RESUMEN
Bone defects are usually difficult to be regenerated due to pathological states or the size of the injury. Researchers are focusing on tissue engineering approaches in order to drive the regenerative events, using stem cells to regenerate bone. The purpose of this study is to evaluate the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) on biologically derived Gallus gallus domesticus-derived demineralized bone particle (GDD) sponge. The sponges were prepared by freeze-drying method using 1, 2, and 3 wt% GDD and cross-linked with glutaraldehyde. The GDD sponge was characterized using scanning electron microscopy, compressive strength, porosity, and Fourier transform infrared. The potential bioactivity of the sponge was evaluated by osteogenic differentiation of BMSCs using 3(4, dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay and quantifying alkaline phosphatase (ALP) activity. in vivo experiments were evaluated through a micro-computerized tomography (µ-CT) and histological assays. The analysis confirmed that an increase in the concentration of the GDD in the sponge leads to a higher bone formation and deposition in rat calvarial defects. Histological assay results were in line with µ-CT. The results reported in this study demonstrated the potential application of GDD sponges as osteoinductor in bone tissue engineering in pathological or nonunion bone defects.
Asunto(s)
Células de la Médula Ósea/metabolismo , Técnicas de Cultivo de Célula , Células Inmovilizadas , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Cráneo , Andamios del Tejido/química , Animales , Células de la Médula Ósea/patología , Células Inmovilizadas/metabolismo , Células Inmovilizadas/patología , Células Inmovilizadas/trasplante , Femenino , Células Madre Mesenquimatosas/patología , Ratas , Ratas Sprague-Dawley , Cráneo/lesiones , Cráneo/metabolismo , Cráneo/patologíaRESUMEN
Corneal endothelial cells (CEnCs) play a fundamental role in maintaining the transparency of the cornea. CEnCs lose their full proliferating capacity when tissue damages occur. The loss in proliferation rate is associated with corneal edema and decrease in visual acuity, leading in severe cases, to blindness. In these situations, a corneal transplant is usually needed to restore the original tissue functions. Tissue engineering is an efficient alternative for the production of implantable films, which can regenerate the tissue functions regulating at the same time the immune-response. In this study, we proposed a stable and transparent film, composed of silk fibroin modified with glycerol (G/SF), as a potential substrate for corneal endothelial cells regeneration. Our results confirmed that G/SF films have a uniform structure, rougher surface and lower thickness respect to the SF film. In vitro tests show that G/SF films can induce a slight increase in CEnCs initial adhesion and proliferation rate if compared with the SF film. Morphology and gene expression evaluations demonstrated that the bioactive effects of silk fibroin were not affected by the presence of glycerol. For this reason, the G/SF films are suitable as CEnCs carrier and promising for the corneal damages treatments.
Asunto(s)
Endotelio Corneal/fisiología , Fibroínas/química , Glicerol/química , Membranas Artificiales , Regeneración , Seda/química , Animales , Materiales Biocompatibles/química , Adhesión Celular , Proliferación Celular , Lesiones de la Cornea/terapia , Regulación de la Expresión Génica , Interacciones Hidrofóbicas e Hidrofílicas , Conejos , Propiedades de Superficie , Ingeniería de Tejidos , Andamios del Tejido/químicaRESUMEN
The purpose of this study is to produce injectable taurine (Tr)-loaded alginate (Agn) hydrogel for age-related macular degeneration (AMD) treatment by inducing the regeneration of RPE (retinal pigment epithelium) cells. Porosity and swelling ratio were measured to evaluate the mechanical properties of the hydrogels, and Fourier transform infrared spectroscopy (FTIR) was used to evaluate the physical and chemical properties. RPE cells extracted from the pigmented epithelium of rabbits were encapsulated in the Tr/Agn hydrogels. Cells proliferation and migration were improved in Tr/Agn hydrogels with an enhanced expression of RPE-specific genes including RPE65, CRALBP, NPR-A, MITF and collagen type I and II. In vivo tests demonstrated the excellent biocompatibility and biodegradability without inflammatory response by the host when implanted with the hydrogel. Moreover, when the Tr/Agn hydrogels were injected into the sub-retinal space, high adhesion of RPE cells and retinal regeneration were confirmed. These results demonstrated a potential role of injectable Tr/Agn hydrogels as potential therapeutic tools for the treatment of retinal diseases, including AMD.
