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1.
Int J Mol Sci ; 22(15)2021 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-34361085

RESUMO

A novel aptamer-based competitive drug screening platform for osteoporosis was devised in which fluorescence-labeled, sclerostin-specific aptamers compete with compounds from selected chemical libraries for the binding of immobilized recombinant human sclerostin to achieve high-throughput screening for potential small-molecule sclerostin inhibitors and to facilitate drug repurposing and drug discovery. Of the 96 selected inhibitors and FDA-approved drugs, six were shown to result in a significant decrease in the fluorescence intensity of the aptamer, suggesting a higher affinity toward sclerostin compared with that of the aptamer. The targets of these potential sclerostin inhibitors were correlated to lipid or bone metabolism, and several of the compounds have already been shown to be potential osteogenic activators, indicating that the aptamer-based competitive drug screening assay offered a potentially reliable strategy for the discovery of target-specific new drugs. The six potential sclerostin inhibitors suppressed the level of both intracellular and/or extracellular sclerostin in mouse osteocyte IDG-SW3 and increased alkaline phosphatase activity in IDG-SW3 cells, human bone marrow-derived mesenchymal stem cells and human fetal osteoblasts hFOB1.19. Potential small-molecule drug candidates obtained in this study are expected to provide new therapeutics for osteoporosis as well as insights into the structure-activity relationship of sclerostin inhibitors for rational drug design.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Aptâmeros de Nucleotídeos/química , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteoblastos/efeitos dos fármacos , Osteócitos/efeitos dos fármacos , Osteoporose/tratamento farmacológico , Bibliotecas de Moléculas Pequenas/farmacologia , Animais , Aptâmeros de Nucleotídeos/isolamento & purificação , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteócitos/citologia , Osteócitos/metabolismo , Osteoporose/metabolismo , Osteoporose/patologia
3.
Artigo em Inglês | MEDLINE | ID: mdl-23737818

RESUMO

This study proposed a novel combination of neural regeneration techniques for the repair of damaged peripheral nerves. A biodegradable nerve conduit containing genipin-cross-linked gelatin was annexed using beta-tricalcium phosphate (TCP) ceramic particles (genipin-gelatin-TCP, GGT) to bridge the transection of a 15 mm sciatic nerve in rats. Two trigger points were irradiated transcutaneously using 660 nm of gallium-aluminum arsenide phosphide (GaAlAsP) via laser diodes for 2 min daily over 10 consecutive days. Walking track analysis showed a significant improvement in sciatic functional index (SFI) (P < 0.01) and pronounced improvement in the toe spreading ability of rats undergoing laser stimulation. Electrophysiological measurements (peak amplitude and area) illustrated by compound muscle action potential (CMAP) curves demonstrated that laser stimulation significantly improved nerve function and reduced muscular atrophy. Histomorphometric assessments revealed that laser stimulation accelerated nerve regeneration over a larger area of neural tissue, resulting in axons of greater diameter and myelin sheaths of greater thickness than that observed in rats treated with nerve conduits alone. Motor function, electrophysiological reactions, muscular reinnervation, and histomorphometric assessments all demonstrate that the proposed therapy accelerated the repair of transected peripheral nerves bridged using a GGT nerve conduit.

4.
J Biomed Mater Res A ; 101(10): 2763-77, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23427146

RESUMO

This study proposes a biodegradable nerve conduit comprising 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) cross-linked gelatin annexed with ß-tricalcium phosphate (ß-TCP) ceramic particles (EDC-gelatin-TCP, EGT). For this study, the EGT-implant site in rats was irradiated using 660-nm GaAlAsP laser diodes (50 mW) for trigger point therapy to investigate the use of low-level laser (LLL) stimulation in the regeneration of a 15-mm transected sciatic nerve. Animals were divided into three groups: a control group undergoing autologous nerve graft (autograft); a sham-irradiated group (EGT), and an experimental group undergoing laser stimulation (EGT/LS). Two trigger points on the surgical incision along the sciatic nerve were irradiated transcutaneously for 2 min daily for 10 consecutive days. Twelve weeks after implantation, walking track analysis showed a significantly higher sciatic functional index (SFI; p < 0.05) and improved toe spreading development in the autograft and EGT/LS groups, compared to the EGT group. In the electrophysiological measurement, the mean recovery index (peak amplitude and area) of the compound muscle action potential curves in the autograft and EGT/LS groups showed significantly improved functional recovery than in the EGT group (p < 0.05). Compared with the EGT group, the autograft and EGT/LS groups showed a reduction in muscular atrophy. Histomorphometric assessments showed that the EGT/LS group had undergone more rapid nerve regeneration than the EGT group. Therefore, motor function, electrophysiological reaction, muscular reinnervation, and histomorphometric assessments demonstrate that LLL therapy can accelerate the repair of a 15-mm transected peripheral nerve in rats after being bridged with the EGT nerve conduit.


