依那西普包埋丝素/普鲁兰水凝胶增强骨髓刺激软骨修复

摘要：2022年3月，重庆第三军医大学(军医大学)西南医院联合外科中心;重庆医科大学医学信息学院(Center for Joint Surgery, Southwest Hospital, Third Military Medical University (Arm

2022年3月，重庆第三军医大学(军医大学)西南医院联合外科中心;重庆医科大学医学信息学院(Center for Joint Surgery, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China,;College of Medical Informatics, Chongqing Medical University,Chongqing, China) Xiaoyuan Gong老师研究团队在《Biomolecules》上发表论文：

“Etanercept embedded silk fibroin/pullulan hydrogel enhance cartilage repair in bone marrow stimulation”

“依那西普包埋丝素/普鲁兰水凝胶增强骨髓刺激软骨修复”

Abstract

Background: Bone marrow stimulation (BMS) is the most used operative treatment in repairing cartilage defect clinically, but always results in fibrocartilage formation, which is easily worn out and needs second therapy. In this study, we prepared an Etanercept (Ept) embedded silk fibroin/pullulan hydrogel to enhance the therapeutic efficacy of BMS.

Methods: Ept was dissolved in silk fibroin (SF)—tyramine substituted carboxymethylated pullulan (PL) solution and enzyme crosslinked to obtain the Ept contained SF/PL hydrogel. The synergistical effect of SF/PL hydrogel and Ept was verified by rabbit osteochondral defect model. The mechanism of Ept in promoting articular cartilage repair was studied on human osteoarthritic chondrocytes (hOACs) and human bone marrow mesenchymal stromal cells (hBMSCs) in vitro, respectively.

Results: At 4 and 8 weeks after implanting the hydrogel into the osteochondral defect of rabbit, histological analysis revealed that the regenerated tissue in Ept + group had higher cellular density with better texture, and the newly formed hyaline cartilage tissue was seamlessly integrated with adjacent native tissue in the Ept + group. In cellular experiments, Ept treatment significantly promoted both gene and protein expression of type II collagen in hOACs, while decreased the protein levels of metalloproteinase (MMP)-13 and a disintegrin and metalloprotease with thrombospondin motifs 5 (ADAMTS5); alcian blue staining, type II collagen and aggrecan stainings showed that addition of Ept significantly reversed the chondrogenesis inhibition effect of tumor necrosis factor alpha (TNF-α) on hBMSCs.

Conclusion: BMS could be augmented by Ept embedded hydrogel, potentially by regulating the catabolic and anabolic dynamics in adjacent chondrocytes and enhancement of BMSCs chondrogenesis.

摘要

背景:骨髓刺激(Bone marrow stimulation, BMS)是临床上修复软骨缺损最常用的手术方法，但常导致纤维软骨形成，易磨损，需二次治疗。本研究制备了依那西普包埋丝素/普鲁兰水凝胶，以提高BMS的治疗效果。

方法:将Ept溶解于丝素(SF) -酪胺取代羧甲基化普鲁兰(PL)溶液中，酶交联得到含SF/PL的Ept水凝胶。通过兔骨软骨缺损模型验证了SF/PL水凝胶与Ept的协同作用。分别在体外培养人骨关节炎软骨细胞(hOACs)和人骨髓间充质基质细胞(hBMSCs)上研究Ept促进关节软骨修复的机制。

结果:将水凝胶植入兔骨软骨缺损后4周和8周，组织学分析显示，Ept +组再生组织细胞密度更高，质地更好，新形成的透明软骨组织与邻近原生组织无缝融合。在细胞实验中，Ept处理显著促进了hOACs中II型胶原的基因和蛋白表达，同时降低了金属蛋白酶(MMP)-13和具有血栓反应蛋白基元5的崩解素和金属蛋白酶(ADAMTS5)的蛋白水平;阿利新蓝染色、II型胶原和聚集蛋白染色显示，添加Ept可显著逆转肿瘤坏死因子α (TNF-α)对hBMSCs的软骨形成抑制作用。

结论:Ept包埋水凝胶可能通过调节邻近软骨细胞的分解代谢和合成代谢动力学以及促进骨髓间充质干细胞的软骨形成来增强骨髓间充质干细胞。

该论文中，hBMSCs和hOACs细胞是使用Ausbian进口特级胎牛血清完成的