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1.
Braz Oral Res ; 33: e092, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31576904

RESUMO

This study evaluated the effect of antimicrobial photodynamic therapy (aPDT) on the endodontic treatment of apical periodontitis (AP). AP was induced in 48 premolars of 6 dogs. After biomechanical preparation, the teeth were divided into 4 groups: Calcium-Hydroxide (CH)/120d and CH/180d: root canals filled with CH-based dressing for 15 days before obturation; aPDT/120d and aPDT/180d: conditioning with phenothiazine photosensitizer (10 mg/mL) for 1 minute and irradiation with diode laser in the same session as obturation. Root filling was performed with AH Plus sealer. After the experimental periods, animals were euthanized and teeth were submitted for histology. HE staining was performed for descriptive analysis of the periapical region, measurement of apical periodontitis and for inflammatory cells, and blood vessels count. Immunohistochemistry was performed for osteopontin (OPN) and alkaline phosphatase (ALP). Data were analyzed statistically by two-way ANOVA and chi-square test (α = 5%). Teeth in Group CH/120d presented only a slightly enlarged periodontal ligament (PL) with advanced repair. Group aPDT/120d presented the PL moderately enlarged, with moderate inflammatory infiltrate and few collagen fibers. The same pattern was observed at 180 days. AP lesions in CH-treated groups were smaller than those in aPDT-treated groups (p < 0.001) with more blood vessels (p < 0.0001), regardless of the evaluation period, without significant differences in the number of inflammatory cells (p > 0.05). CH-treated groups showed significantly more intense immunostaining for ALP and OPN (p < 0.001) in both periods. Although aPDT stimulated angiogenesis and expression of bone formation markers, the two-session endodontic treatment with CH-based dressing promoted better apical periodontitis repair.


Assuntos
Cimentos para Ossos/uso terapêutico , Hidróxido de Cálcio/uso terapêutico , Periodontite Periapical/tratamento farmacológico , Fotoquimioterapia/métodos , Tratamento do Canal Radicular/métodos , Animais , Vasos Sanguíneos/efeitos dos fármacos , Regeneração Óssea/efeitos dos fármacos , Imuno-Histoquímica , Neovascularização Fisiológica/efeitos dos fármacos , Periodontite Periapical/patologia , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do Tratamento
2.
Chem Biodivers ; 16(8): e1900232, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31287621

RESUMO

Vascular endothelial growth factor receptor-2 (VEGFR-2) plays an important role in both vasculogenesis and angiogenesis. Inhibition of VEGFR-2 has been demonstrated as a key method against tumor-associated angiogenesis. Thiazolopyrimidine is an important analog of the purine ring, and we choose the thiazolopyrimidine scaffold as the mother nucleus. Two series of thiazolo[5,4-d]pyrimidine derivatives were synthesized and evaluated for their antiproliferative activity. In HUVEC inhibition assay, compounds 3l (=1-(5-{[2-(4-chlorophenyl)-5-methyl[1,3]thiazolo[5,4-d]pyrimidin-7-yl]amino}pyridin-2-yl)-3-(3,4-dimethylphenyl)urea) and 3m (=1-(5-{[2-(4-chlorophenyl)-5-methyl[1,3]thiazolo[5,4-d]pyrimidin-7-yl]amino}pyridin-2-yl)-3-(4-methoxyphenyl)urea) exhibited the most potent inhibitory effect (IC50 =1.65 and 3.52 µm, respectively). Compound 3l also showed the best potency against VEGFR-2 at 50 µm (98.5 %). These results suggest that further investigation of compound 3l might provide potential angiogenesis inhibitors.


Assuntos
Inibidores da Angiogênese/síntese química , Desenho de Drogas , Pirimidinas/química , Tiazóis/química , Inibidores da Angiogênese/química , Inibidores da Angiogênese/farmacologia , Proliferação de Células/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana , Humanos , Neovascularização Fisiológica/efeitos dos fármacos , Pirimidinas/farmacologia , Relação Estrutura-Atividade , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
3.
Int J Nanomedicine ; 14: 3345-3360, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31190796

RESUMO

Background: Designing a wound dressing that effectively prevents multi-drug-resistant bacterial infection and promotes angiogenesis and re-epithelialization is of great significance for wound management. Methods and results: In this study, a biocompatible composite membrane comprising biomimetic polydopamine-modified eggshell membrane nano/microfibres coated with KR-12 antimicrobial peptide and hyaluronic acid (HA) was developed in an eco-friendly manner. The physicochemical properties of the composite membrane were thoroughly characterized, and the results showed that the surface hydrophilicity and water absorption ability of the composite membrane were improved after the successive conjugation of the HA and the KR-12 peptide. Furthermore, the in vitrobiological results revealed that the composite membrane had excellent antibacterial activity against Gram-positive Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA) and Gram-negative Escherichia coli, and it could prevent MRSA biofilm formation on its surface. Additionally, it promoted the proliferation of keratinocytes and human umbilical vein endothelial cells and increased the secretion of VEGF. Finally, an in vivo animal study indicated that the composite membrane could promote wound healing via accelerating angiogenesis and re-epithelialization, which were demonstrated by the enhanced expression of angiogenetic markers (CD31 and VEGF) and keratinocyte proliferation marker (PCNA), respectively. Conclusion: These results indicated that the composite membrane is a potential candidate of wound dressings.


