重庆医学2025，Vol.54Issue(12)：2782-2791,10.DOI:10.3969/j.issn.1671-8348.2025.12.011
搭载BMP-2多肽的生物矿化胶原凝胶促进BMSCs成骨分化的研究
Study on osteogenic differentiation of BMSCs promoted by a bio-mineralized collagen gel loaded with BMP-2 peptide
LIU Bangding 1TANG Yongliang 1LI Ni 2REN Bo1
作者信息
- 1. Department of Orthopedics Ⅱ,Xi'an Central Hospital,Xi'an,Shaanxi 710004,China
- 2. Health Examination Center,Xi'an Chest Hospital,Xi'an,Shaanxi 710004,China
- 折叠
摘要
Abstract
Objective To synthesize a bio-mineralized collagen gel scaffold based on nano-hydroxyapa-tite(nHA)and collagen,and load with bone morphogenetic protein-2(BMP-2)peptide,to explore its sus-tained-release function and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs cells,male SD rats,and New Zealand big-eared white rabbits were divided into the blank group(no intervention with bio-mineralized collagen gel),the control group(intervention with bio-mineralized collagen gel without BMP-2),and the experimental group(intervention with bio-mineralized collagen gel load-ed with BMP-2)according to their intervention materials.Prepared bio-mineralized collagen gel loaded with BMP-2.Rabbit BMSCs were extracted from female New Zealand milk rabbits.The BMSCs were co-cultured with bio-mineralized collagen gel and grouped.The characterization of BMSCs in each group was observed by electron scanning microscope.The bio-mineralized collagen gel was injected into both sides of the subcutane-ous tissue on the back of rats and grouped.The degradation of the subcutaneous gel and the skin inflammation in each group of rats were observed.The proliferation of BMSCs in each group was detected by CCK-8 meth-od.The alkaline phosphatase(ALP)activity of BMSCs in each group was qualitatively and quantitatively ana-lyzed by ALP chromogenic kit.Live cell/dead cell immunofluorescence staining was performed after co-culture for 1 and 3 days,respectively.Alizarin red staining was used to evaluate the mineralization of calcium nodules in BMSCs of each group.qPCR was used to detect the mRNA expression levels of osteogenesis-related genes RUNX2 and OCN in BMSCs of each group.Immunofluorescence staining was used to detect the protein ex-pression levels of RUNX2 and OCN in BMSCs of each group.The critical bone defect model of the radius in vivo was established using male New Zealand white rabbits with big ears.The corresponding biomateralized collagen was implanted according to the groups.The bone regeneration of the radius in the white rabbits with big ears was detected by micro-CT.The data of bone volume fraction(BV/TV)and trabecular bone thickness(Tb.Th)were obtained and compared.HE staining and Masson staining were used to evaluate the new bone structure,material residue,inflammatory response and collagen fiber maturity of the radius in each group of rabbits.Results In vitro experiments showed that on the 7th day of co-culture,compared with the blank group,the proliferation rate of BMSCs in the control group and the experimental group was significantly in-creased(P<0.01).Compared with the blank group,the ALP activity in the experimental group and the con-trol group was significantly increased(P<0.001),and the activity in the experimental group was higher than that in the control group(P<0.001).Alizarin red staining results showed that compared with the blank group,the number of calcium nodules in the experimental group and the control group was significantly in-creased(P<0.001),and the number in the experimental group was more than that in the control group(P<0.01).The qPCR results showed that the expression levels of OCN and RUNX2 mRNA in the experimental group were significantly higher than those in the control group and the blank group,and the immunofluores-cence staining results showed the same trend.In vivo experiments,the results of HE and Masson staining showed that the in vivo osteogenic performance of the experimental group was significantly higher than that of the blank group and the control group(P<0.001).The BV/TV and Tb.Th in the experimental group were sig-nificantly higher than those in the blank group and the control group(P<0.001).Conclusion The bio-mineralized collagen gel loaded with BMP-2 polypeptide can promote the proliferation of BMSCs in vitro and the osteogen-ic differentiation of BMSCs at the same time.关键词
骨髓间充质干细胞/胶原/骨形态发生蛋白-2/缓释/骨再生Key words
bone marrow mesenchymal stem cells/collagen/bone morphogenetic protein-2/sustained release/bone regeneration分类
医药卫生引用本文复制引用
LIU Bangding,TANG Yongliang,LI Ni,REN Bo..搭载BMP-2多肽的生物矿化胶原凝胶促进BMSCs成骨分化的研究[J].重庆医学,2025,54(12):2782-2791,10.
