Metabolomics integrated network pharmacology reveals the mechanism of Ma-Mu-Ran Antidiarrheal Capsules on acute enteritis mice.

Acute enteritis (AE) is a type of digestive disease caused by biochemical factors that irritate the intestinal tract or pathogenic bacteria that infect it. In China, Ma-Mu-Ran Antidiarrheal Capsules (MMRAC) have been applied against diarrhea caused by AE and bacillary dysentery for many years, but the underlying mechanisms of their beneficial effects are not known. In the present study, network pharmacology and metabolomics were performed to clarify the active ingredients of MMRAC and explore the specific mechanism of MMRAC on AE mice. A total of 43 active components of MMRAC with 87 anti-AE target genes were identified, and these target genes were enriched in IL-17 and HIF-1 signaling pathways. Integration analysis revealed that purine metabolism was the critical metabolic pathway by which MMRAC exerted its therapeutic effect against AE. Specifically, MAPK14, MMP9, PTGS2, HIF1A, EGLN1, NOS2 were the pivotal targets of MMRAC for the treatment of AE, and Western blot analysis revealed MMRAC to decrease protein levels of these pro-inflammatory signaling molecules. According to molecular docking, these key targets have a strong affinity with the MMRAC compounds. Collectively, MMRAC relieved the colon inflammation of AE mice via regulating inflammatory signaling pathways to reduce hypoxia and improved energy metabolism.

Integrated network pharmacology and metabolomics analyses of the mechanism underlying the efficacy of Ma-Mu-Ran Antidiarrheal Capsules against dextran sulfate sodium-induced ulcerative colitis.

The current study utilizes a comprehensive network pharmacology and metabolomics analysis to investigate the mechanism of action of Ma-Mu-Ran Antidiarrheal Capsules (MMRAC) for the treatment of ulcerative colitis (UC). In this study, we established a mouse model of UC using dextran sulfate sodium. Colonic tissues were collected from mice and then subjected to hematoxylin and eosin staining, as well as histopathological analysis, to assess the therapeutic effect of MMRAC. Furthermore, we assessed the mechanisms through which MMRAC combats UC by employing integrated metabolomics and network pharmacology strategies. Lastly, we validated the key targets identified through western blot and molecular docking. An integrated network of metabolomics and network pharmacology was constructed using Cytoscape to identify eight endogenous metabolites involved in the therapeutic action of MMRAC on UC. Further comprehensive analyses were focused on four key targets and their associated core metabolites and pathways. The results of western blot and molecular docking demonstrated that MMRAC could modulate key targets and their expression levels. The cumulative results indicated that MMRAC restored intestinal function in UC, reduced inflammatory responses, and alleviated oxidative stress by influencing the methionine and cysteine metabolic pathways, as well as the urea cycle. In addition, it had an impact on arginine, proline, glutamate, aspartate, and asparagine metabolic pathways and their associated targets.

Anti-osteoporotic activity of puerarin 6"-O-xyloside on ovariectomized mice and its potential mechanism.

CONTEXT: Osteoporosis is one of the most common bone diseases, and radix of Pueraria lobata (Willd.) Ohwi possesses an obvious therapeutical effect on postmenopausal osteoporosis. OBJECTIVE: This study investigates the anti-osteoporotic activity of the puerarin 6"-O-xyloside (PXY) on ovariectomized mice and its related mechanism. MATERIALS AND METHODS: Osteoporotic mice model was established by ovariectomy (OVX). A total of 50 mice were divided into five groups (n = 10): sham, OVX group, PXY treatment groups (20, 40, and 60 mg/kg/d, i.p.). After 12 weeks' treatment, body weights were recorded. Then, mice were sacrificed, and serum samples were collected to determine the blood calcium, blood phosphorus, alkaline phosphatase (ALP), and osteoprotegerin (OPG) concentrations and uterine index was assayed. The thigh-bones of mice were collected to evaluate histopathological changes. In the in vitro experiment, the effect of PXY on osteoblasts' proliferation was evaluated and western blotting was performed to determine expressions of OPG and the receptor activators of NF-κB ligand (RANKL), as well as the ratio of OPG/RANKL. RESULTS: PXY (40 and 60 mg/kg/d, i.p.) obviously decreased body weights and increased uterine index of OVX (p < 0.05), and improved osteoporotic syndromes of OVX mice; PXY also significantly increased the concentrations of blood calcium, blood phosphorus, ALP, and OPG of OVX mice (p < 0.05); moreover, PXY obviously up-regulated the ratio of OPG/RANKL (p < 0.05). CONCLUSION: Our results demonstrated that the puerarin 6"-O-xyloside possesses significant anti-osteoporotic activity on ovariectomy mice.

