Efficacy of calcium hydroxide-loaded poly(lactic-co-glycolic acid) biodegradable nanoparticles as an intracanal medicament against endodontopathogenic microorganisms in a multi-species biofilm model.

This study aimed to evaluate the antimicrobial activity of calcium hydroxide-loaded poly(lactic-co-glycolic acid) nanoparticles (CH-loaded PLGA NPs) on multi-species biofilms. Human root blocks were prepared (n = 40), and multi-species suspensions of Candida albicans, Enterococcus faecalis and Streptococcus gordonii were incubated within the root canals for 21 days. Canals (n = 10/group) were then medicated with saline solution (negative control), chlorhexidine (positive control), calcium hydroxide and CH-loaded PLGA NPs for 7 days. Samples taken from the 0.1 mm root canal dentin were collected, and cell growth was detected by culture on BHI agar. The viable cell count of the Ca(OH)2, chlorhexidine gel and CH-loaded PLGA NPs group was significantly lower than the normal saline group (p < 0.001). CH-loaded PLGA NPs demonstrated a significant lower viable cell than Ca(OH)2 (p < 0.001); it has potential as a medicament for endodontic therapy.

The Effects of Calcium Hydroxide-loaded Poly(Lactic-co-glycolic Acid) Biodegradable Nanoparticles in the ex vivo External Inflammatory Root Resorption Model.

AIM: To evaluate the calcium ions (Ca2+) diffusion of calcium hydroxide-loaded poly(lactic-co-glycolic acid) biodegradable nanoparticles [Ca(OH)2-loaded PLGA NPs] compared with conventional Ca(OH)2 in a simulated external root resorption ex vivo model using inductively coupled plasma mass spectrometry (ICP-MS). MATERIALS AND METHODS: Thirty human mandibular premolars were prepared by sectioning the root segments to create roots measuring 10 mm from the anatomical apex. The root canals were instrumented and irrigated. The external root surface cavities were created. The specimens were randomly divided into the following three groups: Poly(lactic-co-glycolic acid) (PLGA; control group, n = 10), conventional calcium hydroxide [Ca(OH)2] (Metapaste, n = 10), and Ca(OH)2-loaded PLGA NPs [15% Ca(OH)2, n = 10]. The intracanal materials were placed in the root canals, and the teeth were stored in phosphate-buffered saline at 37°C. The release of Ca2+ was measured at 7, 30, and 60 days using ICP-MS. RESULTS: Both Ca(OH)2-loaded PLGA NPs and Metapaste groups exhibited higher levels of Ca2+ release compared to the PLGA group at all time points. During the initial 7-day period, the Ca(OH)2-loaded PLGA NPs exhibited a significantly greater release of Ca2+ compared to Metapaste. From day 7 to day 30, Metapaste displayed a significantly higher release of Ca2+ than the Ca(OH)2-loaded PLGA NPs, but it experienced a subsequent decline in Ca2+ release after the 30-day period. After the 30-day mark, the Ca(OH)2-loaded PLGA NPs once again exhibited a significantly higher release of Ca2+ compared to Metapaste. CONCLUSION: The Ca(OH)2-loaded PLGA NPs exhibited sustained release of Ca2+ that exceeded conventional Ca(OH)2, particularly during the first week, demonstrating a greater amount of Ca2+ release. CLINICAL SIGNIFICANCE: The utilization of Ca(OH)2-loaded PLGA NPs as an intracanal medication for external inflammatory root resorption provided sustained release and had the potential to enhance the efficacy of inhibiting root resorption more effectively than conventional Ca(OH)2.

Schomburginones A‒J, geranylated benzophenones from the leaves of Garcinia schomburgkiana and their cytotoxic and anti-inflammatory activities.

Ten undescribed benzophenones, schomburginones A-J, together with 14 known analogs were isolated from the leaves of Garcinia schomburgkiana, an edible plant native to the Indochina region. The structures of the undescribed compounds were elucidated by NMR combined with HRMS spectroscopy, while their absolute configurations were determined using ECD and single-crystal X-ray diffraction analysis. The isolated metabolites represent benzophenone derivatives containing a modified monoterpene unit, including tri- and tetracyclic skeletons, which are rarely found in genus Garcinia. The cytotoxic evaluation on three cancerous cell lines demonstrated that schomburginone G, schomburginone H, and 3-geranyl-2,4,6-trihydroxybenzophenone were active against HeLa cells with IC50 values in the range of 12.2-15.7 µM, respectively, and selective compared to the non-cancerous L929 cells (SI > 3.5). In addition, the three cytotoxic compounds together with clusiacyclol A showed significant NO inhibitory activity in RAW 264.7 macrophage cells over 85% inhibition without obvious cytotoxicity at a final concentration of 100 µM. The promising activities of these compounds in cytotoxic and anti-inflammatory assays make them attractive for further study in the development of anticancer drugs.

