RESUMO
PURPOSE/AIM OF THE STUDY: The ultimate goal of periodontal treatment is to regenerate the lost periodontal tissues. The interest in nanomaterials in dentistry is growing rapidly and has focused on improvements in various biomedical applications, such as periodontal regeneration and periodontal tissue engineering. To enhance periodontal tissue regeneration, hydroxyapatite (HA) was used in conjunction with other scaffold materials, such as Poly lactic-co-glycolic-acid (PLGA) and collagen (C). The main target of this study was to compare the effects of nano and macrostructures of the tissue scaffolds on cell behavior in vitro for periodontal tissue engineering. MATERIALS AND METHODS: Nanofibrillar and macroporous-spongious composite tissue scaffolds were produced using PLGA/C/HA. Subgroups with BMP-2 signal molecule and without HA were also created. The scaffolds were characterized by FTIR, SEM/EDX techniques, and mechanical tests. The scaffolds were compared in the periodontal ligament (PDL) and MCT3-E1 cell cultures. The cell behaviors; adhesions by SEM, proliferation by WST-1, differentiation by ALP and mineralization with Alizarin Red Tests were determined. RESULTS: Cell adhesion and mineralization were higher in the nanofibrillar scaffolds compared to the macroporous-spongious scaffolds. Macroporous-spongious scaffolds seemed better for the proliferation of PDL cells and differentiation of MC3T3-E1-preosteoblastic cells, while nanofibrillar scaffolds were more convenient for the differentiation of PDL cells and proliferation of MC3T3-E1-preosteoblastic cells. CONCLUSIONS: In general, nanofibrillar scaffolds showed more favorable results in cell behaviors, compared to the macroporous-spongious scaffolds, and mostly, BMP-2 and HA promoted the activities of the cells.
Assuntos
Engenharia Tecidual , Alicerces Teciduais , Proliferação de Células , Durapatita/química , Ácido Láctico , Osteogênese , Ligamento Periodontal , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
OBJECTIVE: To test the surface properties and in vitro effects of a new sequential release system on MC3T3-E1 cells for improved osseointegration. BACKGROUND: BMP6-loaded anodized titanium coated with PDGF containing silk fibroin (SF) may improve osseointegration. METHODS: Titanium surfaces were electrochemically anodized, and SF layer was covered via electrospinning. Five experimental groups (unanodized Ti (Ti), anodized Ti (AnTi), anodized + BMP6-loaded Ti (AnTi-BMP6), anodized + BMP6 loaded + silk fibroin-coated Ti (AnTi-BMP6-SF), and anodized + BMP6-loaded + silk fibroin with PDGF-coated Ti (AnTi-BMP6-PDGF-SF)) were tested. After SEM characterization, contact angle analysis, and FTIR analysis, the amount of released PDGF and BMP6 was detected using ELISA. Cell proliferation (XTT), mineralization, and gene expression (RUNX2 and ALPL) were also evaluated. RESULTS: After successful anodization and loading of PDGF and BMP6, contact angle measurements showed hydrophobicity for TiO2 and hydrophilicity for protein-adsorbed surfaces. In FTIR, protein-containing surfaces exhibited amide-I, amide-II, and amide-III bands at 1600 cm-1 -1700 cm-1 , 1520 cm-1 -1540 cm-1 , and 1220 cm-1 -1300 cm-1 spectrum levels with a significant peak in BMP6- and/or SF-loaded groups at 1100 cm-1 . PDGF release and BMP6 release were delayed, and relatively slower release was detected in SF-coated surfaces. Higher MC3T3-E1 proliferation and mineralization and lower gene expression of RUNX2 and ALPL were detected in AnTi-BMP6-PDGF-SF toward day 28. CONCLUSION: The new system revealed a high potential for an improved early osseointegration period by means of a better factor release curve and contribution to the osteoblastic cell proliferation, mineralization, and associated gene expression.