[Influence on the expression of RHOA and ROCK-II after global cerebral ischemia-reperfusion in hippocampus by shenxiong huayu capsule].

OBJECTIVE: To investigate the effect of Shenxiong Huayu capsule on the expression of hippocampal CA1 recombinant protein A (small GTP binding protein A, RHOA) and ROCK-2 (RHO associated protein kinase-2, ROCK-II). METHODS: Clean SD male rats (n=96), divided into three groups with 32 rats for each group, gavage was applied 7 days before modeling until the morning of the day to put to death. The groups included the normal control group (normal saline), global cerebral ischemia model group (normal saline) and Shenxiong Huayu capsule+global cerebral ischemia group (Shenxiong Huayu capsule 0.048 g/kg, was dissolved in 0.5 mL double distilled water, once a day, orally 0.3 mL/100 g). Modified Pulsinelli four-vessel occlusion model was constructed in global cerebral ischemia model and Shenxiong Huayu treatment groups and at 1, 3, 7, 14 d after successful modeling, water maze learning test was applied to evaluate the memory abilities of different groups, histopathological changes in HE staining, expression and protein content of RHOA and ROCK-II in immunohistochemical staining and Western blot was observed. RESULTS: At each time point, escape latency in model group was prolonged (P<0.05) when compared with that in normal control group, and that in Shenxiong Huayu was shorter (P<0.05) than that of model group, but still longer (P<0.05) than that of normal control group. HE staining showed that, compared with the normal group, model hippocampal CA1 reduced gradually from 1 d to 14 d; an increased survival neurons (P<0.05) in Shenxiong Huayu treatment group at each time points was observed, but still less than that in normal group (P<0.05); immunohistochemistry and Western blot analysis demonstrated that the expression of RHOA and ROCK-II in normal control group was not obvious, in model group was decreased after an initial increasing, and that in Shenxiong Huayu treatment group was lower than that of model group (P<0.05), but still higher than that in normal group (P<0.05). CONCLUSION: Shenxiong Huayu capsule improve neuronal damage induced by global ischemia, decreased the expression of hippocampal CA1 region of RHOA and ROCK-II.

[Effect of shenxiong huayu capsule on cerebral ischemia/reperfusion injury and the expression of GAP43 in hippocampal CA1 of rats].

OBJECTIVE: To explore the effect of Shenxiong Huayu Capsule (SHC) on the cerebral ischemia-reperfusion (IR) injury and the expression of growth associated protein 43 (GAP43) after total cerebral IR in the hippocampal CA1 region of rats. METHODS: Totally 100 male adult SD rats were randomly divided into five groups, i.e., the control group, the model group, the group A (by taking SHC once daily), the group B (by taking SHC twice daily), and the group C (by taking SHC thrice daily), 20 in each group. The total IR model was prepared by improved Pulsinelli's 4-vessel occlusion method. Morphological changes of the hippocampal CA1 region were observed by HE staining at day 1, 3, 7, and 14. The expression of GAP43 in the hippocampal CA1 region was detected using immunohistochemical assay at day 1, 3, 7, and 14. Meanwhile, the behavioral score was determined. The expression of GAP43 in the hippocampal CA1 region was detected using Western blot at day 14. RESULTS: Compared with the control group, the expression of GAP43 increased in the model group, the behavioral score was elevated, degenerated neurons increased, and survival neurons decreased in the model group (all P < 0.05). Compared with the model group, the expression of GAP-43 increased (with the most significant difference seen in the group C, P < 0.01), the behavioral score significantly decreased, degenerated neurons decreased, and survival neurons increased in each HSC group (all P < 0.05). Survival neurons obviously increased at day 14, of which, most number of survival neurons and highest contents of GAP43 protein could be seen in the group C, showing statistical difference when compared with those of the group A and the group B (P < 0.01). CONCLUSION: SHC had protective effect on total cerebral IR in the hippocampal CA1, which might be associated with increased expression of GAP43.

Effect of Heparan Sulfate on Vasculogenesis and Dentinogenesis of Dental Pulp Stem Cells.

