RESUMO
INTRODUCTION: Dry eye disease (DED) is a multifactor-induced disease accompanied by increased osmolarity of the tear film and inflammation of the ocular surface. Traditional anti-inflammation agent corticosteroids applied in DED treatment could result in high intraocular pressure, especially in long-term treatment. Therefore, we explored a nano drug that aimed to block the formation pathway of DED which had anti-inflammatory, sustained release, and good biocompatibility characteristics in this study. METHODS: We prepared a novel nanomedicine (Tet-ATS@PLGA) by the thin film dispersion-hydration ultrasonic method and detected its nanostructure, particle size, and zeta potential. Flow cytometry was used to detect the cell survival rate of each group after 24 h of drug treatment on inflammed Statens Seruminstitut Rabbit Corneal (SIRC) cells. Observed and recorded corneal epithelial staining, tear film rupture time, and Schirmer test to detect tear secretion on the ocular surface of rabbits. The corneal epithelial thickness, morphology, and number of bulbar conjunctival goblet cells were recorded by H&E staining. Finally, we detected the expression of VEGF, IL-1ß, PGE2, and TNF-α by cellular immunofluorescence staining and enzyme-linked immunosorbent assay (ELISA). RESULTS: The encapsulation efficiency and drug loading of Tet-ATS@PLGA were 79.85% and 32.47%, respectively. At eye surface temperature, Tet can easily release from Tet-ATS@PLGA while that it was difficult to release at storage temperature and room temperature. After 2 weeks medication, Tet-ATS@PLGA can effectively improve the tear film rupture time and tear secretion time in a DED model (p <0.05). Compared with the normal group (62.34 ± 4.86 mm), the thickness of corneal epithelium in ATS (29.47 ± 3.21 mm), Tet-ATS (46.23 ± 2.87 mm), and Tet-ATS@PLGA (55.76 ± 3.95 mm) gradually increased. Furthermore, the flow cytometry indicated that Tet-ATS@PLGA can effectively promote the apoptosis of inflammatory SIRC cells, and the cellular immunofluorescence and ELISA experiments showed that the expression intensity of inflammatory factors such as VEGF, IL-1ß, PGE2, and TNF-α decreased in this process. Interestingly, Tet also had the effect of reducing intraocular pressure. CONCLUSION: Tet-ATS@PLGA can effectively promote the apoptosis of inflammatory corneal epithelial cells, thus inhibiting the expression of inflammatory factors to block the formation of DED and improve the secretion of tear on the ocular surface.
Assuntos
Síndromes do Olho Seco , Nanopartículas , Animais , Coelhos , Ácido Poliglicólico/análise , Ácido Poliglicólico/metabolismo , Ácido Poliglicólico/uso terapêutico , Fator de Necrose Tumoral alfa , Dinoprostona/análise , Dinoprostona/metabolismo , Dinoprostona/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/metabolismo , Síndromes do Olho Seco/diagnóstico , Lágrimas/metabolismo , Córnea/metabolismo , Anti-Inflamatórios/uso terapêutico , Nanopartículas/químicaRESUMO
BACKGROUND: Advanced stage cancer treatments are often invasive and painful-typically comprised of surgery, chemotherapy, and/or radiation treatment. Low transport efficiency during systemic chemotherapy may require high chemotherapeutic doses to effectively target cancerous tissue, resulting in systemic toxicity. Nanotherapeutic platforms have been proposed as an alternative to more safely and effectively deliver therapeutic agents directly to tumor sites. However, cellular internalization and tumor penetration are often diametrically opposed, with limited access to tumor regions distal from vasculature, due to irregular tissue morphologies. To address these transport challenges, nanoparticles (NPs) are often surface-modified with ligands to enhance transport and longevity after localized or systemic administration. Here, we evaluate stealth polyethylene-glycol (PEG), cell-penetrating (MPG), and CPP-stealth (MPG/PEG) poly(lactic-co-glycolic-acid) (PLGA) NP co-treatment strategies in 3D cell culture representing hypo-vascularized tissue. RESULTS: Smaller, more regularly-shaped avascular tissue was generated using the hanging drop (HD) method, while more irregularly-shaped masses were formed with the liquid overlay (LO) technique. To compare NP distribution differences within the same type of tissue as a function of different cancer types, we selected HeLa, cervical epithelial adenocarcinoma cells; CaSki, cervical epidermoid carcinoma cells; and SiHa, grade II cervical squamous cell carcinoma cells. In HD tumors, enhanced distribution relative to unmodified NPs was measured for MPG and PEG NPs in HeLa, and for all modified NPs in SiHa spheroids. In LO tumors, the greatest distribution was observed for MPG and MPG/PEG NPs in HeLa, and for PEG and MPG/PEG NPs in SiHa spheroids. CONCLUSIONS: Pre-clinical evaluation of PLGA-modified NP distribution into hypo-vascularized tumor tissue may benefit from considering tissue morphology in addition to cancer type.