Proteomics and Molecular Docking Analyses Reveal the Bio-Chemical and Molecular Mechanism Underlying the Hypolipidemic Activity of Nano-Liposomal Bioactive Peptides in 3T3-L1 Adipocytes.

Obesity is a global health concern. Physical activities and eating nutrient-rich functional foods can prevent obesity. In this study, nano-liposomal encapsulated bioactive peptides (BPs) were developed to reduce cellular lipids. The peptide sequence NH2-PCGVPMLTVAEQAQ-CO2H was chemically synthesized. The limited membrane permeability of the BPs was improved by encapsulating the BPs with a nano-liposomal carrier, which was produced by thin-layer formation. The nano-liposomal BPs had a diameter of ~157 nm and were monodispersed in solution. The encapsulation capacity was 61.2 ± 3.2%. The nano-liposomal BPs had no significant cytotoxicity on the tested cells, keratinocytes, fibroblasts, and adipocytes. The in vitro hypolipidemic activity significantly promoted the breakdown of triglycerides (TGs). Lipid droplet staining was correlated with TG content. Proteomics analysis identified 2418 differentially expressed proteins. The nano-liposomal BPs affected various biochemical pathways beyond lipolysis. The nano-liposomal BP treatment decreased the fatty acid synthase expression by 17.41 ± 1.17%. HDOCK revealed that the BPs inhibited fatty acid synthase (FAS) at the thioesterase domain. The HDOCK score of the BPs was lower than that of orlistat, a known obesity drug, indicating stronger binding. Proteomics and molecular docking analyses confirmed that the nano-liposomal BPs were suitable for use in functional foods to prevent obesity.

Discovery of a Multifunctional Octapeptide from Lingzhi with Antioxidant and Tyrosinase Inhibitory Activity.

Ganoderma lucidum or Lingzhi is a fungus species widely known as a traditional medicine. Exploring the beneficial peptides by hydrolysis using pepsin and trypsin has been extensively performed to identify new bioactive natural products. A multifunctional peptide that expresses potential scavenging activity and tyrosinase inhibition is valuable in therapeutic and cosmetic applications. This study aimed to identify and investigate the effects of a novel multifunctional peptide from Lingzhi on the melanogenic enzymes in melanoma cells by a targeted-proteomics approach. The multifunctional peptide was de novo sequenced by LC-MS/MS to be NH2-PVRSSNCA-CO2H (octapeptide). This sequence was chemically synthesized by solid-phase peptide synthesis (SPPS). The antioxidant ability of the synthesized octapeptide was measured by the DPPH, ABTS, and FRAP assays. The results showed that the peptide exhibited an antioxidant activity equal to 0.121 ± 0.01 mg equivalent to ascorbic acid, 0.173 ± 0.03 mg equivalent to gallic acid, and 2.21 ± 0.23 mM equivalent to FeSO4, respectively, which is comparable to these well-known antioxidants. The proteomics approach identified a total of 5804 proteins and several pathways involved in the effects of the octapeptide in melanoma cells. Targeted proteomics revealed three specific proteins associated with pigmentation including Rab29, Dct, and Tyrp1. The Rab29 and Dct were upregulated whereas Tyrp1 was downregulated in the octapeptide treatment group. These findings could be used in the understanding of the molecular functions of the multifunctional octapeptide on melanogenic enzymes, supporting its potential as a therapeutic and cosmetic ingredient.

Study of the Lipolysis Effect of Nanoliposome-Encapsulated Ganoderma lucidum Protein Hydrolysates on Adipocyte Cells Using Proteomics Approach.

Excessive lipid accumulation is a serious condition. Therefore, we aimed at developing safe strategies using natural hypolipidemic products. Lingzhi is an edible fungus and potential lipid suppression stimulant. To use Lingzhi as a functional hyperlipidemic ingredient, response surface methodology (RSM) was conducted to optimize the time (X1) and enzyme usage (X2) for the hydrolysate preparation with the highest degree of hydrolysis (DH) and % yield. We encapsulated the hydrolysates using nanoscale liposomes and used proteomics to study how these nano-liposomal hydrolysates could affect lipid accumulation in adipocyte cells. RSM analysis revealed X1 at 8.63 h and X2 at 0.93% provided the highest values of DH and % yields were 33.99% and 5.70%. The hydrolysates were loaded into liposome particles that were monodispersed. The loaded nano-liposomal particles did not significantly affect cell survival rates. The triglyceride (TG) breakdown in adipocytes showed a higher TG increase compared to the control. Lipid staining level upon the liposome treatment was lower than that of the control. Proteomics revealed 3425 proteins affected by the liposome treatment, the main proteins being TSSK5, SMU1, GRM7, and KLC4, associated with various biological functions besides lipolysis. The nano-liposomal Linzghi hydrolysate might serve as novel functional ingredients in the treatment and prevention of obesity.