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Paolo Bellavite - One of the best experts on this subject based on the ideXlab platform.
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Fibronectin Gene Up-regulation by Arnica montana in Human Macrophages: Validation by Real-Time Polymerase Chain Reaction Assay.
Homeopathy : the journal of the Faculty of Homeopathy, 2020Co-Authors: Marta Marzotto, Fabio Arruda-silva, Paolo BellaviteAbstract:Background and Aim Arnica montana L. (Arnica m.) is a popular traditional medicine, used for its therapeutic properties in healing traumas, but little is known about its biological action on tissue formation and repair. This new work tested the effects of Arnica m. homeopathic dilutions on human macrophages, key cells in tissue defence and repair. Materials and Methods Macrophages derived from the THP-1 cell line were differentiated with interleukin-4 to induce a ‘wound-healing’-like phenotype, and treated with various dilutions of Arnica m. centesimal (100 times) dilutions (2c, 3c, 5c, 9c, and 15c) or control solvent for 24 hours. RNA samples from cultured cells were analysed by real-time quantitative polymerase chain reaction in five separate experiments. Results Arnica montana at the 2c dilution (final concentration of sesquiterpene lactones in cell culture = 10−8 mol/L) significantly stimulated the expression of three genes which code for regulatory proteins of the extracellular matrix, namely FN1 (fibronectin 1, % increase of 21.8 ± standard error of the mean 4.6), low-density lipoprotein-receptor-related protein 1 (% increase of 33.4 ± 6.1) and heparan sulphate proteoglycan 2 (% increase of 21.6 ± 9.1). Among these genes, the most quantitatively expressed was FN1. In addition, FN1, unlike other candidate genes, was upregulated in cells treated with higher dilutions/dynamisations (3c, 5c, and 15c) of Arnica m. Conclusion The results support evidence that the extracellular matrix is a potential therapeutic target of Arnica m., with positive effects on cell adhesion and migration during tissue development and healing.
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Arnica montana Stimulates Extracellular Matrix Gene Expression in a Macrophage Cell Line Differentiated to Wound-Healing Phenotype
PloS one, 2016Co-Authors: Marta Marzotto, Clara Bonafini, Debora Olioso, Anna Baruzzi, Laura Bettinetti, Francesca Di Leva, Elisabetta Galbiati, Paolo BellaviteAbstract:Arnica montana (Arnica m.) is used for its purported anti-inflammatory and tissue healing actions after trauma, bruises, or tissue injuries, but its cellular and molecular mechanisms are largely unknown. This work tested Arnica m. effects on gene expression using an in vitro model of macrophages polarized towards a "wound-healing" phenotype. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. centesimal (c) dilutions 2c, 3c, 5c, 9c, 15c or Control. Total RNA was isolated and cDNA libraries were sequenced with a NextSeq500 sequencer. Genes with significantly positive (up-regulated) or negative (down-regulated) fold changes were defined as differentially expressed genes (DEGs). A total of 20 DEGs were identified in Arnica m. 2c treated cells. Of these, 7 genes were up-regulated and 13 were down-regulated. The most significantly up-regulated function concerned 4 genes with a conserved site of epidermal growth factor-like region (p
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Arnica montana stimulates extracellular matrix gene expression in a macrophage cell line differentiated to wound healing phenotype
PLOS ONE, 2016Co-Authors: Marta Marzotto, Clara Bonafini, Debora Olioso, Anna Baruzzi, Laura Bettinetti, Francesca Di Leva, Elisabetta Galbiati, Paolo BellaviteAbstract:Arnica montana (Arnica m.) is used for its purported anti-inflammatory and tissue healing actions after trauma, bruises, or tissue injuries, but its cellular and molecular mechanisms are largely unknown. This work tested Arnica m. effects on gene expression using an in vitro model of macrophages polarized towards a "wound-healing" phenotype. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. centesimal (c) dilutions 2c, 3c, 5c, 9c, 15c or Control. Total RNA was isolated and cDNA libraries were sequenced with a NextSeq500 sequencer. Genes with significantly positive (up-regulated) or negative (down-regulated) fold changes were defined as differentially expressed genes (DEGs). A total of 20 DEGs were identified in Arnica m. 2c treated cells. Of these, 7 genes were up-regulated and 13 were down-regulated. The most significantly up-regulated function concerned 4 genes with a conserved site of epidermal growth factor-like region (p<0.001) and three genes of proteinaceous extracellular matrix, including heparin sulphate proteoglycan 2 (HSPG2), fibrillin 2 (FBN2), and fibronectin (FN1) (p<0.01). Protein assay confirmed a statistically significant increase of fibronectin production (p<0.05). The down-regulated transcripts derived from mitochondrial genes coding for some components of electron transport chain. The same groups of genes were also regulated by increasing dilutions of Arnica m. (3c, 5c, 9c, 15c), although with a lower effect size. We further tested the healing potential of Arnica m. 2c in a scratch model of wound closure based on the motility of bone marrow-derived macrophages and found evidence of an accelerating effect on cell migration in this system. The results of this work, taken together, provide new insights into the action of Arnica m. in tissue healing and repair, and identify extracellular matrix regulation by macrophages as a therapeutic target.