Assuntos
Portadores de Fármacos/metabolismo , Ácido Láctico/metabolismo , Nanopartículas/metabolismo , Neoplasias/irrigação sanguínea , Polietilenoglicóis/metabolismo , Ácido Poliglicólico/metabolismo , Técnicas de Cultura de Células/métodos , Linhagem Celular Tumoral , Portadores de Fármacos/análise , Células HeLa , Humanos , Ácido Láctico/análise , Nanopartículas/análise , Neoplasias/metabolismo , Polietilenoglicóis/análise , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Esferoides Celulares , Células Tumorais CultivadasRESUMO
Therapeutic nanoparticles (NPs) approved for clinical use in solid tumor therapy provide only modest improvements in patient survival, in part due to physiological barriers that limit delivery of the particles throughout the entire tumor. Here, we explore the thresholds for NP size and surface poly(ethylene glycol) (PEG) density for penetration within tumor tissue extracellular matrix (ECM). We found that NPs as large as 62nm, but less than 110nm in diameter, diffused rapidly within a tumor ECM preparation (Matrigel) and breast tumor xenograft slices ex vivo. Studies of PEG-density revealed that increasing PEG density enhanced NP diffusion and that PEG density below a critical value led to adhesion of NP to ECM. Non-specific binding of NPs to tumor ECM components was assessed by surface plasmon resonance (SPR), which revealed excellent correlation with the particle diffusion results. Intravital microscopy of NP spread in breast tumor tissue confirmed a significant difference in tumor tissue penetration between the 62 and 110nm PEG-coated NPs, as well as between PEG-coated and uncoated NPs. SPR assays also revealed that Abraxane, an FDA-approved non-PEGylated NP formulation used for cancer therapy, binds to tumor ECM. Our results establish limitations on the size and surface PEG density parameters required to achieve uniform and broad dispersion within tumor tissue and highlight the utility of SPR as a high throughput method to screen NPs for tumor penetration.
Assuntos
Portadores de Fármacos/metabolismo , Nanopartículas/metabolismo , Neoplasias/metabolismo , Polietilenoglicóis/metabolismo , Paclitaxel Ligado a Albumina/administração & dosagem , Paclitaxel Ligado a Albumina/metabolismo , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/metabolismo , Mama/efeitos dos fármacos , Mama/metabolismo , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Colágeno/metabolismo , Difusão , Doxorrubicina/administração & dosagem , Doxorrubicina/análogos & derivados , Doxorrubicina/metabolismo , Portadores de Fármacos/análise , Combinação de Medicamentos , Feminino , Humanos , Ácido Láctico/análise , Ácido Láctico/metabolismo , Laminina/metabolismo , Camundongos , Camundongos Nus , Nanopartículas/análise , Neoplasias/tratamento farmacológico , Tamanho da Partícula , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/análise , Ácido Poliglicólico/análise , Ácido Poliglicólico/metabolismo , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Proteoglicanas/metabolismo , Propriedades de SuperfícieRESUMO
Poly-lactide-co-glycolide (PLGA) is one of the few polymers approved by the US Food and Drug Administration as a carrier for drug administration in humans; therefore, it is one of the most used materials in the formulation of polymeric nanoparticles (NPs) for therapeutic purposes. Because the cellular uptake of polymeric NPs is a hot topic in the nanomedicine field, the development of techniques able to ensure incontrovertible evidence of the presence of NPs in the cells plays a key role in gaining understanding of their therapeutic potential. On the strength of this premise, this article aims to evaluate the application of synchrotron radiation-based Fourier transform infrared spectroscopy (SR-FTIR) spectromicroscopy and SR X-ray fluorescence (SR-XRF) microscopy in the study of the in vitro interaction of PLGA NPs with cells. To reach this goal, we used PLGA NPs, sized around 200 nm and loaded with superparamagnetic iron oxide NPs (PLGA-IO-NPs; Fe3O4; size, 10-15 nm). After exposing human mesothelial (MeT5A) cells to PLGA-IO-NPs (0.1 mg/mL), the cells were analyzed after fixation both by SR-FTIR spectromicroscopy and SR-XRF microscopy setups. SR-FTIR-SM enabled the detection of PLGA NPs at single-cell level, allowing polymer detection inside the biological matrix by the characteristic band in the 1,700-2,000 cm(-1) region. The precise PLGA IR-signature (1,750 cm(-1) centered pick) also was clearly evident within an area of high amide density. SR-XRF microscopy performed on the same cells investigated under SR-FTIR microscopy allowed us to put in evidence the Fe presence in the cells and to emphasize the intracellular localization of the PLGA-IO-NPs. These findings suggest that SR-FTIR and SR-XRF techniques could be two valuable tools to follow the PLGA NPs' fate in in vitro studies on cell cultures.
