Department of Molecular Science and Technology, Ajou University, Suwon 443-749, Republic of Korea.
Cell Therapy Center, Ajou University Medical Center, Suwon 443-749, Republic of Korea.
Acta Biomater. 2018 Jul 1;74:192-206. doi: 10.1016/j.actbio.2018.05.032. Epub 2018 May 21.
In this work, we chose cartilage acellular matrix (CAM) as a promising antiadhesive material because CAM effectively inhibits the formation of blood vessels, and we used electrospinning to prepare antiadhesive barriers. Additionally, we synthesized N-hydroxysuccinimide (NHS)-poly(caprolactone-co-lactide-co-glycolide)-NHS (MP) copolymers (to tune degradation) as a cross-linking agent for CAM. This is the first report on the development of electrospun cross-linked (Cx) CAM/MP (CA/P) nanofiber (NF) (Cx-CA/P-NF) with a tunable degradation period as an antiadhesive barrier. Compared with the CA/P-NF before cross-linking, the electrospun Cx-CA/P-NF after cross-linking showed different biodegradation. Cx-CA/P-NF significantly inhibited the in vitro attachment and proliferation of human umbilical vein endothelial cells (HUVECs), as confirmed by an MTT assay and scanning electron microscopy images. Cx-CA/P-NFs implanted between a surgically damaged peritoneal wall and cecum gradually degraded in 7 days; this process was monitored by NIR imaging. The in vivo evaluation of the anti-tissue adhesive effect of Cx-CA/P-NFs revealed little adhesion, few blood vessels, and negligible inflammation at 7 days determined by hematoxylin and eosin staining. ED1 staining of Cx-CA/P-NFs showed infiltration of few macrophages because of the inflammatory response to the Cx-CA/P-NF as compared with an untreated injury model. Additionally, Cx-CA/P-NFs significantly suppressed the formation of blood vessels between the peritoneal wall and cecum, according to CD31 staining. Overall, Cx-CA/P-NFs yielded little adhesion, infiltration by macrophages, or formation of blood vessels in a postoperative antiadhesion assay. Thus, it is reasonable to conclude that the Cx-CA/P-NF designed herein successfully works as an antiadhesive barrier with a tunable degradation period.
The cartilage acellular matrix (CAM) can inhibit the formation of fibrous tissue bridges and blood vessels between the tissue at an injured site and the surrounding healthy tissues. However, CAM has not been rigorously investigated as an antiadhesive barrier. In this manuscript, the cross-linked CAM nanofiber (Cx-CA/P-NF) designed herein successfully works as an antiadhesive barrier. Cx-CA/P-NFs yielded little adhesion, infiltration by macrophages, or formation of blood vessels in a postoperative antiadhesion assay. Moreover, we demonstrated the suitable properties of Cx-CA/P-NF such as easy cross-linking by maintaining the antiadhesive properties, controllable biodegradation, and in vivo antiadhesive effect of Cx-CA/P-NF.
本研究选择脱细胞软骨基质（CAM）作为一种有前途的抗粘连材料，因为 CAM 能有效抑制血管形成，我们使用静电纺丝来制备抗粘连屏障。此外，我们合成了 N-羟基琥珀酰亚胺（NHS）-聚（己内酯-co-丙交酯-co-乙交酯）-NHS（MP）共聚物（用于调节降解）作为 CAM 的交联剂。这是首次报道开发具有可调降解期的交联（Cx）CAM/MP（CA/P）纳米纤维（NF）（Cx-CA/P-NF）作为抗粘连屏障。与交联前的 CA/P-NF 相比，交联后的静电纺 Cx-CA/P-NF 表现出不同的生物降解性。MTT 检测和扫描电子显微镜图像证实，Cx-CA/P-NF 显著抑制人脐静脉内皮细胞（HUVEC）的体外黏附和增殖。Cx-CA/P-NF 植入手术损伤的腹膜壁和盲肠之间，7 天内逐渐降解；这一过程通过近红外成像进行监测。通过苏木精和伊红染色，在体内评估 Cx-CA/P-NF 的抗组织粘连效果，结果显示 7 天时粘连少、血管少、炎症轻微。与未处理的损伤模型相比，Cx-CA/P-NF 的 ED1 染色显示巨噬细胞浸润较少，这是由于对 Cx-CA/P-NF 的炎症反应所致。此外，Cx-CA/P-NF 的 CD31 染色显示，其显著抑制了腹膜壁和盲肠之间血管的形成。总之，在术后抗粘连试验中，Cx-CA/P-NF 引起的粘连、巨噬细胞浸润和血管形成均较少。因此，可以合理地得出结论，本研究设计的 Cx-CA/P-NF 成功用作具有可调降解期的抗粘连屏障。