Assuntos
Regeneração Tecidual Guiada , Terapia com Luz de Baixa Intensidade , Regeneração Nervosa/efeitos da radiação , Nervo Isquiático/efeitos da radiação , Nervo Isquiático/cirurgia , Animais , Autoenxertos/efeitos dos fármacos , Autoenxertos/efeitos da radiação , Materiais Biocompatíveis/farmacologia , Fenômenos Eletrofisiológicos/efeitos dos fármacos , Fenômenos Eletrofisiológicos/efeitos da radiação , Imuno-Histoquímica , Atrofia Muscular/patologia , Atrofia Muscular/fisiopatologia , Bainha de Mielina/metabolismo , Regeneração Nervosa/efeitos dos fármacos , Tetróxido de Ósmio/metabolismo , Cuidados Pós-Operatórios , Implantação de Prótese , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica/efeitos dos fármacos , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/fisiopatologia
5.
Chang Gung Med J ; 34(2): 205-12, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21539763

RESUMO

BACKGROUND: Nausea and vomiting are common chief postoperative complaints. The clinical literature indicates that postoperative nausea and vomiting (PONV) is common after orthopedic surgery. This study examines the clinical therapeutic efficacy of Ondansetron injected intravenously before the end of shoulder arthroscopy as antiemetic prophylaxis to help reduce the incidence of PONV. METHODS: Participants were identified through retrospective chart review and patients undergoing shoulder arthroscopy performed by the same orthopedic surgeon at the same hospital from 2005 to 2009 were analyzed. Subjects were classified into two groups based on whether Ondansetron was given. Differences in the incidence of PONV among the two groups were compared. Basic patient information, anesthesia records, and surgical records were obtained, as well as records on PONV, postoperative pain intensity, and postoperative analgesic injections within 24 hours after surgery. RESULTS: The study involved 90 patients. The Group A contained 34 patients who did not receive Ondansetron, and the Group B contained 56 patients who were given Ondansetron. Analytical results for the postoperative 24 hour period showed a significant difference in the incidence of vomiting between the two groups, with a lower incidence (p < 0.05) for the. Group B. However there was no significant difference in the incidence of nausea between the two groups in the same postoperative 24 hour period, although there was a trend of a lower incidence in the Group B (p = 0.17). The overall incidence of PONV during the 24-hour period was lower in the Group B (14%) than the Group A (32%), and the Group B demonstrated lower pain intensity and lower analgesic injection needs. CONCLUSION: Routine intravenous injection of Ondansetron 30 minutes before completion of shoulder arthroscopy can reduce the incidence of vomiting and overall PONV in patients. Additionally, the patients using Ondansetron demonstrated lower pain intensity and lower analgesic injection needs than the control group.


Assuntos
Antieméticos/uso terapêutico , Artroscopia , Ondansetron/uso terapêutico , Náusea e Vômito Pós-Operatórios/prevenção & controle , Antagonistas do Receptor 5-HT3 de Serotonina/uso terapêutico , Ombro/cirurgia , Adulto , Idoso , Feminino , Humanos , Injeções Intravenosas , Masculino , Pessoa de Meia-Idade
6.
Tissue Eng Part A ; 17(11-12): 1549-60, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21284540

RESUMO

Human placenta-derived mesenchymal stem cells (hPMSCs) represent a promising source of stem cells. The application of hPMSCs in cartilage tissue engineering, however, was less reported. In this study, hPMSCs were grown in a three-dimensional (3D) environment for cartilage tissue formation in vitro. To select proper scaffolds for 3D culture of mesenchymal stem cells (MSCs), rat adipose-derived MSCs were initially employed to optimize the composition and condition of the 3D environment. The suitability of a poly(D,L-lactide-co-glycolide) (PLGA) precision scaffold previously developed for seeding and culture of primary chondrocytes was tested for MSCs. It was established that MSCs had to be embedded in alginate gel before seeded in the PLGA precision scaffold for cartilage-like tissue formation. The inclusion of nano-sized calcium-deficient hydroxyapatite (nCDHA) and/or a recombinant protein containing arginine-glycine-aspartate (RGD) into the alginate gel enhanced the chondrogenesis for both rat adipose-derived MSCs and hPMSCs. The amount of extracellular matrix such as glycosaminoglycan and type II collagen accumulated during a period of 21 days was found to be the greatest for hPMSCs embedded in the alginate/nCDHA/RGD gel and injected and cultivated in the precision scaffold. Also, histological analyses revealed the lacunae formation and extracellular matrix production from the seeded hPMSCs. Comparing human bone marrow-derived MSCs (hBMSCs) and hPMSCs grown in the previous composite scaffolds, the secretion of glycosaminoglycan was twice as higher for hPMSCs as that for hBMSCs. It was concluded that the alginate/nCDHA/RGD mixed gel in the aforementioned system could provide a 3D environment for the chondrogenesis of hPMSCs, and the PLGA precision scaffold could provide the dimensional stability of the whole construct. This study also suggested that hPMSCs, when grown in a suitable scaffold, may be a good source of stem cells for building up the tissue-engineered cartilage.