Assuntos
Células Epiteliais/química , Células Epiteliais/citologia , Ácido Láctico/análise , Microscopia de Fluorescência/métodos , Imagem Molecular/métodos , Nanopartículas/análise , Ácido Poliglicólico/análise , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Linhagem Celular , Humanos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Estatística como Assunto , Frações Subcelulares/química , Frações Subcelulares/ultraestrutura , SíncrotronsRESUMO
PURPOSE: To investigate accelerated blood clearance (ABC) induction upon repeated injections of PLGA-PEG nanoparticles as a commonly used polymeric drug carrier. METHODS: Etoposide-loaded PLGA-PEG NPs were developed and administered as the test dose to rats pre-injected with various NP treatments at certain time intervals. Pharmacokinetic parameters of etoposide and production of anti-PEG IgM antibody were evaluated. RESULTS: A notable ABC effect was induced by a wide range of polymer doses (0.1 to 20 mg) of empty NPs, accompanied by IgM secretion. However, a further increase in polymer dose resulted not only in the abrogation of the observed ABC induction but also in distinctly a higher value for AUC of the NPs relative to the control. The data from the PEG-negative group verified the fundamental role of PEG for ABC induction. The first injection of etoposide-containing PEGylated nanoparticles (a cell cycle phase-specific drug) produced a strong ABC phenomenon. Three sequential administrations of etoposide-loaded NPs abolished ABC, although a high level of IgM was still detected, which suggests saturation with insignificant poisoning of immune cells. CONCLUSION: The presented results demonstrate the importance of clinical evaluations for PLGA-PEG nanocarriers that consider the administration schedule in multiple drug delivery, particularly in cancer chemotherapy.
Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Antineoplásicos Fitogênicos/sangue , Portadores de Fármacos/metabolismo , Etoposídeo/administração & dosagem , Etoposídeo/sangue , Ácido Láctico/metabolismo , Polietilenoglicóis/metabolismo , Ácido Poliglicólico/metabolismo , Animais , Anticorpos/análise , Portadores de Fármacos/administração & dosagem , Portadores de Fármacos/análise , Imunoglobulina M/imunologia , Injeções Intravenosas , Ácido Láctico/administração & dosagem , Ácido Láctico/análise , Masculino , Nanopartículas/administração & dosagem , Nanopartículas/análise , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/análise , Ácido Poliglicólico/administração & dosagem , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Ratos , Ratos WistarRESUMO
OBJECTIVE: To fabricate the antibiotic-releasing coatings on TiO(2) nanotube surfaces for wide applications of implant and bone plate in medical and dental surgery, the optimal deposition time of amoxicillin/PLGA solution simultaneously performing non-toxicity and a high bactericidal effect for preventing early implant failures was found. MATERIALS AND METHODS: FE-SEM, ESD and FT-IR were used for confirming deposition of amoxicillin/PLGA on the TiO(2) surface. Also, the elution of amoxicillin/PLGA in a TiO(2) nanotube surface was measured by a UV-VIS spectrophotometer. The bactericidal effect of amoxicillin on the TiO(2) nanotube surface was evaluated by using Staphylococcus aureus (S. aureus). The cytotoxicity and cell proliferation were observed by WST assay using MC3T3-E1 osteoblast cells. RESULTS: The results indicated that the TiO(2) nanotube surface controlled by electro-spray deposition time with amoxicillin/PLGA solution could provide a high bactericidal effect against S. aureus by the bactericidal effect of amoxicillin, as well as good osteoblast cell proliferation at the TiO(2) nanotube surface without toxicity. CONCLUSIONS: This study used electro-spray deposition (ESD) methodology to obtain amoxicillin deposition in nanotube structures of TiO(2) and found the optimal deposition time of amoxicillin/PLGA solution simultaneously performing non-toxicity and a high bactericidal effect for preventing early implant failures.
