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1.
Sci Rep ; 14(1): 8250, 2024 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-38589494

RESUMO

Personalized, ultra-fractionated stereotactic adaptive radiotherapy (PULSAR) is designed to administer tumoricidal doses in a pulsed mode with extended intervals, spanning weeks or months. This approach leverages longer intervals to adapt the treatment plan based on tumor changes and enhance immune-modulated effects. In this investigation, we seek to elucidate the potential synergy between combined PULSAR and PD-L1 blockade immunotherapy using experimental data from a Lewis Lung Carcinoma (LLC) syngeneic murine cancer model. Employing a long short-term memory (LSTM) recurrent neural network (RNN) model, we simulated the treatment response by treating irradiation and anti-PD-L1 as external stimuli occurring in a temporal sequence. Our findings demonstrate that: (1) The model can simulate tumor growth by integrating various parameters such as timing and dose, and (2) The model provides mechanistic interpretations of a "causal relationship" in combined treatment, offering a completely novel perspective. The model can be utilized for in-silico modeling, facilitating exploration of innovative treatment combinations to optimize therapeutic outcomes. Advanced modeling techniques, coupled with additional efforts in biomarker identification, may deepen our understanding of the biological mechanisms underlying the combined treatment.


Assuntos
DEAE-Dextrano , Radiocirurgia , Animais , Camundongos , Imunoterapia/métodos , Redes Neurais de Computação , Terapia Combinada , Antígeno B7-H1
2.
Lancet ; 403(10432): 1141-1152, 2024 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-38461841

RESUMO

BACKGROUND: Intravitreal aflibercept 8 mg could improve treatment outcomes and provide sustained disease control in patients with neovascular age-related macular degeneration (nAMD), with extended dosing compared with aflibercept 2 mg. METHODS: PULSAR is a phase 3, randomised, three-group, double-masked, non-inferiority, 96-week trial conducted across 223 sites worldwide. Adults with nAMD were randomised 1:1:1 to aflibercept 8 mg every 12 weeks (8q12), aflibercept 8 mg every 16 weeks (8q16), or aflibercept 2 mg every 8 weeks (2q8), following three initial monthly doses in all groups. From week 16, patients in the aflibercept 8 mg groups had their dosing interval shortened if pre-specified dose regimen modification criteria denoting disease activity were met. The primary endpoint was change from baseline in best-corrected visual acuity (BCVA) at week 48. All patients with at least one dose of study treatment were included in the efficacy and safety analyses. This trial is registered with ClinicalTrials.gov (NCT04423718) and is ongoing. FINDINGS: Of 1011 patients randomised to aflibercept 8q12 (n=336), 8q16 (n=338), or 2q8 (n=337) between Aug 11, 2020, and July 30, 2021, 1009 patients received study treatment (aflibercept 8q12 n=335; aflibercept 8q16 n=338; and aflibercept 2q8 n=336). Aflibercept 8q12 and 8q16 showed non-inferior BCVA gains versus aflibercept 2q8 (mean BCVA change from baseline +6·7 [SD 12·6] and +6·2 [11·7] vs +7·6 [12·2] letters). The least squares mean differences between aflibercept 8q12 versus 2q8 and 8q16 versus 2q8, respectively, were -0·97 (95% CI -2·87 to 0·92) and -1·14 (-2·97 to 0·69) letters (non-inferiority margin at 4 letters). The incidence of ocular adverse events in the study eye was similar across groups (aflibercept 8q12 n=129 [39%]; aflibercept 8q16 n=127 [38%]; and aflibercept 2q8 n=130 [39%]). INTERPRETATION: Aflibercept 8 mg showed efficacy and safety with extended dosing intervals, which has the potential to improve the management of patients with nAMD. FUNDING: Bayer AG and Regeneron Pharmaceuticals.


Assuntos
Inibidores da Angiogênese , Degeneração Macular , Adulto , Humanos , Inibidores da Angiogênese/efeitos adversos , DEAE-Dextrano , Degeneração Macular/tratamento farmacológico , Receptores de Fatores de Crescimento do Endotélio Vascular/uso terapêutico , Proteínas Recombinantes de Fusão/efeitos adversos , Resultado do Tratamento
3.
Mol Biotechnol ; 65(4): 544-555, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35999479

RESUMO

Genetic engineering of mesenchymal stromal cells (MSCs) is a tool widely used to explore MSC properties in vitro and in vivo. Lentiviral infection with the use of polycations as an adjuvant is a method that is commonly used to generate stably transduced cells. However, it is known that some polycations can negatively affect primary MSCs and to date, no study has explored the effect of different polycations on the transduction efficiency and properties of all main types of MSCs, namely those derived from umbilical cord, bone marrow and adipose tissue. Here we explore a range of polycations, using transduction protocols with and without spinoculation, to produce stably transduced MSCs from these three tissue sources. We identified that an overnight incubation with diethylaminoethyl-dextran (DEAE-Dextran) is the protocol associated with the best transduction efficiency without compromising the viability of the cells, and which worked consistently with lentiviral particles encoding for different transgenes. Transduced and sorted MSC populations revealed no significant changes in proliferation, morphology and expression of MSC markers compared to naïve MSCs. Following this study, we conclude that DEAE-Dextran is a polycation that can be successfully used to enhance the transduction of MSCs from all major tissue sources.