Assuntos
Cartilagem/fisiologia , Condrogênese , Células-Tronco Mesenquimais/citologia , Placenta/citologia , Engenharia Tecidual/métodos , Tecidos Suporte/química , Alginatos/farmacologia , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Cartilagem/efeitos dos fármacos , Contagem de Células , Células Cultivadas , Condrogênese/efeitos dos fármacos , Colágeno Tipo II/metabolismo , Feminino , Ácido Glucurônico/farmacologia , Glicosaminoglicanos/metabolismo , Ácidos Hexurônicos/farmacologia , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Microesferas , Gravidez , Ratos , Coloração e Rotulagem
7.
Photomed Laser Surg ; 28 Suppl 2: S37-42, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20932129

RESUMO

OBJECTIVE: In this study, the possible effect of low-level laser (LLL) on improving the adhesion of endothelial cells (ECs) to a biomaterial substrate was evaluated. BACKGROUND DATA: Despite the numerous studies regarding the effects of LLL on biologic systems, the influence of LLL on the binding between cells and materials was rarely investigated. MATERIALS AND METHODS: A low-power He-Ne laser apparatus with a continuous wavelength of 632.8 nm (a maximum power output of 50 mW) was used. The average irradiation energy on cells was 1.18 J/cm(2). Cell morphology and the concentrations of nitric oxide and calcium after laser exposure were measured. Biomedical grade poly(carbonate)urethane (PU) was synthesized and used to prepare microporous vascular grafts. ECs exposed to laser were harvested and seeded on the PU grafts. No further exposure was given. RESULTS: LLL could change the morphology and increase the matrix secretion of ECs, and such effects persisted when preexposed cells were harvested and seeded to another substrate. The number of ECs attached on the biomaterial substrate was not affected. Preexposed ECs on the PU graft, however, were, on average, more resistant to flushing (i.e., greater cell retention). CONCLUSION: ECs were pretreated with LLL before being seeded onto the PU biomaterial vascular grafts. The retention of LLL-preexposed ECs on the graft surface was enhanced, but not as significantly as that of ECs preexposed to low-intensity ultrasound.


Assuntos
Células Endoteliais/efeitos da radiação , Endotélio Vascular/citologia , Terapia com Luz de Baixa Intensidade , Engenharia Tecidual , Animais , Materiais Biocompatíveis , Prótese Vascular , Bovinos , Adesão Celular/efeitos da radiação , Teste de Materiais , Óxido Nítrico/química , Poliuretanos , Tecidos Suporte
8.
J Biomed Mater Res B Appl Biomater ; 95(1): 207-17, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20737555

RESUMO

To modulate the mechanical properties of nerve guide conduit for surgical manipulation, this study develops a biodegradable composite containing genipin cross-linked gelatin annexed with ß-tricalcium phosphate ceramic particles as a nerve guide material. The conduit was dark bluish and round with a rough and compact outer surface compared to the genipin cross-linked gelatin conduit (without ß-tricalcium phosphate). Water uptake and swelling tests indicate that the conduit noticeably increases the stability in water, and the hydrated conduit does not collapse and stenose. The conduit has a sufficiently high level of mechanical properties to serve as a nerve guide. After subcutaneous implantation on the dorsal side of a rat, the degraded conduit only evokes a mild tissue response, and the formation of a very thin fibrous capsule surrounds the conduit. This paper assesses the effectiveness of the conduit as a guidance channel when we use it to repair a 10 mm gap in the rat's sciatic nerve. The experimental results show no gross inflammatory reactions of the peripheral nerve tissues at the implantation site in either group. In overall gross examination, the diameter of the intratubular and newly formed nerve fibers in the conduits exceeds that of the silicone tubes during the implantation period. The quantitative results indicate the superiority of the conduits over the silicone tubes. This study microscopically observes the nerve regeneration in the tissue section at the middle region of all implanted conduits. Therefore, the histomorphometric assessment demonstrates that the conduit could be a candidate for peripheral nerve repair.