Assuntos
Amoxicilina/administração & dosagem , Antibacterianos/administração & dosagem , Materiais Revestidos Biocompatíveis/administração & dosagem , Implantação Dentária Endóssea/métodos , Implantes Dentários , Ácido Láctico/administração & dosagem , Ácido Poliglicólico/administração & dosagem , Infecções Estafilocócicas/prevenção & controle , Células 3T3 , Amoxicilina/análise , Animais , Antibacterianos/análise , Placas Ósseas/microbiologia , Adesão Celular , Proliferação de Células , Materiais Revestidos Biocompatíveis/análise , Implantes Dentários/microbiologia , Combinação de Medicamentos , Ácido Láctico/análise , Camundongos , Nanotubos , Osteoblastos , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Espectrofotometria , Espectroscopia de Infravermelho com Transformada de Fourier , Staphylococcus aureus/efeitos dos fármacos , Propriedades de Superfície , TitânioRESUMO
OBJECTIVE: The selection of the degradation pattern of suture materials is critical in uvulopalatopharyngeal surgery because of the complex oral environment such as the constant presence of saliva; the microbial accumulation; and the functions related to speech, mastication, and swallowing. The aim of the study was to identify appropriate suture materials for uvulopalatopharyngeal surgery by analyzing the clinical and biomechanical results obtained using four different absorbable suture materials. STUDY DESIGN: A randomized open-label trial. SETTING: Tertiary referral center. SUBJECTS AND METHODS: Eighty patients with obstructive sleep apnea syndrome who had undergone microdebrider-assisted extended uvulopalatoplasty were randomized into the four suture groups: sutures of polyglycolide (Dexon) and copolymers of polyglycolide and other degradable polymers (Polysorb, Monocryl, Maxon). Handling characteristics, surgical outcomes, Young's modulus reduction, morphologic changes by scanning electron microscope, and saliva absorption test were analyzed. RESULTS: Monocryl (poly(glycolide-co-ε-caprolactone)) had the optimal handling characteristics and exhibited two-stage degradation: sufficient tensile strength was maintained initially to allow wound healing, and then the suture degraded rapidly to avoid irritation. In contrast, the hydrolytic degradation of Dexon, Polysorb, and Maxon seemed to involve only an approximately single-stage process in the oral environment. Moreover, a large amount of debris was observed among filaments in Dexon and Polysorb, whereas only some scattered debris accumulated on Monocryl and Maxon under scanning electron microscope. CONCLUSION: Poly(glycolide-co-ε-caprolactone) has good handling characteristics and degrades in two stages. It is a suitable suture material for uvulopalatopharyngeal surgery.
