Increased Alkaline Phosphatase Activity Research Articles

Blood biochemical parameters in mice under the action of polyphosphate esters and their complexes with antibiotics

Complexes of polyphosphate esters with antibiotics were developed in Lviv Polytechnic National University together with scientists of Institute of Animal Biology NAAS to reduce the negative impact of antibiotics on the animal body. The conducted experiments allow assessing the effect of antibiotics, polyphosphate esters and complexes of polyphosphate esters with antibiotics on the body of laboratory animals based on biochemical markers of hepato- and nephrotoxicity. The antibiotics were administered in average daily therapeutic doses. It was found that the physiological state of mice and their blood biochemical indicators were within physiological normal values after the administration of polyphosphate ester P4 and complexes of polyphosphate ester P4+antibiotics (amoxicillin, oxytetracycline, and doxycycline). At the same time, intramuscular administration of polyphosphate ester P6 and complexes of P6+antibiotics have a certain negative effect on mice, which is manifested by changes in the activity of marker enzymes: aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP). We found an increase in AST and ALT activities. P6+amoxicillin and P6+oxytetracycline complexes increased ALP activity. Complexes P4+antibiotics decreased ALP. Blood urea content decreased after the administration of polyphosphate ester P6 by 38.5%, P6+oxytetracycline by 26.9%, P6+doxycillin by 21.8%. P6+amoxicillin complex caused a significant increase by 237% in the concentration of creatinine in the blood of mice. The changes of blood creatinine concentration of other experimental groups fell within normal physiological range. Conducted studies of blood biochemical characteristics of mice under the action of new complexes of nanobiopolymer transporters with antibiotics ensured the selection of antibacterial drugs with low toxicity.

Dexamethasone and doxycycline functionalized nanoparticles enhance osteogenic properties of titanium surfaces.

To evaluate the effect of doxycycline and dexamethasone doped nanoparticles covering titanium surfaces, on osteoblasts proliferation and differentiation. Doxycycline and dexamethasone doped polymeric nanoparticles were applied on titanium discs (Ti-DoxNPs and Ti-DexNPs). Undoped NPs and uncovered Ti discs were used as control. Human MG-63 osteoblast-like cells were cultured. Osteoblasts proliferation was tested by MTT assay. Alkaline phosphatase activity was analyzed. Differentiation gene expression was assessed by real-time quantitative polymerase chain reaction. Scanning Electron Microscopy was performed to assess osteoblasts morphology. Mean comparisons were conducted by ANOVA and Wilcoxon or Tukey tests (p<0.05). No differences in osteoblasts proliferation were found. Osteoblasts grown on Ti-DoxNPs significantly increased alkaline phosphatase activity. Doxycycline and dexamethasone nanoparticles produced an over-expression of the main osteogenic proliferative genes (TGF-β1, TGF-βR1 and TGF-βR2). The expression of Runx-2 was up-regulated. The osteogenic proteins (AP, OSX and OPG) were also overexpressed on osteoblasts cultured on Ti-DoxNPs and Ti-DexNPs. The OPG/RANKL ratio was the highest when DoxNPs were present (75-fold increase with respect to the control group). DexNPs also produced a significantly higher OPG/RANKL ratio with respect to the control (20 times higher). Osteoblasts grown on titanium discs were mainly flat and polygonal in shape, with inter-cellular connections. In contrast, osteoblasts cultured on Ti-DoxNPs or Ti-DexNPs were found to be spindle-shaped and had abundant secretions on their surfaces. DoxNPs and DexNPs were able to stimulate osteoblasts differentiation when applied on titanium surfaces, being considered potential inducers of osteogenic environment when performing regenerative procedures around titanium dental implants.