INTRODUCTION: Heparan sulfate (HS) is a major component of dental pulp tissue. We previously reported that inhibiting HS biosynthesis impedes endothelial differentiation of dental pulp stem cells (DPSCs). However, the underlying mechanisms by which exogenous HS induces DPSC differentiation and pulp tissue regeneration remain unknown. This study explores the impact of exogenous HS on vasculogenesis and dentinogenesis of DPSCs both in vitro and in vivo. METHODS: Human-derived DPSCs were cultured in endothelial and odontogenic differentiation media and treated with HS. Endothelial differentiation of DPSCs was investigated by real-time polymerase chain reaction and capillary sprouting assay. Odontogenic differentiation was assessed through real-time polymerase chain reaction and detection of mineralized dentin-like deposition. Additionally, the influence of HS on pulp tissue was assessed with a direct pulp capping model, in which HS was delivered to exposed pulp tissue in rats. Gelatin sponges were loaded with either phosphate-buffered saline or 101-102 µg/mL HS and placed onto the pulp tissue. Following a 28-day period, tissues were investigated by histological analysis and micro-computed tomography imaging. RESULTS: HS treatment markedly increased expression levels of key endothelial and odontogenic genes, enhanced the formation of capillary-like structures, and promoted the deposition of mineralized matrices. Treatment of exposed pulp tissue with HS in the in vivo pulp capping study induced formation of capillaries and reparative dentin. CONCLUSIONS: Exogenous HS effectively promoted vasculogenesis and dentinogenesis of DPSCs in vitro and induced reparative dentin formation in vivo, highlighting its therapeutic potential for pulp capping treatment.

A directional total variation minimization algorithm for isotropic resolution in digital breast tomosynthesis.

An optimization-based image reconstruction algorithm is developed for contrast enhanced digital breast tomosynthesis (DBT) using dual-energy scanning. The algorithm minimizes directional total variation (TV) with a data discrepancy and non-negativity constraints. Iodinated contrast agent (ICA) imaging is performed by reconstructing images from dual-energy DBT data followed by weighted subtraction. Physical DBT data is acquired with a Siemens Mammomat scanner of a structured breast phantom with ICA inserts. Results are shown for both directional TV minimization and filtered back-projection for reference. It is seen that directional TV is able to substantially reduce depth blur for the ICA objects.

Platelet-to-Lymphocyte Ratio (PLR), Neutrophil-to-Lymphocyte Ratio (NLR), Monocyte-to-Lymphocyte Ratio (MLR), and Eosinophil-to-Lymphocyte Ratio (ELR) as Biomarkers in Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease (AECOPD).

Purpose: The study comprehensively evaluated the prognostic roles of the platelet-to-lymphocyte ratio (PLR), neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), basophil-to-lymphocyte ratio (BLR), and eosinophil-to-lymphocyte ratio (ELR) in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Patients and Methods: Six hundred and nineteen patients with AECOPD and 300 healthy volunteers were retrospectively included into the study. The clinical characteristics of the patients with AECOPD and the complete blood counts (CBCs) of the healthy volunteers were collected. The associations of PLR, NLR, MLR, BLR, and ELR with airflow limitation, hospital length of stay (LOS), C-reactive protein (CRP), and in-hospital mortality in patients with AECOPD were analyzed. Results: Compared with the healthy volunteers, PLR, NLR, MLR, BLR, and ELR were all elevated in COPD patients under stable condition. PLR, NLR, MLR, and BLR were further elevated while ELR was lowered during exacerbation. In the patients with AECOPD, PLR, NLR, and MLR were positively correlated with hospital LOS as well as CRP. In contrast, ELR was negatively correlated with hospital LOS as well as CRP. Elevated PLR, NLR, and MLR were all associated with more severe airflow limitation in AECOPD. Elevated PLR, NLR, and MLR were all associated with increased in-hospital mortality while elevated ELR was associated with decreased in-hospital mortality. Binary logistic regression analysis showed that smoking history, FEV1% predicted, pneumonia, pulmonary heart disease (PHD), uric acid (UA), albumin, and MLR were significant independent predictors ofin-hospital mortality. These predictors along with ELR were used to construct a nomogram for predicting in-hospital mortality in AECOPD. The nomogram had a C-index of 0.850 (95% CI: 0.799-0.901), and the calibration curve, decision curve analysis (DCA), and clinical impact curve (CIC) further demonstrated its good predictive value and clinical applicability. Conclusion: In summary, PLR, NLR, MLR, and ELR served as useful biomarkers in patients with AECOPD.

