VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats
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 repai...
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Elsevier
2023-04-01
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Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844023018339 |
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author | Wanqiu Xu Xiaohang Xu Lihong Yao Bing Xue Hualei Xi Xiaofang Cao Guiyan Piao Song Lin Xiumei Wang |
author_facet | Wanqiu Xu Xiaohang Xu Lihong Yao Bing Xue Hualei Xi Xiaofang Cao Guiyan Piao Song Lin Xiumei Wang |
author_sort | Wanqiu Xu |
collection | DOAJ |
description | 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. |
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language | English |
last_indexed | 2024-04-09T15:19:21Z |
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spelling | doaj.art-790835dec5844febb05a2cfe139e92652023-04-29T14:50:25ZengElsevierHeliyon2405-84402023-04-0194e14626VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in ratsWanqiu Xu0Xiaohang Xu1Lihong Yao2Bing Xue3Hualei Xi4Xiaofang Cao5Guiyan Piao6Song Lin7Xiumei Wang8Department of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaDepartment of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaCorresponding author. Department of Dentistry, The Second Affiliated Hospital of Harbin Medical University, 148 Baojian Road, Harbin, 150001, China.; Department of Dentistry, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150001, ChinaObjective: 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.http://www.sciencedirect.com/science/article/pii/S2405844023018339rDPSCsVEGFANeural differentiationFacial nerve defectsSU5416Neural tissue engineering |
spellingShingle | Wanqiu Xu Xiaohang Xu Lihong Yao Bing Xue Hualei Xi Xiaofang Cao Guiyan Piao Song Lin Xiumei Wang VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats Heliyon rDPSCs VEGFA Neural differentiation Facial nerve defects SU5416 Neural tissue engineering |
title | VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats |
title_full | VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats |
title_fullStr | VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats |
title_full_unstemmed | VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats |
title_short | VEGFA-modified DPSCs combined with LC-YE-PLGA NGCs promote facial nerve injury repair in rats |
title_sort | vegfa modified dpscs combined with lc ye plga ngcs promote facial nerve injury repair in rats |
topic | rDPSCs VEGFA Neural differentiation Facial nerve defects SU5416 Neural tissue engineering |
url | http://www.sciencedirect.com/science/article/pii/S2405844023018339 |
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