Assuntos
Osseointegração , Fator de Crescimento Derivado de Plaquetas , Titânio , Animais , Proliferação de Células , Camundongos , Fator de Crescimento Derivado de Plaquetas/farmacologia , Propriedades de SuperfícieRESUMO
Aim: In this study, we aimed to develop a polycationic non-viral carrier for the delivery of the reprogramming factors to the L929 fibroblast cell.Methods: We have prepared (3-hydroxybutyrate-co-3-hydroxyhexanoate) PHBHHx-based nanoparticles with the solvent diffusion method. Cytotoxicity of PXNs was determined via MTT assay. Transfection efficiency was evaluated via screening GFP expression by fluorescence microscopy. The expression of reprogramming factors (Oct4, Klf4, and Sox2) was determined by RT-qPCR.Results: PXNs with 32.9 ± 0.41 mV zeta potential and 177.6 ± 0.80 nm size were used for transfection of L929 Fbroblast cells. The percentage of cell viability of PXN were between 91.8%(±2.9) and 42.1%(±1.3). The transfection efficiency was found as 71.6%(±3,5). According to RT-qPCR data, the rate of transfection factors was significantly increased after the 11th cycle compared to non-transfected cells. Based on these results, it can be concluded that newly developed PXN is thought to be an effective tool for reprogramming cells.
Assuntos
Caproatos/química , Nanopartículas/química , Reprogramação Celular , Expressão Gênica , Proteínas de Fluorescência Verde , Humanos , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/biossíntese , Fatores de Transcrição Kruppel-Like/genética , Fator 3 de Transcrição de Octâmero/biossíntese , Fator 3 de Transcrição de Octâmero/genética , Tamanho da Partícula , Paxilina/genética , Fatores de Transcrição SOXB1/biossíntese , Fatores de Transcrição SOXB1/genética , Sais de Tetrazólio/farmacologia , Tiazóis/farmacologia , Transfecção/métodosRESUMO
OBJECTIVES: A number of chemical and biological factors, including mesenchymal stem cells (MSCs), have been developed to enhance nerve regeneration by introduction through a variety of nerve conduits. This study was designed to assess the efficacy of using chitosan-coated poly-3-hydroxybutyrate (PHB) nerve conduits seeded with human bone marrow-derived MSCs (hMSC-bm) to augment repair in an experimental rat model of sciatic nerve injury. METHODS: A total of 30 rats were randomly assigned to one of three groups (n = 10). In each rat, a 10 mm segment of the sciatic nerve was removed and was replaced by a chitosan-coated PHB conduit seeded with hMSC-bm (PHB/chitosan-hMSC-bm group), a chitosan-coated PHB conduit (PHB/chitosan group), or an autograft (autograft group) as the control. The results were evaluated 8 weeks postoperatively by observation, electromyography and histologic examination with light microscopy and immunostaining. RESULTS: Histologic examination showed that both PHB/chitosan-hMSC-bm conduits and PHB/chitosan conduits led the damaged axons through the injured area. When the effects were compared, the results with the PHB/chitosan-hMSC-bm conduits were superior to those with the PHB/chitosan conduits (p < 0.05) but not as successful as with the autologous nerve grafts (p < 0.05). CONCLUSION: PHB/chitosan-hMSC-bm nerve conduits may be a useful artificial guide for nerve regeneration.
Assuntos
Materiais Biocompatíveis/uso terapêutico , Quitosana/uso terapêutico , Hidroxibutiratos/uso terapêutico , Células-Tronco Mesenquimais/fisiologia , Regeneração Nervosa/efeitos dos fármacos , Poliésteres/uso terapêutico , Neuropatia Ciática/cirurgia , Análise de Variância , Animais , Modelos Animais de Doenças , Eletromiografia , Potencial Evocado Motor/efeitos dos fármacos , Potencial Evocado Motor/fisiologia , Masculino , Proibitinas , Ratos , Ratos Wistar , Caminhada/fisiologiaRESUMO
In the present study, culture conditions of Streptococcus equi was optimized through Box-Behnken experimental design for hyaluronic acid production. About 0.87 gL-1 of hyaluronic acid was produced under the determined conditions and optimal conditions were found as 38.42 °C, 24 hr and 250 rpm. The validity and practicability of this statistical optimization strategy were confirmed relation between predicted and experimental values. The hyaluronic acid obtained under optimal conditions was characterized. The effects of different conditions such as ultraviolet light, temperature and enzymatic degradation on hyaluronic acid produced under optimal conditions were determined. 118 °C for 32 min of autoclaved HA sample included 63.09 µg mL-1 of d-glucuronic acid, which is about two-fold of enzymatic effect. Cytotoxicity of hyaluronic acid on human dermal cells (HUVEC, HaCaT), L929 and THP-1 cells was studied. In vitro effect on pro or anti-inflammatory cytokine release of THP-1 cells was determined. Although it varies depending on the concentration, cytotoxicity of hyaluronic acid is between 5 and 30%. However, it varies depending on the concentration of hyaluronic acid, TNF-α release was not much increased compared to control study. Consequently, purification procedure is necessary to develop and it is worth developing the bacterial hyaluronic acid.
