载槲皮素明胶微球对MC3T3-E1增殖和分化的影响

doi:10.13591/j.cnki.kqyx.2024.07.003

Abstract

Abstract:

Objective This study prepared gelatin three-dimensional porous microspheres and investigated the feasibility of gelatin three-dimensional porous microspheres loaded with quercetin (G-quercetin) as bone tissue scaffold material. Methods Porous gelatin microspheres were prepared by emulsification and loaded with quercetin by lyophilization. Scanning electron microscopy was used to observe morphology of the microspheres. The cytotoxicity of G-quercetin microspheres and their effects on the adhesion, proliferation and differentiation of mouse embryonic osteoblast precursor cells (MC3T3-E1) were detected by immunofluorescence staining, live/dead cell staining and CCK-8 assay, alkaline phosphatase (ALP) staining and alizarin red staining. RT-PCR was used to detect the transcriptional levels of osteoblast-related cytokines such as Runx-2, ALP, OPN and OCN. Results The scanning electron microscopy results showed that the prepared three-dimensional microporous material loaded with quercetin gelatin had a porous structure. Cell adhesion showed that the cells could spread well on the surface of the microspheres. Compared with the control group, the results of live/dead cell staining and CCK-8 detection showed that the microspheres had no significant cytotoxicity (P>0.05). Compared with the control group, G-quercetin microspheres showed an increase in ALP expression and mineralization in vitro. PCR results also showed a significant increase in Runx-2, ALP, OCN, OPN (P<0.05). Conclusion The G-quercetin porous microspheres prepared in this experiment have good biocompatibility and can promote the osteogenic differentiation of MC3T3-E1 in vitro. It is expected to be used as a new scaffold material for bone tissue engineering.

Key words: gelatin, quercetin, microspheres, osteogenic differentiation, bone tissue engineering, mouse embryonic osteoblast precursor cells (MC3T3-E1)

CLC Number:

R318.08

DONG Weijie, SU Tingshu, XIN Xianzhen. Effects of quercetin loaded gelatin microspheres on proliferation and differentiation of MC3T3-E1[J]. Stomatology, 2024, 44(7): 494-499.

