Anti-inflammation and anti-apoptosis effects of growth arrest-specific protein 6 in acute liver injury induced by LPS/D-GalN in mice

Abstract Purpose To investigate the effect of growth arrest-specific protein 6 (Gas6) on acute liver injury in mice and related mechanisms. Methods Thirty C57BL/6 (6-8 weeks old) mice were randomly divided into control, LPS/D-GalN, and LPS/D-GalN+Gas6 groups (10 mice in each group). The LPS/D-GalN group was intraperitoneally administered with LPS (0.25 mg/Kg) and D-GalN (400 mg/Kg) for 5h. The LPS/D-GalN+Gas6 group was intraperitoneally administered with rmGas6 one hour before intraperitoneal application of LPS/D-GalN. All subjects were sacrificed at 5 h for blood and tissue analysis. The expression of protein and mRNA was assessed by western blotting and RT-PCR, respectively. Results Compared with the control group, AST, ALT, IL-1β, TNF-α, IL-6 IL-10, MPO activity were increased in the LPS/D-GalN group. However, they were significantly inhibited by Gas6. Gas6 markedly suppressed the expression of apoptosis-related protein induced by LPS/D-GalN. Moreover, Gas6 attenuated the activation of the NF-κB signaling pathway in acute liver injury induced by LPS/D-GalN. Conclusions Gas6 alleviates acute liver injury in mice through regulating NF-κB signaling pathways. Gas6 can be a potential therapeutic agent in treating LPS/D-GalN-induced acute liver injury in the future.

Uso de la Fibrina Rica en Plaquetas Inyectable (i-PRF) en Defectos Infra Óseos en Terapia Periodontal no Quirúrgica. Reporte de Dos Casos / Use of Injectable Platelet-Rich Fibrin (i-PRF) in Infra Osseous Defects in Non-Surgical Periodontal Therapy. Report of Two Cases

RESUMEN: Entre los concentrados plaquetarios de segunda generación, ha suscitado creciente interés, el uso de fibrina rica en plaquetas y leucocitos inyectable (i-PRF); que se obtiene a partir de la centrifugación inmediata de sangre venosa del propio individuo, y que aporta concentraciones elevadas de factor de crecimiento vascular endotelial, factor de crecimiento transformante beta, y factor de crecimiento derivado de plaquetas, entre otras proteínas que inician y coordinan el proceso reparativo. Su nula citotoxicidad y consistencia líquida abren un nuevo campo de estudio y experimentación en el ámbito de la Cirugía Oral y de la Periodoncia, como sustancia para irrigar. El objetivo de este manuscrito fue reportar el uso del i-PRF como irrigador subgingival en el tratamiento periodontal convencional de defectos infra óseos con 6 meses de seguimiento. En ambos casos, se verificó un efecto positivo de irrigación, lo que abre el debate al uso de productos farmacéuticos tradicionales como la clorhexidina versus preparados autólogos sin efectos adversos reportados a la fecha.

ABSTRACT: Second generation platelet concentrates include the use of injectable platelet-rich fibrin (i-PRF), which has generated increasing interest because it is derived from immediate centrifugation of venous blood from the patients themselves. It provides high concentrations of vascular endothelial growth factor, transforming growth factor beta, and platelet-derived growth factor, among other proteins that initiate and coordinate the healing process. Its null cytotoxicity and liquid consistency has opened new research lines in the field of oral surgery and periodontics, as an irrigation substance. The aim of this manuscript was to report the use of i-PRF, as a subgingival irrigator in conventional periodontal treatment of infra osseous defects, with six months follow-up. In both cases, a positive effect of irrigation was confirmed. These findings, open the debate as regards the use of traditional pharmaceutical products (such as chlorhexidine), versus autonomous preparations without adverse effects reported to date.