RESUMO
Vitamin D3 (VitD3) has several beneficial effects on many metabolic pathways such as immunity system, bone development. The aim of the study, encapsulation of VitD3 with solid lipids, determine encapsulation efficiency and biocompatibility of nanoparticles. Therefore, VitD3-loaded solid lipid nanoparticles (SLNPs) were developed by optimising ratios of VitD3, stearic acid, beeswax and sodium dodecyl sulphate (SDS). Thermal stability, degradation profile, crystallinity rate, encapsulation efficiency and release profile of SLNPs were determined. Cytotoxicity of SLNPs on HaCaT, L929 and HUVEC cells were investigated. Negatively charged and VitD3-loaded nanoparticles with diameters between 30 and 60 nm were obtained. SLNPs containing up to 5.1 mg VitD3 per 10 mg powder samples were obtained. Cell proliferations were stimulated after exposure with VitD3-loaded SLNPs. Besides, inflammatory response after exposure to VitD3-loaded SLNPs was evaluated via determining IL10 and TNF-alpha levels on THP-1 cells. According to the results, no inflammatory response was observed.
Assuntos
Colecalciferol/farmacologia , Interleucina-10/metabolismo , Lipídeos/química , Nanopartículas/química , Fator de Necrose Tumoral alfa/metabolismo , Materiais Biocompatíveis/química , Linhagem Celular , Proliferação de Células , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , HumanosRESUMO
This study is designed to evaluate the treatment effect of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) and human mesenchymal stem cells (hMSC) on axonal regeneration in experimental rat sciatic nerve damage, and compare the results of this modality with autologous nerve grafting. In Spraque-Dawley albino rats, 10-mm-long experimental nerve gaps were created. Three groups were constituted, the gap was repaired with autologous nerve graft (autograft group), PHBHHx nerve graft alone (PHBHHx alone group), and PHBHHx nerve graft with hMSCs inside (PHBHHx with hMSC group), respectively. The results were evaluated with functional recovery, electrophysiological evaluation, and histological evaluation either with light microscopy and transmission electron microscopy for axonal regeneration and myelin formation. In functional evaluation, autograft and PHBHHx with hMSC groups showed functional improvement with time, whereas PHBHHx alone group did not. Electrophysiological evaluation showed better results in autograft and PHBHHx with hMSC groups when compared to PHBHHx alone group. There was no statistical difference between autograft and PHBHHx with hMSC groups. Histological evaluation showed regenerated axons in each group. Autograft group was better than the others, and PHBHHx with hMSC group was better than PHBHHx alone group both for axonal regeneration and myelin formation. This study showed that the nerve grafts which were prepared from PHBHHx with oriented nanofiber three-dimensional surfaces aided to nerve regeneration, either used alone or with hMSC. PHBHHx provided better nerve regeneration when used with hMSCs inside than alone, and reached the same statistical treatment effect in functional evaluation and electrophysiological evaluation when compared to autografting.
Assuntos
Ácido 3-Hidroxibutírico/farmacologia , Ácido 3-Hidroxibutírico/uso terapêutico , Caproatos/farmacologia , Caproatos/uso terapêutico , Transplante de Células-Tronco Mesenquimais , Regeneração Nervosa/efeitos dos fármacos , Neuropatia Ciática/tratamento farmacológico , Neuropatia Ciática/cirurgia , Animais , Axônios/patologia , Axônios/ultraestrutura , Células Cultivadas , Modelos Animais de Doenças , Eletromiografia , Potencial Evocado Motor/efeitos dos fármacos , Feminino , Humanos , Locomoção/efeitos dos fármacos , Locomoção/fisiologia , Células-Tronco Mesenquimais/fisiologia , Células-Tronco Mesenquimais/ultraestrutura , Microscopia Eletrônica de Varredura , Músculo Esquelético/patologia , Músculo Esquelético/fisiopatologia , Fibras Nervosas Mielinizadas/patologia , Fibras Nervosas Mielinizadas/ultraestrutura , Regeneração Nervosa/fisiologia , Ratos , Ratos Sprague-DawleyRESUMO
Diabetic Retinopathy (DRP) is a disease consisting of all the structural and functional changes that develop in the retinal layer of the eye due to diabetes. DRP is the most important cause of blindness between the ages of 20-74 in the world, and the most successful standard treatment option in the treatment of DRP is intravitreal injections. To synthesize acetazolamide loaded nanoparticles to be applied intravitreal treatment of DRP and to examine thein vitroefficacy of the nanoparticles. ACZ loaded PHBV nanoparticles (PHBV-ACZ NPs) formulations were prepared. Nanoparticles with a particle size of 253.20 ± 0.55 nm. A DRP model was established and characterized in HRMEC cells. The effect of the nanoparticles on permeability has been investigated and carrier proteins in BRB due to the development of DRP has been investigated. To establish thein vitroDRP model, HRMEC was stimulated with Recombinant human 165 Vascular Endothelial Growth Factor (VEGF), thereby temporarily reducing the expression levels of endothelial junction proteins, increasing the number of intercellular spaces in the monolayers of HRMECs. It was determined that after the cells were exposed to Carbonic anhydrase inhibitors (CAI) loaded nanoparticles, permeability decreased and protein expression increased.
