polysaccharide, a novel soluble dietary fiber made via enzymatic conversion of starch

Dietary fibers are at the forefront of nutritional research because they positively contribute to human health. Much of our processed foods contain, however, only small quantities of dietary fiber, because their addition often negatively affects the taste, texture, and mouth feel. There is thus an urge for novel types of dietary fibers that do not cause unwanted sensory effects when applied as ingredient, while still positively contributing to the health of consumers. Here, we report the generation and characterization of a novel type of soluble dietary fiber with prebiotic properties, derived from starch via enzymatic modification, yielding isomalto/malto-polysaccharides (IMMPs), which consist of linear (α1 → 6)-glucan chains attached to the nonreducing ends of starch fragments. The applied Lactobacillus reuteri 121 GTFB 4,6-α-glucanotransferase enzyme synthesizes these molecules by transferring the nonreducing glucose moiety of an (α1 → 4)-glucan chain to the nonreducing end of another (α1 → 4)-α-glucan chain, forming an (α1 → 6)-glycosidic linkage. Once elongated in this way, the molecule becomes a better acceptor substrate and is then further elongated with (α1 → 6)-linked glucose residues in a linear way. Comparison of 30 starches, maltodextrins, and α-glucans of various botanical sources, demonstrated that substrates with long and linear (α1 → 4)-glucan chains deliver products with the highest percentage of (α1 → 6) linkages, up to 92%. In vitro experiments, serving as model of the digestive power of the gastrointestinal tract, revealed that the IMMPs, or more precisely the IMMP fraction rich in (α1 → 6) linkages, will largely pass the small intestine undigested and therefore end up in the large intestine. IMMPs are a novel type of dietary fiber that may have health promoting activity

Immune-instructive copolymer scaffolds using plant-derived nanoparticles to promote bone regeneration

BACKGROUND: Age-driven immune signals cause a state of chronic low-grade inflammation and in consequence affect bone healing and cause challenges for clinicians when repairing critical-sized bone defects in elderly patients. METHODS: Poly(l-lactide-co-ɛ-caprolactone) (PLCA) scaffolds are functionalized with plant-derived nanoparticles from potato, rhamnogalacturonan-I (RG-I), to investigate their ability to modulate inflammation in vitro in neutrophils and macrophages at gene and protein levels. The scaffolds’ early and late host response at gene, protein and histological levels is tested in vivo in a subcutaneous rat model and their potential to promote bone regeneration in an aged rodent was tested in a critical-sized calvaria bone defect. Significant differences were tested using one-way ANOVA, followed by a multiple-comparison Tukey’s test with a p value ≤ 0.05 considered significant. RESULTS: Gene expressions revealed PLCA scaffold functionalized with plant-derived RG-I with a relatively higher amount of galactose than arabinose (potato dearabinated (PA)) to reduce the inflammatory state stimulated by bacterial LPS in neutrophils and macrophages in vitro. LPS-stimulated neutrophils show a significantly decreased intracellular accumulation of galectin-3 in the presence of PA functionalization compared to Control (unmodified PLCA scaffolds). The in vivo gene and protein expressions revealed comparable results to in vitro. The host response is modulated towards anti-inflammatory/ healing at early and late time points at gene and protein levels. A reduced foreign body reaction and fibrous capsule formation is observed when PLCA scaffolds functionalized with PA were implanted in vivo subcutaneously. PLCA scaffolds functionalized with PA modulated the cytokine and chemokine expressions in vivo during early and late inflammatory phases. PLCA scaffolds functionalized with PA implanted in calvaria defects of aged rats downregulating pro-inflammatory gene markers while promoting osteogenic markers after 2 weeks in vivo. CONCLUSION: We have shown that PLCA scaffolds functionalized with plant-derived RG-I with a relatively higher amount of galactose play a role in the modulation of inflammatory responses both in vitro and in vivo subcutaneously and promote the initiation of bone formation in a critical-sized bone defect of an aged rodent. Our study addresses the increasing demand in bone tissue engineering for immunomodulatory 3D scaffolds that promote osteogenesis and modulate immune responses. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s41232-022-00196-9

Isomalto/Malto-Polysaccharide, A Novel Soluble Dietary Fiber Made Via Enzymatic Conversion of Starch

Dietary fibers are at the forefront of nutritional research because they positively contribute to human health. Much of our processed foods contain, however, only small quantities of dietary fiber, because their addition often negatively affects the taste, texture, and mouth feel. There is thus an urge for novel types of dietary fibers that do not cause unwanted sensory effects when applied as ingredient, while still positively contributing to the health of consumers. Here, we report the generation and characterization of a novel type of soluble dietary fiber with prebiotic properties, derived from starch via enzymatic modification, yielding isomalto/malto-polysaccharides (IMMPs), which consist of linear (α1 → 6)-glucan chains attached to the nonreducing ends of starch fragments. The applied <i>Lactobacillus reuteri</i> 121 GTFB 4,6-α-glucanotransferase enzyme synthesizes these molecules by transferring the nonreducing glucose moiety of an (α1 → 4)-glucan chain to the nonreducing end of another (α1 → 4)-α-glucan chain, forming an (α1 → 6)-glycosidic linkage. Once elongated in this way, the molecule becomes a better acceptor substrate and is then further elongated with (α1 → 6)-linked glucose residues in a linear way. Comparison of 30 starches, maltodextrins, and α-glucans of various botanical sources, demonstrated that substrates with long and linear (α1 → 4)-glucan chains deliver products with the highest percentage of (α1 → 6) linkages, up to 92%. In vitro experiments, serving as model of the digestive power of the gastrointestinal tract, revealed that the IMMPs, or more precisely the IMMP fraction rich in (α1 → 6) linkages, will largely pass the small intestine undigested and therefore end up in the large intestine. IMMPs are a novel type of dietary fiber that may have health promoting activity