DNA改性胶原支架促进口腔黏膜缺损愈合的研究  

Study on DNA-modified collagen scaffolds to promote healing of oral mucosal defects

作　　者：党高鹏 汪亦菲 王雨竹 宋婧涵 白鹊 李之婷 牛丽娜 顾俊婷 DANG Gaopeng;WANG Yifei;WANG Yuzhu;SONG Jinghan;BAI Que;LI Zhiting;NIU Li'na;GU Junting(State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,National Clinical Research Center for Oral Diseases,Shaanxi Key Laboratory of Stomatology,Department of Prosthodontics,School of Stomatology,Air Force Medical University,Xi'an 710032,China;Stomatology Center,Taihe Hospital,Hubei University of Medicine,Shiyan 442000,China)

机构地区：[1]口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔医学重点实验室,空军军医大学口腔医院修复科,陕西西安710032 [2]湖北医药学院附属太和医院口腔医学中心,湖北十堰442000

出　　处：《空军军医大学学报》2024年第10期1104-1109,共6页Journal of Air Force Medical University

基　　金：国家自然科学基金(82325012,82301043);国家口腔疾病临床研究中心资助课题计划项目(LCA202004);国家资助博士后研究人员计划项目(GZC20233582);陕西省科协青年人才托举计划项目(20240304);陕西省创新能力支撑计划项目(2020TD-033)。

摘　　要：目的构建DNA修饰的胶原(DNA-Col)支架,探究改性后胶原(Col)生物相容性与促口腔黏膜软组织缺损愈合的性能。方法通过将Col材料浸泡于含DNA和磷酸基团活化剂的缓冲液中进行化学交联,以此构建出DNA-Col。通过甲基绿染色对DNA-Col上交联的DNA进行表征,并进一步利用衰减全反射-傅里叶变换红外光谱对DNA与Col的交联机制进行初步探索。之后,利用扫描电镜观察DNA-Col的表面形貌。在评价DNA-Col的生物相容性方面,采用了溶血实验、CCK-8细胞增殖实验和活/死细胞染色实验。随后在体外将DNA-Col与人口腔成纤维细胞共培养,通过qRT-PCR检测转化生长因子-β(TGF-β)、成纤维细胞生长因子-1(FGF-1)的mRNA表达水平,分析DNA-Col促进口腔黏膜缺损愈合的潜力。最后,在大鼠腭部黏膜缺损模型中检测DNA-Col的促愈合能力,并于术后10 d对各组黏膜愈合情况进行组织切片分析。结果DNA-Col保持了Col的特征性结构且生物相容性良好。人口腔成纤维细胞与DNA-Col共培养后,TGF-β、FGF-1等与细胞增殖迁移相关基因表达显著升高,提示DNA-Col具有促口腔黏膜愈合的潜力。大鼠腭部黏膜缺损模型显示DNA-Col组在术后10 d时创面愈合率可达(97.04±2.07)%,显著高于普通Col组[(79.38±5.76)%,P<0.01]。结论DNA-Col具有良好的生物相容性以及促口腔黏膜软组织缺损愈合的性能。Objective To construct a DNA-modified collagen(DNA-Col)scaffold to investigate the biocompatibility of the modified collagen(Col)and its performance in promoting the healing of oral mucosal soft tissue defects.Methods DNA-Col was constructed by chemical cross-linking methods by soaking Col materials in a buffer containing DNA and phosphate group activators.DNA crosslinked on DNA-Col was characterized by methyl green staining,and the cross-linking mechanism between DNA and Col was preliminarily explored by attenuated total reflection-Fourier transform infrared spectroscopy.The surface morphology of DNA-Col was observed by scanning electron microscopy.The biocompatibility of DNA-Col was evaluated by hemolysis test,CCK-8 cell proliferation assay and live/dead staining test.DNA-Col was then co-cultured with human oral fibroblasts in vitro,and the mRNA expression levels of transforming growth factor-β(TGF-β)and fibroblast growth factor-1(FGF-1)were detected by qRT-PCR,so as to analyze the potential of DNA-Col in promoting the healing of oral mucosal defects.Finally,the healing ability of DNA-Col was detected in the rat palatal mucosal defect model,and the mucosal healing of each group was analyzed by tissue sections 10 d after surgery.Results DNA-Col maintained the characteristic structure of Col and exhibited good biocompatibility.After co-culture of human oral fibroblasts with DNA-Col,the expressions of TGF-β,FGF-1 and other genes related to cell proliferation and migration were significantly increased,indicating that DNA-Col has the potential to promote oral mucosal healing.The rat palatal mucosal defect model showed that the wound healing rate of DNA-Col group was(97.04±2.07)%at 10 d after surgery,which was significantly higher than that of Col group[(79.38±5.76)%,P<0.01].Conclusion DNA-Col exhibits excellent biocompatibility and has the ability to promote the healing of oral mucosal soft tissue defects.

关 键 词：胶原支架 DNA 口腔黏膜 软组织增量 生物相容性 

分 类 号：R318[医药卫生—生物医学工程]