Assuntos
Teste de Materiais/métodos , Ácido Poliglicólico/análise , Polímeros/análise , Apneia Obstrutiva do Sono/cirurgia , Técnicas de Sutura/instrumentação , Suturas , Adulto , Fenômenos Biomecânicos , Feminino , Humanos , Masculino , Microscopia Eletrônica de Varredura , Procedimentos Cirúrgicos Bucais/métodos , Procedimentos Cirúrgicos Otorrinolaringológicos/métodos , Estudos Prospectivos , Reprodutibilidade dos TestesRESUMO
The present investigation was aimed to develop and explore the prospective of engineered PLGA nanoparticles as vehicles for targeted delivery of 5-fluorouracil (5-FU). Nanoparticles of 5-FU-loaded hyaluronic acid-poly(ethylene glycol)-poly(lactide-co-glycolide) (HA-PEG-PLGA-FU) copolymer were prepared and characterized by FTIR, NMR, transmission electron microscopy, particle size analysis, DSC, and X-ray diffractometer measurement studies. The nanoparticulate formulation was evaluated for in vitro release, hemolytic toxicity, and hematological toxicity. Cytotoxicity studies were performed on Ehrlich ascites tumor (EAT) cell lines using MTT cell proliferation assay. Biodistribution studies of 99m Tc labeled formulation were conducted on EAT-bearing mice. The in vivo tumor inhibition study was also performed after i.v. administration of HA-PEG-PLGA-FU nanoparticles. The HA conjugated formulation was found to be less hemolytic but more cytotoxic as compared to free drug. The hematological data suggested that HA-PEG-PLGA-FU formulation was less immunogenic compared to plain drug. The tissue distribution studies displayed that HA-PEG-PLGA-FU were able to deliver a higher concentration of 5-FU in the tumor mass. In addition, the HA-PEG-PLGA-FU nanoparticles reduced tumor volume significantly in comparison with 5-FU. Thus, it was concluded that the conjugation of HA imparts targetability to the formulation, and enhanced permeation and retention effect ruled out its access to the non-tumor tissues, at the same time favored selective entry in tumors, thereby reducing the side-effects both in vitro and in vivo.
Assuntos
Sistemas de Liberação de Medicamentos , Fluoruracila/administração & dosagem , Ácido Hialurônico/administração & dosagem , Ácido Láctico , Nanopartículas , Ácido Poliglicólico , Animais , Materiais Biocompatíveis/administração & dosagem , Carcinoma de Ehrlich/tratamento farmacológico , Carcinoma de Ehrlich/metabolismo , Linhagem Celular Tumoral , Química Farmacêutica , Portadores de Fármacos/administração & dosagem , Fluoruracila/farmacocinética , Ácido Hialurônico/química , Ácido Láctico/administração & dosagem , Ácido Láctico/análise , Ácido Láctico/química , Imageamento por Ressonância Magnética , Camundongos , Microscopia Eletrônica de Transmissão , Nanopartículas/administração & dosagem , Nanopartículas/análise , Nanopartículas/química , Tamanho da Partícula , Ácido Poliglicólico/administração & dosagem , Ácido Poliglicólico/análise , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Espectroscopia de Infravermelho com Transformada de Fourier , Testes de Toxicidade , Difração de Raios XRESUMO
Efficient and specific delivery of antisenses (ASs) and protection of the sequences from degradation are critical factors for effective therapy. Sustained release nanoparticles (NP) offer increased resistance to nuclease degradation, increased amounts of AS uptake, and the possibility of control in dosing and sustained duration of AS administration. The biodegradable and biocompatible poly(D,L-lactic-co-glycolic acid) copolymer (PLGA) was utilized to encapsulate AS directed against osteopontin (OPN), which is a promising therapeutic target in mammary carcinoma. Whole body biodistribution of OPN AS NP was evaluated in comparison to naked AS, in intact and mammary carcinoma metastasis model bearing rats. Naked and NP encapsulated AS exhibited different biodistribution profiles. AS NP, in contrast to naked AS, tended to accumulate mostly in the spleen, liver, and at the tumor inoculation site. Drug levels in intact organs were negligible. The elimination of naked AS was faster, due to rapid degradation of the unprotected sequence. It is concluded that AS NP protect the AS from degradation, provide efficient AS delivery to the tumor tissue, and minimize AS accumulation in intact organs due to the AS sustained release profile as well as the favorable NP physicochemical properties.
