从牙本质基质中提取的碱性蛋白质作为用于软骨修复和再生的生物活性分子混合物。
Alkali-extracted proteins from the tooth dentin matrix as a mixture of bioactive molecules for cartilage repair and regeneration.
作者信息
Wang Sainan, Mao Sicong, Huang Guibin, Jia Peipei, Dong Yanmei, Zheng Jinxuan
机构信息
Department of Cariology and Endodontology, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing, 100081, China.
Department of General Dentistry, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing, 100081, China.
出版信息
Regen Ther. 2024 Jul 4;26:407-414. doi: 10.1016/j.reth.2024.06.015. eCollection 2024 Jun.
INTRODUCTION
Dentin matrix extracted protein (DMEP) is a mixture of proteins extracted from the organic matrix of a natural demineralized dentin matrix that is rich in a variety of growth factors. However, the effect of DMEP on cartilage regeneration is unclear. The aim of this study was to investigate the efficacy of DMEP extracted via a novel alkali conditioning method in promoting cartilage regeneration.
METHODS
Alkali-extracted DMEP (a-DMEP) was obtained from human dentin fragments using pH 10 bicarbonate buffer. The concentration of chondrogenesis-related growth factors in a-DMEP was measured via enzyme-linked immunosorbent assay (ELISA). Human bone marrow mesenchymal stem cells (hBMMSCs) in pellet form were induced with a-DMEP. Alcian blue and Safranin O staining were performed to detect cartilage matrix formation, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to assess chondrogenic-related gene expression in the pellets. Rabbit articular osteochondral defects were implanted with collagen and a-DMEP. Cartilage regeneration was assessed with histological staining 4 weeks after surgery.
RESULTS
Compared with traditional neutral-extracted DMEP, a-DMEP significantly increased the levels of transforming growth factor beta 1(TGF-β1), insulin-like growth factor-1(IGF-1) and basic fibroblast growth factor (bFGF). After coculture with hBMMSC pellets, a-DMEP significantly promoted the expression of the collagen type II alpha 1(COL2A1) and aggrecan (ACAN) genes and the formation of cartilage extracellular matrix in cell pellets. Moreover, compared with equivalent amounts of exogenous human recombinant TGF-β1, a-DMEP had a stronger chondrogenic ability. a-DMEP induced osteochondral regeneration with hyaline cartilage-like structures.
CONCLUSIONS
Our results showed that a-DMEP, a compound of various proteins derived from natural tissues, is a promising material for cartilage repair and regeneration.
引言
牙本质基质提取蛋白（DMEP）是从天然脱矿牙本质基质的有机基质中提取的蛋白质混合物，富含多种生长因子。然而，DMEP对软骨再生的影响尚不清楚。本研究旨在探讨通过一种新型碱处理方法提取的DMEP促进软骨再生的效果。
方法
使用pH 10的碳酸氢盐缓冲液从人牙本质碎片中获得碱提取的DMEP（a-DMEP）。通过酶联免疫吸附测定（ELISA）测量a-DMEP中软骨生成相关生长因子的浓度。用a-DMEP诱导呈丸粒形式的人骨髓间充质干细胞（hBMMSCs）。进行阿尔辛蓝和番红O染色以检测软骨基质形成，并使用定量实时聚合酶链反应（qRT-PCR）评估丸粒中软骨生成相关基因的表达。将胶原蛋白和a-DMEP植入兔关节软骨骨缺损处。术后4周通过组织学染色评估软骨再生情况。
结果
与传统中性提取的DMEP相比，a-DMEP显著提高了转化生长因子β1（TGF-β1）、胰岛素样生长因子-1（IGF-1）和碱性成纤维细胞生长因子（bFGF）的水平。与hBMMSC丸粒共培养后，a-DMEP显著促进了Ⅱ型胶原蛋白α1（COL2A1）和聚集蛋白聚糖（ACAN）基因的表达以及细胞丸粒中软骨细胞外基质的形成。此外，与等量的外源性人重组TGF-β1相比，a-DMEP具有更强的软骨生成能力。a-DMEP诱导软骨骨再生形成透明软骨样结构。
结论
我们的结果表明，a-DMEP作为一种源自天然组织的多种蛋白质的复合物，是一种有前景的软骨修复和再生材料。
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