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In vitro analysis of the potential cartilage implant bacterial nanocellulose using the bovine cartilage punch model

机译:使用牛软骨冲床模型的潜在软骨植入细菌纳米纤维的体外分析

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Biocompatible bacterial nanocellulose (BNC) shows high potential as wound dressing and dura mater replacement, and even for the development of blood vessel or cartilage implants. Thus, the regenerative capacity of BNC implants was analyzed using a standardized bovine cartilage punch model. Cartilage rings with an outer diameter of 6 mmand an inner defect diameter of 2 mm were derived from the trochlear groove (femur-patellar articulation site). BNC implants were cultured inside the cartilage rings for up to 12 weeks. Cartilage-BNC-constructs were then evaluated by histology (hematoxylin/eosin; safranin O), immunohistology (aggrecan, collagens 1 and 2), and for protein content, mRNA expression, and push-out force of the implants. Cartilage-BNC-constructs displayed vital chondrocytes (= 90% until week 9; 80% until 12 weeks), preserved matrix integrity during culture, limited loss of matrix-bound proteoglycan from 'host' cartilage or cartilage-BNC-interface, and constant release of proteoglycans into the culture supernatant. In addition, the content of the matrix protein collagen 2 in cartilage and cartilage-BNC-interface was approximately constant over time (with very limited quantities of collagen 1). Interestingly, BNC implants showed: (1) cell colonization of the implant; (2) progressively increasing mRNA levels for the proteoglycan aggrecan and collagen 2 (max. fivefold); and (3) significantly increasing push-out forces during culture (max. 1.6-fold). Retained tissue integrity and progressively increasing chondrogenic differentiation in implant and carti-lage-implant-interface suggest beginning cartilage regeneration in the BNC in the present model and indicate a high potential of BNC as a cartilage replacement material. Thus, the present model appears suitable to predict the in vivo performance of cartilage replacementmaterials (e.g., BNC) for tissue engineering.
机译:生物相容性细菌纳米纤维素(BNC)显示出高潜力作为伤口敷料和硬脑膜替代品,甚至用于开发血管或软骨植入物。因此,使用标准化的牛软骨冲头模型分析了BNC植入物的再生能力。外径为6mmAnd的软骨环,内部缺陷直径为2mm(股骨 - 髌骨铰接位点)。 BNC植入物在软骨环内培养长达12周。然后通过组织学(苏木精/曙红; Safranin O),免疫组织学(蛋白,胶原1和2)和用于植入物的蛋白质含量,mRNA表达和推出力来评估软骨-BNC构建体。软骨-BNC构建体展示了重要的软骨细胞(& = 90%直到第9周;& 80%直到12周),培养过程中保留的基质完整性,来自“宿主”软骨或软骨-BNC的基质结合蛋白多糖的损失有限蛋白多糖的界面,恒定释放到培养上清液中。此外,在软骨和软骨-BNC界面中的基质蛋白胶原2的含量随时间近似恒定(具有非常有限的胶原1)。有趣的是,BNC植入物显示:(1)植入物的细胞定植; (2)促蛋白生成蛋白酶蛋白和胶原蛋白2的mRNA水平逐渐增加(最大五倍); (3)在培养期间显着增加推出力(最大1.6倍)。保留的组织完整性和逐渐增加植入物和Casti-leg-incormant-interfactions-interface的软骨内分化表明在本模型中开始于BNC中的软骨再生,并表示BNC作为软骨置换材料的高潜力。因此,本模型似乎适于预测组织工程的软骨替代材料(例如,BNC)的体内性能。

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