Shifts of active microbial community structure and functions in constructed wetlands responded to continuous decreasing temperature in winter

Important functions of constructed wetland related to biogeochemical processes are mediated by soil microbes and low-temperature damage is the main limiting factor for microbes in winter. However, the response thresholds for active microbial community and enzyme activities to continuous decreases in temperature remain unclear. In this study, total 90 soil samples were collected every week over a 6-week period to track the dynamics of four enzymes involved in cycles of C, N, P and active bacterial community as field soil temperature decreased continuously from 6.62 °C to 0.55 °C. Enzyme activity changed suddenly when the temperature decreased to 4.83 °C, the nitrite reductase activity reduced by 36.2%, while alkaline phosphatase activity is increased by 396%. The cellulase and urease were only marginally influenced by cold stress. Decreased nitrite reductase activities corresponded with loss of nir-type denitrifiers important for nitrite reduction. For cold stress, N-related bacteria were sensitive species. Whereas increased alkaline phosphatase activity may be due to the fact that P-related bacteria were opportunistic species. Key functional taxa connected with degradation of cellulose promoted species coexistence and microbial network stability. The lower and upper temperature thresholds for community change were 4.85 °C and 6.30 °C, respectively. Collectively, these results revealed that microbial taxa involved in C, N and P cycling respond differently to continuous decreases in temperature and higher than 4.85 °C is an ideal environment to prevent loss of microbial diversity and functions in winter, providing a scientific reference for the targeted isolation and cultivation of key microbial taxa in rhizosphere soil and adjusting temperature range to improve the purification capacity of wetlands during low temperature periods.

Osteogenesis promotion by injectable methacryloylated gelatin containing psoralen and its bacteriostatic properties.

The treatment of periodontitis focuses on controlling the progression of inflammation, reducing plaque accumulation, and promoting bone tissue reconstruction. Among them, the reconstruction of irregular bone resorption caused by periodontitis is a long-standing challenge. At present, the local drug treatment of periodontitis is mainly anti-inflammatory and antibacterial drugs. In this study, psoralen (Pso), a Chinese herbal medicine with anti-inflammatory, antibacterial, and osteogenic effects, was selected for the local treatment of periodontitis. Meanwhile, an injectable methacrylate gelatin (GelMA) platform loading with Pso was constructed. Pso-GelMA had the properties of fluidity, light cohesion, self-healing, and slow release, which could be better used in the deep and narrow structure of the periodontal pocket, and greatly increased the effectiveness of local drug delivery. The pore size of Gelma hydrogel did not change after loading Pso by SEM. In vitro, Pso-GelMA effectively upregulated the expression of osteogenic genes and proteins, increased alkaline phosphatase activity, promoted the mineralisation of rat bone marrow mesenchymal stem cells (BMSCs) extracellular matrix, and had significant antibacterial effects on Staphylococcus aureus and Fusobacterium nucleatum. Therefore, Pso-GelMA has immense promise in the adjuvant treatment of periodontitis.

Particulate beta-tricalcium phosphate and hydroxyapatite doped with silver promote invitro osteoblast differentiation in MC3T3-E1 cells.

Calcium phosphates including β-tricalcium phosphate (β-TCP) and hydroxyapatite (HAp) have been widely used for bone regeneration application because of their high osteoconductive activities. In addition, various kinds of inorganic ions enhance differentiation, proliferation, and mineralization of osteoblasts. However, information about the effects of silver-doped β-TCP [β-TCP (Ag)] and HAp [HAp (Ag)] particles on osteogenic differentiation is not available yet. We focused on the impact of β-TCP (Ag) and HAp (Ag) particles on the osteogenic differentiation of MC3T3-E1 osteoblast precursor cells. MC3T3-E1 osteoblast precursor cells were pre-treated by β-TCP (Ag) or HAp (Ag). And then the medium was changed to differentiation medium. Subsequently, osteoblast differentiation-related markers were determined. We found that treatment with β-TCP (Ag) or HAp (Ag) particles increased alkaline phosphatase activity in MC3T3-E1 cells. Expression of osteoblast differentiation-related genes also increased after treatment with β-TCP (Ag) or HAp (Ag) particles, a response thought to be regulated by zinc finger-containing transcription factor osterix. The ratio of the receptor activator of nuclear factor kappa-B ligand (RANKL) to osteoprotegerin (OPG) was decreased by β-TCP (Ag) and HAp (Ag) particles. Silver doping of β-TCP and HAp particles is effective for bone regeneration.