Assuntos
Antineoplásicos/farmacocinética , Elementos Antissenso (Genética)/farmacocinética , Neoplasias Ósseas , Carcinoma , Portadores de Fármacos/farmacocinética , Neoplasias Mamárias Experimentais , Nanopartículas/química , Animais , Antineoplásicos/análise , Antineoplásicos/sangue , Antineoplásicos/urina , Elementos Antissenso (Genética)/análise , Elementos Antissenso (Genética)/sangue , Elementos Antissenso (Genética)/urina , Neoplasias Ósseas/sangue , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/secundário , Neoplasias Ósseas/urina , Carcinoma/sangue , Carcinoma/tratamento farmacológico , Carcinoma/secundário , Carcinoma/urina , Linhagem Celular Tumoral , Preparações de Ação Retardada/análise , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Preparações de Ação Retardada/uso terapêutico , Portadores de Fármacos/análise , Sistemas de Liberação de Medicamentos , Feminino , Humanos , Ácido Láctico/análise , Ácido Láctico/química , Ácido Láctico/uso terapêutico , Masculino , Neoplasias Mamárias Experimentais/sangue , Neoplasias Mamárias Experimentais/tratamento farmacológico , Neoplasias Mamárias Experimentais/urina , Nanopartículas/análise , Nanopartículas/uso terapêutico , Osteopontina/genética , Tamanho da Partícula , Ácido Poliglicólico/análise , Ácido Poliglicólico/química , Ácido Poliglicólico/uso terapêutico , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Ratos , Ratos Nus , Distribuição Tecidual , Ensaios Antitumorais Modelo de Xenoenxerto/métodosRESUMO
The present study had two main objectives. First, was to compare the immune stimulatory effect of two synthetic lipid A analogues (7-acyl lipid A and pentaerythritol-based lipid A (PET lipid A)) on maturation/stimulation of bone marrow derived dendritic cells (DCs). Our second objective was to develop a liquid chromatography/mass spectrometry (LC-MS) method for the quantitative analysis of lipid A-based vaccine adjuvants. Treatment of immature DCs with 7-acyl lipid A and PET lipid A up regulated the surface expression of CD86 and CD40 molecules, and also induced similar profile of pro-inflammatory cytokine secretion. LC-MS analyses were performed using a Waters Micromass ZQ 4000 spectrometer, coupled to a Waters 2795 separations module with an autosampler. Calibration curves with R(2)>0.999 were constructed over the concentration range of 1.25-20 microg/ml for the solution of 7-acyl lipid A and PET lipid A. The method was tested in a 3 day validation protocol. The accuracy of the assay at different concentrations tested ranged from 89 to 108% and from 92 to 107% for 7-acyl lipid A and PET lipid A, respectively. The limit of quantification for both 7-acyl lipid A and PET lipid A was 1.25 microg/ml (signal/noise (S/N)) ratio >15:1. The sensitivity of the method (the limit of detection) was 0.35 and 0.15 ng for 7-acyl lipid A and PET lipid A, respectively (S/N ratio between 4:1 or 3:1). As a preliminary application, this method has been successfully applied to the determination of 7-acyl lipid A and PET lipid A content in poly (D,L-lactic-co-glycolic acid) nanoparticles (PLGA-NP).
Assuntos
Adjuvantes Imunológicos/análise , Ácido Láctico/análise , Lipídeo A/análise , Nanopartículas/análise , Ácido Poliglicólico/análise , Polímeros/análise , Animais , Antígeno B7-2/biossíntese , Células da Medula Óssea/fisiologia , Antígenos CD40/biossíntese , Química Farmacêutica , Cromatografia Líquida , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Células Dendríticas/fisiologia , Composição de Medicamentos , Citometria de Fluxo , Lipídeo A/análogos & derivados , Espectrometria de Massas , Camundongos , Camundongos Endogâmicos C57BL , Fenótipo , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Propilenoglicóis/química , Padrões de Referência , Reprodutibilidade dos Testes , Soluções , Regulação para Cima/efeitos dos fármacosRESUMO
The purpose of this study is to examine the effect of electron-beam (e-beam) radiation on the hydrolytic degradation of poly(lactide-co-glycolide) (PLGA) films. PLGA films were irradiated and observed to undergo radiation-induced degradation through chain scission, as observed from a drop in its average molecular weight with radiation dose. Irradiated (5, 10 and 20 Mrad) and non-irradiated (0 Mrad) samples of PLGA were subsequently hydrolytically degraded in phosphate-buffered saline solution at 37.0 degrees C over a span of 12 weeks. It was observed that the natural logarithmic molecular weight (lnMn) of PLGA decreases linearly with hydrolytic degradation time. The rate of water uptake is higher for samples irradiated at higher radiation dose (e.g. 20 Mrad) and subsequently causing an earlier onset of mass loss. It is postulated that the increase in water uptake is due to the presence of more hydrophilic end groups, which results in the formation of microcavities because of an increase in osmotic pressure. A relationship between radiation dose and the rate of hydrolytic degradation of PLGA films, through its molecular weight was also established. This relationship allows a more accurate and precise control of the life span of PLGA through the use of e-beam radiation.