Assuntos
Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Casca de Ovo/química , Ácido Hialurônico/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Peptídeos/farmacologia , Reepitelização/efeitos dos fármacos , Animais , Antibacterianos/farmacologia , Galinhas , Escherichia coli/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Camundongos , Testes de Sensibilidade Microbiana , Peptídeos/química , Porosidade , Staphylococcus aureus/efeitos dos fármacos
4.
BMC Complement Altern Med ; 19(1): 115, 2019 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-31159783

RESUMO

BACKGROUND: Skin burn wound is a notable medical burden worldwide. Rapid and effective treatment of burnt skin is vital to fasten wound closure and healing properly. Amniotic graft and Aloe vera are widely used as wound managing biomaterials. Sophisticated processing, high cost, availability, and the requirement of medics for transplantation limit the application of amnion grafts. We aim to prepare a novel gel from amnion combined with the Aloe vera extract for burn wound healing which overcome the limitations of graft. METHODS: Two percent human amniotic membrane (AM), Aloe vera (AV) and AM+AV gels were prepared. In vitro cytotoxicity, biocompatibility, cell attachment, proliferation, wound healing scratch assays were performed in presence of the distinct gels. After skin irritation study, second-degree burns were induced on dorsal region of Wistar rats; and gels were applied to observe the healing potential in vivo. Besides, macroscopical measurement of wound contraction and re-epithelialization; gel treated skin was histologically investigated by Hematoxylin and eosin (H&E) staining. Finally, quantitative assessment of angiogenesis, inflammation, and epithelialization was done. RESULTS: The gels were tested to be non-cytotoxic to nauplii and compatible with human blood and skin cells. Media containing 500 µg/mL AM+AV gel were observed to promote HaCaT and HFF1 cells attachment and proliferation. In vitro scratch assay demonstrated that AM+AV significantly accelerated wound closure through migration of HaCaT cells. No erythema and edema were observed in skin irritation experiments confirming the applicability of the gels. AV and AM+AV groups showed significantly accelerated wound closure through re-epithelialization and wound contraction with P < 0.01. Macroscopically, AM and AM+AV treated wound recovery rates were 87 and 90% respectively with P < 0.05. Histology analysis revealed significant epitheliazation and angiogenesis in AM+AV treated rats compared to control (P < 0.05). AM+AV treated wounds had thicker regenerated epidermis, increased number of blood vessels, and greater number of proliferating keratinocytes within the epidermis. CONCLUSION: We demonstrated that a gel consisting of a combination of amnion and Aloe vera extract has high efficacy as a burn wound healing product. Amniotic membrane combined with the carrier Aloe vera in gel format is easy to produce and to apply.


Assuntos
Âmnio , Queimaduras/tratamento farmacológico , Preparações de Plantas/uso terapêutico , Animais , Artemia , Linhagem Celular , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , Neovascularização Fisiológica/efeitos dos fármacos , Fitoterapia , Preparações de Plantas/farmacologia , Ratos Wistar , Reepitelização/efeitos dos fármacos
5.
Parasit Vectors ; 12(1): 315, 2019 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-31234915

RESUMO

BACKGROUND: Angiogenesis can occur under pathological conditions when stimuli such as inflammation, vascular obstruction or hypoxia exist. These stimuli are present in cardiopulmonary dirofilariosis (Dirofilaria immitis). The aim of this study was to analyze the capacity of D. immitis antigens to modify the expression of angiogenic factors and trigger the formation of pseudocapillaries (tube-like structures) in an in vitro model of endothelial cells. METHODS: The expression of VEGF-A, sFlt, mEndoglin and sEndoglin in cultures of canine microvascular endothelial cells stimulated with extract of adult worms of D. immitis obtained from an untreated dog (DiSA) and from a dog treated for 15 days with doxycycline (tDiSA), was determined by using commercial kits. The capacity of pseudocapillary formation was evaluated analyzing cell connections and cell groups in Matrigel cell cultures stimulated with DiSA and tDiSA. In both cases non-stimulated cultures were used as controls. RESULTS: First, we demonstrated that worms obtained from the dog treated with doxycycline showed a significantly lower amount of Wolbachia (less than 60%) than worms removed from the untreated dog. Only DiSA was able to significantly increase the expression of the proangiogenic factor VEGF-A in the endotelial cells cultures. None of the D. immitis extracts modified the expression of sFlt. tDiSA extract was able to modify the expression of the endoglins, significantly decreasing the expression of the pro-angiogenic mEndoglin and increasing the anti-angiogenic sEndoglin. The formation of pseudocapillaries was negatively influenced by tDiSA, which reduced the organization and number of cellular connections. CONCLUSIONS: The ability of antigens from adult D. immitis worms to modify the expression of pro and anti-angiogenic factors in endotelial cell cultures was demonstrated, as well as the trend to form pseudocapillaries in vitro. The capacity of stimulation may be linked to the amount of Wolbachia present in the antigenic extracts.