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response to commentary Arnica montana effects on gene expression in a human macrophage cell line evaluation by quantitative real time pcr
Frontiers in Immunology, 2016Co-Authors: Debora Olioso, Clara Bonafini, Marta Marzotto, Maurizio Brizzi, Paolo BellaviteAbstract:We recently investigated the effects of Arnica montana (Arnica) on the THP-1 myelomonocytic cell line, differentiated by phorbol-myristate acetate and IL-4 in the wound healing phenotype (M2) (1). Our study was the object of a commentary by Chirumbolo and Bjorklund (2), based on some recalculations and on their extrapolations from the values of SEM. Even though we had correctly reported the dosage of sesquiterpenes – reference substances for Arnica pharmacopeia – the authors inferred that this information was insufficient and they recalculated the doses in terms of helenalin: “in the starting 30% alcoholic preparation of Arnica (1c), a percentage of 0.036% sesquiterpene lactones, should correspond to 0.72 μg, i.e., 72 ng in the dilution 2c.” This calculation is flawed because our dose was referred to the mother tincture and not to the 1c dilution. Moreover, helenalin is not the only active ingredient in Arnica, which contains many other substances (3, 4). Finally, 0.72 μg in 1c are not 72 ng in 2c but 7.2 ng.
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Effectiveness and Safety of Arnica montana in Post-Surgical Setting, Pain and Inflammation.
American journal of therapeutics, 2016Co-Authors: Tommaso Iannitti, Paolo Bellavite, Julio Cesar Morales-medina, Valentina Rottigni, Beniamino PalmieriAbstract:Arnica montana has been widely used as a homeopathic remedy for the treatment of several inflammatory conditions in pain management and postoperative settings. This review gives an overview of the therapeutic use of Arnica montana in the above-mentioned fields also focusing on its mechanisms of action learned from animal models and in vitro studies. Arnica montana is more effective than placebo when used for the treatment of several conditions including post-traumatic and postoperative pain, edema, and ecchymosis. However, its dosages and preparations used have produced substantial differences in the clinical outcome. Cumulative evidence suggests that Arnica montana may represent a valid alternative to non-steroidal anti-inflammatory drugs, at least when treating some specific conditions.
Marta Marzotto - One of the best experts on this subject based on the ideXlab platform.
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Fibronectin Gene Up-regulation by Arnica montana in Human Macrophages: Validation by Real-Time Polymerase Chain Reaction Assay.
Homeopathy : the journal of the Faculty of Homeopathy, 2020Co-Authors: Marta Marzotto, Fabio Arruda-silva, Paolo BellaviteAbstract:Background and Aim Arnica montana L. (Arnica m.) is a popular traditional medicine, used for its therapeutic properties in healing traumas, but little is known about its biological action on tissue formation and repair. This new work tested the effects of Arnica m. homeopathic dilutions on human macrophages, key cells in tissue defence and repair. Materials and Methods Macrophages derived from the THP-1 cell line were differentiated with interleukin-4 to induce a ‘wound-healing’-like phenotype, and treated with various dilutions of Arnica m. centesimal (100 times) dilutions (2c, 3c, 5c, 9c, and 15c) or control solvent for 24 hours. RNA samples from cultured cells were analysed by real-time quantitative polymerase chain reaction in five separate experiments. Results Arnica montana at the 2c dilution (final concentration of sesquiterpene lactones in cell culture = 10−8 mol/L) significantly stimulated the expression of three genes which code for regulatory proteins of the extracellular matrix, namely FN1 (fibronectin 1, % increase of 21.8 ± standard error of the mean 4.6), low-density lipoprotein-receptor-related protein 1 (% increase of 33.4 ± 6.1) and heparan sulphate proteoglycan 2 (% increase of 21.6 ± 9.1). Among these genes, the most quantitatively expressed was FN1. In addition, FN1, unlike other candidate genes, was upregulated in cells treated with higher dilutions/dynamisations (3c, 5c, and 15c) of Arnica m. Conclusion The results support evidence that the extracellular matrix is a potential therapeutic target of Arnica m., with positive effects on cell adhesion and migration during tissue development and healing.