Assuntos
Diabetes Mellitus , Retinopatia Diabética , Nanopartículas , Humanos , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso , Retinopatia Diabética/tratamento farmacológico , Retinopatia Diabética/etiologia , Retinopatia Diabética/metabolismo , Inibidores da Anidrase Carbônica , Fator A de Crescimento do Endotélio Vascular , Nanopartículas/química , PoliésteresRESUMO
Alginate (ALG) is a biocompatible and biodegradable polymer. Mechanical weakness is one of the main problems for the alginate-based scaffolds. Various plasticizer additives or modifications tested to improve the mechanical properties. In the presented study, ALG plasticized with triacetin (TA), and tributyl citrate (TBC) than tested on bone healing. In the presented study, the alginate modified with triacetin or tributyl citrate. In-vitro, and in-vivo efficiency of the scaffolds tested on bone tissue regeneration. Scaffolds fabricated by solvent casting, and physicochemical characterizations performed. Monocytes (THP-1) cultured with scaffolds, and macrophage-released cytokines was determined. In-vivo efficacy of the scaffolds was tested in the rat drill hole model. Alginate and tributyl citrate-modified scaffolds have no cytotoxic effect on osteoblastic cells (MC-3T3). Tributyl citrate modification increased tumor necrosis factor-alpha (TNF-alpha) level but did not increase interleukin -1 beta (IL-1 beta) level. In vivo studies showed that osteoblastic growth was significant in alginate and triacetin-modified scaffolds. However, the best values for osteoclastic activity and osteoid tissue formation seen in the triacetin modification. The results demonstrated that the modified alginate scaffolds were more successful than non-modified alginate scaffolds and can used as long-term bone repairing treatments.
Assuntos
Alginatos , Alicerces Teciduais , Ratos , Animais , Alicerces Teciduais/química , Alginatos/farmacologia , Alginatos/química , Engenharia Tecidual/métodos , Diáfises , Triacetina/farmacologia , Regeneração Óssea , FêmurRESUMO
Laccase is an important enzyme used in many industries because of its multi-substrate catalyst. New immobilization agents are excellent tools for enhancing the abilities of this enzyme. In this study, immobilization of laccase on silica microparticles with NH2 (S-NH2) surface modification to use in dye removal applications was aimed. The yield of immobilization by this method was found to be 93.93 ± 2.86% under optimum conditions. In addition, this newly created immobilized enzyme was adapted to a decolorization application with 87.56 ± 1.60% efficiency. Silica microparticles with NH2 (S-NH2) surface modification were used for laccase immobilization and this immobilized laccase had quite good potential. Besides, Random Amplified Polymorphic DNA (RAPD) analysis in evaluating the toxicity of the decolorization process was utilized. After amplification with two RAPD primers, decreased toxicity of dye in this study was observed. This study showed that RAPD analysis in toxicity testing could be accepted as an alternative and practical method that this approach will contribute to the literature in terms of providing fast and reliable results. The use of amine-modified surface silica microparticles for laccase immobilization and RAPD for toxicity testing is a crucial aspect of our investigation.