Assuntos
Antígenos de Helmintos/farmacologia , Dirofilaria immitis/química , Células Endoteliais/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Animais , Antígenos de Bactérias/farmacologia , Capilares/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Dirofilaria immitis/microbiologia , Cães , Inflamação , Wolbachia/química , Wolbachia/genética
6.
Transplant Proc ; 51(5): 1458-1462, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31155179

RESUMO

BACKGROUND: Subcutaneous pockets provide an extrahepatic transplant site for islet grafting to treat type 1 diabetes. However, a hypoxic environment may cause central necrosis to islets and lead to graft failure. Our previous studies focused on a pre-treated subcutaneous site with basic fibroblast growth factor (bFGF) for the formation of vascular bed. In addition to neovascularization, bFGF was also shown to protect islets against oxidative stress and chemical-induced damage in vitro. Accordingly, we propose that subcutaneous islet transplantation with a bFGF-slow releasing device simultaneously can improve islet survival in vivo. METHODS: A bFGF-impregnated collagen sheet was implanted in the right back of a streptozotocin-induced diabetic mouse for neovascularization. After 10 days, the sheet was removed and the rat islet-embedding gel within the immune-isolation device was transplanted (2-time operation [OP]). In another group, the diabetic mice received bFGF-impregnated gel with rat islets within the immune-isolation device simultaneously (1-time OP). RESULTS: Diabetic mice in 2-time OP group experienced a decrease in their non-fasting blood glucose level for a period of 10 days, and the glucose levels were lower than those of untreated diabetic mice post-implantation. However, the mice in the 1-time OP group remained hyperglycemic post-operation and showed no improvements in body weight or the area under curve in intraperitoneal glucose tolerance test. Furthermore, mice in the 2-time OP had relatively higher serum insulin levels with improved renal and metabolic biomarkers. CONCLUSION: Our findings suggest that bFGF had no beneficial effect on a 1-time operation in subcutaneous islet transplantation.


Assuntos
Diabetes Mellitus Experimental/cirurgia , Fator 2 de Crescimento de Fibroblastos/administração & dosagem , Transplante das Ilhotas Pancreáticas/instrumentação , Transplante das Ilhotas Pancreáticas/métodos , Tela Subcutânea , Animais , Glicemia , Diabetes Mellitus Experimental/metabolismo , Modelos Animais de Doenças , Xenoenxertos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neovascularização Fisiológica/efeitos dos fármacos , Ratos , Ratos Endogâmicos Lew , Tela Subcutânea/cirurgia
7.
Environ Pollut ; 251: 493-501, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31103009

RESUMO

17α-ethinyl estradiol (EE2) is a synthetic compound widely used in the generation of contraceptive pills. EE2 is present in the urine of women taking contraceptives and its presence has been confirmed at increasing concentrations contaminating rivers all over the world. Because of this cycle, it can entry the human food chain when watering plants. A negative influence of EE2 on fertility and reproductive capacity of wildlife was already suggested. The short-term impact of exposure to contaminating EE2 on pregnancy outcome has not been addressed. Pregnant mice were exposed to either 0.005 µg (concentrations found in water) or 5 µg EE2/kg (contraceptive dose) body weight/day from gestation day 1-7 by oral gavage. Control mice received a 0.1% ethanol solution. High frequency ultrasound imaging was used to follow-up fetal and placental growth in vivo. Doppler measurements were utilized to analyze blood flow parameters in uterine and umbilical arteries. Mice were sacrificed at gd5, 10, and 14. We show that most fetuses of mothers exposed to the high EE2 dose die intrauterine at gd10, with implantation sizes beginning to be smaller already at gd8. Mothers exposed to the low EE2 dose show an impaired remodeling of the spiral arteries, a higher placental weight and pups that are large for gestational age. The insulin-like growth factor system that regulates fetal and placental growth and development is affected by the EE2 treatment. Our results show that a short-term exposure to EE2 during early pregnancy has severe consequences for fetal growth and survival depending on the dose. Exposition to synthetic estrogens affects placenta growth and angiogenesis. These findings urge to the study of mechanisms dysregulated upon environmental exposition to estrogens.


Assuntos
Etinilestradiol/toxicidade , Desenvolvimento Fetal/efeitos dos fármacos , Exposição Materna/efeitos adversos , Poluentes Químicos da Água/toxicidade , Animais , Relação Dose-Resposta a Droga , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Neovascularização Fisiológica/efeitos dos fármacos , Placenta/irrigação sanguínea , Placenta/efeitos dos fármacos , Gravidez , Resultado da Gravidez , Análise de Sobrevida
8.
Mar Drugs ; 17(5)2019 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-31035725

RESUMO

Melanoma is one of the most malignant and aggressive types of cancer worldwide. Fibroblast growth factor 2 (FGF2) is one of the critical regulators of melanoma angiogenesis and metastasis; thus, it might be an effective anti-cancer strategy to explore FGF2-targeting drug candidates from existing drugs. In this study, we evaluate the effect of the marine drug propylene glycol alginate sodium sulfate (PSS) on FGF2-mediated angiogenesis and invasion. The data shows that FGF2 selectively bound to PSS with high affinity. PSS inhibited FGF2-mediated angiogenesis in a rat aortic ring model and suppressed FGF2-mediated invasion, but not the migration of murine melanoma B16-F10 cells. The further mechanism study indicates that PSS decreased the expression of activated matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9), and also suppressed their activity. In addition, PSS was found to decrease the level of Vimentin in B16-F10 cells, which is known to participate in the epithelial-mesenchymal transition. Notably, PSS did not elicit any changes in cancer cell viability. Based on the results above, we conclude that PSS might be a potential drug to regulate the tumor microenvironment in order to facilitate the recovery of melanoma patients.


