
收稿日期: 2025-02-28
录用日期: 2025-07-18
网络出版日期: 2025-08-28
基金资助
上海交通大学医学院大学生创新训练计划第18期(1824504)
Application of single-cell RNA sequencing in bone regeneration
Received date: 2025-02-28
Accepted date: 2025-07-18
Online published: 2025-08-28
Supported by
The 18th Innovation Training Program of Shanghai Jiao Tong University School of Medicine(1824504)
骨再生是恢复骨稳态的关键,涉及多种细胞的有序协作,过程复杂,类型多样。理清各阶段分子机制以开发促骨再生新方案是该学科的发展方向。传统高通量测序常以整体转录组为研究对象,丢失细胞水平信息。单细胞RNA测序技术通过反映单个细胞中RNA表达水平,使分析亚群异质性成为可能。基于已绘制的单细胞图谱,还可根据实际需要结合特定算法模拟细胞分化轨迹,助力更深入的机制研究。借助该技术,已在不同类型的长骨、颅颌面骨再生过程中识别出关键细胞亚群,鉴定了特征性细胞标志物与潜在疾病诊断指标,对比了炎症、衰老等背景下的再生差异,探究了部分引导骨再生术的成骨机制,揭示了不同材料促骨再生差异性。该文综述了单细胞RNA测序在长骨、颅颌面骨再生及骨组织工程中的应用,总结了其在解析细胞异质性、基因调控和微环境作用方面的贡献,梳理通过测序已鉴定的核心细胞亚群与功能,指出当前研究的局限性,并展望了该技术未来在骨再生中的应用前景,有望为后续相关研究提供全新视角。
黄紫晗 , 黄心智 . 单细胞RNA测序在骨再生研究中的应用[J]. 上海交通大学学报(医学版), 2025 , 45(8) : 1053 -1058 . DOI: 10.3969/j.issn.1674-8115.2025.08.013
Bone regeneration is pivotal for restoring bone homeostasis, involving the coordinated collaboration of diverse cell types in a complex and heterogeneous process. Elucidating the molecular mechanisms at each stage to develop novel bone regeneration strategies represents a key direction in this field. Traditional high-throughput sequencing examines bulk transcriptomes, losing cellular-level resolution. Single-cell RNA sequencing (scRNA-seq) technology enables the analysis of subpopulation heterogeneity by revealing RNA expression profiles at the single-cell level. Based on single-cell atlases, researchers can further employ specific algorithms to simulate cellular differentiation trajectories, facilitating more profound mechanistic investigations. Utilizing this technology, critical cell subpopulations involved in long bone and craniofacial bone regeneration have been identified, characteristic cellular markers and potential diagnostic indicators have been defined, regenerative differences under inflammatory or aging conditions have been compared, the osteogenic mechanisms involved in guided bone regeneration procedures have been explored, and the differential bone-promoting effects of various biomaterials have been revealed. This review summarizes the applications of scRNA-seq in long bone and craniofacial bone regeneration, as well as in bone tissue engineering. It highlights its contributions in deciphering cellular heterogeneity, gene regulation, and microenvironmental interactions, consolidates key cell subpopulations and their functions identified through sequencing, and discusses current research limitations. Furthermore, it outlines future prospects for this technology in bone regeneration research, offering new perspectives for subsequent studies.
| [1] | TANG F C, BARBACIORU C, WANG Y Z, et al. mRNA-Seq whole-transcriptome analysis of a single cell[J]. Nat Methods, 2009, 6(5): 377-382. |
| [2] | WANG T, WANG L, ZHANG L P, et al. Single-cell RNA sequencing in orthopedic research[J]. Bone Res, 2023, 11(1): 10. |
| [3] | GU Y Y, HU Y, ZHANG H, et al. Single-cell RNA sequencing in osteoarthritis[J]. Cell Prolif, 2023, 56(12): e13517. |
| [4] | SAUL D, KHOSLA S. Fracture healing in the setting of endocrine diseases, aging, and cellular senescence[J]. Endocr Rev, 2022, 43(6): 984-1002. |
| [5] | NICHOLSON J A, MAKARAM N, SIMPSON A, et al. Fracture nonunion in long bones: a literature review of risk factors and surgical management[J]. Injury, 2021, 52(Suppl 2): S3-S11. |
| [6] | ZHANG H, WANG R, WANG G, et al. Single-cell RNA sequencing reveals B cells are important regulators in fracture healing[J]. Front Endocrinol, 2021, 12: 666140. |
| [7] | LU Y N, LUO Y, ZHANG Q, et al. Decoding the immune landscape following hip fracture in elderly patients: unveiling temporal dynamics through single-cell RNA sequencing[J]. Immun Ageing, 2023, 20(1): 54. |
| [8] | AVIN K G, DOMINGUEZ J M 2nd, CHEN N X, et al. Single-cell RNAseq provides insight into altered immune cell populations in human fracture nonunions[J]. J Orthop Res, 2023, 41(5): 1060-1069. |
| [9] | TANG W, LI Z W, MIAO G Q, et al. Single-cell RNA sequencing reveals transcriptional changes in the cartilage of subchondral insufficiency fracture of the knee[J]. J Inflamm Res, 2022, 15: 6105-6112. |
| [10] | YAO L, LU J, ZHONG L, et al. Activin A marks a novel progenitor cell population during fracture healing and reveals a therapeutic strategy[J]. eLife, 2023, 12: e89822. |
| [11] | HAO R C, LI Z L, WANG F Y, et al. Single-cell transcriptomic analysis identifies a highly replicating Cd168+ skeletal stem/progenitor cell population in mouse long bones[J]. J Genet Genom, 2023, 50(9): 702-712. |