Assuntos
Aminas , Lacase , Lacase/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Enzimas Imobilizadas , Dióxido de Silício , CorantesRESUMO
To improve poor water solubility of cyclosporine A (CsA), hydroxypropyl-beta-cyclodextrin (HPßCD) was incorporated into the nanoparticle formulation. Solid complexes of CsA with HPßCD in different ratios were prepared by the kneading method. CsA containing alone or in combination with HPßCD in poly-lactide-co-glycolide (P-CsA or P-CsA-HPßCD) nanoparticles were prepared by the emulsification solvent evaporation method. The mean size of CsA-loaded NPs was found to be approximately 220 nm. The solubility of CsA was significantly improved and the phase solubility diagram of CsA-HPßCD systems showed an A(L) type phase. Nanoparticles showed high CsA encapsulation efficiency (88%) and production yield (89%). Release rate was increased by the presence of HPßCD and total cumulative release ranged from 75% to 96% in 24 h. In vitro cytotoxicity study assay resulted in a low toxicity for all types of nanoparticles. After 6 h incubation period, the cellular uptake was found at 33% and 32% for P-CsA and P-HPßCD-CsA nanoparticles, respectively.
Assuntos
Ciclodextrinas , Ciclosporina , Ácido Láctico , Nanopartículas/química , Ácido Poliglicólico , Administração Oftálmica , Animais , Linhagem Celular , Ciclodextrinas/química , Ciclodextrinas/farmacocinética , Ciclodextrinas/farmacologia , Ciclosporina/química , Ciclosporina/farmacocinética , Ciclosporina/farmacologia , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Preparações de Ação Retardada/farmacologia , Avaliação Pré-Clínica de Medicamentos , Ácido Láctico/química , Ácido Láctico/farmacocinética , Ácido Láctico/farmacologia , Camundongos , Tamanho da Partícula , Ácido Poliglicólico/química , Ácido Poliglicólico/farmacocinética , Ácido Poliglicólico/farmacologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico , SolubilidadeRESUMO
Graphene is a material with various application potentials Graphene is a unique material with superiorities and has been applied in various fields for different purposes. Although studies on the utility of graphene oxide in the biomedical field are available, no evaluation has yet been done regarding the utility of sulfur doped (S-doped) graphene. The study focuses on the effect of blending the poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) membrane with sulfur heteroatom doped graphene and the evaluation of biological responses to S-doped graphene/PHBHHx. PHBHHx membranes were blended with 1%, 0.5%, 0.1% (w/v) S-doped graphene. The morphological (SEM and Microscopy), chemical (FTIR and Raman spectroscopy), and surface area (BET) characterizations of S-doped graphene/PHBHHx membranes were performed. The presence of S groups on the surface was determined with the EDS results. Besides, the swelling profile and biodegradation tendency of the membranes were evaluated. The differentiation of protein adhesion, cell viability, cell adhesion, and cell proliferation by the increasing content of S-doped graphene was examined. The contact angle analysis revealed that modification of PHBHHx with S-doped Graphene reduced the free surface energy of PHBHHx membranes. Blending with S-doped Graphene has decreased the polarity of the PHBHHx membrane. The protein adsorption on the PHBHHx membrane was determined as 10.12 ± 0.247 mg/ml. Protein absorption on 1%, 0.5% and 0.1% S-doped graphene/PHBHHx membranes were determined as 11.34 ± 0.551 mg/ml, 9.91 ± 0.294 mg/ml and 9.48 ± 0.093 mg/ml, respectively. The cell attachment to the surface decreased with the increasing amount of S-doped graphene, however, PHBHHx membranes with graphene did not affect cytotoxicity. S-doped graphene blended PHBHHx membrane seems like a suitable patch for biomedical treatments as a hydrophobic membrane where less cell adhesion and proliferation are required like the prevention of peritoneal adhesion.