Assuntos
DEAE-Dextrano , Células-Tronco Mesenquimais , Humanos , Transdução Genética , DEAE-Dextrano/metabolismo , Lentivirus/genética , Vetores Genéticos/genética , Fenótipo , Proliferação de Células , Diferenciação Celular , Células Cultivadas
4.
J Mater Chem B ; 9(47): 9784-9793, 2021 12 08.
Artigo em Inglês | MEDLINE | ID: mdl-34820677

RESUMO

Liquid coacervate microdroplets have been widely explored as membrane-free compartment protocells for cargo delivery in therapeutic applications. In this study, coacervate protocells were developed as gene carriers for transfection of nitric oxide synthase (NOS) and overproduction of nitric oxide (NO) for killing of cancer cells. The coacervate microdroplet protocells were formed via the liquid-liquid phase separation of oppositely charged diethylaminoethyl-dextran/polyacrylic acids. The coacervate microdroplet protocells were found to facilitate gene transfection, which was demonstrated by cell imaging of the internalized coacervate microdroplets containing plasmids of enhanced green fluorescent protein. Due to their high transfection capability, the coacervate protocells were subsequently utilized for the delivery of NOS plasmids (pNOS). The cellular internalization of pNOS-containing coacervate carriers was found to result in high NOS expression coupled with NO overproduction, which then induced cell apoptosis and decreased cell viability. The cell apoptosis is associated with NO-mediated mitochondrial damage. The enhanced gene transfection was attributed to coacervate microdroplets' unique high sequestration capability and liquid-like fluidity. Overall, the incorporation of genes in coacervate microdroplets was demonstrated as a viable and novel strategy for the development of cargo biocarriers for biomedical applications.


Assuntos
Apoptose/efeitos dos fármacos , Células Artificiais/química , DNA/farmacologia , Portadores de Fármacos/química , Óxido Nítrico/metabolismo , Resinas Acrílicas/química , Linhagem Celular Tumoral , DEAE-Dextrano/química , DNA/genética , Proteínas de Fluorescência Verde/genética , Humanos , Óxido Nítrico Sintase/genética , Plasmídeos , Transfecção/métodos
5.
J Biomed Mater Res B Appl Biomater ; 109(4): 527-537, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32945122

RESUMO

Presently, most of anticancer drugs are high toxic for normal cells and, and as a result, they have severe side effects. Moreover, most of the formulations are lipophilic and have poor selectivity. To overcome these limitations, various drug delivery systems could be proposed. The aim of the current study was to fabricate novel polysaccharide nanocontainers (NC) by one-step ultrasonication technique and to evaluate their accumulation efficacy and cytotoxicity in 2D (monolayer culture) and 3D (tumor spheroids) in vitro models. NC with mean sizes in a range of 340-420 nm with the core-shell structure are synthetized and characterized. The NC shell is composed from diethylaminoethyl dextran/xanthan gum polyelectrolyte complex, while the hydrophobic core was loaded with the lipophilic anticancer drug thymoquinone. To enhance NC accumulation in human breast adenocarcinoma MCF-7 cells, the NC surface was modified with poly-L-lysine (PLL) or polyethylene glycol. Cell uptake of the NC loaded with Nile Red into the tumor cells was investigated by laser scanning confocal microscopy, fluorescent flow cytometry and fluorimetry. Modification of the NC with PLL allowed to obtain the optimal drug delivery system with maximal cytotoxicity, which was tested by MTT-test. The developed NC are promising for lipophilic anticancer drug delivery.


Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Benzoquinonas/administração & dosagem , Embalagem de Medicamentos/instrumentação , Sistemas de Liberação de Fármacos por Nanopartículas , Antineoplásicos Fitogênicos/química , Benzoquinonas/química , Técnicas de Cultura de Células em Três Dimensões , DEAE-Dextrano , Emulsões , Feminino , Citometria de Fluxo , Fluorometria , Humanos , Interações Hidrofóbicas e Hidrofílicas , Técnicas In Vitro , Células MCF-7 , Microscopia Confocal , Oxazinas/análise , Polietilenoglicóis , Polilisina , Polissacarídeos Bacterianos , Sonicação , Esferoides Celulares/efeitos dos fármacos
6.
Biomaterials ; 192: 612-620, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30502967

RESUMO

mRNA pharmaceuticals represent a new class of therapeutics, with applications, in cancer vaccination, tumour therapy and protein substitution. Formulations are required to deliver messenger RNA (mRNA) to the target sites where induction of genetic transfection following receptor mediated cell uptake & translation is required. In the current study, the cationic polysaccharide diethylaminoethylen (DEAE) - Dextran was selected as a model system carrier for the investigation of polyplex nanoparticle formation together with mRNA as a function of the molar ratio of the components. The structure of the mRNA/Dextran colloids was investigated as a function of the polymer-to-mRNA ratio and correlated with the biological activity determined by cellular transfection with luciferase coding mRNA. Dynamic light scattering (DLS), small angle x-ray scattering (SAXS), and small angle neutron scattering (SANS) with deuterium contrast variation were used to achieve structural insight into the systems. Similarly to previously investigated lipid based systems, colloidally stable particles with confined size were obtained with either excess of positive or negative charge. Highest activity was obtained with positive charge excess. From the scattering experiments information on the internal organization inside the polymer/mRNA systems was derived. Indication for the presence of structural elements in the length scale of ten to 20 nm were found in the excess of dextran, which could be due to either excess or particulate polymer. Information on the molecular organization of the mRNA nanoparticle products may provide a valuable basis for defining critical quality attributes of drug products for pharmaceutical application.


Assuntos
DEAE-Dextrano/química , Sistemas de Liberação de Medicamentos , RNA Mensageiro/química , Células Dendríticas/metabolismo , Heparina/metabolismo , Humanos , Tamanho da Partícula , Espalhamento a Baixo Ângulo , Eletricidade Estática , Difração de Raios X
7.
Integr Biol (Camb) ; 10(9): 549-554, 2018 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-30140840

RESUMO

A DEAE-dextran-MMA copolymer (DDMC)-paclitaxel (PTX) conjugate was prepared using PTX as the guest and DDMC as the host. The resistance of B16F10 melanoma cells to PTX was confirmed, while the DDMC-PTX conjugate showed excellent anticancer activity that followed the Hill equation. The robustness in the tumor microenvironment of the allosteric system was confirmed via BIBO stability. This feedback control system, explained via a transfer function, was very stable and showed the sustainability of the system via a loop, and it showed superior anti-cancer activity without drug resistance from cancer cells. The block diagram of this signal system in the tumor microenvironment used its loop transfer function G(s) and the dN(s) of the external force. This indicial response is an ideal one without a time lag for the outlet response. The cell death rate of DDMC-PTX is more dependent on the Hill coefficient n than on the Michaelis constant Km. This means that this supermolecular reaction with tubulin follows an "induced fit model".


Assuntos
Melanoma/tratamento farmacológico , Paclitaxel/administração & dosagem , Zalcitabina/análogos & derivados , Sítio Alostérico , Animais , Antineoplásicos Fitogênicos/administração & dosagem , Linhagem Celular Tumoral , DEAE-Dextrano/química , Feminino , Humanos , Cinética , Melanoma Experimental , Camundongos , Camundongos Endogâmicos C57BL , Transplante de Neoplasias , Tamanho da Partícula , Transdução de Sinais , Microambiente Tumoral , Zalcitabina/administração & dosagem
8.
Pharmacol Rep ; 70(3): 549-557, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29677525

RESUMO

BACKGROUND: Angiogenesis, the physiological process involving growth of new blood vessels from preexisting vessels, is essential for organ growth and repair. However, the imbalance in angiogenesis contributes to copious pathologies including cancer. Preceding the development of anti-angiogenic or proangiogenic agents, its evaluation is equally imperative; hence, precise and adequate models required. Valid mammalian models are expensive, time-consuming and not easy to set up, instigating legal and ethical aspects making it necessary to establish models with satisfactory activity and limited drawbacks. METHODS: We investigated the activity of DEAE-Dextran on diversified models viz. in vitro cell migration assay, ex vivo aortic ring assay, in vitro chick yolk sac membrane assay and in vivo matrigel plug xenograft model corroborating its anti-angiogenic potential and establishing the best means of evaluation. RESULTS: Assorted models were reproducible and correlative to one another. DEAE-Dextran exhibited excellent anti-angiogenic effect in cell migration assay over a duration of 24h compared to the vehicle control fibroblast cell line and aortic ring possessed an alleviated rate of sprouting when treated with DEAE-Dextran with contrast to vehicle control aorta. Similarly, decreased vascular density was observed in DEAE-Dextran treated chick embryos implicating potency of the ß-interferon inducer. Augmenting to these results, the matrigel plugs also mitigated vascular net as well as reduced levels of angiogenic marker CD31. CONCLUSION: Substantially, DEAE-Dextran leads to anti-tumor activity through anti-angiogenic action and a combination of in vitro and in vivo model is vital for the judgement of anti-angiogenic potential since an in vitro model exempts mammalian-culture considerations.