LncRNA LINC00205 stimulates osteoporosis and contributes to spinal fracture through the regulation of the miR-26b-5p/KMT2C axis

BackgroundOsteoporosis (OP) is a common bone disease marked by decreased bone strength. Increasing evidence suggests that long non-coding RNA (lncRNAs) play important roles in the occurrence and progression of OP. This study aimed to investigate the role and mechanism of LINC00205 in the osteogenic differentiation of human mesenchymal stem cells (hMSCs) and OP.MethodsBone tissue samples were obtained from healthy controls and patients with osteoporosis with a spinal fracture (OP-Frx) or without a spinal fracture (OP-no-Frx). HMSCs were cultured and induced to undergo osteogenic differentiation. The expression of LINC00205, lysine (K)-specific methyltransferase 2C (KMT2C), and miR-26b-5p in bone tissues and cells was evaluated using western blotting and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The effects of LINC00205, miR-26b-5p, and KMT2C on calcium deposition, alkaline phosphatase (ALP) activity, and mRNA levels of the osteogenic differentiation marker genes [ALP, osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2)] were investigated using alizarin red S staining, an ALP activity assay, and qRT-PCR, respectively. Dual-luciferase reporter assay was performed to ascertain the binding relationship between miR-26b-5p and LINC00205/KMT2C.ResultsLINC00205 and KMT2C were upregulated in patients with OP-Frx and OP-no-Frx, whereas miR-26b-5p was downregulated. Furthermore, LINC00205 and KMT2C expression decreased, whereas that of miR-26b-5p increased over time from day 7 to 21 of the osteogenic differentiation of hMSCs. The knockdown of LINC00205 and KMT2C significantly increased ALP activity, calcium deposition, and the expression of RUNX2, ALP, and OCN. In contrast, the inhibition of miR-26b-5p yielded the opposite result. These data suggest that LINC00205 inhibits the osteogenic differentiation of hMSCs by modulating the miR-26b-5p/KMT2C signaling axis.ConclusionLINC00205 promotes OP and is involved in spinal fractures. LINC00205 is also a potential negative regulator of the osteogenic differentiation of hMSCs.

Effect of Rhodiola rosea Rhizome and Punica granatum Fruit Peel on the Metabolic Processes and Physiological Activity of Rats Fed with Excessively Fat Diet.

Rhodiola rosea (golden root) and Punica granatum (pomegranate), as well as a number of other species of medicinal plants, exert an array of biological effects: adaptogenic, antioxidant and anti-inflammatory. However, there are not enough contemporary studies of their influence on metabolic processes, especially in cases of imbalanced diet. Lipid dysregulation is the main reason for many diseases, including obesity, cardiovascular disorders, non-alcoholic fatty liver disease, hypertension, atherosclerosis and insulin resistance. Recently, a growing amount of evidence has suggested the positive effects of certain natural nutrients on lipid metabolism. The work aims to define the general effect of golden root rhizome and pomegranate peel on physiological activity and metabolic processes in model animals fed with excessively fat diet. This study is relevant for the development of drugs and food additives for treatment and prophylaxis of metabolism disorders. In a 30-day experiment, we determined the effect of golden root (Rhodiola rosea L.) rhizome and pomegranate (Punica granatum L.) peel on the physiological activity and metabolic processes of 24 laboratory rats consuming a high-fat diet. The physical activity was evaluated according to the mass gain of animals and change in the relative mass of the internal organs, and also the functional conditions of the central nervous system, as demonstrated by the indicators of the locomotor activity and emotional status, determined in the open field test. The influence on the metabolic processes was revealed by biochemical and clinical blood analyses. Body mass of rats fed with golden root (R. rosea) reached 125.8 % of the initial body mass; when fed on pomegranate (P. granatum), it reached 123.9 %; and the control group reached only 111.5 % of the initial body mass. The rhizomes of R. rosea in the diet of male rats during the month of the experiment did not cause significant changes in the relative organ mass, and the pomegranate peel fruits led to a decrease in the thymus relative mass, as well as liver and brain. R. rosea rhizomes in the rats' diet led to an increase in the activity of alkaline phosphatase, and also to a decrease in the concentration of urea and urea nitrogen. Diet supplemented with R. rosea also contributed to a strong decrease in plasma concentrations of bilirubin and triglycerides (up to 57.0 % compared with the concentration in the control group). The rhizomes of R. rosea contributed to an unreliable decrease in the atherogenicity index. The pomegranate peel also greatly increased alkaline phosphatase activity and reduced plasma triglyceride concentrations. In addition, in rats consuming the peel of P. granatum, blood glucose concentration decreased. Under the influence of P. granatum, a strong increase in the atherogenic index of plasma (up to 518.6 % of the control) was observed because of a decrease in the concentration of high-density lipoproteins (up to 57.1 %) and a simultaneous elevation of the concentration of low-density lipoproteins (up to 158.3 % of the control). Open field test between groups remained without significant changes. The results indicate that the rhizome of R. rosea and peel of P. granatum are safe as food additives to high-fat diet and did not cause pathological changes and side effects, and at the same time significantly influence the metabolic processes (lipid and carbohydrate). Our study theoretically substantiates the use of R. rosea rhizome and P. granatum peel for the production of nutraceutical and pharmacological products for the correction of metabolic disorders of people and animals. Doses and periods of their application require further research.