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Arnica montana Stimulates Extracellular Matrix Gene Expression in a Macrophage Cell Line Differentiated to Wound-Healing Phenotype
PloS one, 2016Co-Authors: Marta Marzotto, Clara Bonafini, Debora Olioso, Anna Baruzzi, Laura Bettinetti, Francesca Di Leva, Elisabetta Galbiati, Paolo BellaviteAbstract:Arnica montana (Arnica m.) is used for its purported anti-inflammatory and tissue healing actions after trauma, bruises, or tissue injuries, but its cellular and molecular mechanisms are largely unknown. This work tested Arnica m. effects on gene expression using an in vitro model of macrophages polarized towards a "wound-healing" phenotype. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. centesimal (c) dilutions 2c, 3c, 5c, 9c, 15c or Control. Total RNA was isolated and cDNA libraries were sequenced with a NextSeq500 sequencer. Genes with significantly positive (up-regulated) or negative (down-regulated) fold changes were defined as differentially expressed genes (DEGs). A total of 20 DEGs were identified in Arnica m. 2c treated cells. Of these, 7 genes were up-regulated and 13 were down-regulated. The most significantly up-regulated function concerned 4 genes with a conserved site of epidermal growth factor-like region (p
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Arnica montana stimulates extracellular matrix gene expression in a macrophage cell line differentiated to wound healing phenotype
PLOS ONE, 2016Co-Authors: Marta Marzotto, Clara Bonafini, Debora Olioso, Anna Baruzzi, Laura Bettinetti, Francesca Di Leva, Elisabetta Galbiati, Paolo BellaviteAbstract:Arnica montana (Arnica m.) is used for its purported anti-inflammatory and tissue healing actions after trauma, bruises, or tissue injuries, but its cellular and molecular mechanisms are largely unknown. This work tested Arnica m. effects on gene expression using an in vitro model of macrophages polarized towards a "wound-healing" phenotype. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. centesimal (c) dilutions 2c, 3c, 5c, 9c, 15c or Control. Total RNA was isolated and cDNA libraries were sequenced with a NextSeq500 sequencer. Genes with significantly positive (up-regulated) or negative (down-regulated) fold changes were defined as differentially expressed genes (DEGs). A total of 20 DEGs were identified in Arnica m. 2c treated cells. Of these, 7 genes were up-regulated and 13 were down-regulated. The most significantly up-regulated function concerned 4 genes with a conserved site of epidermal growth factor-like region (p<0.001) and three genes of proteinaceous extracellular matrix, including heparin sulphate proteoglycan 2 (HSPG2), fibrillin 2 (FBN2), and fibronectin (FN1) (p<0.01). Protein assay confirmed a statistically significant increase of fibronectin production (p<0.05). The down-regulated transcripts derived from mitochondrial genes coding for some components of electron transport chain. The same groups of genes were also regulated by increasing dilutions of Arnica m. (3c, 5c, 9c, 15c), although with a lower effect size. We further tested the healing potential of Arnica m. 2c in a scratch model of wound closure based on the motility of bone marrow-derived macrophages and found evidence of an accelerating effect on cell migration in this system. The results of this work, taken together, provide new insights into the action of Arnica m. in tissue healing and repair, and identify extracellular matrix regulation by macrophages as a therapeutic target.
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response to commentary Arnica montana effects on gene expression in a human macrophage cell line evaluation by quantitative real time pcr
Frontiers in Immunology, 2016Co-Authors: Debora Olioso, Clara Bonafini, Marta Marzotto, Maurizio Brizzi, Paolo BellaviteAbstract:We recently investigated the effects of Arnica montana (Arnica) on the THP-1 myelomonocytic cell line, differentiated by phorbol-myristate acetate and IL-4 in the wound healing phenotype (M2) (1). Our study was the object of a commentary by Chirumbolo and Bjorklund (2), based on some recalculations and on their extrapolations from the values of SEM. Even though we had correctly reported the dosage of sesquiterpenes – reference substances for Arnica pharmacopeia – the authors inferred that this information was insufficient and they recalculated the doses in terms of helenalin: “in the starting 30% alcoholic preparation of Arnica (1c), a percentage of 0.036% sesquiterpene lactones, should correspond to 0.72 μg, i.e., 72 ng in the dilution 2c.” This calculation is flawed because our dose was referred to the mother tincture and not to the 1c dilution. Moreover, helenalin is not the only active ingredient in Arnica, which contains many other substances (3, 4). Finally, 0.72 μg in 1c are not 72 ng in 2c but 7.2 ng.
