Nanofiber matrix formulations for the delivery of Exendin-4 for tendon regeneration: In vitro and in vivo assessment.
Nanofiber matrix formulations for the delivery of Exendin-4 for tendon regeneration: In vitro and in vivo assessment.
复制标题
纳米纤维基质制剂用于递送Exendin-4用于肌腱再生：体外和体内评估。
DOI：
10.1016/j.bioactmat.2023.01.013
复制
发表时间：
2023-07
影响因子：
18.9
通讯作者：
Kumbar, Sangamesh G.
中科院分区：
文献类型：
--
作者：
Abdulmalik, Sama;Gallo, Jack;Nip, Jonathan;Katebifar, Sara;Arul, Michael;Lebaschi, Amir;Munch, Lucas N.;Bartly, Jenna M.;Choudhary, Shilpa;Kalajzic, Ivo;Banasavadi-Siddegowdae, Yeshavanth Kumar;Nukavarapu, Syam P.;Kumbar, Sangamesh G.
肌腱和韧带损伤是最常见的肌肉骨骼损伤，不仅影响生活质量，还会造成巨大的经济负担。肌腱/韧带损伤的手术干预利用生物和/或工程移植物来重建受损组织，但这些方法存在局限性。工程基质相较于生物移植物具有更优的物理化学性质，但缺乏促进组织愈合的理想生物活性。虽然加入药物可以增强生物活性，但较大的基质表面积和疏水性可能导致药物不受控制的突释和/或由于结合而不完全释放。为了克服这些局限性，我们在肌腱损伤模型中评估了使用一种增强型纳米纤维基质递送一种肽生长因子（艾塞那肽 - 4；Ex - 4）的情况。为了克服由于聚己内酯（PCL）基质疏水性导致的药物表面结合（这种疏水性原本预期会增强细胞 - 材料相互作用），我们将PCL和醋酸纤维素（CA）混合，并电纺出纤维直径在600到1000纳米之间的纳米纤维基质。为了避免突释并保护药物，我们将Ex - 4封装在埃洛石纳米管（HNTs）的开放内腔中，用聚合物混合物密封HNT管端，并在电纺前将载有Ex - 4的HNTs混入聚合物混合物中。这使得突释从约75%降低到约40%，但没有改变基质形态、纤维直径或拉伸性能。我们通过在基质表面培养人骨髓间充质干细胞（hMSCs）21天并测量肌腱分化情况来评估Ex - 4纳米纤维制剂的生物活性，并与仅在基础培养基中的纳米纤维基质进行比较。令人惊讶的是，我们观察到与对照组相比，Ex - 4纳米纤维基质加速了hMSC的增殖速率，并提高了硫酸化糖胺聚糖、肌腱相关基因（Scx、Mkx和Tnmd）以及细胞外基质相关基因（Col - I、Col - III和Dcn）的水平。然后，我们通过组织学、标志物表达、功能性步态分析和力学测试评估了Ex - 4纳米纤维基质在大鼠跟腱全层缺损中的安全性和有效性。我们的分析证实，与单独的纳米纤维基质相比，Ex - 4纳米纤维基质促进了肌腱愈合并减少了纤维软骨的形成。这些发现表明Ex - 4是肌腱组织工程中一种潜在的有价值的工具。
用于递送艾塞那肽 - 4（Ex - 4）以促进全层肌腱缺损愈合的生物活性纳米纤维基质配方。
生物活性纳米纤维基质在体外加速了人骨髓间充质干细胞（MSCs）的增殖速率，并提高了硫酸化糖胺聚糖、肌腱相关基因（Scx、Mkx和Tnmd）以及细胞外基质相关基因（Col - I、Col - III和Dcn）的水平。
生物活性基质通过降低白细胞介素 - 6（IL - 6）水平并提高白细胞介素 - 10（IL - 10）水平，在全身和局部促进抗炎作用。
针对跟腱功能指数（AFI）的DigiGait分析证实，使用含或不含大鼠MSCs的生物活性基质治疗的组具有更好的功能恢复和力学性能。
基质能够为小分子和生长因子提供可定制的释放曲线，以促进软组织愈合和减少瘢痕。
Tendon and ligament injuries are the most common musculoskeletal injuries, which not only impact the quality of life but result in a massive economic burden. Surgical interventions for tendon/ligament injuries utilize biological and/or engineered grafts to reconstruct damaged tissue, but these have limitations. Engineered matrices confer superior physicochemical properties over biological grafts but lack desirable bioactivity to promote tissue healing. While incorporating drugs can enhance bioactivity, large matrix surface areas and hydrophobicity can lead to uncontrolled burst release and/or incomplete release due to binding. To overcome these limitations, we evaluated the delivery of a peptide growth factor (exendin-4; Ex-4) using an enhanced nanofiber matrix in a tendon injury model. To overcome drug surface binding due to matrix hydrophobicity of poly(caprolactone) (PCL)—which would be expected to enhance cell-material interactions—we blended PCL and cellulose acetate (CA) and electrospun nanofiber matrices