Biological properties of lithium-containing surface pre-reacted glass fillers as direct pulp-capping cements.

OBJECTIVE: Surface pre-reacted glass fillers (S-PRG) can release different types of ions and in our previous study, we modified these fillers with lithium chloride (S-PRG/Li-100 mM) to induce reparative dentin formation by activating the Wnt/ß-catenin signaling pathway. Here, we assessed the biological performance of S-PRG/Li-100 mM and compared it with that of mineral trioxide aggregate (MTA) and S-PRG without additives. METHODS: In vivo studies were conducted on male Wistar rats using Masson's trichrome staining in pulp-capped molars. The test materials were implanted subcutaneously to evaluate their capacity for vascularization and biocompatibility. The ability of the test materials to form apatite was tested by immersing them in simulated body fluid. Rhodamine-B staining was conducted to assess their sealing ability in bovine teeth, while their antibacterial activity was evaluated against Streptococcus mutans and Lactobacillus casei in terms of colony-forming units and by live/dead staining. RESULTS: Masson's trichrome staining and tissue-implantation tests confirmed the biocompatibility of S-PRG/Li-100 mM and it was similar to that of MTA and S-PRG; inflammation regression was observed 14 days after operation in the subcutaneous tissues. S-PRG/Li-100 mM promoted the formation of apatite on its surface. Both the S-PRG groups showed higher sealing capability and bactericidal/bacteriostatic activity against oral bacterial biofilms than MTA. SIGNIFICANCE: Lithium-containing surface pre-reacted glass cements exhibit better antibacterial and sealing capabilities than MTA, suggesting their potential as high-performance direct pulp-capping materials.

α-Terthienyl induces prostate cancer cell death through inhibiting androgen receptor expression.

Prostate cancer is a disease that often occurs in elderly men. Androgen receptor signaling pathway runs through the occurrence and development of prostate cancer. Thereby, targeting androgen receptor is a crucial strategy for the treatment of prostate cancer. α-Terthienyl, which has been used as photosensitive activator and insecticide, is a natural compound rich in marigold. In the present study, we found α- terthienyl could inhibit the cell viability of four prostate cancer cell lines, especially on LNCaP and 22Rv1 cells which endogenously express androgen receptor. Then we proved that it could inhibit the proliferation of prostate cancer cells and induce apoptosis of prostate cancer cells by plate clone formation assay and flow cytometry respectively. Furthermore, we found α-terthienyl could inhibit androgen receptor nuclear translocation, reduce androgen receptor expression, reduce the mRNA and protein expression of androgen receptor target genes (KLK3, TMPRSS2, PCA3) and nuclear proliferation antigen Ki67 and PCNA. In addition, it inhibited the expression and phosphorylation of Akt protein while increasing the expression of tumor suppressor p27. Besides, we constructed a mouse xenograft prostate cancer model and confirmed that α-terthienyl also inhibited the growth of prostate cancer in vivo. In conclusively, α-terthienyl played an anti-prostate cancer role by inhibiting both the expression of androgen receptor and the transduction of its signal pathway, suggesting that it is a promising natural small molecule for the treatment of prostate cancer.

Investigation of the allergens in 2,316 children with allergic rhinitis from Guangdong, China.

