藏红花素水凝胶对软骨细胞与MC3T3-E1细胞的影响  

Effect of crocin hydrogel on chondrocytes and MC3T3-E1 cells

作　　者：尹航 宋奎 Yin Hang;Song Kui(School of Physical Education,Wuhan Business University,Wuhan 430056,Hubei Province,China;First Affiliated Hospital of Jishou University,Jishou 416000,Hunan Province,China)

机构地区：[1]武汉商学院体育学院,湖北省武汉市430056 [2]吉首大学第一附属医院,湖南省吉首市416000

出　　处：《中国组织工程研究》2025年第34期7293-7300,共8页Chinese Journal of Tissue Engineering Research

基　　金：湖南省自然科学基金项目(2023JJ30609),项目负责人:宋奎。

摘　　要：背景:研究发现,藏红花素可抑制骨关节炎软骨细胞的凋亡、减轻骨关节炎大鼠炎症反应,促进骨质疏松大鼠骨形成并抑制骨丢失,在软骨与骨损伤中具有巨大的应用潜力。目的:观察藏红花素水凝胶对软骨细胞、MC3T3-E1细胞的影响。方法:将不同浓度(0,15,20,25 mmol/L)的藏红花素溶液分别加入氧化透明质酸-氧化硫酸软骨素-Ⅱ型胶原混合溶液中,采用席夫碱交联方式制备藏红花素水凝胶,随后制备4种水凝胶浸提液。将4种藏红花素水凝胶浸提液分别与人软骨细胞共培养,检测细胞糖胺聚糖合成,CCK-8法检测细胞增殖,Transwell小室实验检测细胞迁移,RT-qPCR检测细胞SOX9、Ⅱ型胶原、聚集蛋白聚糖mRNA表达。将4种藏红花素水凝胶浸提液分别与MC3T3-E1细胞共培养,CCK-8法检测细胞增殖,RT-qPCR检测成骨诱导分化后细胞RUNX2、Ⅰ型胶原、碱性磷酸酶、骨钙素mRNA表达,成骨诱导分化后的碱性磷酸酶活性与矿化结节形成。结果与结论:①CCK-8检测结果显示,15,20 mmol/L藏红花素水凝胶浸提液促进软骨细胞的增殖,25 mmol/L藏红花素水凝胶浸提液抑制软骨细胞的增殖。15,20,25 mmol/L藏红花素水凝胶浸提液均可促进软骨细胞的迁移,提升软骨细胞SOX9、Ⅱ型胶原、聚集蛋白聚糖mRNA表达与糖胺聚糖合成,并且呈现剂量依赖性。②15,20,25 mmol/L藏红花素水凝胶浸提液均可促进MC3T3-E1细胞的增殖,提高成骨诱导分化后细胞RUNX2、Ⅰ型胶原、碱性磷酸酶、骨钙素mRNA表达,并且具有剂量依赖性。15,20,25 mmol/L藏红花素水凝胶浸提液可提高成骨诱导分化后MC3T3-E1细胞碱性磷酸酶活性与矿化结节形成,其中以20 mmol/L藏红花素水凝胶浸提液的提高效果最显著。③结果表明,藏红花素水凝胶可促进软骨细胞的增殖、迁移及软骨细胞外基质的合成,促进MC3T3-E1细胞的增殖与成骨分化,其中20 mmol/L藏红花素水凝胶的综合效果较�BACKGROUND:Studies have found that crocin can inhibit the apoptosis of osteoarthritis chondrocytes,reduce the inflammatory response of osteoarthritis rats,promote bone formation and inhibit bone loss in osteoporotic rats,and has great application potential in cartilage and bone damage.OBJECTIVE:To observe the effects of crocin hydrogel on chondrocytes and MC3T3-E1 cells.METHODS:Crocin hydrogels were prepared by Schiff base crosslinking method by adding different concentrations of crocin solution(0,15,20,and 25 mmol/L)into oxidized hyaluronic acid-oxidized chondroitin sulfate-type Ⅱ collagen mixed solution,and then four kinds of hydrogel extracts were prepared.Human chondrocytes were cultured with four kinds of crocin hydrogel extracts to detect glycosaminoglycan synthesis.Cell proliferation was detected by CCK-8 assay.Cell migration was detected by Transwell chamber assay.RT-qPCR was used to detect the mRNA expression of SOX9,type Ⅱ collagen,and aggrecan in cells.Four kinds of crocin hydrogel extracts were co-cultured with MC3T3-E1 cells,respectively.Cell proliferation was detected by CCK-8 assay.RT-qPCR was used to detect the mRNA expression of RUNX2,type I collagen,alkaline phosphatase,and osteocalcin in cells.and the alkaline phosphatase activity and mineralized nodule formation after osteogenic differentiation.RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that 15 and 20 mmol/L crocin hydrogel extract promoted the proliferation of chondrocytes,while 25 mmol/L crocin hydrogel extract inhibited the proliferation of chondrocytes.15,20,and 25 mmol/L crocin hydrogel extracts could all promote the migration of chondrocytes,increase the expression of SOX9,type Ⅱ collagen,aggrecan mRNA and glycosaminoglycan synthesis of chondrocytes in a dose-dependent manner.(2)15,20,and 25 mmol/L crocin hydrogel extract could promote the proliferation of MC3T3-E1 cells and increase the expression of RUNX2,type I collagen,alkaline phosphatase,and osteocalcin mRNA in cells after osteogenic differentiation in a dose-depend

关 键 词：藏红花素 水凝胶 软骨细胞 MC3T3-E1细胞 细胞增殖 细胞迁移 细胞分化 工程化软骨材料 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R684