Green fluorescent proteins engineered for cartilage-targeted drug delivery: Insights for transport into highly charged avascular tissues.
Green fluorescent proteins engineered for cartilage-targeted drug delivery: Insights for transport into highly charged avascular tissues.
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DOI：
10.1016/j.biomaterials.2018.08.050
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发表时间：
2018-11
期刊：
影响因子：
14
通讯作者：
Grodzinsky AJ
中科院分区：
文献类型：
--
作者：
Krishnan Y;Rees HA;Rossitto CP;Kim SE;Hung HK;Frank EH;Olsen BD;Liu DR;Hammond PT;Grodzinsky AJ
骨关节炎（OA）是关节炎的最常见形式，是一种多因素疾病，主要影响软骨以及其他关节组织，如软骨下骨。由于软骨的无血管性质和关节内递送的药物通过滑膜的快速清除，缺乏有效的药物递送仍然是开发用于OA的疾病调节药物的主要挑战。阳离子传递载体通过与带负电荷的基质蛋白聚糖相互作用，可以显著增强药物在软骨中的摄取、渗透和保留。在这项研究中，我们使用了“增压”的绿色荧光蛋白（GFP），工程具有广泛的净正电荷和表面电荷分布，以表征在隔离的其他因素，如载体的大小和形状的载体电荷运输到软骨的影响。我们定量了GFP变体在活的人膝关节软骨和牛软骨外植体中的摄取、软骨渗透程度和细胞摄取。基于这些结果，我们确定了位于软骨细胞外基质以及常驻活软骨细胞内的潜在药物靶标的GFP载体的最佳净电荷。这些阳离子GFP在药物递送所需的剂量下在测量的细胞活力和代谢、软骨细胞生物合成和基质降解方面对软骨没有不利影响。除了量化GFP吸收的动力学之外，我们还开发了一种用于GFP变体运输的预测数学模型，这些变体在软骨中表现出最高的吸收和渗透。该模型进一步用于预测GFP在放大到体内应用（例如关节内注射到人膝盖中）期间的转运行为。从这项研究中获得的见解为软骨靶向递送系统的开发奠定了基础，以防止软骨退化，改善组织再生并减少可能导致受OA影响的其他关节组织退化的炎症。
Osteoarthritis (OA), the most common form of arthritis, is a multi-factorial disease that primarily affects cartilage as well as other joint tissues such as subchondral bone. The lack of effective drug delivery, due to the avascular nature of cartilage and the rapid clearance of intra-articularly delivered drugs via the synovium, remains a major challenge in the development of disease mod- ifying drugs for OA. Cationic delivery carriers can significantly enhance the uptake, penetration and retention of drugs in cartilage by interacting with negatively charged matrix proteoglycans. In this study, we used “supercharged” green fluorescent proteins (GFPs), engineered to have a wide range of net positive charge and surface charge distributions, to characterize the effects of carrier charge on transport into cartilage in isolation of other factors such as carrier size and shape. We quantified the uptake, extent of cartilage penetration and cellular uptake of the GFP variants into living human knee cartilage and bovine cartilage explants. Based on these results, we identified optimal net charges of GFP carriers for potential drug targets located within cartilage extracellular matrix as well as the resident live chondrocytes. These cationic GFPs did not have adverse effects on cartilage in terms of measured cell viability and metabolism, cartilage cell biosynthesis and matrix degradation at doses needed for drug delivery. In addition to quantifying the kinetics of GFP uptake, we developed a predictive mathematical model for transport of the GFP variants that exhibited the highest uptake and penetration into cartilage. This model was further used to predict the transport behavior of GFPs during scale-up to in vivo applications such as intra-articular injection into human knees. The insights gained from this study set the stage for development of cartilage-targeted delivery systems to prevent cartilage degeneration, improve tissue regeneration and reduce inflammation that may cause degradation of other joint tissues affected by OA.
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影响因子：
8
作者：
Fröhlich E
通讯作者：
Fröhlich E
影响因子：
33.7
作者：
Bajpayee, Ambika G.;Grodzinsky, Alan J.
通讯作者：
Grodzinsky, Alan J.
影响因子：
6.2
作者：
Lam, Christopher N.;Yao, Helen;Olsen, Bradley D.
通讯作者：
Olsen, Bradley D.
影响因子：
7
作者：
Bajpayee AG;Quadir MA;Hammond PT;Grodzinsky AJ
通讯作者：
Grodzinsky AJ
DOI：
10.1016/0304-4165(86)90306-5
发表时间：
1986-09-04
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
FARNDALE, RW;BUTTLE, DJ;BARRETT, AJ
通讯作者：
BARRETT, AJ