VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats

Wanqiu Xu; Xiaohang Xu; Lihong Yao; Bing Xue; Hualei Xi; Xiaofang Cao; Guiyan Piao; Song Lin; Xiumei Wang

doi:10.1016/j.heliyon.2023.e14626

VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats

Heliyon. 2023 Mar 28;9(4):e14626. doi: 10.1016/j.heliyon.2023.e14626. eCollection 2023 Apr.

Authors

Wanqiu Xu¹, Xiaohang Xu¹, Lihong Yao¹, Bing Xue¹, Hualei Xi¹, Xiaofang Cao¹, Guiyan Piao¹, Song Lin¹, Xiumei Wang¹

Affiliation

¹ Department of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.

Abstract

Objective: The aim of this research was to investigate the effect of vascular endothelial growth factor A (VEGFA)-overexpressing rat dental pulp stem cells (rDPSCs) combined with laminin-coated and yarn-encapsulated poly(_l-lactide-co-glycolide) (PLGA) nerve guidance conduit (LC-YE-PLGA NGC) in repairing 10 mm facial nerve injury in rats.

Study design: rDPSCs isolated from rat mandibular central incisor were cultured and identified in vitro and further transfected with the lentiviral vectors (Lv-VEGFA). To investigate the role and mechanisms of VEGFA in neurogenic differentiation in vitro, semaxanib (SU5416), Cell Counting Kit-8 (CCK-8), real-time quantitative polymerase chain reaction (qPCR) and Western blotting were performed. Ten-millimeter facial nerve defect models in rats were established and bridged by LC-YE-PLGA NGCs. The repair effects were detected by transmission electron microscopy (TEM), compound muscle action potential (CMAP), immunohistochemistry and immunofluorescence.

Results: Extracted cells exhibited spindle-shaped morphology, presented typical markers (CD44⁺CD90⁺CD34^-CD45^-), and presented multidirectional differentiation potential. The DPSCs with VEGFA overexpression were constructed successfully. VEGFA enhanced the proliferation and neural differentiation ability of rDPSCs, and the expression of neuron-specific enolase (NSE) and βIII-tubulin was increased. However, these trends were reversed with the addition of SU5416. This suggests that VEGFA mediates the above effects mainly through vascular endothelial growth factor receptor 2 (VEGFR2) binding. The LC-YE-NGC basically meet the requirements of facial nerve repair. For the in vivo experiment, the CMAP latency period was shorter in DPSCS-VEGFA-NGC group in comparison with other experimental groups, while the amplitude was increased. Such functional recovery correlated well with an increase in histological improvement. Further study suggested that VEGFA-modified DPSCs could increase the myelin number, thickness and axon diameter of facial nerve. NSE, βIII-tubulin and S100 fluorescence intensity and immunohistochemical staining intensity were significantly enhanced.

Conclusion: VEGFA-modified rDPSCs combined with LC-YE-PLGA NGCs have certain advantages in the growth and functional recovery of facial nerves in rats.

Keywords: Facial nerve defects; Neural differentiation; Neural tissue engineering; SU5416; VEGFA; rDPSCs.