Allergic rhinitis (AR) is one of the popular childhood diseases, bringing physical and metal burdens to the children and their families. The study was performed to detect common allergens eliciting AR in children, to investigate the prevalence of allergens in different age and gender cohorts, and to provide a reliable basis for clinical prevention and treatment of AR during childhood. We measured serum-specific IgE and performed inhalant and ingestion allergen examinations in 2,316 children with AR, in collaboration with BioSciTec GmbH. The prevalence of different allergens was determined according to gender, age, severity, and season. Among the 2,316 AR cases, the top five inhalant allergens were Dermatophagoides pteronyssinus (1,674 cases, 72.3%), Dermatophagoides farinae (1,520 cases, 65.6%), Blomia tropicalis (1,477 cases, 63.8%), Cockroach (602 cases, 26.0%), and Dog hair (602 cases, 26.0%). The top five ingestive allergens were Milk (1,111 cases, 48.0%), Egg white (543 cases, 23.4%), Shrimp/Crab (425 cases, 18.4%), Beef/Mutton (422 cases, 18.2%), and Egg yold (329 cases, 14.2%). AR severity analyses showed that 50.9% (1,180 cases) of D. pteronyssinus allergies were above level three, 47.9% (1,109 cases) of D. farinae allergies were above level three, only 23.3% (539 cases) of B. tropicalis allergies were level three, and B. tropicalis allergies were mainly of level 2. Other AR-inducing allergens mainly produced level one or two reactions. Regarding ingestion allergens, 7.9% (183 cases) of milk allergies and 4.7% (108 cases) of Shrimp/Crab allergies were above level three, and other allergens induced AR mainly of level one or two. The study investigated the major allergens eliciting AR in children from Guangdong, China, assessed the prevalence and severity among cohorts regarding age, gender, and season, and produced essential information on childhood AR, laying important references for AR prevention and treatment in the future.

Analysis of regulating activities of 5'-epiequisetin on proliferation, apoptosis, and migration of prostate cancer cells in vitro and in vivo.

Advanced prostate cancer has a poor prognosis, and it is urgent to develop new effective drugs. 5'-Epiequisetin is a tetramic acid derivative which was isolated from a marine sponge-derived fungus Fusarium equiseti in our previous study. In this study, 5'-epiequisetin showed cytotoxicity against four prostate cancer cell lines, namely, LNCaP, 22Rv1, DU145, and PC-3 cells, with the lowest IC50 value of 4.43 ± 0.24 µM in PC-3 cells. Further studies showed that it could dramatically regulate the clonal colony formation, apoptosis, and migration of PC-3 cells. In addition, flow cytometry data showed that 5'-epiequisetin could block the cell cycle at the G1 phase. Proteome profiler array and Western blot revealed that 5'-epiequisetin could regulate the expression of proteins responsible for cell proliferation, apoptosis, and migration. 5'-Epiequisetin regulated the expression of PI3K, Akt, phosphorylated Akt, and proteins which control the cell cycle. Meanwhile, 5'-epiequisetin upregulated expression of DR5 and cleave-caspase 3, which play important roles in the process of apoptosis. Moreover, when DR5 was silenced by small interfering RNA, the proportion of apoptotic cells induced by 5'-epiequisetin remarkably declined. In addition, 5'-epiequisetin downregulated the expression of survivin which plays a key role in the process of survival and apoptosis. 5'-Epiequisetin also impacted beta-catenin and cadherins, which were associated with cell migration. In addition, 5'-Epiequisetin significantly inhibited the progression of prostate cancer in mice, accompanied by regulating the protein expression of DR5, caspase 8, survivin, and cadherins in vivo. Taken together, these findings indicated that 5'-epiequisetin showed an anti-prostate cancer effect by inducing apoptosis and inhibiting cell proliferation and migration both in vitro and in vivo, suggesting a promising lead compound for the pharmacotherapy of prostate cancer.

Biological Evaluation of the Effect of Root Canal Sealers Using a Rat Model.

Gutta-percha points and root canal sealers have been used for decades in endodontics for root canal obturation. With techniques such as single cone methods, the amount of sealer is larger, making their properties more critical. However, relatively few reports have comprehensively evaluated their biological effects. To this end, we evaluated three types of sealers, zinc oxide-fatty acid-, bio-glass- and methacrylate resin-containing sealers were considered. Their biological effects were evaluated using a rat subcutaneous implantation model. Each sealer was loaded inside a Teflon tube and implanted subcutaneously in the backs of rats. Inflammatory cells were observed around all samples 7 days after implantation and reduced after 28 days. Our results revealed that all samples were in contact with the subcutaneous tissue surrounding the sealer. Additionally, Ca and P accumulation was observed in only the bio-glass-containing sealer. Furthermore, each of the three sealers exhibited unique immune and inflammatory modulatory effects. In particular, bio-glass and methacrylate resin sealers were found to induce variable gene expression in adjacent subcutaneous tissues related to angiogenesis, wound healing, muscle tissue, and surrounding subcutaneous tissue. These results may help to understand the biological impacts of root canal sealers on surrounding biological tissues, guiding future research and comparisons with new generations of materials.

Autologous fat transplantation for the treatment of abdominal wall scar adhesions after cesarean section.

