Charge based intra-cartilage delivery of single dose dexamethasone using Avidin nano-carriers suppresses cytokine-induced catabolism long term.
Charge based intra-cartilage delivery of single dose dexamethasone using Avidin nano-carriers suppresses cytokine-induced catabolism long term.
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DOI：
10.1016/j.joca.2015.07.010
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发表时间：
2016-01
影响因子：
7
通讯作者：
Grodzinsky AJ
中科院分区：
文献类型：
--
作者：
Bajpayee AG;Quadir MA;Hammond PT;Grodzinsky AJ
抗生物素蛋白具有靶向软骨内药物递送的理想特性：其小尺寸和最佳正电荷使得能够快速穿透全层软骨和静电结合相互作用，从而在体内产生长半衰期。在此，我们将抗生物素蛋白与地塞米松（DEX）结合，并测试了以下假设：单剂量抗生物素蛋白递送的DEX可以改善与创伤后OA相关的受尼古丁攻击的软骨中的分解代谢作用。使用快速（酯）和缓慢的pH敏感性释放（腙）接头将抗生物素蛋白与DEX共价缀合。使用3 H-DEX-抗生物素蛋白（闪烁计数）表征这些缀合物的DEX释放动力学。用IL-1α处理的软骨外植体与或不与抗生物素蛋白-DEX缀合物一起培养，并与可溶性DEX进行比较。硫酸化糖胺聚糖（sGAG）的损失和生物合成速率分别使用DMMB测定和35 S-掺入。使用荧光染色测量软骨细胞活力。在生理缓冲液条件下，酯接头从抗生物素蛋白释放DEX的速度显著快于腙。抗生物素蛋白-DEX单次给药可在3周内抑制马槟榔诱导的sGAG丢失，挽救IL-1α诱导的细胞死亡，并恢复sGAG合成水平，而不会引起细胞毒性。与单剂量可溶性DEX相比，1：1组合（快：慢）的两种抗生物素蛋白-DEX缀合物具有最显著的生物活性，可溶性DEX具有较短的寿命效应，因此需要在整个培养期间连续补充以改善分解代谢效应。软骨内药物递送仍然不足，因为药物从关节迅速清除，需要多次注射或在滑液中持续释放高剂量。单剂量的抗生物素蛋白偶联药物能够以低组织内剂量在软骨内快速摄取和持续递送，并且可能最大限度地减少不必要的药物暴露于其他关节组织。
Avidin exhibits ideal characteristics for targeted intra-cartilage drug delivery: its small size and optimal positive charge enable rapid penetration through full-thickness cartilage and electrostatic binding interactions that give long half-lives in-vivo. Here we conjugated Avidin with dexamethasone (DEX) and tested the hypothesis that single-dose Avidin-delivered DEX can ameliorate catabolic effects in cytokine-challenged cartilage relevant to post-traumatic OA. Avidin was covalently conjugated with DEX using fast (ester) and slow, pH-sensitive release (hydrazone) linkers. DEX release kinetics from these conjugates was characterized using 3H-DEX-Avidin (scintillation counting). Cartilage explants treated with IL-1α were cultured with or without Avidin-DEX conjugates and compared to soluble DEX. Sulfated-glycosaminoglycan (sGAG) loss and biosynthesis rates were measured using DMMB assay and 35S-incorporation, respectively. Chondrocyte viability was measured using fluorescence staining. Ester linker released DEX from Avidin significantly faster than hydrazone under physiological buffer conditions. Single dose Avidin-DEX suppressed cytokine-induced sGAG loss over 3-weeks, rescued IL-1α-induced cell death, and restored sGAG synthesis levels without causing cytotoxicity. The two Avidin-DEX conjugates in 1:1 combination (fast:slow) had the most prominent bioactivity compared to single dose soluble-DEX, which had a shorter-lived effect and thus needed continuous replenishment throughout the culture period to ameliorate catabolic effects. Intra-cartilage drug delivery remains inadequate as drugs rapidly clear from the joint, requiring multiple injections or sustained release of high doses in synovial fluid. A single dose of Avidin-conjugated drug enables rapid uptake and sustained delivery inside cartilage at low intratissue doses, and potentially can minimize unwanted drug exposure to other joint tissues.