Commercial dishes with gelatin-free microstructured inserts for elongated stem cell self-renewal and pluripotency

Here, we report the scalable fabrication of 2i-functionalized micro-pyramid-array (μPyA/+2i) inserts for use in commercial multi-well plates, as the alternative cultivation platform for maintaining long-term self-renewal and pluripotency of multiple mESCs and mouse induced pluripotent stem cells. Relevant evidence including cell morphology characterization increased alkaline phosphatase activity, high expression of mESC self-renewal markers, decreased levels of differentiation-associated markers, and high proportion of self-renewal marker cells are provided. Further studies demonstrated that μPyA/+2i could cause a higher cell density in mESC colony, and induce gene expression changes. Subsequent studies showed that μPyA/+2i can influence the cytoskeleton and promote cell adhesion through Cldn-7 upregulation. In summary, these μPyA/+2i inserts offer flexible and gelatin-free micro-envriomnets to maintain long-term self-renewal and pluripotency of mESCs. Enabled by the microstructured inserst, the facile stem cell manipulation and transfer among culture dishes will broaden stem cells both in routine and translational applications.

Effects of Dental Pulp Stem Cell Preconditioning on Osteogenesis using Conditioned Media of Probiotics Bacteria

Stem cells are used to treat numerous diseases; however, their lifespan is rather short. Factors such as probiotics affect and improve various cell lineage efficacies. The aim of this study was to investigate the effects of probiotics-conditioned media on dental pulp stem cell potentials in osteogenesis. The experiment was initiated by culturing Lactobacillus casei and Lactobacillus acidophilus probiotics as well as DPS-7 cells. Bacterial supernatants were separated and concentrated as the conditioned media. The DPS-7 cells were treated with various concentrations of the conditioned media. Furthermore, MTT assay and alkaline phosphatase activity were used. The mRNA expression of three genes (bFGF, EGF-β and BMP-2) involved in osteogenesis was analyzed using a real-time polymerase chain reaction. The response of dental pulp stem cells to probiotics preconditioning promoted cell proliferation, increased alkaline phosphatase activity and upregulated bFGF and BMP-2 gene expression. Increased expression was significant for BMP-2 and moderate for bFGF; however, it was non-significant for EGF-β. The use of the two probiotics was the most effective. In general, synergism of the combined probiotics preconditioning induces differentiation of DPS-7 cells into osteoblasts most effectively.

Commensal Bacillus siamensis LF4 induces antimicrobial peptides expression via TLRs and NLRs signaling pathways in intestinal epithelial cells of Lateolabrax maculatus

Antimicrobial peptides (AMPs) play an important role in modulating intestinal microbiota, and our previous study showed that autochthonous Baccilus siamensis LF4 could shape the intestinal microbiota of spotted seabass (Lateolabrax maculatus). In the present study, a spotted seabass intestinal epithelial cells (IECs) model was used to investigate whether autochthonous B. siamensis LF4 could modulate the expression of AMPs in IECs. And then, the IECs were treated with active, heat-inactivated LF4 and its supernatant to illustrate their AMPs inducing effects and the possible signal transduction mechanisms. The results showed that after 3 h of incubation with 108 CFU/mL B. siamensis LF4, lactate dehydrogenase (LDH), glutamic oxaloacetic transaminase (GOT), glutamic propylic transaminase (GPT) activities in supernatant decreased significantly and obtained minimum values, while supernatant alkaline phosphatase (AKP) activity, β-defensin protein level and IECs Na+/K+-ATPase activity, AMPs (β-defensin, hepcidin-1, NK-lysin, piscidin-5) genes expression increased significantly and obtained maximum values (P < 0.05). Further study demonstrated that the active, heat-inactivated LF4 and its supernatant treatments could effectively decrease the LDH, GOT, and GPT activities in IECs supernatant, increase AKP activity and β-defensin (except LF4 supernatant treatment) protein level in IECs supernatant and Na+/K+-ATPase and AMPs genes expression in IECs. Treatment with active and heat-inactivated B. siamensis LF4 resulted in significantly up-regulated the expressions of TLR1, TLR2, TLR3, TLR5, NOD1, NOD2, TIRAP, MyD88, IRAK1, IRAK4, TRAF6, TAB1, TAB2, ERK, JNK, p38, AP-1, IKKα, IKKβ and NF-κB genes. Treatment with B. siamensis LF4 supernatant also resulted in up-regulated these genes, but not the genes (ERK, JNK, p38, and AP-1) in MAPKs pathway. In summary, active, heat-inactivated and supernatant of B. siamensis LF4 can efficiently induce AMPs expression through activating the TLRs/NLRs-MyD88-dependent signaling, active and heat-inactivated LF4 activated both the downstream MAPKs and NF-κB pathways, while LF4 supernatant only activated NF-κB pathway.