Cesarean section results in scarring, which usually leads to adhesion between the subcutaneous fat and the abdominal wall muscle. The present study aimed to evaluate the therapeutic effect of autologous fat grafting on scar adhesion to the abdominal wall after cesarean section. Thirty-six patients with scar adhesion to the abdominal wall after cesarean section were recruited and treated between October 2013 and December 2015. The adhesion between the subcutaneous fat and the abdominal wall muscle was carefully separated through a small incision in the original scar to form multiple subcutaneous tunnels. Aspirated fat was injected into the scar lesion and subcutaneous tunnels, and the wound was then sutured. The clinical outcome was evaluated by comparing the pretreatment and 1-year posttreatment photographs and Patient and Observer Scar Assessment Scale (POSAS) scores. All patients had a marked improvement in the appearance, texture, and depression of the scar during 12 months of follow-up. The 1-year posttreatment POSAS scores for the color, pain, pruritus, hardness, fullness, mobility, and appearance of the scar were significantly decreased compared with the pretreatment scores. Hematoxylin-eosin staining revealed adipocyte-like cells in treated scar tissue specimens obtained 1 year after treatment. None of the patients reported severe adverse reactions. Autologous fat grafting combined with adhesion release may be a good treatment option for abdominal wall scarring after cesarean section. This method is minimally invasive and effective in achieving good functional and esthetic outcomes.

Two cases of hand, foot and mouth disease caused by enterovirus A71 after vaccination.

BACKGROUND: Enterovirus A71 (EVA71) is one of the main pathogens causing hand, foot and mouth disease (HFMD). In China, the proportion of cases of HFMD caused by EVA71 is known to be significantly lower following EVA71 vaccination; however, infection with EVA71 can still occur after vaccination. METHODS: The complete genomic sequences of EVA71-KM18A and KM18B (from two rare cases of EVA71 infection following vaccination) were obtained. Phylogenetic analysis, nucleotide mutation analysis, recombinant analysis and comparative analysis of amino acid mutations were performed. RESULTS: Phylogenetic analysis determined that the EVA71 strains belonged to the C4a subgenotype. The KM18A and KM18B strains were highly similar to the vaccine strains. For the KM18B strain, there were some obvious homologous recombination signals in the 5'non-coding region, region 2A, region 2C and region 3D. Amino acid mutations were observed in the SP55 (position 729) and 71-6 (position 500) conformational neutralizing epitopes of the KM18A and KM18B strains. CONCLUSIONS: These amino acid mutations may affect the SP55 and 71-6 conformational neutralizing epitopes and change their spatial conformation, thereby weakening vaccine effectiveness.

Performance of a Biodegradable Composite with Hydroxyapatite as a Scaffold in Pulp Tissue Repair.

Vital pulp therapy is an important endodontic treatment. Strategies using growth factors and biological molecules are effective in developing pulp capping materials based on wound healing by the dentin-pulp complex. Our group developed biodegradable viscoelastic polymer materials for tissue-engineered medical devices. The polymer contents help overcome the poor fracture toughness of hydroxyapatite (HAp)-facilitated osteogenic differentiation of pulp cells. However, the composition of this novel polymer remained unclear. This study evaluated a novel polymer composite, P(CL-co-DLLA) and HAp, as a direct pulp capping carrier for biological molecules. The biocompatibility of the novel polymer composite was evaluated by determining the cytotoxicity and proliferation of human dental stem cells in vitro. The novel polymer composite with BMP-2, which reportedly induced tertiary dentin, was tested as a direct pulp capping material in a rat model. Cytotoxicity and proliferation assays revealed that the biocompatibility of the novel polymer composite was similar to that of the control. The novel polymer composite with BMP-2-induced tertiary dentin, similar to hydraulic calcium-silicate cement, in the direct pulp capping model. The BMP-2 composite upregulated wound healing-related gene expression compared to the novel polymer composite alone. Therefore, we suggest that novel polymer composites could be effective carriers for pulp capping.

Lithium-containing surface pre-reacted glass fillers enhance hDPSC functions and induce reparative dentin formation in a rat pulp capping model through activation of Wnt/ß-catenin signaling.