Assuntos
Alginatos/farmacologia , Fator 2 de Crescimento de Fibroblastos/metabolismo , Melanoma Experimental/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Neoplasias Cutâneas/tratamento farmacológico , Alginatos/uso terapêutico , Animais , Aorta/efeitos dos fármacos , Organismos Aquáticos/química , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Embrião de Galinha , Membrana Corioalantoide , Avaliação Pré-Clínica de Medicamentos , Transição Epitelial-Mesenquimal , Humanos , Laminaria/química , Melanoma Experimental/irrigação sanguínea , Melanoma Experimental/patologia , Camundongos , Invasividade Neoplásica/patologia , Invasividade Neoplásica/prevenção & controle , Neovascularização Patológica/patologia , Neovascularização Fisiológica/efeitos dos fármacos , Técnicas de Cultura de Órgãos , Ratos , Neoplasias Cutâneas/irrigação sanguínea , Neoplasias Cutâneas/patologia , Microambiente Tumoral/efeitos dos fármacos
9.
Int J Mol Sci ; 20(9)2019 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-31071929

RESUMO

(1) The beneficial effects of hydrogen sulfide (H2S) on the cardiovascular and nervous system have recently been re-evaluated. It has been shown that lanthionine, a side product of H2S biosynthesis, previously used as a marker for H2S production, is dramatically increased in circulation in uremia, while H2S release is impaired. Thus, lanthionine could be classified as a novel uremic toxin. Our research was aimed at defining the mechanism(s) for lanthionine toxicity. (2) The effect of lanthionine on H2S release was tested by a novel lead acetate strip test (LAST) in EA.hy926 cell cultures. Effects of glutathione, as a redox agent, were assayed. Levels of sulfane sulfur were evaluated using the SSP4 probe and flow cytometry. Protein content and glutathionylation were analyzed by Western Blotting and immunoprecipitation, respectively. Gene expression and miRNA levels were assessed by qPCR. (3) We demonstrated that, in endothelial cells, lanthionine hampers H2S release; reduces protein content and glutathionylation of transsulfuration enzyme cystathionine-ß-synthase; modifies the expression of miR-200c and miR-423; lowers expression of vascular endothelial growth factor VEGF; increases Ca2+ levels. (4) Lanthionine-induced alterations in cell cultures, which involve both sulfur amino acid metabolism and calcium homeostasis, are consistent with uremic dysfunctional characteristics and further support the uremic toxin role of this amino acid.


Assuntos
Alanina/análogos & derivados , Cálcio/metabolismo , Insuficiência Renal Crônica/tratamento farmacológico , Sulfetos/farmacologia , Uremia/tratamento farmacológico , Alanina/química , Alanina/farmacologia , Aminoácidos Sulfúricos/efeitos dos fármacos , Aminoácidos Sulfúricos/metabolismo , Linhagem Celular , Cistationina beta-Sintase/genética , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Humanos , Sulfeto de Hidrogênio/metabolismo , MicroRNAs/genética , Neovascularização Fisiológica/efeitos dos fármacos , Neovascularização Fisiológica/genética , Oxirredução , Insuficiência Renal Crônica/genética , Insuficiência Renal Crônica/metabolismo , Sulfetos/química , Uremia/genética , Uremia/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética
10.
Chem Biol Interact ; 308: 1-10, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31071337

RESUMO

Diarylheptanoids display an array of biological and pharmacological properties. We previously reported the synthesis of a diarylheptanoid Alpinoid c and a series of its derivatives, evaluated their cytotoxicity against various human cancer cells. We found some of these derivatives were significantly more potent than Alpinoid c in preventing the proliferation of various cancer cell lines. Among these, (S, E)-1-(3, 4 dimethoxyphenyl)-6-hydroxy-7-phenylhept-4-en-3-one (DPHP) showed most potent cytotoxicity against COLO205 cells, however, the mechanism by which DPHP prevents the growth of these colon cancer cells remains unknown. In the current study, we investigated the molecular mechanism of DPHP on colon cancer cells. DPHP inhibited the proliferation of COLO205 (IC50 7.01 ±â€¯0.62 µM) and A549 (IC50 20.03 ±â€¯3.11 µM) cells more specifically than normal human colon epithelial cell line NCM460 (IC50 55.6 ±â€¯4.02 µM). In COLO205 cells, DPHP induced cell shrinkage, membrane blebbing, chromatin condensation, phosphatidylserine externalization, and an accumulation of cells at sub-G1 phase. Further analysis these cells treated with DPHP revealed a decrease in mitochondrial membrane potential, an increase in Bax/Bcl2 ratio, the release of cytochrome c, activation of caspases -9, -3/7, and cleavage of the poly-ADP-ribose polymerase. DPHP treatment resulted in inhibition of hypoxia induced VEGF downstream signaling pathway in COLO205 cells is concurrent with inhibition of angiogenesis in CAM. Based on these data we suggest that DPHP significantly induced apoptosis possibly via intrinsic mitochondrial apoptosis pathway and inhibited angiogenesis. Our study suggests DPHP could be a therapeutic agent in treating colon cancer.


