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1.
PLoS One ; 10(9): e0137017, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26379273

RESUMEN

The biotechnology to immobilize biomolecules on material surfaces has been developed vigorously due to its high potentials in medical applications. In this study, a simple and effective method was designed to immobilize biomolecules via amine-N-hydroxysuccinimide (NHS) ester conjugation reaction using functionalized poly-p-xylylene coating on material surfaces. The NHS ester functionalized coating is synthesized via chemical vapor deposition, a facile and solvent-less method, creating a surface which is ready to perform a one-step conjugation reaction. Bone morphogenetic protein 2 (BMP-2) is immobilized onto material surfaces by this coating method, forming an osteogenic environment. The immobilization process is controlled at a low temperature which does not damage proteins. This modified surface induces differentiation of preosteoblast into osteoblast, manifested by alkaline phosphatase (ALP) activity assay, Alizarin Red S (ARS) staining and the expression of osteogenic gene markers, Alpl and Bglap3. With this coating technology, immobilization of growth factors onto material surface can be achieved more simply and more effectively.


Asunto(s)
Proteína Morfogenética Ósea 2/química , Adhesión Celular/efectos de los fármacos , Osteoblastos/citología , Osteogénesis/efectos de los fármacos , Propiedades de Superficie/efectos de los fármacos , Células 3T3 , Fosfatasa Alcalina/biosíntesis , Fosfatasa Alcalina/metabolismo , Animales , Antraquinonas/metabolismo , Diferenciación Celular/efectos de los fármacos , Línea Celular , Ratones , Osteoblastos/metabolismo , Osteogénesis/fisiología , Polímeros/química , Succinimidas/química
2.
ACS Appl Mater Interfaces ; 6(24): 21906-10, 2014 Dec 24.
Artículo en Inglés | MEDLINE | ID: mdl-25434778

RESUMEN

Protein molecules immobilized on biomaterial surfaces are performed based on oriented conjugation or replaced mimicking peptides. The sustainable immobilization of growth factor proteins using functionalized parylene coatings is demonstrated in this study. Site-specific and nonspecific immobilization approaches are realized to conjugate bone morphogenetic protein (BMP-2). The binding affinities and conformational changes of BMP-2 are confirmed by QCM and SPR characterizations. Osteoinduction of stem cells is examined by ALP activity on the BMP-2 modified surfaces. Finally, immobilizations and equally sustained biological functions of vascular endothelial growth factor (VEGF) and a mimicking peptide of KLTWQELYQLKYKG (QK) are also examined and confirmed.


Asunto(s)
Péptidos y Proteínas de Señalización Intercelular/química , Polímeros/química , Xilenos/química , Animales , Bovinos , Células Cultivadas , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo
3.
J Mater Chem B ; 2(48): 8496-8503, 2014 Dec 28.
Artículo en Inglés | MEDLINE | ID: mdl-32262208

RESUMEN

Advanced antibacterial surfaces are designed based on covalently attached antibacterial agents, avoiding potential side effects associated with overdosed or eluted agents. The technique is widely applicable regardless of the underlying substrate material. In addition, antibacterial surfaces are effective against the early stages of bacterial adhesion and can significantly reduce the formation of biofilm, without compromising biocompatibility. Here, this concept was realized by employing a benzoyl-functionalized parylene coating. The antibacterial agent chlorhexidine was used as a proof of concept. Chlorhexidine was immobilized by reaction with photoactivated benzoyl-functionalized surfaces, including titanium alloy, stainless steel, polyether ether ketone, polymethyl methacrylate, and polystyrene. A low concentration of chlorhexidine (1.4 ± 0.08 nmol cm-2) covalently bound to surfaces rendered them sufficiently resistant to an Enterobacter cloacae inoculum and its adherent biofilm. Compared to unmodified surfaces, up to a 30-fold reduction in bacterial attachment was achieved with this coating technology. The immobilization of chlorhexidine was verified with infrared reflection absorption spectroscopy (IRRAS) and X-ray photoelectron spectroscopy (XPS), and a leaching test was performed to confirm that the chlorhexidine molecules were not dislodged. Cell compatibility was examined by culturing fibroblasts and osteoblasts on the modified surfaces, revealing greater than 93% cell viability. This coating technology may be broadly applicable for a wide range of other antibacterial agents and allow the design of new biomaterials.

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