Ary Gomes Da Silva - One of the best experts on this subject based on the ideXlab platform.
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a clinical trial with brazilian Arnica solidago chilensis meyen glycolic extract in the treatment of tendonitis of flexor and extensor tendons of wrist and hand
Phytotherapy Research, 2015Co-Authors: Ary Gomes Da Silva, Elbe Rodrigues Machado, Leonardo Mendes De Almeida, Ricardo Marcelo Menezes Nunes, Patricia Caldeira Pena Giesbrecht, Regina Mamed Costa, Helber B Costa, Wanderson Romao, Ricardo Machado KusterAbstract:One of the Brazilian Arnicas, Solidago chilensis Meyen, is a species of the Asteraceae family. This plant is known by this common name because it shares remarkably similar organoleptic properties with the genus Arnica L., also within the family Asteraceae. We examined the effectiveness of the S. chilensis fluid extract used externally for treating tendinitis of flexor and extensor tendons of wrist and hand in placebo-controlled double-blind clinical pharmacological studies. This study was approved by the Ethical Committee for Scientific Research in Human Beings at University Vila Velha-UVV. Two daily skin applications on the arm skin of a gel cream containing a 5% glycolic plant extract were administered to eight volunteers for 21 days. Among the volunteers, one of their arms was used as the placebo group, and the other one was used as a test group. Statistical data analyses demonstrated a significant reduction in the perception of pain in the arms in the test group, when it was compared to those receiving only the placebo. Copyright © 2015 John Wiley & Sons, Ltd.
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evaluation of an extract of brazilian Arnica solidago chilensis meyen asteraceae in treating lumbago
Phytotherapy Research, 2010Co-Authors: Ary Gomes Da Silva, Carlos P G De Sousa, Jaeder Koehler, Joelder Fontana, Alexandre Gabriel Christo, Rejan R GuedesbruniAbstract:One of the Brazilian Arnicas, Solidago chilensis Meyen, is a species of the family Asteraceae. This plant is known by this common name because it shares very similar organoleptic properties with the genus Arnica L., also within the family Asteraceae, that comprises approximately 30 European species of perennial, herbaceous plants. The effectiveness of a fluid extract of S. chilensis used externally for treating lumbago was examined in placebo-controlled double-blind clinical pharmacological studies. Two daily skin applications of a gel containing a 5% extract in glycol were administered for 15 days to ten volunteers in a placebo group and to an equal number in a test group. Statistical analyses of the results demonstrated a significant reduction in the perception of pain and a significant increase in the flexibility of patients in the test group as compared with those receiving only the placebo.
Debora Olioso - One of the best experts on this subject based on the ideXlab platform.
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Arnica montana Stimulates Extracellular Matrix Gene Expression in a Macrophage Cell Line Differentiated to Wound-Healing Phenotype
PloS one, 2016Co-Authors: Marta Marzotto, Clara Bonafini, Debora Olioso, Anna Baruzzi, Laura Bettinetti, Francesca Di Leva, Elisabetta Galbiati, Paolo BellaviteAbstract:Arnica montana (Arnica m.) is used for its purported anti-inflammatory and tissue healing actions after trauma, bruises, or tissue injuries, but its cellular and molecular mechanisms are largely unknown. This work tested Arnica m. effects on gene expression using an in vitro model of macrophages polarized towards a "wound-healing" phenotype. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. centesimal (c) dilutions 2c, 3c, 5c, 9c, 15c or Control. Total RNA was isolated and cDNA libraries were sequenced with a NextSeq500 sequencer. Genes with significantly positive (up-regulated) or negative (down-regulated) fold changes were defined as differentially expressed genes (DEGs). A total of 20 DEGs were identified in Arnica m. 2c treated cells. Of these, 7 genes were up-regulated and 13 were down-regulated. The most significantly up-regulated function concerned 4 genes with a conserved site of epidermal growth factor-like region (p
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Arnica montana stimulates extracellular matrix gene expression in a macrophage cell line differentiated to wound healing phenotype
