Department of Orthopedic, Kaohsiung Veterans General Hospital, Kaohsiung City, Taiwan, Republic of China.
Graduate Institute of Medical Science, National Defense Medical Center, Taipei City, Taiwan, Republic of China.
J Tissue Eng Regen Med. 2021 Feb;15(2):163-175. doi: 10.1002/term.3162. Epub 2020 Dec 15.
At present, no definitive treatment for articular cartilage defects has been perfected. Most of the previous treatments involved multiple drilling and microfracture over defect sites with repair-related substances, which poses a limited therapeutic effect. End-stage therapy includes artificial knee joint replacement. In this study, we prepared a novel decellularized natural cartilage scaffold from porcine articular cartilage by supercritical CO extraction technology and three-dimensional (3D) composites made using decellularized porcine cartilage graft (dPCG) as scaffolds, platelet-rich plasma (PRP), thrombin as signals and chondrocytes as cells for the treatment of articular cartilage defects. In this study, in vitro and in vivo cartilage regeneration and the expression of chondrogenic markers were examined. Decellularized cartilage graft (dPCG) was evaluated for the extent of cell and DNA removal. Residual cartilage ECM structure was confirmed to be type II collagen by SDS PAGE and immunostaining. The new 3D composite with dPCG (100 mg and 2 × 10 chondrocytes) scaffold promotes chondrogenic marker expression in vitro. We found that the in vivo 3D composite implanted cartilage defect showed significant regeneration relative to the blank and control implant. Immunohistochemical staining showed increase of expression including Collagen type II and aggrecan in 3D composite both in vitro and in vivo studies. In this study, the bioengineered 3D composite by combining dPCG scaffold, chondrocytes, and PRP facilitated the chondrogenic marker expression in both in vitro and in vivo models with accelerated cartilage regeneration. This might serve the purpose of clinical treatment of large focal articular cartilage defects in humans in the near future.
目前，尚未完善针对关节软骨缺损的明确治疗方法。大多数既往治疗方法涉及在缺陷部位多次钻孔和微骨折，并结合修复相关物质，其疗效有限。终末期治疗包括人工膝关节置换。在本研究中，我们采用超临界 CO2 萃取技术，从猪关节软骨中制备了一种新型去细胞天然软骨支架，并采用去细胞猪软骨移植物（dPCG）作为支架、富含血小板的血浆（PRP）、凝血酶作为信号以及软骨细胞作为细胞，制备了 3D 复合材料，用于治疗关节软骨缺损。本研究检测了体外和体内软骨再生以及软骨形成标志物的表达情况。评估了去细胞软骨移植物（dPCG）的细胞和 DNA 去除程度。通过 SDS-PAGE 和免疫染色证实残留的软骨 ECM 结构为 II 型胶原。具有 dPCG（100mg 和 2×10 个软骨细胞）支架的新型 3D 复合材料可促进体外软骨形成标志物的表达。我们发现，与空白和对照植入物相比，体内植入的 3D 复合材料植入的软骨缺损具有显著的再生。免疫组织化学染色显示，在体外和体内研究中，3D 复合材料中的 Collagen type II 和 aggrecan 表达增加。在本研究中，通过结合 dPCG 支架、软骨细胞和 PRP 的生物工程 3D 复合材料，在体外和体内模型中均促进了软骨形成标志物的表达，加速了软骨再生。这可能为未来不久在人类中治疗大的局灶性关节软骨缺损提供临床治疗方法。