Assuntos
Inibidores da Angiogênese/farmacologia , DEAE-Dextrano/farmacologia , Interferon beta/metabolismo , Neovascularização Patológica/tratamento farmacológico , Neovascularização Fisiológica/efeitos dos fármacos , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Linhagem Celular , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Embrião de Galinha , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Células HEK293 , Humanos , Indutores de Interferon/farmacologia , Camundongos , Neovascularização Patológica/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
9.
Eur J Pharm Biopharm ; 122: 37-48, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29031923

RESUMO

Triple negative breast cancer revolution has identified a plethora of therapeutic targets making it apparent that a single target for its treatment could be rare hence creating an urge to develop robust technologies for combination drug therapy. Paclitaxel, hailed as the most significant advancement in chemotherapy faces several underpinnings due to its low solubility and permeability. Advancing research has demonstrated the role of interferons in cancer. DEAE-Dextran, an emerging molecule with evidence of interferon induction was utilized in the present study to develop a nanoformulation in conjugation with paclitaxel to target multiple therapeutic pathways, with diminution of paclitaxel adverse effects and develop a specific targeted nano system. Evidently, it was demonstrated that DEAE-Dextran coated nanoformulation portrays significant synergistic cytotoxicity in the various cell lines. Moreover, overcoming the activation of ROS by paclitaxel, the combination drug therapy more effectively inhibited ROS through ß-interferon induction. The nanoformulation was further conjugated to FITC for internalization studies which subsequently indicated maximum cellular uptake at 60min post treatment demonstrated by green fluorescence from FITC lighting up the nuclear membrane. Precisely, the mechanistic approach of nuclear-targeted nanoformulation was evaluated by in vivo xenograft studies which showed a synergistic release of ß-interferon at the target organ. Moreover, the combination nanoformulation inculcated multiple mechanistic approaches through VEGF and NOTCH1 inhibition along with dual ß and γ-interferon overexpression. Overall, the combination therapy may be a promising multifunctional nanomaterial for intranuclear drug delivery in TNBC.


Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , DEAE-Dextrano/química , Nanopartículas/química , Paclitaxel/administração & dosagem , Receptor Notch1/antagonistas & inibidores , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Animais , Antineoplásicos Fitogênicos/química , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/química , Linhagem Celular , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos/métodos , Feminino , Células HEK293 , Humanos , Interferons/metabolismo , Células MCF-7 , Camundongos , Camundongos Endogâmicos BALB C , Paclitaxel/química , Neoplasias de Mama Triplo Negativas/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
10.
Regul Toxicol Pharmacol ; 88: 262-272, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28694171

RESUMO

Cancer has emerged as a global threat with challenges for safe chemotherapeutics. Most of the currently available anti-cancer drugs exhibit significant toxicity. Amongst novel agents, interferons have exhibited anti-proliferative and cytoprotective roles. However, due to stability drawbacks of interferons, we have identified an interferon inducer DEAE-Dextran, which resolves the stability issues. Based on the previous history of toxicity pertaining to the current chemotherapeutic agents, it is equally essential to determine the safety of DEAE-Dextran. In the present study, repeated dose 28 day oral toxicity of DEAE-Dextran has been evaluated in accordance to OECD-407. We found absence of any CNS behavioral changes related to self-mutilation, walking backwards, aggressiveness on handling or tonic-clonic seizures during the 28 day study. Neither the motor activity nor grip strength was altered during the treatment duration with DEAE-Dextran implying absence of any effect on the skeletal muscles. Interestingly, we also found that treatment with DEAE-Dextran did not present any significant cardiac, hepatic, renal, gastrointestinal, lymphatic or reproductive system toxicity or alteration in the body's normal physiology based upon the various organ function tests. Henceforth, it may be concluded that DEAE-Dextran is a safe anti-cancer agent devoid of any sub-acute toxicity.