Assuntos
Implantes Absorvíveis , Materiais Biocompatíveis/química , Materiais Biocompatíveis/efeitos da radiação , Ácido Láctico/química , Ácido Láctico/efeitos da radiação , Teste de Materiais/métodos , Ácido Poliglicólico/química , Ácido Poliglicólico/efeitos da radiação , Polímeros/química , Polímeros/efeitos da radiação , Absorção , Materiais Biocompatíveis/análise , Líquidos Corporais/química , Relação Dose-Resposta à Radiação , Elétrons , Hidrólise , Ácido Láctico/análise , Peso Molecular , Permeabilidade/efeitos da radiação , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros/análise , Porosidade/efeitos da radiação , Doses de Radiação , Propriedades de SuperfícieRESUMO
In total, approximately 400 million people worldwide suffer from urinary bladder cancer (Nat Biotechnol 17 (1999) 149). When radical cysectomy is required as treatment, a replacement material is clearly necessitated. For this purpose, three-dimensional poly(lactic-co-glycolic acid) (PLGA) scaffolds were constructed using solvent casting and salt leaching processes. These scaffolds were manipulated to possess nano-dimensional surface features by soaking in sodium hydroxide at select concentrations and for various periods of time. Human bladder smooth muscle cells were then seeded onto these nano-dimensional scaffolds; adhesion and longer-term cell growth experiments were performed for either 4 h, or 1, 3, and 5 days, respectively. Additionally, collagen and elastin production was quantified following each experiment. In all cases, control cells were placed in an incubator and subjected to normal atmospheric pressure, while experimental cells were placed in a pressure chamber and subjected to a sustained pressure of 10 cm H(2)O. Results of this study provided evidence that porous, nano-dimensional polymeric scaffolds enhanced cell adhesion and growth, while also promoting increased elastin and collagen production. Moreover, in general, exposure to pressure did not alter cellular adhesion, growth, or extracellular matrix protein production, which suggests that the scaffolds and their resident cells will fair well in the complex mechanical environment of the bladder wall. In combination, these results provide evidence that the nano-dimensional PLGA scaffolds created in this research are promising as the next generation of bladder wall replacement materials.
Assuntos
Bioprótese , Ácido Láctico/química , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/fisiologia , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Ácido Poliglicólico/química , Polímeros/química , Engenharia Tecidual/métodos , Bexiga Urinária/citologia , Bexiga Urinária/crescimento & desenvolvimento , Materiais Biocompatíveis/química , Adesão Celular/fisiologia , Técnicas de Cultura de Células/métodos , Proliferação de Células , Sobrevivência Celular/fisiologia , Células Cultivadas , Humanos , Ácido Láctico/análise , Teste de Materiais , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros/análise , Propriedades de Superfície , Bexiga Urinária/irrigação sanguíneaRESUMO
A novel biologically benign technique was developed to produce three-dimensional tissue engineering scaffolds with well-defined structure. Photolithography was used to design and pattern a planar scaffold skeletal structure on a photoresist (SU-8), and a variety of microembossing processes including sacrificial layer embossing and bilayer embossing were developed to transfer the skeletal pattern to the poly(DL-lactide-co-glycolide) substrate as scaffold skeletons. Subcritical carbon dioxide was then introduced to assemble these skeletons to a three-dimensional scaffold at a low temperature. Compared with conventional scaffolds, which have a broad pore size distribution and varying pore geometry, these microfabricated scaffolds have a uniform and well-defined geometry and structure. This uniformity of structural parameters allows for the studies of cell attachment, spreading, and proliferation in scaffolds in a controlled and logical manner. The cytocompatibility of these microfabricated scaffolds was tested by seeding three different cell lines with different morphologies and growth patterns into these scaffolds. All three cell lines attached well to the scaffolds and grew to high densities as observed with scanning electron microscopy. This study demonstrates a controllable method to fabricate tissue scaffolds with a well-defined 3D architecture that can be used to better elucidate the effect of structure parameters such as pore geometry and pore size on tissue growth in 3D scaffolds.