Propofol mediates bone metastasis by regulating PC-derived exosomal miR-142-3p

In this study we investigated the role of propofol in mediating prostate cancer (PCa) bone metastasis through regulating exosomal factors derived from PCa. We isolated exosomes from PCa cells and co-cultured them with mesenchymal stem cells (MSCs). PCa-derived exosomes increased calcium deposition of MSCs and upregulated ALPL'Alkaline phosphatase, tissue-nonspecific isozyme) and BGLAP (Bone Gamma-Carboxyglutamate Protein) expression. Propofol treatment reduced alkaline phosphatase (ALP) activity, and ALPL and BGLAP expression that was induced by PCa-derived exosomes in MSCs. miRNAs present in cancer cell-derived exosomes increased osteogenesis in these cells. We evaluated miRNA expression in PCa cells after treatment with propofol, and found that miR-142-3p was upregulated in PCa cells. Furthermore, we transfected MSCs with miR-142-3p mimics or inhibitors and revealed that miR-142-3p mimics reduced calcium deposition and downregulated ALP activity, and ALPL and BGLAP levels, while miR-142-3p inhibitors increased calcium deposition and increased ALP activity, and ALPL and BGLAP levels. Finally, we determined that MSCs co-cultured with PCa-derived exosomes and transfected with miR-142-3p mimic exhibited reduced calcium deposition and lower ALP activity, and expression of ALPL and BGLAP. These data demonstrate that propofol inhibits osteogenic differentiation and mineralization of MSCs induced by PCa-derived exosomes by regulation of miR-142-3p levels.

Niobium-containing bioactive glasses modulate alkaline phosphatase activity during bone repair.

This study aimed to evaluate the pre-clinical behavior of niobium-containing bioactive glasses (BAGNb) by their ability to promote bone repair and regulate alkaline phosphatase (ALP) levels in an animal model. BAGNbs were produced as powders and as scaffolds and surgically implanted in the femur of male rats (Wistar lineage n=10). Glasses without Nb (BAG) were produced and implanted as well. The Autogenous Bone (AB) was used as a control. After 15, 30, and 60 days of surgical implantation, blood serum samples were collected to quantify ALP activity, and femurs were removed to assess bone repair. Bone samples were histologically processed and stained with H&E to quantify the % new bone into defects. No postoperative complications were identified. Early-stage repair (15 days) resulted in increased ALP activity for all groups, with increased values ​​for powdered BAGNb. The maturation of the new bone led to a reduction in serum ALP levels. Histological sections showed the formation of immature bone tissue and vascularization with the progression of bone deposition to mature and functional tissue over time. BAG powder showed less new bone formation in 15 days, while the analysis at 30 and 60 days showed no difference between groups (p > .05). Niobium-containing bioactive glasses safely and successfully induced bone repair in vivo. The modulation of ALP activity may be a pathway to describe the ability of niobium-containing materials to contribute to new bone formation.