Assuntos
Apoptose/efeitos dos fármacos , Diarileptanoides/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Células A549 , Animais , Caspase 3/metabolismo , Caspase 9/metabolismo , Linhagem Celular Tumoral , Embrião de Galinha , Diarileptanoides/química , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína X Associada a bcl-2/metabolismo
11.
Int J Nanomedicine ; 14: 3015-3026, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31118619

RESUMO

Purpose: The combination of a bone graft with a barrier membrane is the classic method for guided bone regeneration (GBR) treatment. However, the insufficient osteoinductivity of currently-available barrier membranes and the consequent limited bone regeneration often inhibit the efficacy of bone repair. In this study, we utilized the piezoelectric properties of biomaterials to enhance the osteoinductivity of barrier membranes. Methods: A flexible nanocomposite membrane mimicking the piezoelectric properties of natural bone was utilized as the barrier membrane. Its therapeutic efficacy in repairing critical-sized rabbit mandible defects in combination with xenogenic grafts of deproteinized bovine bone (DBB) was explored. The nanocomposite membranes were fabricated with a homogeneous distribution of piezoelectric BaTiO3 nanoparticles (BTO NPs) embedded within a poly(vinylidene fluoridetrifluoroethylene) (P(VDF-TrFE)) matrix. Results: The piezoelectric coefficient of the polarized nanocomposite membranes was close to that of human bone. The piezoelectric coefficient of the polarized nanocomposite membranes was highly stable, with more than 90% of the original piezoelectric coefficient (d33) remaining up to 28 days after immersion in culture medium. Compared with commercially-available polytetrafluoroethylene (PTFE) membranes, the polarized BTO/P(VDF-TrFE) nanocomposite membranes exhibited higher osteoinductivity (assessed by immunofluorescence staining for runt-related transcription factor 2 (RUNX-2) expression) and induced significantly earlier neovascularization and complete mature bone-structure formation within the rabbit mandible critical-sized defects after implantation with DBB Bio-Oss® granules. Conclusion: Our findings thus demonstrated that the piezoelectric BTO/P(VDF-TrFE) nanocomposite membranes might be suitable for enhancing the clinical efficacy of GBR.


Assuntos
Materiais Biomiméticos/farmacologia , Transplante Ósseo , Eletricidade , Membranas Artificiais , Nanocompostos/química , Osteogênese , Proteínas/isolamento & purificação , Animais , Compostos de Bário/farmacologia , Regeneração Óssea/efeitos dos fármacos , Bovinos , Diferenciação Celular/efeitos dos fármacos , Humanos , Mandíbula/efeitos dos fármacos , Mandíbula/patologia , Mandíbula/cirurgia , Nanocompostos/ultraestrutura , Neovascularização Fisiológica/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Implantação de Prótese , Coelhos , Ratos , Titânio/farmacologia
12.
Life Sci ; 225: 98-106, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30959026

RESUMO

AIMS: Arterial insufficiency ulcers are frequent complications of peripheral artery disease and infection or long-term neglect of the ulcer can eventually lead to amputation of the affected body part. An ischemic environment, caused by interrupted blood flow, affects the supply of nutrients and elongates the inflammation period, inducing tissue degeneration. Thus, the modulation of neovascularization and inflammation could be an ideal therapeutic strategy for ischemic wound healing. This study aimed to elucidate whether systemically administered substance P (SP) could promote ischemic wound repair in mice by restoring blood perfusion and suppressing inflammation. MAIN METHODS: The effects of SP were assessed by analyzing wound size, blood flow, epidermal and dermal layer regeneration, vessel formation, and the inflammatory cytokine profiles in a hind-limb ischemia non-clinical mouse model. KEY FINDINGS: SP-treated mice exhibited dramatically rapid wound healing and restoration of blood flow within the ischemic zone, compared with saline-treated mice. Notably, SP-treated mice showed enhanced pericyte-covered vasculature compared to saline-treated mice. Moreover, anti-inflammatory effects were detected in mice in the SP-treated group, including suppression of inflammation-mediated spleen enlargement, reduction of tumor necrosis factor-alpha, and promotion of circulatory interleukin-10 levels. SIGNIFICANCE: These results suggest that SP could be a possible therapeutic candidate for patients with peripheral artery disease, including those with ischemic ulcers.