Assuntos
Ácido 3-Hidroxibutírico/farmacologia , Bactérias/química , Fibroblastos/citologia , Grafite/farmacologia , Adsorção , Animais , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Camundongos , Soroalbumina Bovina/química , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Termogravimetria , Água/químicaRESUMO
Induced pluripotent stem cells (IPSC) are preferred as an alternative source for regenerative medicine, disease modeling, and drug screening due to their unique properties. As seen from the previous studies in the literature, most of the vector systems to transfer reprogramming factors are viral-based and have some well-known limitations. This study aims to develop a non-viral vector system for the transfection of reprogramming factors. Cationic stearamide lipid nanoparticles (CSLN) were prepared via the solvent diffusion method. The obtained CSLNs were used for the delivery of plasmid DNA (pDNA) encoding Oct3/4, Sox2, Klf4, and GFP to fibroblast cell lines. The optimization studies, for zeta potential and particle size of the conjugate, was performed to achieve high cell viability. CSLN63 with 36.5±0.06â mV zeta potential and 173.6±13.91â nm size was used for the transfection of Fibroblast cells. The transfection efficiency was observed by following GFP expression and was found as 70 %±0.11. The expression of the Oct4, Sox2, Klf4 was determined by RT-qPCR; an increase was observed after the 12th cycle in Klf4 (Ct averages: 13,41), Sox2 (Ct averages; 12,4), Oct4 (Ct average; 13,77). The tendency of colonization was observed. The upregulation efficiency of Oct4 and SSEA-1 with CSLN and another non-viral vector designed for the transportation of Yamanaka factors developed in our lab previously were compared with flow cytometer analysis.
Assuntos
DNA/genética , Portadores de Fármacos/química , Nanopartículas/química , Plasmídeos/genética , Ácidos Esteáricos/química , Animais , Linhagem Celular , Reprogramação Celular/genética , Portadores de Fármacos/toxicidade , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Fator 4 Semelhante a Kruppel , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Camundongos , Nanopartículas/toxicidade , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/metabolismo , Tamanho da Partícula , Fatores de Transcrição SOXB1/genética , Fatores de Transcrição SOXB1/metabolismo , Ácidos Esteáricos/toxicidade , Transfecção/métodosRESUMO
BACKGROUND: Bioengineering products can help bone tissue regeneration. OBJECTIVE: There is an ongoing research for more effective biomaterials in bone regeneration. Chitosan (Ch) grafted stearic acid (Ch-g-Sa) polymer was synthesized and its usability as a putty was evaluated in this study. METHODS: The chemical structure of Ch-g-Sa polymer was investigated using Proton nuclear magnetic resonance (H-NMR) and Fourier-transformed infrared spectroscopy-attenuated total reflectance (FTIR-ATR). Thermal properties of Ch-g-Sa polymer were determined by thermal gravimetric analysis (TGA). Putties containing nano-hydroxyapatite were prepared and in-vitro degradation properties and viscosity of the putties were determined. RESULTS: The cytotoxicity, oxidation effect and osteogenic potential of the putties were investigated on MC3T3 cells while the inflammatory effect of the putties was studied on THP-1 cells. For the determination of the osteogenic effect of the putties, ALP and RUNX2 gene expression of MC3T3 cells were studied. CONCLUSION: Ch-g-Sa/HA putties are promising biomaterials for bone tissue regeneration.
Assuntos
Osso e Ossos/efeitos dos fármacos , Quitosana/administração & dosagem , Osteogênese/efeitos dos fármacos , Ácidos Esteáricos/administração & dosagem , Engenharia Tecidual/métodos , Materiais Biocompatíveis/administração & dosagem , Estabilidade de Medicamentos , Durapatita/administração & dosagem , Durapatita/química , Humanos , Mediadores da Inflamação/metabolismo , Oxirredução/efeitos dos fármacos , Polímeros , Espectroscopia de Prótons por Ressonância Magnética , Espectroscopia de Infravermelho com Transformada de Fourier , Células THP-1 , ViscosidadeRESUMO
Purpose: The aim of this study was to design naproxen sodium (NS)-containing, biomimetic, porous poly(lactide-co-glycolide) (PLGA) scaffolds for regeneration of damaged corneal epithelium. Methods: NS-incorporated PLGA scaffolds were prepared using the emulsion freeze-drying method and then coated with collagen or poly-l-lysine. Porosity measurements of the scaffolds were performed by the gas adsorption/desorption method and the scaffolds demonstrated highly porous, open-cellular pore structures with pore sizes from 150 to 200 µm. Results: The drug loading efficiency of scaffolds was found to be higher than 84%, and about 90%-98% of NS was released at the end of 7 days with a fast drug release rate at the initial period of time and then in a slow and sustained manner. The corneal epithelial cells were isolated from New Zealand white rabbits. The obtained cells were seeded onto scaffolds and continued to increase during the time period of the study, indicating that the scaffolds might promote corneal epithelial cell proliferation without causing toxic effects for at least 10 days. Conclusions: The NS-loaded PLGA scaffolds exhibited a combination of controlled drug release and biomimetic properties that might be attractive for use in treatment of corneal damage both for controlled release and biomedical applications.