Assuntos
Astrocitoma/patologia , Materiais Biocompatíveis/química , Dióxido de Carbono/química , Técnicas de Cultura de Células/métodos , Ácido Láctico/química , Neoplasias Mamárias Animais/patologia , Fotografação/métodos , Ácido Poliglicólico/química , Polímeros/química , Animais , Adesão Celular/fisiologia , Proliferação de Células , Tamanho Celular , Células Cultivadas , Cricetinae , Cricetulus , Humanos , Ácido Láctico/análise , Teste de Materiais , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros/análise , Propriedades de Superfície , Engenharia Tecidual/métodosRESUMO
This study evaluated cellular uptake of polymeric nanoparticles by using Caco-2 cells, a human colon adenocarcinoma cell line, as an in vitro model with the aim to apply nanoparticles of biodegradable polymers for oral chemotherapy. The feasibility was demonstrated by showing the localization and quantification of the cell uptake of fluorescent polystyrene nanoparticles of standard size and poly(lactic-co-glycolic acid) (PLGA) nanoparticles coated with polyvinyl alcohol (PVA) or vitamin E TPGS. Coumarin-6 loaded PLGA nanoparticles were prepared by a modified solvent extraction/evaporation method and characterized by laser light scattering for size and size distribution, scanning electron microscopy (SEM) for surface morphology, zeta-potential for surface charge, and spectrofluorometry for fluorescent molecule release from the nanoparticles. The effects of particle size and particle surface coating on the cellular uptake of the nanoparticles were quantified by spectrofluorometric measurement. Cellular uptake of vitamin E TPGS-coated PLGA nanoparticles showed 1.4 folds higher than that of PVA-coated PLGA nanoparticles and 4-6 folds higher than that of nude polystyrene nanoparticles. Images of confocal laser scanning microscopy, cryo-SEM and transmission electron microscopy clearly evidenced the internalization of nanoparticles by the Caco-2 cells, showing that surface modification of PLGA nanoparticles with vitamin E TPGS notably improved the cellular uptake. It is highly feasible for nanoparticles of biodegradable polymers to be applied to promote oral chemotherapy.
Assuntos
Antineoplásicos/administração & dosagem , Endocitose/efeitos dos fármacos , Ácido Láctico/química , Nanotubos/química , Nanotubos/ultraestrutura , Ácido Poliglicólico/química , Polímeros/química , Vitamina E/administração & dosagem , Administração Oral , Antineoplásicos/química , Células CACO-2 , Materiais Revestidos Biocompatíveis/administração & dosagem , Materiais Revestidos Biocompatíveis/química , Difusão , Humanos , Ácido Láctico/análise , Teste de Materiais , Tamanho da Partícula , Ácido Poliglicólico/análise , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Polímeros/análise , Vitamina E/químicaRESUMO
A quantitative determination of peptides incorporated into poly(d,l-lactide-co-glycolide) microspheres by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was accomplished in a single step without pretreatment for extracting the peptide from the microsphere. The conventional extraction methods often underestimate the actual amount of peptide because of incomplete extraction from the microspheres or loss during the procedures. In this study, the microspheres dissolved in acetonitrile containing 0.1% trifluoroacetic acid were mixed with matrix solution containing the internal standard, and the peptide content was directly determined by MALDI-TOF MS. The drug content values determined by MALDI-TOF MS in both the leuprolide- and salmon calcitonin-incorporated microspheres were closer to the theoretical contents than those determined by the conventional extraction method. This method using MALDI-TOF MS could be a good alternative to time-consuming and less-accurate conventional methods.