Assuntos
Antineoplásicos/toxicidade , DEAE-Dextrano/toxicidade , Administração Oral , Animais , Comportamento Animal/efeitos dos fármacos , Sistema Nervoso Central/efeitos dos fármacos , DEAE-Dextrano/administração & dosagem , Camundongos , Segurança , Fatores de Tempo , Testes de Toxicidade Subaguda
11.
Sci Rep ; 6: 29197, 2016 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-27380727

RESUMO

Gene therapy is a promising technique for the treatment of various diseases. The development of minimally toxic and highly efficient non-viral gene delivery vectors is the most challenging undertaking in the field of gene therapy. Here, we developed dimethyldioctadecylammonium bromide (DODAB)-nanoceria (CeO2) hybrids as a new class of non-viral gene delivery vectors. These DODAB-modified CeO2 nanoparticles (CeO2/DODAB) could effectively compact the pDNA, allowing for highly efficient gene transfection into the selected cell lines. The CeO2/DODAB nanovectors were also found to be non-toxic and did not induce ROS formation as well as any stress responsive and pro-survival signaling pathways. The overall vector performance of CeO2/DODAB nanohybrids was comparable with lipofectamine and DOTAP, and higher than calcium phosphate and DEAE-dextran for transfecting small plasmids. The increased cellular uptake of the nanovector/DNA complexes through clathrin- and caveolae-mediated endocytosis and subsequent release from the endosomes further support the increased gene transfection efficiency of the CeO2/DODAB vectors. Besides, CeO2/DODAB nanovectors could transfect genes in vivo without any sign of toxicity. Taken together, this new nano-vector has the potential to be used for gene delivery in biomedical applications.


Assuntos
Cério/química , Endocitose , Técnicas de Transferência de Genes , Lipídeos/química , Nanopartículas/química , Cátions , Linhagem Celular Tumoral , DEAE-Dextrano , DNA/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Hidrodinâmica , Tamanho da Partícula , Plasmídeos/metabolismo , Compostos de Amônio Quaternário , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Transfecção
12.
Diagn Pathol ; 9: 195, 2014 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-25394479

RESUMO

BACKGROUND: Chronic obstructive pulmonary disease presents with two different phenotypes: chronic bronchitis and emphysema with parenchymal destruction. Decreased expression of vascular endothelial growth factor and increased endothelial cell apoptosis are considered major factors for emphysema. Stem cells have the ability of vascular regeneration and function as a repair mechanism for the damaged endothelial cells. Currently, minimally invasive interventional procedures such as placement of valves, bio-foam or coils are performed in order to improve the disturbed mechanical function in emphysema patients. However, these procedures cannot restore functional lung tissue. Additionally stem cell instillation into the parenchyma has been used in clinical studies aiming to improve overall respiratory function and quality of life. METHODS: In our current experiment we induced emphysema with a DDMC non-viral vector in BALBC mice and simultaneously instilled stem cells testing the hyposthesis that they might have a protective role against the development of emphysema. The mice were divided into four groups: a) control, b) 50.000 cells, c) 75.000 and d) 100.000 cells. RESULTS: Lung pathological findings revealed that all treatment groups had less damage compared to the control group. Additionally, we observed that emphysema lesions were less around vessels in an area of 10 µm. CONCLUSIONS: Our findings indicate that stem cell instillation can have a regenerative role if applied upon a tissue scaffold with vessel around. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_195.


Assuntos
DEAE-Dextrano , Pulmão/irrigação sanguínea , Pulmão/patologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Enfisema Pulmonar/prevenção & controle , Regeneração , Animais , Células Cultivadas , Modelos Animais de Doenças , Genes Reporter , Humanos , Pulmão/metabolismo , Células-Tronco Mesenquimais/metabolismo , Camundongos Endogâmicos BALB C , Enfisema Pulmonar/induzido quimicamente , Enfisema Pulmonar/metabolismo , Enfisema Pulmonar/patologia , Transfecção
13.
Gene Ther ; 21(2): 158-67, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24285215

RESUMO

Lung cancer still remains to be challenged by novel treatment modalities. Novel locally targeted routes of administration are a methodology to enhance treatment and reduce side effects. Intratumoral gene therapy is a method for local treatment and could be used either in early-stage lung cancer before surgery or at advanced stages as palliative care. Novel non-viral vectors are also in demand for efficient gene transfection to target local cancer tissue and at the same time protect the normal tissue. In the current study, C57BL/6 mice were divided into three groups: (a) control, (b) intravenous and (c) intatumoral gene therapy. The novel 2-Diethylaminoethyl-Dextran Methyl Methacrylate Copolymer Non-Viral Vector (Ryujyu Science Corporation) was conjugated with plasmid pSicop53 from the company Addgene for the first time. The aim of the study was to evaluate the safety and efficacy of targeted gene therapy in a Lewis lung cancer model. Indeed, although the pharmacokinetics of the different administration modalities differs, the intratumoral administration presented increased survival and decreased distant metastasis. Intratumoral gene therapy could be considered as an efficient local therapy for lung cancer.


