RESUMO
Limited osteointegration of orthopedic implants with surrounding tissues has been the leading issue until the failure of orthopedic implants in the long term, which could be induced by multiple factors, including infection, limited abilities for bone formation and remodeling, and an overstressed reactive oxygen species (ROS) environment around implants. To address this challenge, a multifunctional coating composed of tannic acid (TA), nanohydroxyapatite (nHA) and gelatin (Gel) was fabricated by a layer-by-layer (LBL) technique, into which TA, nHA, and Gel were integrated, and their respective functions were utilized to synergistically promote osteogenesis. The fabrication process of (TA@nHA/Gel)n coatings and related bio-multifunctionalities were thoroughly investigated by various techniques. We found that the (TA@nHA/Gel)n coatings showed strong antioxidant activity and accelerated cellular attachment in the early stage and proliferation in the long term, largely enhancing osteogenesis in vitro and promoting bone formation in vivo. We believe our findings will guide the design of orthopedic implants in the future, and the strategy developed here could pave the way for multifunctional orthopedic implant coating and protein-related coatings with various potential applications, including biosensors, catalysis, tissue engineering, and life science.
Assuntos
Antioxidantes , Osteogênese , Antioxidantes/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Gelatina , Próteses e Implantes , Taninos , TitânioRESUMO
There is a substantial global market for orthopedic implants, but these implants still face the problem of a high failure rate in the short and long term after implantation due to the complex physiological conditions in the body. The use of multifunctional coatings on orthopedic implants has been proposed as an effective way to overcome a range of difficulties. Here, a multifunctional (TA@HA/Lys)n coating composed of tannic acid (TA), hydroxyapatite (HA), and lysozyme (Lys) was fabricated in a layer-by-layer (LBL) manner, where TA deposited onto HA firmly stuck Lys and HA together. The deposition of TA onto HA, the growth of (TA@HA/Lys)n, and multiple related biofunctionalities were thoroughly investigated. Our data demonstrated that such a hybrid coating displayed antibacterial and antioxidant effects, and also facilitated the rapid attachment of cells [both mouse embryo osteoblast precursor cells (MC3T3-E1) and dental pulp stem cells (DPSCs)] in the early stage and their proliferation over a long period. This accelerated osteogenesis in vitro and promoted bone formation in vivo. We believe that our findings and the developed strategy here could pave the way for multifunctional coatings not only on orthopedic implants, but also for additional applications in catalysts, sensors, tissue engineering, etc.
RESUMO
Rumen of ruminant animals is known as a natural reactor involved in highly efficient lignocelluloses degradation. Rumen fibrolytic microbes have attracted an increasing attention for their potential value in biofuel research. Studies on rumen microbes have traditionally entailed the isolation of fibrolytic bacteria and subsequent analysis of fibrolytic enzymes. Developments in genomic and metagenomic approaches have made it possible to isolate directly genes and gene clusters encoding fibrolytic activities from rumen samples, permitting a global analysis of mechanisms of degradation of lignocellulose in rumen. Research in this field shows that lignocellulose degradation in rumen is a complex process involving a number of different microbes and is effected by a huge array of hydrolytic enzymes in a concerted fashion. This review briefly summarizes results from recent studies, especially metagenomic studies, on lignocellulose degradation in rumen.
Assuntos
Bactérias/metabolismo , Celulase/genética , Glicosídeo Hidrolases/genética , Lignina/metabolismo , Rúmen/microbiologia , Animais , Bactérias/genética , Biodegradação Ambiental , Cromossomos Artificiais Bacterianos , Biblioteca Gênica , MetagenômicaRESUMO
The rumen hosts one of the most efficient microbial systems for degrading plant cell walls, yet the predominant cellulolytic proteins and fibrolytic mechanism(s) remain elusive. Here we investigated the cellulolytic microbiome of the yak rumen by using a combination of metagenome-based and bacterial artificial chromosome (BAC)-based functional screening approaches. Totally 223 fibrolytic BAC clones were pyrosequenced and 10,070 ORFs were identified. Among them 150 were annotated as the glycoside hydrolase (GH) genes for fibrolytic proteins, and the majority (69%) of them were clustered or linked with genes encoding related functions. Among the 35 fibrolytic contigs of >10 Kb in length, 25 were derived from Bacteroidetes and four from Firmicutes. Coverage analysis indicated that the fibrolytic genes on most Bacteroidetes-contigs were abundantly represented in the metagenomic sequences, and they were frequently linked with genes encoding SusC/SusD-type outer-membrane proteins. GH5, GH9, and GH10 cellulase/hemicellulase genes were predominant, but no GH48 exocellulase gene was found. Most (85%) of the cellulase and hemicellulase proteins possessed a signal peptide; only a few carried carbohydrate-binding modules, and no cellulosomal domains were detected. These findings suggest that the SucC/SucD-involving mechanism, instead of one based on cellulosomes or the free-enzyme system, serves a major role in lignocellulose degradation in yak rumen. Genes encoding an endoglucanase of a novel GH5 subfamily occurred frequently in the metagenome, and the recombinant proteins encoded by the genes displayed moderate Avicelase in addition to endoglucanase activities, suggesting their important contribution to lignocellulose degradation in the exocellulase-scarce rumen.