with fiber diameters ranging from 600 to 1000 nm. To avoid burst release and protect the drug, we encapsulated Ex-4 in the open lumen of halloysite nanotubes (HNTs), sealed the HNT tube endings with a polymer blend, and mixed Ex-4-loaded HNTs into the polymer mixture before electrospinning. This reduced burst release from ∼75% to ∼40%, but did not alter matrix morphology, fiber diameter, or tensile properties. We evaluated the bioactivity of the Ex-4 nanofiber formulation by culturing human mesenchymal stem cells (hMSCs) on matrix surfaces for 21 days and measuring tenogenic differentiation, compared with nanofiber matrices in basal media alone. Strikingly, we observed that Ex-4 nanofiber matrices accelerated the hMSC proliferation rate and elevated levels of sulfated glycosaminoglycan, tendon-related genes (Scx, Mkx, and Tnmd), and ECM-related genes (Col-I, Col-III, and Dcn), compared to control. We then assessed the safety and efficacy of Ex-4 nanofiber matrices in a full-thickness rat Achilles tendon defect with histology, marker expression, functional walking track analysis, and mechanical testing. Our analysis confirmed that Ex-4 nanofiber matrices enhanced tendon healing and reduced fibrocartilage formation versus nanofiber matrices alone. These findings implicate Ex-4 as a potentially valuable tool for tendon tissue engineering. Bioactive nanofiber matrix formulation for the delivery of exendin-4 (Ex-4) for full-thickness tendon defect healing. Bioactive nanofiber matrices accelerated in vitro human mesenchymal stem cells (MSCs) proliferation rate and elevated levels of sulfated glycosaminoglycan, tendon-related genes (Scx, Mkx, and Tnmd), and ECM-related genes (Col-I, Col-III, and Dcn). Bioactive matrix promoted anti-inflammatory effects systemically and locally by reducing the level of IL-6 and increasing IL-10. DigiGait analysis for Achilles functional index (AFI) confirmed superior functional recovery and mechanical properties for the groups treated with a bioactive matrix with and without rat MSCs. Matrices enable tailorable release profiles for small molecules and growth factors to promote soft tissue healing and scar reduction.
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DOI：
10.1002/jbmr.3199
发表时间：
2017-09
期刊：
Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
影响因子：
--
作者：
Asahara H;Inui M;Lotz MK
通讯作者：
Lotz MK
影响因子：
12.4
作者：
Chen H;Wang G;Lang L;Jacobson O;Kiesewetter DO;Liu Y;Ma Y;Zhang X;Wu H;Zhu L;Niu G;Chen X
通讯作者：
Chen X
影响因子：
14
作者：
Banks, Jessica M.;Mozdzen, Laura C.;Harley, Brendan A. C.;Bailey, Ryan C.
通讯作者：
Bailey, Ryan C.
影响因子：
5.4
作者：
Bianchi E;Ruggeri M;Rossi S;Vigani B;Miele D;Bonferoni MC;Sandri G;Ferrari F
通讯作者：
Ferrari F
影响因子：
14
作者：
Czaplewski, Sarah K.;Tsai, Tsung-Lin;Li, Wan-Ju
通讯作者：
Li, Wan-Ju