Assuntos
Antineoplásicos/efeitos adversos , Carcinoma Pulmonar de Lewis/terapia , DEAE-Dextrano/efeitos adversos , Metilmetacrilato/efeitos adversos , Metástase Neoplásica/terapia , Proteína Supressora de Tumor p53/metabolismo , Administração Intravenosa , Animais , Antineoplásicos/administração & dosagem , Carcinoma Pulmonar de Lewis/patologia , Linhagem Celular Tumoral , DEAE-Dextrano/administração & dosagem , Terapia Genética/efeitos adversos , Terapia Genética/métodos , Metilmetacrilato/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Plasmídeos/administração & dosagem
14.
Gene Ther ; 20(10): 1022-8, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23719068

RESUMO

Revealing the lung tumor genome has directed the current treatment strategies toward targeted therapy. First line treatments targeting the genome of lung tumor cells have been approved and are on the market. However, they are limited by the small number of patients with the current investigated genetic mutations. Novel treatment administration modalities have been also investigated in an effort to increase the local drug deposition and disease control. In the current study, we investigated the safety of the new nonviral vector 2-diethylaminoethyl-dextran methyl methacrylate copolymer (DDMC; Ryujyu Science), which belongs to the 2-diethylaminoethyl-dextran family by aerosol administration. Thirty male BALBC mice, 2 month old, were included and divided into three groups. However, pathological findings indicated severe emphysema within three aerosol sessions. In addition, the CytoViva technique was applied for the first time to display the nonviral particles within the pulmonary tissue and emphysema lesions, and a spectral library of the nonviral vector was also established. Although our results in BALBC mice prevented us from further investigation of the DDMC nonviral vector as a vehicle for gene therapy, further investigation in animals with larger airways is warranted to properly evaluate the safety of the vector.


Assuntos
DEAE-Dextrano/toxicidade , Enfisema/induzido quimicamente , Terapia Genética , Pulmão/patologia , Metilmetacrilato/toxicidade , Administração por Inalação , Animais , DEAE-Dextrano/administração & dosagem , Terapia Genética/efeitos adversos , Terapia Genética/métodos , Neoplasias Pulmonares/terapia , Masculino , Metilmetacrilato/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C , Distribuição Aleatória
15.
J Control Release ; 162(2): 364-72, 2012 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-22846985

RESUMO

Electrostatic interactions between polycations and polyanions are being explored to fabricate polyelectrolyte complexes (PEC) that could entrap and regulate the release of a wide range of biomolecules. Here, we report the in vivo application of PEC shells fabricated from three different polycations: poly-l-ornithine (PLO), poly-l-arginine (PLA) and DEAE-dextran (DEAE-D) to condense heparin on the surface of alginate microbeads and further control the delivery of recombinant human bone morphogenetic protein 2 (rhBMP-2) in spinal fusion application. We observed large differences in the behavior of PEC shells fabricated from the cationic polyamino acids (PLO and PLA) when compared to the cationic polysaccharide, DEAE-D. Whereas DEAE-D-based PEC shells eroded and released rhBMP-2 over 2 days in vitro, PLO- and PLA-based shells retained at least 60% of loaded rhBMP-2 after 3 weeks of incubation in phosphate-buffered saline. In vivo implantation in a rat model of posterolateral spinal fusion revealed robust bone formation in the PLO- and PLA-based PEC shell groups. This resulted in a significantly enhanced mechanical stability of the fused segments. However, bone induction and biomechanical stability of spine segments implanted with DEAE-D-based carriers were significantly inferior to both PLO- and PLA-based PEC shell groups (p<0.01). From these results, we conclude that PEC shells incorporating native heparin could be used for growth factor delivery in functional bone tissue engineering application and that PLA- and PLO-based complexes could represent superior options to DEAE-D for loading and in vivo delivery of bioactive BMP-2 in this approach.


Assuntos
Proteína Morfogenética Óssea 2/administração & dosagem , Portadores de Fármacos/administração & dosagem , Heparina/administração & dosagem , Fator de Crescimento Transformador beta/administração & dosagem , Alginatos/química , Fosfatase Alcalina/metabolismo , Animais , Proteína Morfogenética Óssea 2/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , DEAE-Dextrano/administração & dosagem , DEAE-Dextrano/química , Portadores de Fármacos/química , Ácido Glucurônico/química , Heparina/química , Ácidos Hexurônicos/química , Masculino , Camundongos , Microesferas , Osteogênese/efeitos dos fármacos , Peptídeos/administração & dosagem , Peptídeos/química , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/química , Coluna Vertebral , Fator de Crescimento Transformador beta/química
16.
Skin Res Technol ; 16(3): 265-9, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20636993