Asunto(s)
Alginatos/química , Hidrogeles/química , Regeneración , Epitelio Pigmentado de la Retina/fisiología , Taurina/química , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Fuerza Compresiva , Citocinas/metabolismo , Ratones Desnudos , Porosidad , Conejos , Regeneración/efectos de los fármacos , Epitelio Pigmentado de la Retina/citología , Epitelio Pigmentado de la Retina/patología , Taurina/farmacología , Ingeniería de TejidosRESUMEN
The retinal pigment epithelium (RPE) plays a significant role in retaining structural integrity of eye. Factors such as reduction in cell regeneration due to aging and physical injury pose a major hurdle in RPE regeneration. In this study, we exploited the use of alginate (AGT) incorporated with Curcumin (CCI) forming a hydrogel based system CCI/AGT. The fabricated hydrogel could anchor RPE cell in it. In vitro cell analysis revealed that the CCI/AGT hydrogel shows good biocompatibility, enhanced cell growth ability and higher ECM formation compared to the pure AGT hydrogel. In particular, the presence of CCI in the hydrogels enhances the cells proliferation of the 23% respect to the pure alginate. Also the expression of crucial genes for retina functions and matrix production were positively affected by CCI presence, with an increment of 45% for RPE65, 32% for CRALBP and 26% for Collagen type 1. In vitro tests demonstrated the potential application of CCI/AGT hydrogels for transplantation under the sub-retinal space acting as a cell delivery vehicle and also their capability to provide an appropriate environment for RPE regeneration. These results suggest that CCI/AGT hydrogel could be translated into a potential surgical graft for biological implantation of retinal tissue engineering.
Asunto(s)
Alginatos/farmacología , Curcumina/farmacología , Hidrogeles/farmacología , Regeneración , Epitelio Pigmentado de la Retina/fisiología , Animales , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Condrocitos/citología , Condrocitos/efectos de los fármacos , Ácido Glucurónico/farmacología , Ácidos Hexurónicos/farmacología , Porosidad , ARN Mensajero/genética , ARN Mensajero/metabolismo , Conejos , Regeneración/efectos de los fármacos , Epitelio Pigmentado de la Retina/efectos de los fármacos , Epitelio Pigmentado de la Retina/ultraestructura , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de SuperficieRESUMEN
BACKGROUND AND OBJECTIVES: The purpose was to establish the test-retest reliability of word recognition score (WRS) using Korean standard monosyllabic word lists for adults (KS-MWL-A) recently developed based on the international standard for speech audiometry (ISO 8253-3:2012). SUBJECTS AND METHODS: Subjects consisted of 159 adults aged to 18 to 25 years with normal hearing sensitivity. WRSs were obtained in 2 dB steps from the level of speech recognition thresholds to the level of 86% correct responses or greater. After one or two weeks, retest was performed. Correlation, confidence interval (CI) and prediction interval (PI) were calculated for the reliability. RESULTS: Correlation coefficients were 0.88 for 50 test words, 0.76 for 25 and 0.61 for 10 words. Results also showed that 95% CIs and PIs were narrower for 25 and 50 test words than those for 10 test words. CONCLUSIONS: Korean WRS using the KS-MWL-A has high reliability for 25 and 50 test words, but relatively low for 10 words. It suggested that 95% CIs for each test words would be criteria for significant differences in WRS for groups and 95% PIs at each score of WRS could be utilized for a considerable difference for each individual at retest.