Chitosan/Hydroxyapatite Scaffolds with P28 as a Promising Osteoinductive Scaffold for Bone Healing Applications

Despite bone’s inherent ability to heal, large bone defects remain a major clinical concern. This study proposes an off-the-shelf treatment combining chitosan/hydroxyapatite (CS/HAp) scaffolds, covalently linked with either bone morphogenetic protein-2 (BMP-2) or its related peptide P28 via a UV crosslinking process. Although covalently binding the growth factors was reported as a great alternative to the conventionally physical adsorption and encapsulation methods, this method presents the risk of altering the molecular activity and interaction of the growth factors. Therefore, alkaline phosphatase (ALP) activity and alizarin red staining (ARS) with a quantitative cetylpyridinium chloride (CPC) assay were conducted to validate that our photo-crosslinking fabrication method did not interfere with the functionality of the growth factors. The ALP activity of C2C12 with 100 µg/mL P28 was found to be comparable to 0.5 µg/mL BMP-2 after two weeks, where 0.001 U/mL was recorded for both treatments. The C2C12 cultured with CS/HAp/BMP-2 and CS/HAp/P28 scaffolds also showed an increased ALP activity compared to the negative control. ARS-CPC assay presented the highest optical density in 0.3 µg/mL BMP-2 and 50 µg/mL P28, while the highest intensity of ARS was observed in C2C12 cultured with CS/HAp/BMP-2 and CS/HAp/P28 scaffolds compared to the negative controls. The osteoconductive capability of this delivery system was then investigated through a rat femoral condyle defect model, where the new bone mineral density and the bone volume increased for all CS/HAp scaffolds compared to the collagen sponge control treatment. The histological assessment showed a favourable bone regeneration efficacy of the CS/HAp/P28 compared to the CS/HAp/BMP-2 treatment, thus showing the use of CS/HAp scaffolds with P28 as a promising osteoinductive scaffold for bone healing applications.

Response of cyanobacterial mats to ambient phosphate fluctuations: phosphorus cycling, polyphosphate accumulation and stoichiometric flexibility

Cyanobacterial mats inhabit a variety of aquatic habitats, including the most extreme environments on Earth. They can thrive in a wide range of phosphorus (P) levels and are thus important players for ecosystem primary production and P cycling at the sediment-water interface. Polyphosphate (polyP), the major microbial P storage molecule, is assigned a critical role in compensating for phosphate fluctuations in planktonic cyanobacteria, but little is known about potentially analogous mechanisms of mat-forming cyanobacteria. To investigate acclimation strategies of cyanobacterial mats to fluctuating phosphate concentrations, laboratory batch experiments were conducted, in which the cosmopolitan mat-forming, marine cyanobacterium Sodalinema stali was exposed to low dissolved P concentrations, followed by a P pulse. Our results show that the cyanobacteria dynamically adjusted cellular P content to ambient phosphate concentrations and that they had accumulated polyP during periods of high phosphate availability, which was subsequently recycled to sustain growth during phosphate scarcity. However, following the depletion of dispensable cellular P sources, including polyP, we observed a reallocation of P contained in DNA into polyP, accompanied by increasing alkaline phosphatase activity. This suggests a change of the metabolic focus from growth towards maintenance and the attempt to acquire organic P, which would be naturally contained in the sediment. P overplus uptake following a simulated P pulse further suggests that Sodalinema-dominated mats exhibit elaborated mechanisms to cope with severe P fluctuations to overcome unfavourable environmental conditions, and potentially modulate critical P fluxes in the aquatic cycle.

Effects of Supplementation of Zinc, Manganese, or Copper and Different Phytase Levels in Serum and Bone Acid and Alkaline Phosphatases of Broiler Chicks

The knowledge of biological trace minerals and phytase requirements for modern broiler genotypes is not established and the pressure to reduce their usage in animal feeding due to environmental issues is increasing. Here, the alkaline phosphatase (ALP) and tartarate-resistant acid phosphatase (TRAP) of the tibia and serum of broilers fed with diets containing various levels of phytase and reduced levels of zinc, manganese, and copper was evaluated. The experiment was performed using 1,200 male Cobb broilers raised according to standard commercial husbandry techniques. Data were analyzed as a 4×3 factorial arrangement with four concentrations of zinc (0.34, 0.49, 0.64, and 0.79 ppm), manganese (0.18, 0.43, 0.68, and 0.93 ppm), or copper (0.09, 0.12, 0.15, and 0.18 ppm) and three concentrations of phytase (0, 500, and 1,000 FTU/kg) for age periods of 1-21 and 36-42 days. While the dietary supplementation with copper did not induce a significant effect in bone tissue biochemical markers, serum TRAP activity of 42-day old broilers increased with higher copper levels. Increasing dietary zinc levels linearly increased ALP activity in tibia growth, suggesting that zinc is essential for longitudinal bone growth. Phytase significantly promoted the increase of TRAP and ALP activities, suggesting that manganese increased growth plate activity, accelerated calcification, and remodeled the newly formed tissue into trabecular bone. Although not every enzymatic activity was affected by the treatments, the phytase use, along with trace minerals, improved the animal response to the rapid growth required nowadays and provided the nutrients for adequate bone metabolism.