RESUMO

BACKGROUND: Dermal fillers are used widely; some have a permanent effect, whereas others are temporary. The aim of this study is to describe the ultrasonographic features of permanent and temporary fillers injected into patients for cosmetic purposes. MATERIALS AND METHODS: Between December 2006 and April 2009, 36 subjects, aged 25-45, who had received lips or nasolabial fold filler augmentation, were enrolled for a high-frequency sonographic examination by a blinded investigator. The criteria for exclusion were a history of autoimmunity, infection, neoplastic diseases or episodes of local reactions to the injected filler. Twenty patients underwent a sonographic exam after the injection of a temporary filler (collagen or hyaluronic acid) by FRG; the rest were enrolled among patients seeking a consultation for further cosmetic reasons, but had been treated with an identifiable filler before. RESULTS: It was always possible to identify the filler at the site of injection. Seldom was it possible to discover a silent inflammatory reaction, otherwise unsuspected. The sonographic images differed according to the temporary or the permanent nature of the filler. CONCLUSION: Ultrasonography has proved to be a useful, non-invasive tool for the identification of the presence and type of the filler injected.


Assuntos
Materiais Biocompatíveis/administração & dosagem , Colágeno/administração & dosagem , Técnicas Cosméticas/instrumentação , Derme/diagnóstico por imagem , Ultrassonografia/métodos , Resinas Acrílicas/administração & dosagem , Resinas Acrílicas/farmacocinética , Adulto , Materiais Biocompatíveis/farmacocinética , Colágeno/farmacocinética , DEAE-Dextrano/administração & dosagem , DEAE-Dextrano/farmacocinética , Derme/metabolismo , Derme/patologia , Humanos , Ácido Hialurônico/administração & dosagem , Ácido Hialurônico/análogos & derivados , Ácido Hialurônico/farmacocinética , Hidrogéis/administração & dosagem , Hidrogéis/farmacocinética , Pessoa de Meia-Idade , Envelhecimento da Pele/efeitos dos fármacos , Envelhecimento da Pele/patologia , Gordura Subcutânea/diagnóstico por imagem , Gordura Subcutânea/metabolismo , Gordura Subcutânea/patologia
17.
Exp Cell Res ; 316(9): 1523-34, 2010 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-20302858

RESUMO

DNA helicases are important in the regulation of DNA transaction and thereby various cellular functions. In this study, we developed a cost-effective multiple DNA transfection assay with DEAE-dextran reagent and analyzed the promoter activities of the human DNA helicases. The 5'-flanking regions of the human DNA helicase-encoding genes were isolated and subcloned into luciferase (Luc) expression plasmids. They were coated onto 96-well plate and used for co-transfection with a renilla-Luc expression vector into various cells, and dual-Luc assays were performed. The profiles of promoter activities were dependent on cell lines used. Among these human DNA helicase genes, XPB, RecQL5, and RTEL promoters were activated during TPA-induced HL-60 cell differentiation. Interestingly, duplicated ets (GGAA) elements are commonly located around the transcription start sites of these genes. The duplicated GGAA motifs are also found in the promoters of DNA replication/repair synthesis factor genes including PARG, ATR, TERC, and Rb1. Mutation analyses suggested that the duplicated GGAA-motifs are necessary for the basal promoter activity in various cells and some of them positively respond to TPA in HL-60 cells. TPA-induced response of 44-bp in the RTEL promoter was attenuated by co-transfection of the PU.1 expression vector. These findings suggest that the duplicated ets motifs regulate DNA-repair associated gene expressions during macrophage-like differentiation of HL-60 cells.


Assuntos
DNA Helicases/genética , DNA Helicases/metabolismo , Regiões Promotoras Genéticas/genética , Proteínas Proto-Oncogênicas c-ets/metabolismo , Elementos Reguladores de Transcrição/genética , Antineoplásicos/farmacologia , Western Blotting , Carcinógenos/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , DEAE-Dextrano , Humanos , Luciferases/metabolismo , Macrófagos/citologia , Macrófagos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Acetato de Tetradecanoilforbol/farmacologia , Ativação Transcricional , Transfecção , Tretinoína/farmacologia
18.
J Drug Target ; 18(3): 168-78, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20136463

RESUMO

Our aim was to develop a novel liposomal drug delivery system containing dextrans to reduce undesirable retention of antineoplastic agents and thus alleviate local tissue damage. At the cell level, diethylaminoethyl-dextran (DEAE-Dx) showed the strongest inhibiting effect on liposome uptake by macrophages among tested dextrans. The distribution of radiolabeled liposomes mixed with dextrans in injection site and draining lymph node was investigated in rats after subcutaneous injection. DEAE-Dx substantially reduced the undesired local retention and promoted the draining of liposome into lymphatics, which was further confirmed by confocal microscopy images revealing the substantial prevention of rhodamine B-labelled liposome sequestration by macrophages in normal lymph node in rats. Pharmacokinetic data indicated the accelerated drainage of liposome through lymphatics back to systemic circulation by mixing with DEAE-Dx. In the toxicological study in rabbits, DEAE-Dx alleviated the local tissue damage caused by liposomal doxorubicin. In conclusion, dextrans, particularly DEAE-Dx, could efficiently enhanced liposomes drainage into lymphatics, which proves themselves as promising adjuvants for lymphatic-targeted liposomal drug delivery system.