Treatment of venous ulcers with growth factors: systematic review and meta-analysis / Tratamiento de úlceras venosas confactores de crecimiento: revisión sistemática y metanálisis / Tratamento de úlceras venosas com fatores de crescimento: revisão sistemática e metanálise

ABSTRACT Objective: To identify evidence about the effects of growth factor application on venous ulcer healing. Method: Systematic review and meta-analysis, including Randomized Clinical Trials. Searches: Ovid MEDLINE, EMBASE, CINAHL, Cochrane CENTRAL, LILACS, Web of Science, Digital Library of Theses and Dissertations; Google Scholar and list of references. Results: 802 participants were recruited from the 10 included studies: 472 in the intervention group (growth factors) and 330 as control. The relative risk for the complete healing outcome was 1.06 [95% CI 0.92-1.22], p = 0.41. Participants who received Platelet-Rich Plasma and Epidermal Growth Factor showed a slight tendency to achieve complete healing, but without statistical relevance (p <0.05). Most of the studies were classified as moderate risk of bias. Conclusion: The effect of the application of growth factors for complete healing in venous ulcers is not clear, and clinical trials with methodological quality are required for more accurate recommendations.

RESUMEN Objetivo: Identificar evidencias acerca de los efectos de la aplicación de factores de crecimientoenlacicatrización de úlceras venosas. Método: Revisión sistemática y metanálisis, incluyendo Ensayos Clínicos aleatorizados. Búsquedas: Ovid MEDLINE, EMBASE, CINAHL, Cochrane CENTRAL, LILACS, Web of Science, Biblioteca Digital de Tesis y Disertaciones; Google Académico y lista de referencias Resultados: 802 participantes fueron reclutados por los 10 estudios incluidos: 472 en el grupo intervención (factores de crecimiento) y 330 como control. El riesgo relativo para el desenlace de cicatrización completa fue de 1,06 [IC95% 0,92-1,22], p = 0.41. Los participantes que recibieron Plasma Rico en Plaquetas y Factor de Crecimiento Epidérmico presentaron una ligera tendencia a alcanzar una cicatrización completa, pero sin relevancia estadística (p <0.05). La mayoría de los estudios se clasificaron como moderado riesgo de sesgo. Conclusión: El efecto de la aplicación de factores de crecimiento para cicatrización completa en úlceras venosas no está claro, siendo necesarios ensayos clínicos con calidad metodológica para recomendaciones más precisas.

RESUMO Objetivo: Identificar evidências acerca dos efeitos da aplicação de fatores de crescimento na cicatrização de úlceras venosas. Método: Revisão sistemática e metanálise, incluindo Ensaios Clínicos Randomizados. Buscas: Ovid MEDLINE, EMBASE, CINAHL, Cochrane CENTRAL, LILACS, Web of Science, Biblioteca Digital de Teses e Dissertações; Google Acadêmico e lista de referências. Resultados: 802 participantes foram recrutados pelos 10 estudos incluídos: 472 no grupo intervenção (fatores de crescimento) e 330 como controle. O risco relativo para o desfecho de cicatrização completa foi de 1,06 [IC95% 0,92-1,22], p=0.41. Os participantes que receberam Plasma Rico em Plaquetas e Fator de Crescimento Epidérmico apresentaram uma ligeira tendência a alcançar cicatrização completa, porém sem relevância estatística (p<0.05). A maioria dos estudos foi classificada como moderado risco de viés. Conclusão: O efeito da aplicação de fatores de crescimento para cicatrização completa em úlceras venosas não está claro, sendo necessários ensaios clínicos com qualidade metodológica para recomendações mais precisas.

Comparing osteogenic effects between concentrated growth factors and the acellular dermal matrix

Abstract Concentrated growth factor (CGF) is an autogenuous product that contains highly concentrated number of platelets and can be derived from venous blood by selective centrifugation. It has been speculated that local growth factors in human platelets (insulinlike growth factor, IGF; transforming growth factor, TGF-b; platelet derived growth factor, PDGF) would enhance healing of grafts and also counteract resorption. The osteogensis effect of CGF and acellular dermal matrix (ADM) for alveolar cleft defects was evaluated in this study. Twenty alveolar cleft patients were divided randomly into two groups. One group underwent guided bone regeneration (GBR) using acellular dermal matrix film combined with alveolar bone grafting using iliac crest bone grafts (GBR group), while the other group underwent alveolar bone grafting combined with CGF (CGF group). Cone beam computed tomography (CBCT) images were obtained at 1 week and 6 months following the procedure. Using Mimics 17.0 software, the bone resorption rate and bone density improvement rate were calculated and compared between the two groups. Although not significant between ADM and CGF in bone resorption rate, the bone density improvement in cases with CGF(61.62 ± 4.728%) was much better than in cases with ADM (27.05 ± 5.607%) (p = 0.0002). Thus, CGF could be recommended to patients with alveolar cleft as a better choice.