Assuntos
Anti-Inflamatórios/farmacocinética , Lesões da Córnea/tratamento farmacológico , Epitélio Corneano/efeitos dos fármacos , Naproxeno/farmacocinética , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Anti-Inflamatórios/administração & dosagem , Biomimética , Proliferação de Células/efeitos dos fármacos , Colágeno/química , Colágeno/metabolismo , Composição de Medicamentos/métodos , Liberação Controlada de Fármacos , Epitélio Corneano/patologia , Naproxeno/administração & dosagem , Polietilenoglicóis/química , Polietilenoglicóis/metabolismo , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacocinética , Polilisina/análogos & derivados , Polilisina/química , Polilisina/metabolismo , Porosidade/efeitos dos fármacos , Coelhos , Regeneração/efeitos dos fármacos , Regeneração/fisiologiaRESUMO
Magnetosomes are specialized organelles arranged in intracellular chains in magnetotactic bacteria. The superparamagnetic property of these magnetite crystals provides potential applications as contrast-enhancing agents for magnetic resonance imaging. In this study, we compared two different nanoparticles that are bacterial magnetosome and HSA-coated iron oxide nanoparticles for targeting breast cancer. Both magnetosomes and HSA-coated iron oxide nanoparticles were chemically conjugated to fluorescent-labeled anti-EGFR antibodies. Antibody-conjugated nanoparticles were able to bind the MDA-MB-231 cell line, as assessed by flow cytometry. To compare the cytotoxic effect of nanoparticles, MTT assay was used, and according to the results, HSA-coated iron oxide nanoparticles were less cytotoxic to breast cancer cells than magnetosomes. Magnetosomes were bound with higher rate to breast cancer cells than HSA-coated iron oxide nanoparticles. While 250 µg/ml of magnetosomes was bound 92 ± 0.2%, 250 µg/ml of HSA-coated iron oxide nanoparticles was bound with a rate of 65 ± 5%. In vivo efficiencies of these nanoparticles on breast cancer generated in nude mice were assessed by MRI imaging. Anti-EGFR-modified nanoparticles provide higher resolution images than unmodified nanoparticles. Also, magnetosome with anti-EGFR produced darker image of the tumor tissue in T2-weighted MRI than HSA-coated iron oxide nanoparticles with anti-EGFR. In vivo MR imaging in a mouse breast cancer model shows effective intratumoral distribution of both nanoparticles in the tumor tissue. However, magnetosome demonstrated higher distribution than HSA-coated iron oxide nanoparticles according to fluorescence microscopy evaluation. According to the results of in vitro and in vivo study results, magnetosomes are promising for targeting and therapy applications of the breast cancer cells.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Materiais Revestidos Biocompatíveis , Meios de Contraste , Imageamento por Ressonância Magnética , Nanopartículas de Magnetita/química , Magnetossomos/química , Magnetospirillum/química , Albumina Sérica Humana , Animais , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Materiais Revestidos Biocompatíveis/química , Meios de Contraste/química , Meios de Contraste/farmacologia , Feminino , Xenoenxertos , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Transplante de Neoplasias , Albumina Sérica Humana/química , Albumina Sérica Humana/farmacologiaRESUMO
Patients with diabetes mellitus have an increased cardiovascular risk due to the abnormality of hemostatic system components. Therefore, hemostasis is an important concept when considering that diabetics are under risk due to potential bleeding complications during surgical operation. The aim of our study was to examine the efficiency of a fabricated nano/microbilayer hemostatic dressing for bleeding control in diabetic patients. For this purpose, we prepared a nano/microbilayer hemostatic dressing that has a porous sublayer, including chitosan (CTS), bacterial cellulose (BC) as basement and active agents in coagulation cascade, such as vitamin K (Vit K), protamine sulfate (PS), and kaolin (Kao) as a filler and an upper layer consisting of silk fibroin (SF) or SF/phosphatidylcholine (PC) blend to achieve complete hemostasis in diabetic rats. Coagulative performances of the prepared hemostatic dressings were examined by the determination of bleeding time, blood loss, and mortality rate through diabetic rat femoral artery injury model. The percent of diabetic rat blood absorption was found to be 247 ± 5% for gauze as opposed to 2214 ± 56% for SF-coated PS/BC/CTS. Vit K-reinforced within 138 s and SF-coated BC/CTS hemostatic dressings within 144 s showed a rapid coagulation time. In vivo coagulation studies demonstrated that hemostatic agent-reinforced BC/CTS hemostatic dressing, especially PS/BC/CTS showed a significant hemostatic plug formation. This study suggests that the high positive charge and porosity give to these hemostatic agents reinforced hemostatic dressings the ability to rapidly swell and to promote the accumulation of red blood cells and platelets through electrostatic interactions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1573-1585, 2017.