Assuntos
Dextranos/farmacologia , Doxorrubicina/farmacocinética , Sistemas de Liberação de Medicamentos , Linfonodos/efeitos dos fármacos , Animais , Antibióticos Antineoplásicos/administração & dosagem , Antibióticos Antineoplásicos/farmacocinética , Antibióticos Antineoplásicos/toxicidade , DEAE-Dextrano/química , DEAE-Dextrano/farmacologia , Dextranos/química , Doxorrubicina/administração & dosagem , Doxorrubicina/toxicidade , Drenagem/métodos , Lipossomos , Linfonodos/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos , Microscopia Confocal , Coelhos , Ratos , Ratos Sprague-Dawley , Rodaminas/administração & dosagem , Rodaminas/farmacocinética
19.
Int J Pharm ; 365(1-2): 61-8, 2009 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-18812217

RESUMO

Microparticles from poly(D,L-lactic-co-glycolic acid) [PLGA] are of steadily rising interest for the delivery of antigens to immune cells and the induction of a long-lasting immune response for vaccination or immunological tumor therapy. However, if the desired vaccine contains only weak antigens and fails to activate the antigen presenting cells (APC), the opposite effect, i.e., the induction of immunotolerance may be observed. Therefore, it was the aim of this study to show the ability of protein loaded PLGA microparticles to additionally carry a specific, surface-coated maturation signal to human dendritic cells (DC), i.e., the most potent APC. Polyinosine-polycytidylic acid [poly(I:C)], a ligand of Toll-like receptor (TLR) 3, was efficiently bound either in a single layer or a multilayer attempt to the surface of diethylaminoethyl dextran modified PLGA microparticles. These particles were effectively phagocytized by DC ex vivo and induced a maturation similar to that achieved with a cytokine cocktail or higher concentrations of soluble poly(I:C). In conclusion, the concept of surface coating of biodegradable microparticles with selected TLR ligands might successfully be used in DC-based cell therapies for cancer or in vaccination trials to induce DC maturation and specifically amplify the immunological response to encapsulated antigens.


Assuntos
Células Dendríticas/imunologia , Ácido Láctico/química , Poli I-C/administração & dosagem , Ácido Poliglicólico/química , Células Cultivadas , DEAE-Dextrano/química , Células Dendríticas/metabolismo , Humanos , Ligantes , Microesferas , Fagocitose , Poli I-C/imunologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , Receptor 3 Toll-Like/metabolismo
20.
Gene Ther ; 14(16): 1243-8, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17525704

RESUMO

Adenoviral vector-mediated gene delivery has been vastly investigated for cystic fibrosis (CF) gene therapy; however, one of its drawbacks is the low efficiency of gene transfer, which is due to basolateral colocalization of viral receptors, immune responses to viral vectors and the presence of a thick mucus layer in the airways of CF patients. Therefore, enhancement of gene transfer can lead to reduction in the viral dosage, which could further reduce the acute toxicity associated with the use of adenoviral vectors. Nacystelyn (NAL) is a mucolytic agent with anti-inflammatory and antioxidant properties, and has been used clinically in CF patients to reduce mucus viscosity in the airways. In this study, we show that pretreatment of the airways with NAL followed by administration of adenoviral vectors in complex with DEAE-Dextran can significantly enhance gene delivery to the airways of mice without any harmful effects. Moreover, NAL pretreatment can reduce the airway inflammation, which is normally observed after delivery of adenoviral particles. Taken together, these results indicate that NAL pretreatment followed by adenoviral vector-mediated gene delivery can be beneficial to CF patients by increasing the efficiency of gene transfer to the airways, and reducing the acute toxicity associated with the administration of adenoviral vectors.


Assuntos
Acetilcisteína/análogos & derivados , Adenoviridae/genética , Expectorantes/uso terapêutico , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Pulmão/metabolismo , Lisina/análogos & derivados , Acetilcisteína/uso terapêutico , Animais , Linhagem Celular , DEAE-Dextrano/administração & dosagem , Expressão Gênica , Óperon Lac , Pulmão/virologia , Lisina/uso terapêutico , Camundongos , Camundongos Endogâmicos C57BL , Coloração e Rotulagem , Transdução Genética/métodos
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