The influence of growth factors on skin wound healing in rats / A influência de fatores de crescimento na cicatrização de feridas cutâneas de ratas

ABSTRACT INTRODUCTION: Healing is a process that restores the physical integrity of body structures. It is a dynamic, complex, multicellular process that involves the extracellular matrix, cytokines, blood cells, and growth factors. Growth factors are proteins that activate and stimulate cell proliferation through the activation of angiogenesis, mitogenesis, and gene transcription, accelerating the healing process. OBJECTIVE: To assess the influence of growth factors on the healing process of wounds made on the backs of female rats compared to the control wound, through macro and microscopy. METHODS: This study used 45 female Wistar rats, in which three wounds were made on the back. The first was the control wound, the second received epithelial growth factor injection, and the third received a combination of factors. Macroscopic and microscopic assessments were performed on the third, seventh, and 15th days of the experiment. For microscopic analysis, hematoxylin-eosin staining was utilized to assess the inflammatory process; vimentin, for assessment of blood vessels and fibroblasts, and Sirius Red for collagen assessment. RESULTS: In the macroscopic assessment, the use of growth factors resulted in faster healing and decrease of granulation tissue on days seven and 15; (80.31% reduction in the control wound vs. 83.24% in the epithelial wound vs. 100% in the mixed wound). Utilizing microscopy, at the three stages of the experiment, there were no significant differences between the three wounds; however, when comparing the day of euthanization for each type of wound, there was a favorable outcome for epithelial and mixed wounds (between the third vs. 15th day, p < 0.001, and in the comparison of the seventh vs. 15th day; p = 0.002 and p = 0.001 for epithelial and mixed wounds, respectively) with a higher number of fibroblasts, angiogenesis, and collagen type I. CONCLUSION: The use of growth factors accelerates healing, stimulates greater angiogenic activity, and accelerates fibroplasia and collagen maturation.

Resumo Introdução: A cicatrização é um processo de restauração da integridade física das estruturas do corpo. É um processo dinâmico, complexo, multicelular que envolve matriz extracelular, citosinas, células sanguíneas e fatores de crescimento. Os fatores de crescimento são proteínas que estimulam e ativam a proliferação celular mediante a ativação da angiogênese, mitogênese, transcrição genética, acelerando o processo de cicatrização. Objetivo: Avaliar a influência dos fatores de crescimento no processo cicatricial de feridas realizadas no dorso de ratas em comparação com a ferida, controle através da macro e microscopia. Método: Foram utilizadas 45 ratas Wistar, submetidas à criação de três feridas no dorso. A primeira controle a segunda com injeção de fator de crescimento epitelial e a terceira com fator misto. As avaliações macroscópicas e microscópicas foram realizadas no 3º, no 7º e no 15º dia do experimento. Para análise microscópica, utilizou-se coloração de Hematoxilina-Eosina para avaliar o processo inflamatório; vimentina, para a avaliação dos vasos e fibroblastos, e Sirius Red, para avaliar o colágeno. Resultados: Na avaliação macroscópica, o uso de fatores de crescimento proporcionou cicatrização mais rápida e diminuição do tecido de granulação no 7º e 15º dia (80,31% de redução na ferida controle vs. 83,24% na ferida epitelial vs. 100% na ferida mista). Na microscopia, nos três momentos do experimento, não foram encontradas diferenças significativas entre as três feridas; entretanto, quando comparados os dias de morte em relação a cada tipo de ferida, observou-se resultado favorável para as feridas epiteliais e mistas (entre 3º × 15º dia apresentou p < 0,001 e na comparação entre 7º × 15º dias; p = 0,002 e p = 0,001 para as feridas epiteliais e mistas) com maior número de fibroblasto, angiogênese e colágeno tipo 1. Conclusão: A utilização de fatores de crescimento acelera a cicatrização, estimula maior atividade angiogênica, acelera a fibroplasia e maturação do colágeno.