Assuntos
Bandagens , Artéria Femoral/lesões , Hemorragia/terapia , Hemostáticos/química , Hemostáticos/farmacologia , Animais , Celulose/química , Celulose/farmacologia , Quitosana/química , Quitosana/farmacologia , Modelos Animais de Doenças , Feminino , Fibroínas/química , Fibroínas/farmacologia , Caulim/química , Caulim/farmacologia , Protaminas/química , Protaminas/farmacologia , Ratos , Ratos Sprague-DawleyRESUMO
The aim of the study is in vitro investigation of the feasibility of surface-modified bacterial nanofibrous poly [(R)-3-hydroxybutyrate] (PHB) graft for bladder reconstruction. In this study, the surface of electrospun bacterial PHB was modified with PEG- or EDA via radio frequency glow discharge method. After plasma modification, contact angle of EDA-modified PHB scaffolds decreased from 110 ± 1.50 to 23 ± 0.5 degree. Interestingly, less calcium oxalate stone deposition was observed on modified PHB scaffolds compared to that of non-modified group. Results of this study show that surface-modified scaffolds not only inhibited calcium oxalate growth but also enhanced the uroepithelial cell viability and proliferation.
Assuntos
Materiais Biocompatíveis/farmacologia , Células Epiteliais/efeitos dos fármacos , Hidroxibutiratos/farmacologia , Regeneração/efeitos dos fármacos , Bexiga Urinária/efeitos dos fármacos , Animais , Materiais Biocompatíveis/isolamento & purificação , Materiais Biocompatíveis/metabolismo , Oxalato de Cálcio/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cupriavidus necator/química , Cupriavidus necator/metabolismo , Células Epiteliais/patologia , Etilenodiaminas/química , Hidroxibutiratos/isolamento & purificação , Hidroxibutiratos/metabolismo , Cálculos Renais/química , Camundongos , Polietilenoglicóis/química , Polimerização , Engenharia Tecidual , Bexiga Urinária/patologiaRESUMO
The conventional method of peripheral nerve gap treatment is autografting. This method is limited. In this study, an aligned nanofibrous graft was formed using microbial polyester, Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). The regenerative effect of the graft was compared with that of autografting in vivo. To determine the regenerative effect, rats were assessed with sciatic nerve functional index, electromyographic evaluation, and histological evaluation. Results found in this study include PHBV grafts stimulated progressive nerve regeneration, although regeneration was not comparable with that of autografting. We conclude that the study results were promising for aligned bacterial polymeric grafts for peripheral nerve regeneration.
Assuntos
Cupriavidus necator/química , Nanofibras/química , Regeneração Nervosa/efeitos dos fármacos , Poliésteres/química , Poliésteres/farmacologia , Alicerces Teciduais/química , Animais , Feminino , Ratos , Ratos Sprague-DawleyRESUMO
Early phase osseointegration is crucial for orthopedic implants. For the improvement of osseointegrative properties of orthopedic implants several surface modification methods such as acid etching, hydroxyapatite (HA) coating and sandblasting can be applied. In this article titanium implants were anodized to possess nanotubular titania structures on the surface. Titania nanotube structures with a 45-50 nm of average inner diameter were obtained and to enhance bioactivity, samples were soaked in 10X simulated body fluid (SBF) for apatite deposition on surface for different time periods (1, 2, 3, 5, 8 hours). Apatitic calcium phosphate deposited surfaces were analyzed with infrared spectrometry and wettability studies. Effect of soaking time on osteoblast cell was investigated by cell viability, alkaline phosphatase activity tests and morphological evaluations. As a result, 3 hours of soaking time was found as the optimum time period (p < 0.005). This in vitro study indicated that soaking in 10X SBF can be a rapid and economical technique to enhance osseointegration of anodized titanium implants however excess and/or uncontrolled HA coating of titania layer limits the bioactive potential of the implant.