Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential

Abstract To evaluate the effects of an endoscopic aqueous environment on the viability and differentiation capacity of autologous bone marrow mesenchymal stem cells (MSCs) enriched with β-tricalcium phosphate (β-TCP). A screening-enrichment-combination circulating system (SECCS) was used to prepare...

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Main Authors: Wugui Chen, Guangfeng Ling, Chengshou Lin, Hengmei Chen, Kongning Chen, Shijie Chen, Yuchao Ye, Chengzhao Liu
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-01340-4
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author Wugui Chen
Guangfeng Ling
Chengshou Lin
Hengmei Chen
Kongning Chen
Shijie Chen
Yuchao Ye
Chengzhao Liu
author_facet Wugui Chen
Guangfeng Ling
Chengshou Lin
Hengmei Chen
Kongning Chen
Shijie Chen
Yuchao Ye
Chengzhao Liu
author_sort Wugui Chen
collection DOAJ
description Abstract To evaluate the effects of an endoscopic aqueous environment on the viability and differentiation capacity of autologous bone marrow mesenchymal stem cells (MSCs) enriched with β-tricalcium phosphate (β-TCP). A screening-enrichment-combination circulating system (SECCS) was used to prepare MSCs/β-TCP from a patient. A simulated aqueous flushing environment for spinal endoscopic surgery was established with different flushing times (0–60 min). Scanning electron microscopy (SEM) was employed to determine the adhesion and condition of MSCs/β-TCP. CCK-8 was used to determine the viability of adherent MSCs. The osteogenic, adipogenic, and chondrogenic differentiation potentials of adherent MSCs were assessed with alizarin red, alkaline phosphatase, Oil Red O, and toluidine blue staining. A subcutaneous implantation mouse model was utilized to evaluate the osteogenic capacity of MSCs/β-TCP under different aqueous flushing conditions using micro-CT, haematoxylin–eosin staining, and Von Kossa staining. SEM revealed sustained MSC adhesion on β-TCP across all irrigation durations (0–60 min), with no differences in adhesion density. However, cell viability declined significantly after 30 min(P < 0.05). Differentiation assays demonstrated time-dependent impairments: osteogenic and adipogenic potentials decreased significantly at 30 min (P < 0.05), chondrogenic morphology disorganized after 15 min, while ALP activity declined only at 60 min (P < 0.05). In vivo, irrigation > 15 min markedly reduced bone formation (P < 0.05) and parallel reductions in mineralized matrix(P < 0.05). In summary, prolonged irrigation (> 15 min) in the spinal endoscopic aqueous environment compromises MSC/β-TCP adhesion and osteogenic capacity. Optimizing surgical efficiency and strictly controlling the irrigation time and pressure are critical in preserving graft performance for successful bone fusion.
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spelling doaj-art-f95c05149c7b450f9cc587f800a930632025-08-20T03:10:35ZengNature PortfolioScientific Reports2045-23222025-06-011511910.1038/s41598-025-01340-4Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potentialWugui Chen0Guangfeng Ling1Chengshou Lin2Hengmei Chen3Kongning Chen4Shijie Chen5Yuchao Ye6Chengzhao Liu7Department of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalDepartment of Orthopedics, Fujian Medical University Affiliated With Mindong HospitalAbstract To evaluate the effects of an endoscopic aqueous environment on the viability and differentiation capacity of autologous bone marrow mesenchymal stem cells (MSCs) enriched with β-tricalcium phosphate (β-TCP). A screening-enrichment-combination circulating system (SECCS) was used to prepare MSCs/β-TCP from a patient. A simulated aqueous flushing environment for spinal endoscopic surgery was established with different flushing times (0–60 min). Scanning electron microscopy (SEM) was employed to determine the adhesion and condition of MSCs/β-TCP. CCK-8 was used to determine the viability of adherent MSCs. The osteogenic, adipogenic, and chondrogenic differentiation potentials of adherent MSCs were assessed with alizarin red, alkaline phosphatase, Oil Red O, and toluidine blue staining. A subcutaneous implantation mouse model was utilized to evaluate the osteogenic capacity of MSCs/β-TCP under different aqueous flushing conditions using micro-CT, haematoxylin–eosin staining, and Von Kossa staining. SEM revealed sustained MSC adhesion on β-TCP across all irrigation durations (0–60 min), with no differences in adhesion density. However, cell viability declined significantly after 30 min(P < 0.05). Differentiation assays demonstrated time-dependent impairments: osteogenic and adipogenic potentials decreased significantly at 30 min (P < 0.05), chondrogenic morphology disorganized after 15 min, while ALP activity declined only at 60 min (P < 0.05). In vivo, irrigation > 15 min markedly reduced bone formation (P < 0.05) and parallel reductions in mineralized matrix(P < 0.05). In summary, prolonged irrigation (> 15 min) in the spinal endoscopic aqueous environment compromises MSC/β-TCP adhesion and osteogenic capacity. Optimizing surgical efficiency and strictly controlling the irrigation time and pressure are critical in preserving graft performance for successful bone fusion.https://doi.org/10.1038/s41598-025-01340-4Endoscopic lumbar fusionAqueous medium environmentMSCsβ-TCPOsteogenesis
spellingShingle Wugui Chen
Guangfeng Ling
Chengshou Lin
Hengmei Chen
Kongning Chen
Shijie Chen
Yuchao Ye
Chengzhao Liu
Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential
Scientific Reports
Endoscopic lumbar fusion
Aqueous medium environment
MSCs
β-TCP
Osteogenesis
title Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential
title_full Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential
title_fullStr Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential
title_full_unstemmed Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential
title_short Prolonged irrigation time in endoscopic aqueous medium impairs MSC/β-TCP adhesion and osteogenic potential
title_sort prolonged irrigation time in endoscopic aqueous medium impairs msc β tcp adhesion and osteogenic potential
topic Endoscopic lumbar fusion
Aqueous medium environment
MSCs
β-TCP
Osteogenesis
url https://doi.org/10.1038/s41598-025-01340-4
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