Figures/Tables 8

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References 40

[1]	Li J, Karim MA, Che H, et al. Deletion of p16 prevents estrogen deficiency-induced osteoporosis by inhibiting oxidative stress and osteocyte senescence[J]. Am J Transl Res, 2020, 12(2): 672-683. pmid: 32194914
[2]	Cauley JA. Estrogen and bone health in men and women[J]. Steroids, 2015, 99(Pt A): 11-15. doi: 10.1016/j.steroids.2014.12.010 pmid: 25555470
[3]	Matsumoto Y, Otsuka F, Takano M, et al. Estrogen and glucocorticoid regulate osteoblast differentiation through the interaction of bone morphogenetic protein-2 and tumor necrosis factor-α in C2C12 cells[J]. Mol Cell Endocrinol, 2010, 325(1/2): 118-127.
[4]	Zhang ND, Han T, Huang BK, et al. Traditional Chinese medicine formulas for the treatment of osteoporosis: Implication for antiosteoporotic drug discovery[J]. J Ethnopharmacol, 2016, 189: 61-80.
[5]	Skibola CF, Smith MT. Potential health impacts of excessive flavonoid intake[J]. Free Radic Biol Med, 2000, 29(3/4): 375-383.
[6]	Yamagata K. Metabolic syndrome: Preventive effects of dietary flavonoids[M]// Studies in Natural Products Chemistry. Amsterdam: Elsevier, 2019: 1-28.
[7]	Kelly GS. Quercetin. Monograph[J]. Altern Med Rev, 2011, 16(2):172-194.
[8]	Shirke SS, Jadhav SR, Jagtap AG. Osteoprotective effect of Phaseolus vulgaris L in ovariectomy-induced osteopenia in rats[J]. Menopause, 2009, 16(3): 589-596. doi: 10.1097/gme.0b013e31818e64c4 pmid: 19169163
[9]	Song JE, Tripathy N, Lee DH, et al. Quercetin inlaid silk fibroin/hydroxyapatite scaffold promotes enhanced osteogenesis[J]. ACS Appl Mater Interfaces, 2018, 10(39): 32955-32964.
[10]	Zhou YN, Wu YQ, Jiang XQ, et al. The effect of quercetin on the osteogenesic differentiation and angiogenic factor expression of bone marrow-derived mesenchymal stem cells[J]. PLoS One, 2015, 10(6): e0129605.
[11]	Wang N, Wang LY, Yang JH, et al. Quercetin promotes osteogenic differentiation and antioxidant responses of mouse bone mesenchymal stem cells through activation of the AMPK/SIRT1 signaling pathway[J]. Phytother Res, 2021, 35(5): 2639-2650. doi: 10.1002/ptr.7010 pmid: 33421256
[12]	牟利民, 张文豪, 陈万吉, 等. 雌激素及雌激素受体与骨关节炎的相关性研究进展[J]. 浙江医学, 2021, 43(24): 2721-2724.
[13]	Jiao DL, Zheng A, Liu Y, et al. Bidirectional differentiation of BMSCs induced by a biomimetic procallus based on a gelatin-reduced graphene oxide reinforced hydrogel for rapid bone regeneration[J]. Bioact Mater, 2020, 6(7): 2011-2028.
[14]	Barty-King CH, Chan CLC, Parker RM, et al. Mechanochromic, structurally colored, and edible hydrogels prepared from hydroxypropyl cellulose and gelatin[J]. Adv Mater, 2021, 33(37): e2102112.
[15]	Park S, Edwards S, Hou SJ, et al. A multi-interpenetrating network (IPN) hydrogel with gelatin and silk fibroin[J]. Biomater Sci, 2019, 7(4): 1276-1280.
[16]	He QY, Huang Y, Wang SY. Hofmeister effect-assisted one step fabrication of ductile and strong gelatin hydrogels[J]. Adv Funct Materials, 2018, 28(5): 1705069.
[17]	周昉, 刘俊, 胡姝颖, 等. 可载药的磁性聚己内酯/明胶微球支架的制备及其体外成骨性能的研究[J]. 口腔医学, 2022, 42(4):289-295.