Assuntos
Materiais Biocompatíveis/química , Fosfatos de Cálcio , Gelatina , Regeneração Tecidual Guiada/métodos , Regeneração Nervosa/efeitos dos fármacos , Implantes Absorvíveis , Animais , Reagentes para Ligações Cruzadas , Implantes Experimentais , Inflamação , Glicosídeos Iridoides , Iridoides , Nervos Periféricos , Ratos , Nervo Isquiático/cirurgia
9.
Macromol Biosci ; 8(10): 932-41, 2008 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-18615456

RESUMO

This study investigates the feasibility of a novel nanocomposite (GC/Ag) of a genipin-crosslinked chitosan (GC) film in which was embedded various amounts of Ag nanoparticles for wound-dressing applications. In situ UV-vis results revealed that adding chitosan solution did not affect the characteristics of Ag nanoparticles. The water uptake ratios and surface hydrophilicity of the GC/Ag nanocomposite were better and the degradation rates slightly lower than those of the pure GC film. The presence of Ag nanoparticles enhanced L929 cell attachment and growth. Its function as an anti-microbial agent in a GC/Ag nanocomposite was assessed for Ag contents of over 100 ppm. In conclusion, silver ions had dual functions--structural reinforcement and provision of antimicrobial properties to a biocompatible polymer.


Assuntos
Anti-Infecciosos/farmacologia , Quitosana/química , Reagentes para Ligações Cruzadas/farmacologia , Iridoides/química , Nanopartículas Metálicas/química , Nanocompostos/química , Prata/química , Animais , Anti-Infecciosos/química , Adesão Celular/efeitos dos fármacos , Contagem de Células , Morte Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Quitosana/farmacologia , Escherichia coli/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/ultraestrutura , Glicosídeos Iridoides , Iridoides/farmacologia , Nanopartículas Metálicas/ultraestrutura , Camundongos , Testes de Sensibilidade Microbiana , Muramidase/metabolismo , Nanocompostos/ultraestrutura , Prata/farmacologia , Espectrofotometria Ultravioleta , Propriedades de Superfície/efeitos dos fármacos , Água/química
10.
J Biomed Mater Res B Appl Biomater ; 76(1): 85-92, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16206256

RESUMO

In spite of the extensive studies regarding the effects of ultrasound on biological systems, the influence of low-intensity ultrasound on endothelial cells has rarely been investigated. In this work, the effect of ultrasound in improving the binding between endothelial cells and biomaterial substrates was evaluated. Based on the results, low-intensity ultrasound could change the morphology and matrix secretion of endothelial cells, and such effects persisted when preexposed cells were seeded to another substrate. The cells preexposed to ultrasound were spread further on the substrate. The actin stress fibers of ultrasound-preexposed cells on RGD-modified surfaces were especially intense and well oriented. Ultrasound could probably activate cellular integrins and subsequently allow RGD to bind them. A much firmer adhesion of ultrasound-preexposed endothelial cells to the biomaterial surface coated with the RGD-containing protein was demonstrated. Finally, polyurethane small-diameter vascular grafts seeded with ultrasound-preexposed endothelial cells showed enhanced cell retention on graft surfaces upon flushing.


Assuntos
Materiais Biocompatíveis , Endotélio Vascular/citologia , Animais , Bovinos , Células Cultivadas , Matriz Extracelular , Propriedades de Superfície
11.
Biomol Eng ; 21(3-5): 99-104, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15567103

RESUMO

The effects of low-intensity ultrasound (US) on biological systems have been investigated extensively; however, the effects of ultrasound stimulation on endothelial cells were rarely studied. In this study, 1 MHz, pulsed 1:4, and four different spatial-average temporal-peak intensities (0.5, 1.0, 1.6, and 2W/cm2) of ultrasound were used to stimulate endothelial cells for 10 min per day. The results showed that ultrasound (intensity 1.6-2.0W/cm2) treatment after 6 days enhanced the nitric oxide (NO) and Ca2+ release from the endothelial cells but did not promote cell growth. In addition, ultrasound stimulation changed the cellular morphology and orientation, and increased extracellular matrix secretion from endothelial cells.


Assuntos
Cálcio/metabolismo , Citoesqueleto/efeitos da radiação , Células Endoteliais/fisiologia , Células Endoteliais/efeitos da radiação , Matriz Extracelular/efeitos da radiação , Óxido Nítrico/metabolismo , Ultrassom , Animais , Bovinos , Proliferação de Células/efeitos da radiação , Células Cultivadas , Citoesqueleto/fisiologia , Citoesqueleto/ultraestrutura , Relação Dose-Resposta à Radiação , Células Endoteliais/citologia , Células Endoteliais/diagnóstico por imagem , Matriz Extracelular/fisiologia , Matriz Extracelular/ultraestrutura , Doses de Radiação , Ultrassonografia
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