Assuntos
Modelos Animais de Doenças , Inflamação/prevenção & controle , Isquemia/complicações , Neovascularização Fisiológica/efeitos dos fármacos , Neurotransmissores/farmacologia , Substância P/farmacologia , Cicatrização/efeitos dos fármacos , Animais , Inflamação/etiologia , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pele/efeitos dos fármacos , Pele/metabolismo
13.
BMC Vet Res ; 15(1): 123, 2019 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-31029157

RESUMO

BACKGROUND: MSCs secretome is under investigation as an alternative to whole-cell-based therapies, since it is enriched of bioactive molecules: growth factors, cytokines and chemokines. Taking into account the translational value of the pig model, the leading aim of the present paper was to characterize the secretome of porcine Vascular Wall-Mesenchymal Stem Cells (pVW-MSCs) and its change in presence of LPS stimulation. Moreover, considering the importance of angiogenesis in regenerative mechanisms, we analysed the effect of pVW-MSCs secretome on in vitro angiogenesis. RESULTS: Our results demonstrated that conditioned medium from unstimulated pVW-MSCs contained high levels of IL-8, GM-CSF, IFN-γ and other immunomodulatory proteins: IL-6 IL-18 IL-4 IL-2 IL-10. LPS modulates pVW-MSCs gene expression and secretome composition, in particular a significant increase of IL-6 and IL-8 was observed; conversely, the amount of GM-CSF, IFN-γ, IL-2, IL-4, IL-10 and IL-18 showed a significant transient decrease with the LPS stimulation. Conditioned medium from unstimulated pVW-MSCs induced in vitro endothelial angiogenesis, which is more evident when the conditioned medium was from LPS stimulated pVW-MSCs. CONCLUSIONS: The lines of evidence here presented shed a light on possible future application of secretome derived by pVW-MSCs on research studies in translational regenerative medicine.


Assuntos
Endotélio Vascular/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Animais , Células Cultivadas , Endotélio Vascular/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Suínos , Transcriptoma
14.
Environ Toxicol Pharmacol ; 69: 112-119, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31026735

RESUMO

In China, cooking oil fume derived fine particulate matter (COF-derived PM2.5) is a principal source of indoor air pollution. Here, we investigated cytotoxicity of COF-derived PM2.5, as well as the roles of VEGF, VEGFR2, MEK1/2, ERK1/2, and mTOR cascade in the inhibitory effects of COF-derived PM2.5, on angiogenesis in human umbilical vein endothelial cells (HUVECs). After exposure to COF-derived PM2.5, cell viability and tube formation, as well as protein and mRNA levels of VEGF, VEGFR2, MEK1/2, ERK1/2, and mTOR in HUVECs were measured. Cell viability and number of tubes reduced dose-dependently after COF-derived PM2.5 and SU5416 treatment. In addition, SU5416 and VEGF significantly affected tube formation. The protein and mRNA levels of VEGF, VEGFR2, MEK1/2, ERK1/2, and mTOR all tended to reduce with the increase of COF-derived PM2.5 concentrations. These findings demonstrate that VEGF, VEGFR2, MEK1/2, ERK1/2, and mTOR play key roles in COF-derived PM2.5 induced inhibition of angiogenesis in HUVECs.


Assuntos
Culinária , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Óleos , Material Particulado/toxicidade , Poluição do Ar em Ambientes Fechados/efeitos adversos , Células Cultivadas , Células Endoteliais da Veia Umbilical Humana/fisiologia , Humanos , Proteínas Quinases Ativadas por Mitógeno/fisiologia , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/fisiologia , Fator A de Crescimento do Endotélio Vascular/fisiologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/fisiologia
15.
Int J Pharm ; 563: 228-236, 2019 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-30959236

RESUMO

Bevacizumab, a vascular endothelial growth factor (VEGF)-targeting drug, is widely used as an off-label therapeutic for age-related macular degeneration (AMD). To reduce the monthly administration frequency, this study investigated microspheres comprising a poly(d, l-lactide-co-glycolide)/poly(cyclohexane-1,4-diyl acetone dimethylene ketal) (PLGA/PCADK) blend that could be loaded with bevacizumab-dextran particles using solid-in-oil-in-water (S/O/W) emulsification. Control microspheres were also prepared through water-in-oil-in-water (W/O/W) emulsification. The structural stability of bevacizumab in the polymer monomers was analyzed using dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), size exclusion chromatography (SEC-HPLC), circular dichroism (CD) and fluorescence spectroscopy. Subsequently, microspheres were prepared and evaluated in terms of their morphology, encapsulation efficiency and release behavior. PLGA/PCADK microspheres prepared by the S/O/W method with <20% PCADK showed a smooth spherical structure with a uniform distribution of bevacizumab. The microspheres exhibited a release behavior comprising a slight initial burst and an increasing total release over 50 days both in vitro and in vivo. Additionally, the microspheres were well tolerated by ocular tissue. Finally, a chorioallantoic membrane (CAM) assay revealed that the bioactivity of bevacizumab was retained by PCADK. In conclusion, these results suggest the potential for bevacizumab-loaded PLGA/PCADK microspheres prepared by the S/O/W method as a means of intravitreal therapy for ocular diseases.


