口腔疾病防治2024，Vol.32Issue(5)：330-340,11.DOI:10.12016/j.issn.2096-1456.2024.05.002
光固化水凝胶负载骨髓间充质干细胞修复大鼠颅骨缺损
Reconstruction of rat calvarial defects utilizing an ultraviolet-cured hydrogel loaded with bone marrow mesen-chymal stem cells
摘要
Abstract
Objective To investigate the osteogenic properties of a methacrylated gelatin(GelMA)/bone marrow mesenchymal stem cells(BMSCs)composite hydrogel applied to the skull defect area of rats and to provide an experi-mental basis for the development of bone regeneration biomaterials.Methods This study was approved by the Animal Ethics Committee of Nanjing University.A novel photocurable composite biohydrogel was developed by constructing photoinitiators[lthium phenyl(2,4,6-trimethylbenzoyl)phosphinate,LAP],GelMA,and BMSCs.The surface morphology and elemental composition of the gel were examined using scanning electron microscopy(SEM)and energy-dispersive X-ray spectroscopy(EDX).The compressive strength of the gel was evaluated using an electronic universal testing ma-chine.After in vitro culture for 1,2,and 5 days,the proliferation of the BMSCs in the hydrogels was assessed using a CCK-8 assay,and their survival and morphology were examined through confocal microscopy.A 5 mm critical bone de-ficiency model was generated in a rat skull.The group receiving composite hydrogel treatment was referred to as the Gel-MA/BMSCs group,whereas the untreated group served as the control group.At the 4th and 8th weeks,micro-CT scans were taken to measure the bone defect area and new bone index,while at the 8th week,skull samples from the defect ar-ea were subjected to H&E staining,van Gieson staining,and Goldner staining to evaluate the quality of bone regenera-tion and new bone formation.Results SEM observed that the solidified GelMA showed a 3D spongy gel network with uniform morphology,the porosity of GelMA was 73.41％and the pore size of GelMA was(28.75±7.13)μm.EDX results showed that C and O were evenly distributed in the network macroporous structure of hydrogel.The hydrogel compres-sion strength was 152 kPa.On the 5th day of GelMA/BMSCs culture,the cellular morphology transitioned from oval to spindle shaped under microscopic observation,accompanied by a significant increase in cell proliferation(159.4％,as determined by the CCK-8 assay).At 4 weeks after surgery,a 3D reconstructed micro-CT image revealed a minimal re-duction in bone defect size within the control group and abundant new bone formation in the GelMA/BMSCs group.At 8 weeks after surgery,no significant changes were observed in the control group's bone defect area,with only limited evi-dence of new bone growth;however,substantial healing of skull defects was evident in the GelMA/BMSCs group.Quan-titative analysis at both the 4-and 8-week examinations indicated significant improvements in the new bone volume(BV),new bone volume/total bone volume(BV/TV),bone surface(BS),and bone surface/total bone volume(BS/TV)in the GelMA/BMSCs group compared to those in the control group(P＜0.05).Histological staining showed continuous and dense formation of bone tissue within the defects in the GelMA/BMSCs group and only sporadic formation of new bone,primarily consisting of fibrous connective tissue,at the defect edge in the control group.Conclusion Photocur-ing hydrogel-based stem cell therapy exhibits favorable biosafety profiles and has potential for clinical application by inducing new bone formation and promoting maturation within rat skull defects.关键词
骨缺损/骨组织工程/支架/水凝胶/GelMA水凝胶/骨髓间充质干细胞/干细胞疗法/光固化/骨再生/骨修复Key words
bone defect/bone tissue engineering/scaffold/hydrogel/GelMA hydrogel/bone marrow mes-enchymal stem cells/stem cell therapy/photocuring/bone regeneration/bone repair分类
医药卫生引用本文复制引用
丁梦,李强,李筱叶,贺敖,戴卓,董衡,牟永斌..光固化水凝胶负载骨髓间充质干细胞修复大鼠颅骨缺损[J].口腔疾病防治,2024,32(5):330-340,11.基金项目
This study was supported by the grants from National Natural Science Foundation of China(No.82301104) （No.82301104）
Jiangsu Province Key Research and Development Project(No.BE2020629) （No.BE2020629）
Natural Science Foundation of Jiangsu Province(No.BK20230160) （No.BK20230160）
Nanjing Health Science and Technology Development Fund Project(No.YKK22182). 国家自然科学基金项目(82301104) （No.YKK22182）
江苏省重点研发计划(社会发展)项目(BE2020629) （社会发展）
江苏省科技计划专项资金/青年基金项目(BK20230160) （BK20230160）
南京市卫生科技发展专项资金项目计划(YKK22182) （YKK22182）