Influence of phenolic compounds of Kangra tea [Camellia sinensis (L) O Kuntze] on bacterial pathogens and indigenous bacterial probiotics of Western Himalayas

Phenolic compounds of nutraceutical importance viz., catechins (C), (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG) were estimated in fresh green tea shoots of Camellia sinensis (L) O Kuntze cultivar. The total polyphenols and total catechins were in the range of 219.90 to 317.81 and 140.83 to 271.39 g/kg, respectively in monthly samples of tea. The values of C, EC, EGC, EGCG and ECG in tea powders as analyzed through high performance liquid chromatography (HPLC) were in the range of 1.560 to 3.661, 13.338 to 27.766, 26.515 to 39.597, 62.903 to 102.168 and 18.969 to 39.469 mg/g, respectively. Effect of tea extracts and standard flavanols against five pathogenic bacteria viz., Listeria monocytogenes (MTCC-839), Pseudomonas aeruginosa (MTCC-741), Bacillus cereus (MTCC-1272), Staphylococcus aureus (MTCC-96) and Escherichia coli (MTCC-443), and eleven indigenous potential bacterial probiotics belonging to genera Enterococcus, Bacillus and Lactobacillus spp. obtained from fermented foods of Western Himalayas, was investigated. EGCG, ECG and EGC exhibited antibacterial activity but, C and EC did not show this activity. Tea extracts having high concentrations of EGCG and ECG were more potent in antibacterial action against bacterial pathogens. Tea extracts and standard flavan-3-ols augmented viability of potential probiotics in an order of EGCG > EGC > ECG > EC > C. Tea extracts and standard flavanols had no antibacterial activity against Escherichia coli (MTCC-443) but, in combination with probiotic culture supernatants, this activity was seen. The Kangra tea thus, exerts antibacterial effect on bacterial pathogens through EGCG, ECG and EGC constituents while stimulatory effect on growth of indigenous potential probiotics.

Individual Variation in Growth Factor Concentrations in Platelet-rich Plasma and Its Influence on Human Mesenchymal Stem Cells / 대한진단검사의학회지

BACKGROUND: The objective of this study was to explore whether individual variations in the concentration of growth factors (GFs) influence the biologic effects of platelet-rich plasma (PRP) on human mesenchymal stem cells (HMSCs). METHODS: The concentrations of 7 representative GFs in activated PRP (aPRP) were measured using ELISA. The effects of PRP on the proliferation and alkaline phosphatase (ALP) activity of HMSCs were examined using several concentrations of aPRP from 3 donors; the relationships between the GF levels and these biologic effects were then evaluated using 10% aPRP from 5 subgroups derived from 39 total donors. HMSCs were cultured in DMEM with the addition of aPRP for 4 or 12 days; then, DNA content and ALP activity were measured. RESULTS: The quantity of DNA increased significantly at a 10% concentration of aPRP, but the ALP activity was suppressed at this concentration of aPRP. The GF concentrations varied among donors, and 5 subgroups of characteristic GF release patterns were identified via cluster analysis. DNA levels differed significantly between groups and tended to be higher in groups with higher concentrations of transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factors (PDGFs). DNA quantity was positively correlated with TGF-beta1 concentration, and was negatively correlated with donor age. ALP activity was negatively correlated with PDGF-BB concentration. CONCLUSIONS: The varying GF concentrations may result in different biologic effects; thus, individual differences in GF levels should be considered for reliable interpretation of the biologic functions and standardized application of PRP.