Assuntos
Inibidores da Angiogênese/administração & dosagem , Bevacizumab/administração & dosagem , Microesferas , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/administração & dosagem , Polímeros/administração & dosagem , Inibidores da Angiogênese/química , Animais , Antineoplásicos Imunológicos/administração & dosagem , Antineoplásicos Imunológicos/química , Bevacizumab/química , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/efeitos dos fármacos , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/química , Dextranos/administração & dosagem , Dextranos/química , Liberação Controlada de Fármacos , Oftalmopatias/tratamento farmacológico , Injeções Intravítreas , Masculino , Neovascularização Fisiológica/efeitos dos fármacos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Polímeros/química , Coelhos
16.
Int J Mol Sci ; 20(8)2019 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-31013775

RESUMO

AqB013 and AqB050 compounds inhibit aquaporin 1 (AQP1), a dual water and ion channel implicated in tumour angiogenesis. We tested AqB013 and AqB050 either as monotherapy or in combination on tube formation of murine endothelial cells (2H-11 and 3B-11) and human umbilical vascular endothelial cells (HUVECs). The mechanism underlying their anti-tubulogenic effect was explored by examining cell viability, induction of apoptosis and migration using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, Annexin V/propidium iodide apoptosis assay and scratch wound assay. Tube formation of all the cell lines was inhibited by AqB013, AqB050 and the combination of the two compounds. The inhibition of 2H-11 and 3B-11 was frequently accompanied by impaired migration, whereas that of HUVEC treated with AqB050 and the combination was associated with reduced cell viability due to apoptosis. AqB013 and AqB050 exhibited an anti-tubulogenic effect through inhibition of AQP1-mediated cell migration and induction of apoptosis. Together with previously reported anti-tumour cell effect of AqB013 and AqB050, our findings support further evaluation of these compounds as potential cancer therapeutics.


Assuntos
Apoptose/efeitos dos fármacos , Aquaporina 1/antagonistas & inibidores , Bumetanida/farmacologia , Movimento Celular/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Bumetanida/análogos & derivados , Sobrevivência Celular/efeitos dos fármacos , Células Endoteliais/metabolismo , Humanos
17.
Mater Sci Eng C Mater Biol Appl ; 100: 236-246, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30948058

RESUMO

Crosslinked 3D porous collagen-polysaccharide scaffolds, prepared by freeze-drying, were modified with bovine platelet lysate (BPL) and evaluated in terms of chemical, physical and biological properties. Natural antibacterial polysaccharides like chitosan, chitin/chitosan-glucan complex and calcium salt of oxidized cellulose (CaOC) incorporated in collagen scaffolds affected not only chemo-physical properties of the composite scaffolds but also improved their biological properties, especially when BPL was presented. Lipophilic BPL formed microspheres in porous scaffolds while reduced by half their swelling ratio. The resistance of collagen sponges to hydrolytic degradation in water depended strongly on chemical crosslinking varying from 60 min to more than one year. According to in-vitro tests, chemically crosslinked scaffolds exhibited a good cellular response, cell-matrix interactions, and biocompatibility of the material. The combination of collagen with natural polysaccharides confirmed a significant positive synergistic effect on cultivation of cells as determined by MTS assay and PicoGreen method, as well as on angiogenesis evaluated by ex ovo Chick Chorioallantoic Membrane (CAM) assay. Contrary, modification only by BLP of pure collagen scaffolds exhibited decreased biocompatibility in comparison to unmodified pure collagen scaffold. We propose that the newly developed crosslinked collagen sponges involving bioactive additives could be used as scaffold for growing cells in systems with low mechanical loading in tissue engineering, especially in dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration.


Assuntos
Plaquetas/metabolismo , Colágeno/farmacologia , Polissacarídeos/farmacologia , Engenharia Tecidual/métodos , Tecidos Suporte/química , Células 3T3 , Animais , Bovinos , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Galinhas , Reagentes para Ligações Cruzadas/química , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Hidrólise , Camundongos , Neovascularização Fisiológica/efeitos dos fármacos , Temperatura Ambiente , Água/química
18.
Biomed Pharmacother ; 114: 108811, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30965235

RESUMO

OBJECTIVES: We aimed to determine whether bone remodeling and vessel formation in the osteochondral unit are suppressed by supplementing with docosahexaenoic acid in anterior cruciate ligament transection (ACLT)-induced rats. METHODS: Twelve-week-old male Sprague Dawley rats were randomized to sham-operated, ACLT-operated and treated with vehicle, or ACLT-operated and treated with DHA groups. Micro-architecture and vasculature in the tibial osteochondral unit were examined by micro-CT, as well as by histomorphometry. To evaluate the effects of DHA in vitro, we conducted functional and expressional assays in RAW264.7 cells and HUVECs. Finally, we used OARSI-modified Mankin criteria and histological analyses to assess the status of the cartilage layer. RESULTS: Microstructural parameters in the osteochondral unit showed that bone mass loss and angiogenesis were less in DHA-treated rats than in vehicle-treated rats. Immunofluorescence-positive cells labeled with TRAP, RANKL, CD31, and endomucin agents in the osteochondral unit of ACLT-operated rats were reduced in the DHA-treated group compared with the vehicle-treated group. Furthermore, the number of TRAP-stained cells, areas of bone resorption pits, and mRNA expression of TRAP, CTSK, MITF, and NFATC1 were reduced in RAW264.7 cells treated with RANKL + DHA compared with those treated with only RANKL. Tube formation, proliferation and migration of HUVECs, and VEGF-C mRNA and VEGFR2 protein expression were inhibited by DHA. The decrease in OARSI score, and MMP-13 and collagen X expression suggested that DHA attenuated cartilage degeneration. CONCLUSIONS: DHA has the ability to restrain bone remodeling and vessel formation in the osteochondral unit, which may contribute to protection of cartilage.


