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发布时间：2019-03-09

论文类型：期刊论文

发表时间：2011-07-11

发表刊物：AFRICAN JOURNAL OF BIOTECHNOLOGY

收录刊物：SCIE、Scopus

卷号：10

期号：34

页面范围：6640-6646

ISSN号：1684-5315

关键字：Bone tissue engineering; osteoblast; respiration rate; in vitro culture; encapsulation

摘要：The oxygen consumption rates of Sprague-Dawley (SD) rat's osteoblasts cultured in different in vitro media were on-line measured by using precision dissolved oxygen and respirometry instrumentation. The osteoblasts, isolated from the calvaria of neonatal SD rat by an enzymatic digestive process of 0.25% trypsin solution and 0.1% type II collagenase, were diluted to 1 x 10(6) cells/ml in fresh media and passaged in T-25 flasks in an incubator. After two (2) passages, the cells were cultured directly in T-flasks and encapsulated medium by calcium alginate microcapsules, respectively. The cells were evaluated through live/dead assay, hematoxylin-eosin (HE) and alkaline phosphatase (ALP) staining. Moreover, Von-Kossa staining and Alizarin Red S staining were carried out for mineralized nodule formation. Following this, the oxygen consumption rates of osteoblasts in the earlier mentioned different cultures were measured on-line. The results showed that the osteoblasts performed well in its major functions after being examined by cellular morphology and viability, growth curve in growth kinetics, HE, ALP, Von-Kossa and Alizarin Red stains, which were capable of being used for further investigation as seed cells in bone tissue engineering. The osteoblasts cultured in static T-flask and encapsulated medium in vitro both proliferated well, and the oxygen consumption rates were 5.56 x 10(-6) and 1.25 x 10(-7) mu mol/(min.cell), respectively. By measuring the oxygen consumption rates of osteoblasts in different cultures in vitro, it would provide significant instruction and model function for further fundamental investigation and clinical application in bone tissue engineering.