Surface pre-reacted glass (S-PRG) fillers are new bioactive molecules used in dental clinic work to fill tooth defects. These fillers release various types of ions (Al+3, BO-3, Na+, SiO3-2, Sr+2 and F-) and exhibit high biocompatibility, antibacterial capability, reduced plaque accumulation, and enhanced osteoblast differentiation. We previously showed that cement of S-PRG fillers could induce tertiary dentin formation in rat models. Previous work also showed that lithium ions can activate the Wnt/ß-catenin signaling pathway in vitro and induce dentin formation in pulpotomized teeth in vivo. In the current study, we sought to enhance the effect of S-PRG cement by incorporating LiCl. We show that treatment of human dental pulp stem cells with eluates from S-PRG/LiCl combination cements leads to an upregulation in cell migration, differentiation, and mineralization in vitro. In pulp-capping animal trials, we found that S-PRG/LiCl cements could induce tertiary dentin formation 28-days post-capping. At 7 days post-capping, we identified both ß-catenin and Axin2 expression using immunofluorescence, indicative of Wnt/ß-catenin signaling activity. In conclusion, S-PRG/LiCl cement is highly effective in promoting human dental pulp stem cells profiles and in enhancing reparative dentin formation in rat teeth through activation of the Wnt/ß-catenin canonical signaling pathway. STATEMENT OF SIGNIFICANCE: This is the first study to assess the behavior of S-PRG fillers containing lithium ions on human dental pulp stem cells. We show that this new combination cement promotes positive cell responses by activating the endogenous Wnt/ß-catenin signaling pathway in the pulp. The Wnt/ß-catenin canonical signaling pathway is involved in many developmental and wound healing processes. The released lithium ions from the S-PRG cement were systematically detected <0.01 mmol/L in our rat model. But it was efficient to induce tertiary dentin formation at the defect site. Since this novel bioactive cement is potentially a promising material for clinical pulp regenerative therapy, future human clinical trials will be needed.

Protein S100-A7 Derived from Digested Dentin Is a Critical Molecule for Dentin Pulp Regeneration.

Dentin consists of inorganic hard tissue and organic dentin matrix components (DMCs). Various kinds of bioactive molecules are included in DMCs and some of them can be released after digestion by endogenous matrix metalloproteinases (MMPs) in the caries region. Digested DMCs induced by MMP20 have been reported to promote pulpal wound healing processes, but the released critical molecules responsible for this phenomenon are unclear. Here, we identified protein S100-A7 as a critical molecule for pulpal healing in digested DMCs by comprehensive proteomic approaches and following pulp capping experiments in rat molars. In addition, immunohistochemical results indicated the specific distribution of S100-A7 and receptor for advanced glycation end-products (RAGE) as receptor for S100-A7 in the early stage of the pulpal healing process, and following accumulation of CD146-positive stem cells in wounded pulp. Our findings indicate that protein S100-A7 released from dentin by MMP20 might play a key role in dentin pulp regeneration.

Surface Pre-Reacted Glass Filler Contributes to Tertiary Dentin Formation through a Mechanism Different Than That of Hydraulic Calcium-Silicate Cement.

The induction of tissue mineralization and the mechanism by which surface pre-reacted glass-ionomer (S-PRG) cement influences pulpal healing remain unclear. We evaluated S-PRG cement-induced tertiary dentin formation in vivo, and its effect on the pulp cell healing process in vitro. Induced tertiary dentin formation was evaluated with micro-computed tomography (µCT) and scanning electron microscopy (SEM). The distribution of elements from the S-PRG cement in pulpal tissue was confirmed by micro-X-ray fluorescence (µXRF). The effects of S-PRG cement on cytotoxicity, proliferation, formation of mineralized nodules, and gene expression in human dental pulp stem cells (hDPSCs) were assessed in vitro. µCT and SEM revealed that S-PRG induced tertiary dentin formation with similar characteristics to that induced by hydraulic calcium-silicate cement (ProRoot mineral trioxide aggregate (MTA)). µXRF showed Sr and Si ion transfer into pulpal tissue from S-PRG cement. Notably, S-PRG cement and MTA showed similar biocompatibility. A co-culture of hDPSCs and S-PRG discs promoted mineralized nodule formation on surrounding cells. Additionally, S-PRG cement regulated the expression of genes related to osteo/dentinogenic differentiation. MTA and S-PRG regulated gene expression in hDPSCs, but the patterns of regulation differed. S-PRG cement upregulated CXCL-12 and TGF-ß1 gene expression. These findings showed that S-PRG and MTA exhibit similar effects on dental pulp through different mechanisms.