PLOS ONE, 2016Co-Authors: Marta Marzotto, Clara Bonafini, Debora Olioso, Anna Baruzzi, Laura Bettinetti, Francesca Di Leva, Elisabetta Galbiati, Paolo BellaviteAbstract:Arnica montana (Arnica m.) is used for its purported anti-inflammatory and tissue healing actions after trauma, bruises, or tissue injuries, but its cellular and molecular mechanisms are largely unknown. This work tested Arnica m. effects on gene expression using an in vitro model of macrophages polarized towards a "wound-healing" phenotype. The monocyte-macrophage human THP-1 cell line was cultured and differentiated with phorbol-myristate acetate and Interleukin-4, then exposed for 24h to Arnica m. centesimal (c) dilutions 2c, 3c, 5c, 9c, 15c or Control. Total RNA was isolated and cDNA libraries were sequenced with a NextSeq500 sequencer. Genes with significantly positive (up-regulated) or negative (down-regulated) fold changes were defined as differentially expressed genes (DEGs). A total of 20 DEGs were identified in Arnica m. 2c treated cells. Of these, 7 genes were up-regulated and 13 were down-regulated. The most significantly up-regulated function concerned 4 genes with a conserved site of epidermal growth factor-like region (p<0.001) and three genes of proteinaceous extracellular matrix, including heparin sulphate proteoglycan 2 (HSPG2), fibrillin 2 (FBN2), and fibronectin (FN1) (p<0.01). Protein assay confirmed a statistically significant increase of fibronectin production (p<0.05). The down-regulated transcripts derived from mitochondrial genes coding for some components of electron transport chain. The same groups of genes were also regulated by increasing dilutions of Arnica m. (3c, 5c, 9c, 15c), although with a lower effect size. We further tested the healing potential of Arnica m. 2c in a scratch model of wound closure based on the motility of bone marrow-derived macrophages and found evidence of an accelerating effect on cell migration in this system. The results of this work, taken together, provide new insights into the action of Arnica m. in tissue healing and repair, and identify extracellular matrix regulation by macrophages as a therapeutic target.
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response to commentary Arnica montana effects on gene expression in a human macrophage cell line evaluation by quantitative real time pcr
Frontiers in Immunology, 2016Co-Authors: Debora Olioso, Clara Bonafini, Marta Marzotto, Maurizio Brizzi, Paolo BellaviteAbstract:We recently investigated the effects of Arnica montana (Arnica) on the THP-1 myelomonocytic cell line, differentiated by phorbol-myristate acetate and IL-4 in the wound healing phenotype (M2) (1). Our study was the object of a commentary by Chirumbolo and Bjorklund (2), based on some recalculations and on their extrapolations from the values of SEM. Even though we had correctly reported the dosage of sesquiterpenes – reference substances for Arnica pharmacopeia – the authors inferred that this information was insufficient and they recalculated the doses in terms of helenalin: “in the starting 30% alcoholic preparation of Arnica (1c), a percentage of 0.036% sesquiterpene lactones, should correspond to 0.72 μg, i.e., 72 ng in the dilution 2c.” This calculation is flawed because our dose was referred to the mother tincture and not to the 1c dilution. Moreover, helenalin is not the only active ingredient in Arnica, which contains many other substances (3, 4). Finally, 0.72 μg in 1c are not 72 ng in 2c but 7.2 ng.
Ricardo Machado Kuster - One of the best experts on this subject based on the ideXlab platform.
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a clinical trial with brazilian Arnica solidago chilensis meyen glycolic extract in the treatment of tendonitis of flexor and extensor tendons of wrist and hand
Phytotherapy Research, 2015Co-Authors: Ary Gomes Da Silva, Elbe Rodrigues Machado, Leonardo Mendes De Almeida, Ricardo Marcelo Menezes Nunes, Patricia Caldeira Pena Giesbrecht, Regina Mamed Costa, Helber B Costa, Wanderson Romao, Ricardo Machado KusterAbstract:One of the Brazilian Arnicas, Solidago chilensis Meyen, is a species of the Asteraceae family. This plant is known by this common name because it shares remarkably similar organoleptic properties with the genus Arnica L., also within the family Asteraceae. We examined the effectiveness of the S. chilensis fluid extract used externally for treating tendinitis of flexor and extensor tendons of wrist and hand in placebo-controlled double-blind clinical pharmacological studies. This study was approved by the Ethical Committee for Scientific Research in Human Beings at University Vila Velha-UVV. Two daily skin applications on the arm skin of a gel cream containing a 5% glycolic plant extract were administered to eight volunteers for 21 days. Among the volunteers, one of their arms was used as the placebo group, and the other one was used as a test group. Statistical data analyses demonstrated a significant reduction in the perception of pain in the arms in the test group, when it was compared to those receiving only the placebo. Copyright © 2015 John Wiley & Sons, Ltd.