Degradable hydrogel fibers encapsulate and deliver metformin and periodontal ligament stem cells for dental and periodontal regeneration.

Human periodontal ligament stem cells (hPDLSCs) are promising cells for dental and periodontal regeneration. This study aimed to develop novel alginate-fibrin fibers that encapsulates hPDLSCs and metformin, to investigate the effect of metformin on the osteogenic differentiation of hPDLSCs, and to determine the regulatory role of the Shh/Gli1 signaling pathway in the metformin-induced osteogenic differentiation of hPDLSCs for the first time. CCK8 assay was used to evaluate hPDLSCs. Alkaline phosphatase (ALP) staining, alizarin red S staining, and the expression of osteogenic genes were evaluated. Metformin and hPDLSCs were encapsulated in alginate-fibrinogen solutions, which were injected to form alginate-fibrin fibers. The activation of Shh/Gli1 signaling pathway was examined using qRT-PCR and western blot. A mechanistic study was conducted by inhibiting the Shh/Gli1 pathway using GANT61. The administration of 50 μM metformin resulted in a significant upregulation of osteogenic gene expression in hPDLSCs by 1.4-fold compared to the osteogenic induction group (P < 0.01), including ALP and runt-related transcription factor-2 (RUNX2). Furthermore, metformin increased ALP activity by 1.7-fold and bone mineral nodule formation by 2.6-fold (P<0.001). We observed that hPDLSCs proliferated with the degradation of alginate-fibrin fibers, and metformin induced their differentiation into the osteogenic lineage. Metformin also promoted the osteogenic differentiation of hPDLSCs by upregulating the Shh/Gli1 signaling pathway by 3- to 6- fold compared to the osteogenic induction group (P<0.001). The osteogenic differentiation ability of hPDLSCs were decreased 1.3- to 1.6-fold when the Shh/Gli1 pathway was inhibited, according to ALP staining and alizarin red S staining (P<0.01). Metformin enhanced the osteogenic differentiation of hPDLSCs via the Shh/Gli1 signaling pathway. Degradable alginate-fibrin hydrogel fibers encapsulating hPDLSCs and metformin have significant potential for use in dental and periodontal tissue engineering applications. Alginate-fibrin fibers encapsulating hPDLSCs and metformin have a great potential for use in the treatment of maxillofacial bone defects caused by trauma, tumors, and tooth extraction. Additionally, they may facilitate the regeneration of periodontal tissue in patients with periodontitis.

Interactions with amyloid beta peptide and acetylcholinesterase increase alkaline phosphatase activity.

Multiple studies have shown that the activity of alkaline phosphatase (AP) increases during Alzheimer's disease (AD). In this paper, using UV-Visible spectroscopy, we show that this increase in activity is due to its interaction with key components of AD such as amyloid β peptide and acetylcholinesterase. Activity increase also occurs due to high concentrations of acetylcholine and choline. These conditions are present in AD or could occur due to drugs used for treating AD.

Periplaneta americana extract promotes osteoblast differentiation of human alveolar bone marrow mesenchymal stem cells.