Assuntos
Remodelação Óssea/efeitos dos fármacos , Ácidos Docosa-Hexaenoicos/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Animais , Reabsorção Óssea/tratamento farmacológico , Reabsorção Óssea/metabolismo , Cartilagem/efeitos dos fármacos , Cartilagem/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Camundongos , Células RAW 264.7 , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fator C de Crescimento do Endotélio Vascular/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
19.
Mater Sci Eng C Mater Biol Appl ; 101: 1-14, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31029302

RESUMO

Tracheal reconstruction remains a major surgical challenge, mainly owing to the scarce of resilient hollow grafts with identifiable vascular pedicle in humans. In this study, we developed a three-layer, elastomeric, trachea-like composite made of poly glycerol sebacate (PGS) and polycaprolactone (PCL), which presented appropriate resilient property, timely degradation and interconnected pores. C shape PCL rings fabricated with selective laser sintering (SLS) techniques are regularly positioned around porous PGS tubes and fixed by PCL electrospinning sheath. Such an elastomeric composite underwent host remodeling including rapid vascularization and tissue infiltration after fascia wrapping. With degrading of PGS, C rings well incorporated into growing fascia and lead to the formation of pedicled tracheal grafts, which attributes to the strong and resilient properties of generated hollow grafts thus enabled orthotopic transplantation in segmental tracheal defect. Progressive remodeling on such vascularized and mechanically stable grafts resulted in epithelium regeneration on luminal side as well as production of adequate amount of collagen and elastin, which warrantees the air passage during breathing. Future study employing large animal models more representative of human tracheal regeneration is warranted before clinical translation. Using fast degrading PGS combined with PCL rings represents a philosophical shift from the prevailing focus on tough grafts in airway reconstruction and may impact regenerative medicine in general.


Assuntos
Prótese Vascular , Elastômeros/farmacologia , Fáscia/irrigação sanguínea , Regeneração/efeitos dos fármacos , Stents , Tecidos Suporte/química , Traqueia/fisiologia , Animais , Implante de Prótese Vascular , Decanoatos/farmacologia , Epitélio/efeitos dos fármacos , Fáscia/efeitos dos fármacos , Glicerol/análogos & derivados , Glicerol/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Poliésteres/farmacologia , Polímeros/farmacologia , Porosidade , Coelhos , Traqueia/efeitos dos fármacos , Traqueia/ultraestrutura
20.
Eur Cell Mater ; 37: 250-264, 2019 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-30963526

RESUMO

Different tissue engineering techniques are used to support rapid vascularisation. A novel technique is the use of platelet-rich fibrin (PRF), an autologous source of growth factors. This study was the first to investigate the influence of PRF matrices, isolated following different centrifugation protocols, on human dermal vascular endothelial cells (ECs) in mono-culture and co-culture with human primary fibroblasts (HFs) as an in vitro model for tissue regeneration. Focus was placed on vascular structure formation and growth factor release. HFs and ECs were cultivated with PRF prepared using a high (710 ×g) or low (44 ×g) relative centrifugation force (RCF) over 14 d. Immunofluorescence staining and immunohistochemistry were used to evaluate the microvascular formation. Cell culture supernatants were collected for evaluation of growth factor release. The results showed a PRF-mediated effect on the induction of angiogenesis in ECs. Microvessel-like structure formation was promoted when ECs were combined with low-RCF PRF as compared to high-RCF PRF or control group. The percentage of vascular lumen area was significantly higher in low-RCF PRF, especially at day 7, which coincided with statistically significantly higher growth factor [vascular endothelial factor (VEGF), transforming growth factor ß1 (TGF-ß1) and platelet derived growth factor (PDGF)] concentration measured in low-RCF PRF as compared to high-RCF PRF or control group. In conclusion, reducing the RCF according to the low-speed centrifugation concept (LSCC) resulted in increased growth factor release and angiogenic structure formation with EC mono-culture, suggesting that PRF may be a highly beneficial therapeutic tool for tissue engineering applications.


Assuntos
Células Endoteliais/metabolismo , Fibroblastos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular , Neovascularização Fisiológica/efeitos dos fármacos , Fibrina Rica em Plaquetas , Técnicas de Cultura de Células , Células Endoteliais/citologia , Fibroblastos/citologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia
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