[18]	Jahangiri L, Kim A, Nishimura I. Effect of ovariectomy on the local residual ridge remodeling[J]. J Prosthet Dent, 1997, 77(4): 435-443. pmid: 9104721
[19]	Saarelainen J, Kiviniemi V, Kröger H, et al. Body mass index and bone loss among postmenopausal women: The 10-year follow-up of the OSTPRE cohort[J]. J Bone Miner Metab, 2012, 30(2): 208-216. doi: 10.1007/s00774-011-0305-5 pmid: 21938384
[20]	Eriksen EF, Robins DA. Teriparatide: A bone formation treatment for osteoporosis[J]. Drugs Today, 2004, 40(11): 935-948.
[21]	黎祺, 王贺, 黄紫君, 等. 雌激素对牙周膜细胞修复重建牙周组织调节机制的研究进展[J]. 口腔疾病防治, 2021, 7(11): 787-792.
[22]	王翔宇, 孙克勤, 王毅, 等. 雌激素水平对大鼠实验性牙周炎模型的影响[J]. 中国现代医学杂志, 2018, 28(25): 1-5.
[23]	Longo M, Brama M, Marino M, et al. Interaction of estrogen receptor alpha with protein kinase C alpha and c-Src in osteoblasts during differentiation[J]. Bone, 2004, 34(1): 100-111. doi: 10.1016/j.bone.2003.09.007 pmid: 14751567
[24]	Tsai MH, Huang GS, Hung YC, et al. Psoralea corylifolia extract ameliorates experimental osteoporosis in ovariectomized rats[J]. Am J Chin Med, 2007, 35(4): 669-680.
[25]	庞新岗. 槲皮素通过ER及BMP信号通路调节骨髓间充质干细胞分化的研究[D]. 南京: 东南大学, 2018.
[26]	张鑫彦, 陈松, 李晓娜. 基于明胶的生物复合型药物载体材料的构建与改性[J]. 医用生物力学, 2021, 36(S1): 399.
[27]	何家辰, 刘畅, 陈迟迟, 等. 制备负载细胞可注射微球及体外评价[J]. 中国组织工程研究, 2022, 26(28): 4483-4488.
[28]	杨雪, 王宝群, 姜晓文, 等. 可生物降解植物多糖止血微球的制备及性能[J]. 中国组织工程研究, 2022, 26(16): 2487-2491.
[29]	肖嘉聪, 麦嘉乐, 张罡瑜, 等. 槲皮素调控关节炎软骨细胞胆固醇代谢的机制研究[J]. 中国骨质疏松杂志, 2022, 28(9): 1336-1341.
[30]	吕方南, 黄洁, 陈剑秋, 等. 载槲皮素PNIPAm纳米凝胶的制备及细胞学研究[J]. 南京中医药大学学报, 2020, 36(2): 197-204.
[31]	米娅莎尔·呼加合买提, 李雪, 吕慧欣, 等. 槲皮素对人口腔上皮细胞再生的生物学效应研究[J]. 口腔医学研究, 2018, 34(9): 991-994. doi: 10.13701/j.cnki.kqyxyj.2018.09.017
[32]	葛超. 被动吸烟对实验性牙周炎大鼠牙周组织的影响及槲皮素的保护作用[D]. 石家庄: 河北医科大学, 2014.
[33]	顾艺婧, 傅稼耀, 武文婧, 等. 槲皮素通过抗骨相关细胞衰老作用治疗雌激素缺乏骨质疏松症的初步研究[J]. 同济大学学报(医学版), 2019, 40(3): 274-280.
[34]	张波, 胡凌云, 苟林, 等. 槲皮素对人骨髓间充质干细胞增殖和成骨分化的影响及分子机制[J]. 中国骨质疏松杂志, 2022, 28(12): 1765-1769, 1776.
[35]	Swioklo S, Watson KA, Williamson EM, et al. Defining key structural determinants for the pro-osteogenic activity of flavonoids[J]. J Nat Prod, 2015, 78(11): 2598-2608. doi: 10.1021/acs.jnatprod.5b00075 pmid: 26517554
[36]	张文静. 槲皮素对炎症状态下人牙周膜干细胞成骨损伤的改善作用及机制探究[D]. 济南: 山东大学, 2021.
[37]	金珂, 吴晓玲, 罗小玲, 等. 雌激素刺激下人牙周膜干细胞成骨分化及通路间的交叉串扰[J]. 中国组织工程研究, 2022, 26(24): 3886-3891.
[38]	李小云, 杨丽, 张荣华. 雌激素受体信号通路介导的槲皮素调控骨髓间充质干细胞成骨及成脂分化研究[J]. 中华中医药杂志, 2020, 35(12): 6011-6014.
[39]	郑红, 唐薇, 角建林, 等. 槲皮素通过促进成骨分化治疗去势骨质疏松症大鼠的分子机制[J]. 中药药理与临床, 2017, 33(5): 16-20.
[40]	汪正明, 王瑞, 金钊锴, 等. 基于网络药理学和分子对接探讨高频调髓中药治疗股骨头坏死的作用机制[J]. 特产研究, 2023, 45(4): 74-81.

基因名称	引物序列(5'→3')
Runx-2	F:ATCCAGCCACCTTCACTTACACC R:GGGACCATTGGGAACTGATAGG
ALP	F:TATGTCTGGAACCGCACTGAAC R:CACTAGCAAGAAGAAGCCTTTGG
OPN	F:GACGGCCGAGGTGATAGCTT R:CATGGCTGGTCTTCCCGTTGC
OCN	F:GCCCTGACTGCATTCTGCCTCT R:TCACCACCTTACTGCCCTCCTG
GAPDH	F:GGCACAGTCAAGGCTGAGAATG R:ATGGTGGTGAAGACGCCAGTA

Effects of quercetin loaded gelatin microspheres on proliferation and differentiation of MC3T3-E1

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