Nowwarote Nunthawan, Chahlaoui Zakaria, Petit Stephane, Duong Lucas T, Dingli Florent, Loew Damarys, Chansaenroj Ajjima, Kornsuthisopon Chatvadee, Osathanon Thanaphum, Ferre Francois Come, Fournier Benjamin P J
Centre de Recherche des Cordeliers, Molecular Oral Pathophysiology, INSERM UMRS 1138, Université Paris Cité, Sorbonne Université, Paris, 75006, France.
Department of Oral Biology, Dental Faculty Garancière, Université Paris Cité, Paris, 75006, France.
BMC Oral Health. 2024 Oct 1;24(1):1166. doi: 10.1186/s12903-024-04882-7.
Decellularized extracellular matrix (dECM) has been proposed as a useful source of biomimetic materials for regenerative medicine due to its biological properties that regulate cell behaviors. The present study aimed to investigate the influence of decellularized ECM derived from dental pulp stem cells (DPSCs) on gingival fibroblast (GF) cell behaviors. Cells were isolated from dental pulp and gingival tissues. ECM was derived from culturing dental pulp stem cells in growth medium supplemented with ascorbic acid. A bioinformatic database of the extracellular matrix was constructed using Metascape. GFs were reseeded onto dECM, and their adhesion, spreading, and organization were subsequently observed. The migration ability of the cells was determined using a scratch assay. Protein expression was evaluated using immunofluorescence staining.
Type 1 collagen and fibronectin were detected on the ECM and dECM derived from DPSCs. Negative phalloidin and nuclei were noted in the dECM. The proteomic database revealed enrichment of several proteins involved in ECM organization, ECM-receptor interaction, and focal adhesion. Compared with those on the controls, the GFs on the dECM exhibited more organized stress fibers. Furthermore, cultured GFs on dECM exhibited significantly enhanced migration and proliferation abilities. Interestingly, GFs seeded on dECM showed upregulation of FN1, ITGB3, and CTNNB1 mRNA levels.
ECM derived from DSPCs generates a crucial microenvironment for regulating GF adhesion, migration and proliferation. Therefore, decellularized ECM from DPSCs could serve as a matrix for oral tissue repair.
由于其调节细胞行为的生物学特性，脱细胞细胞外基质（dECM）已被认为是再生医学中仿生材料的有用来源。本研究旨在探讨牙髓干细胞（DPSCs）来源的脱细胞细胞外基质对牙龈成纤维细胞（GF）细胞行为的影响。细胞从牙髓和牙龈组织中分离出来。细胞外基质通过在添加抗坏血酸的生长培养基中培养牙髓干细胞获得。使用Metascape构建细胞外基质的生物信息数据库。将牙龈成纤维细胞重新接种到脱细胞细胞外基质上，随后观察其黏附、铺展和排列情况。使用划痕试验测定细胞的迁移能力。使用免疫荧光染色评估蛋白质表达。
在牙髓干细胞来源的细胞外基质和脱细胞细胞外基质上检测到I型胶原蛋白和纤连蛋白。在脱细胞细胞外基质中观察到鬼笔环肽阴性和细胞核。蛋白质组数据库显示，几种参与细胞外基质组织、细胞外基质-受体相互作用和粘着斑的蛋白质富集。与对照组相比，脱细胞细胞外基质上的牙龈成纤维细胞表现出更有序的应力纤维。此外，在脱细胞细胞外基质上培养的牙龈成纤维细胞表现出显著增强的迁移和增殖能力。有趣的是，接种在脱细胞细胞外基质上的牙龈成纤维细胞显示FN1、ITGB3和CTNNB1 mRNA水平上调。
牙髓干细胞来源的细胞外基质为调节牙龈成纤维细胞的黏附、迁移和增殖创造了关键的微环境。因此，牙髓干细胞来源的脱细胞细胞外基质可作为口腔组织修复的基质。
