依那西普包埋絲素蛋白/普魯蘭水凝膠增強骨髓刺激中的軟骨修復

2022年12月，重慶第三軍醫大學(陸軍醫大學)西南醫院聯合外科中心，重慶醫科大學醫學信息學院，（Center for Joint Surgery, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China.College of Medical Informatics, Chongqing Medical University,Chongqing, China ）Cheng Chen and Xiaoyuan Gong研究團隊在《Frontiers in Bioengineering and Biotechnology》，標題為：“Etanercept embedded silk fibroin/pullulan hydrogel enhance cartilage repair in bone marrow stimulation”

“依那西普包埋絲素蛋白/普魯蘭水凝膠增強骨髓刺激中的軟骨修復”

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)是臨床上修復軟骨缺損最常用的手術方法，但常導致纖維軟骨形成，易磨損，需二次治療。本研究制備了依那西普包埋絲素/普魯蘭水凝膠，以提高BMS的治療效果。

方法:將Ept溶解于絲素(SF) -酪胺取代羧甲基化普魯蘭(PL)溶液中，酶交聯得到含SF/PL的Ept水凝膠。通過兔骨軟骨缺損模型驗證了SF/PL水凝膠與Ept的協同作用。分別在體外培養人骨關節炎軟骨細胞(hOACs)和人骨髓間充質基質細胞(hBMSCs)上研究Ept促進關節軟骨修復的機制。

結果:將水凝膠植入兔骨軟骨缺損后4周和8周，組織學分析顯示，Ept +組再生組織細胞密度更高，質地更好，新形成的透明軟骨組織與鄰近原生組織無縫融合。在細胞實驗中，Ept處理顯著促進了hOACs中II型膠原的基因和蛋白表達，同時降低了金屬蛋白酶(MMP)-13和具有血栓反應蛋白基元5的崩解素和金屬蛋白酶(ADAMTS5)的蛋白水平;阿利新藍染色、II型膠原和聚集蛋白染色顯示，添加Ept可顯著逆轉腫瘤壞死因子α (TNF-α)對hBMSCs的軟骨形成抑制作用。

結論:Ept包埋水凝膠可能通過調節鄰近軟骨細胞的分解代謝和合成代謝動力學以及促進骨髓間充質干細胞的軟骨形成來增強骨髓間充質干細胞。

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