| [12] | LIU R, JIAO Y R, HUANG M, et al. Mechanosensitive protein polycystin-1 promotes periosteal stem/progenitor cells osteochondral differentiation in fracture healing[J]. Theranostics, 2024, 14(6): 2544-2559. |
| [13] | JULIEN A, KANAGALINGAM A, MARTíNEZ-SARRà E, et al. Direct contribution of skeletal muscle mesenchymal progenitors to bone repair[J]. Nat Commun, 2021, 12(1): 2860. |
| [14] | XIAO D, FANG L, LIU Z T, et al. DNA methylation-mediated Rbpjk suppression protects against fracture nonunion caused by systemic inflammation[J]. J Clin Invest, 2023, 134(3): e168558. |
| [15] | LIU J T, LIN X, MCDAVID A, et al. Molecular signatures distinguish senescent cells from inflammatory cells in aged mouse callus stromal cells[J]. Front Endocrinol (Lausanne), 2023, 14: 1090049. |
| [16] | GALEA G L, ZEIN M R, ALLEN S, et al. Making and shaping endochondral and intramembranous bones[J]. Dev Dyn, 2021, 250(3): 414-449. |
| [17] | LEITCH V D, DUNCAN BASSETT J H, WILLIAMS G R. Role of thyroid hormones in craniofacial development[J]. Nat Rev Endocrinol, 2020, 16(3): 147-164. |
| [18] | LU D Z, ZHANG Y F, LIANG S M, et al. M2 macrophages guide periosteal stromal cell recruitment and initiate bone injury regeneration[J]. Biomedicines, 2024, 12(6): 1205. |
| [19] | NAKAYAMA M, OKADA H, SEKI M, et al. Single-cell RNA sequencing unravels heterogeneity of skeletal progenitors and cell-cell interactions underlying the bone repair process[J]. Regen Ther, 2022, 21: 9-18. |
| [20] | WANG Y Y, QIN Q Z, WANG Z Y, et al. The Mohawk homeobox gene represents a marker and osteo-inhibitory factor in calvarial suture osteoprogenitor cells[J]. Cell Death Dis, 2024, 15(6): 420. |
| [21] | WINTERS R, TATUM S A. Craniofacial distraction osteogenesis[J]. Facial Plast Surg Clin North Am, 2014, 22(4): 653-664. |
| [22] | JIANG W D, ZHU P Q, ZHANG T, et al. PRRX1+MSCs enhance mandibular regeneration during distraction osteogenesis[J]. J Dent Res, 2023, 102(9): 1058-1068. |
| [23] | TEVLIN R, GRIFFIN M, CHEN K, et al. Denervation during mandibular distraction osteogenesis results in impaired bone formation[J]. Sci Rep, 2023, 13(1): 2097. |
| [24] | WENG Y T, WANG H C, WU D, et al. A novel lineage of osteoprogenitor cells with dual epithelial and mesenchymal properties govern maxillofacial bone homeostasis and regeneration after MSFL[J]. Cell Res, 2022, 32(9): 814-830. |
| [25] | ZHOU B O, YUE R, MURPHY M M, et al. Leptin-receptor-expressing mesenchymal stromal cells represent the main source of bone formed by adult bone marrow[J]. Cell Stem Cell, 2014, 15(2): 154-168. |
| [26] | MIZOGUCHI T, PINHO S, AHMED J, et al. Osterix marks distinct waves of primitive and definitive stromal progenitors during bone marrow development[J]. Dev Cell, 2014, 29(3): 340-349. |
| [27] | SHU H S, LIU Y L, TANG X T, et al. Tracing the skeletal progenitor transition during postnatal bone formation[J]. Cell Stem Cell, 2021, 28(12): 2122-2136.e3. |
| [28] | DING Y, MO C, GENG J, et al. Identification of periosteal osteogenic progenitors in jawbone[J]. J Dent Res, 2022, 101(9): 1101-1109. |
| [29] | JIN A, XU H, GAO X, et al. ScRNA-seq reveals a distinct osteogenic progenitor of alveolar bone[J]. J Dent Res, 2023, 102(6): 645-655. |
| [30] | WAN Z Q, BAI X Q, WANG X, et al. Mgp high-expressing MSCs orchestrate the osteoimmune microenvironment of collagen/nanohydroxyapatite-mediated bone regeneration[J]. Adv Sci (Weinh), 2024, 11(23): e2308986. |
| [31] | PAN H, WEI Y X, ZENG C J, et al. Hierarchically assembled nanofiber scaffold guides long bone regeneration by promoting osteogenic/chondrogenic differentiation of endogenous mesenchymal stem cells[J]. Small, 2024, 20(26): e2309868. |
| [32] | WANG X Q, MA C Q, ZHANG X C, et al. Mussel inspired 3D elastomer enabled rapid calvarial bone regeneration through recruiting more osteoprogenitors from the Dura mater[J]. Regen Biomater, 2024, 11: rbae059. |
| [33] | GUO P, LIU X Z, ZHANG P H, et al. A single-cell transcriptome of mesenchymal stromal cells to fabricate bioactive hydroxyapatite materials for bone regeneration[J]. Bioact Mater, 2021, 9: 281-298. |
| [34] | HE Z H, LI H, ZHANG Y Y, et al. Enhanced bone regeneration via endochondral ossification using Exendin-4-modified mesenchymal stem cells[J]. Bioact Mater, 2023, 34: 98-111. |
| [35] | WEI J, BAPTISTA-HON D T, WANG Z, et al. Bioengineered human tissue regeneration and repair using endogenous stem cells[J]. Cell Rep Med, 2023, 4(8): 101156. |
| [36] | SUN J, HU L L, BOK S, et al. A vertebral skeletal stem cell lineage driving metastasis[J]. Nature, 2023, 621(7979): 602-609. |
/
| 〈 |
|
〉 |