Neurotoxicity Screening in a Multipotent Neural Stem Cell Line Established from the Mouse Brain

Neural stem cells (NSCs) have mainly been applied to neurodegeneration in some medically intractable neurologic diseases. In this study, we established a novel NSC line and investigated the cytotoxic responses of NSCs to exogenous neurotoxicants, glutamates and reactive oxygen species (ROS). A multipotent NSC line, B2A1 cells, was established from long-term primary cultures of oligodendrocyte-enriched cells from an adult BALB/c mouse brain. B2A1 cells could be differentiated into neuronal, astrocytic and oligodendroglial lineages. The cells also expressed genotypic mRNA messages for both neural progenitor cells and differentiated neuronoglial cells. B2A1 cells treated with hydrogen peroxide and L-buthionine-(S,R)-sulfoximine underwent 30-40% cell death, while B2A1 cells treated with glutamate and kainate showed 25-35% cell death. Cytopathologic changes consisting of swollen cell bodies, loss of cytoplasmic processes, and nuclear chromatin disintegration, developed after exposure to both ROS and excitotoxic chemicals. These results suggest that B2A1 cells may be useful in the study of NSC biology and may constitute an effective neurotoxicity screening system for ROS and excitotoxic chemicals.

Role of protease inhibitors and acylation stimulating protein in the adipogenesis in 3T3-L1 cells

Treatment of AIDS (HIV) and hepatitis C virus needs protease inhibitors (PI) to prevent viral replication. Uses of PI in therapy are usually associated with a decrease in body weight and dyslipidemia. Acylation stimulating protein (ASP) is a protein synthesized in adipocytes to increase triglycerides biosynthesis, for that the relation of PI and ASP to adipogenesis is tested in this work. ASP expression was increased during 3T3-L1 differentiation and reached a peak at day 8 with cell maturation. Addition of PI during adipocytes differentiation dose dependently and significantly (p < 0.5) inhibited the degree of triglycerides (TG) accumulation. Moreover, presence of ASP (450 ng/mL) in media significantly (p < 0.5) stimulated the degree of TG accumulation and there was additive stimulation for ASP when added with insulin (10 microgram/mL). Finally, when ASP in different doses (Low, 16.7; Medium, 45 and High, 450 ng/mL) incubated with a dose of x150 PI, ASP partially inhibited the PI-inhibited adipogenesis and TG accumulation. The results in this study show that PI inhibit lipids accumulation and confirm role of ASP in TG biosynthesis and adipogenesis.

Regulatory effect of estrogen, progestin and HB-EGF on the expression of HOXA10 gene in Ishikawa cells / 华中科技大学学报（医学）（英德文版）

HOXA10 gene plays an essential role in differentiation of the endometrium and in human reproduction. The aim of this study was to investigate the regulatory effect of sex steroids and HB-EGF on HOXA10 gene in Ishikawa cells. Ishikawa cells were incubated with 17-beta estradiol (10(-8) mol/L), medroxyprogesterone acetate (MPA) (10(-6) mol/L), RU486 (10(-5) mol/L) or HB-EGF (10 ng/mL) for 48 h respectively. The expression of HOXA10 gene was detected by immunofluorescence, reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. Our results showed that either estrogen alone, progestin alone or progestin combined with estrogen could significantly increase the expression of HOXA10 gene 48 h after the treatment (P<0.05). But estrogen combined with progestin and RU486 could inhibit the up-regulation by estrogen and progestin. HB-EGF could elevate the expression of HOXA10 gene 48 h after the treatment (P<0.05). It is concluded that both estrogen and progestin can up-regulate the expression of HOXA10 gene in Ishikawa cells, but RU486 can inhibit the effect and HB-EGF can elevate the expression level of HOXA10 gene.

The effect of connective tissue growth factor on human renal tubular epithelial cell transdifferentiation / 华中科技大学学报（医学）（英德文版）

To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of alpha-smooth muscle actin (alpha-SMA) were assessed by indirect immuno-fluorescence, and the percentage of alpha-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of alpha-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of alpha-SMA were markedly stronger than that in negative controls. The percentages of alpha-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9%, 65.5% vs 2.4%, P<0.01). alpha-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).