This study aims to investigate the effects of Traditional Chinese medicine, Periplaneta americana extract (PAE), on osteoblast differentiation of human alveolar bone marrow-derived mesenchymal stem cells (hABMMSCs). Human alveolar bone marrow-derived mesenchymal stem cells were treated with different concentrations of PAE. Cell Counting Kit-8 (CCK-8) assay and transwell migration assay were conducted to evaluate cell proliferation and migration, respectively. Alkaline phosphatase (ALP) staining, ALP activity assay, and Alizarin red S staining were performed to detect osteogenesis in hABMMSCs. In addition, real-time quantitative polymerase chain reaction (RT-qPCR) and western blot (WB) assay were performed to evaluate expression levels of osteogenic markers. Finally, RNA sequencing analysis and WB were carried out to elucidate the underlying mechanism. A total of 0.1 mg/ml PAE promoted cell proliferation and migration. PAE also increased ALP activity and mineralized nodule formation of hABMMSCs. In addition, PAE upregulated the expression of osteogenesis-related genes (RUNX2, COL1A1, and BGLAP). RNA-sequencing analysis revealed that PAE activated the focal adhesion signaling pathway. Treatment with Defactinib, an inhibitor of FAK, attenuated the effects induced by PAE. PAE could enhance osteoblast differentiation of hABMMSCs through focal adhesion signaling pathway, suggesting a therapeutic potential for the alveolar bone defect.

Supplemental bile acids in low fishmeal diet improved the growth, nutrient utilization of Pacific white shrimp (Litopenaeus vannamei)

This study was conducted to investigate the effects of adding bile acids to low fishmeal diet on the growth, nutrient utilization and serum biochemical parameters of Pacific white shrimp, Litopenaeus vannamei. First, a positive and a negative control diet were formulated to contain 200 g/kg fishmeal (FM20) and 140 g/kg FM (FM14), respectively. Then, bile acids were added to FM14 diet at the inclusion levels of 75 (BA75), 150 (BA150) and 300 (BA300) mg/kg. The five isoproteic and isolipidic diets were fed to shrimp with initial weight of 1.09 ± 0.01 g for 56 days. The results showed no significant difference in survival among all the groups (>90 %). Compared to FM20 group, the weight gain (−12.9 %), body crude protein content, apparent digestibility of dry matter and protein, bile acid content in hepatopancreas and serum were decreased significantly, while feed conversion ratio (FCR) was increased by 0.17 in FM14 group (P < 0.05). The addition of bile acids to FM14diet tended to increase the weight gain, body crude protein content, apparent digestibility of dry matter and protein (P > 0.05), reaching the similar levels to FM20 group (P > 0.05). Especially, the FCR of BA150 group was significantly lower (−0.16) than that of FM14 group (P < 0.05). Compared to FM14 group, the addition of bile acids (75–300 mg/kg) significantly reduced serum glutathione transaminase activity and increased alkaline phosphatase activity (P < 0.05). In addition, the supplementation of 300 mg/kg of bile acids significantly reduced serum glutathione transaminase activity and increased hepatopancreatic protease activity and serum bile acid content (P < 0.05). In conclusion, the addition of 150 mg/kg bile acids to low fishmeal diet (140 g/kg FM) could improve the growth performance and nutrient utilization of white shrimp.

High-fat diet promotes the effect of fructo-oligosaccharides on the colonic luminal environment, including alkaline phosphatase activity in rats

We recently reported that fermentable nondigestible carbohydrates such as oligosaccharides, commonly increase colonic alkaline phosphatase (ALP) activity and the gene expression of Alpi-1, coding for rat intestinal alkaline phosphatase-I isozyme in rats and that the effect of oligosaccharides on colonic ALP activity is affected by the quality of dietary fats. We hypothesized that the amount of dietary fat would modulate the effect of oligosaccharides on colonic ALP and luminal environment in rats. In experiment 1, male Sprague-Dawley rats were fed a low-fat (LF, 5% lard) or high-fat (HF, 30% lard) diet with or without 4% fructo-oligosaccharides (FOS). In experiment 2, they were fed a 2.5%, 7%, 20%, or 40% fat (lard) diet with 4% FOS for 2 weeks. Dietary FOS in the HF diet (HF-FOS) significantly increased ALP activity in the colon and cecal digesta and colonic expression of Alpi-1, but not in the LF diet with FOS groups (LF-FOS). In comparison to the LF-FOS group, the increases in fecal mucins, Lactobacillus ratio, as well as cecal n-butyrate, and the decrease in fecal Clostridium coccoides, were more pronounced in the HF-FOS group. Compared with the 2.5% or 7% fat + FOS diet, the 20% fat + FOS diet significantly increased colonic ALP activity, Alpi-1 expression, and fecal mucins. These factors did not differ significantly between 20% and 40% fat + FOS diets. To exert the maximum effect of FOS on the colonic luminal environment, including ALP activity in rats, significantly more fat may be required than that contained present a LF diet.