单细胞RNA测序在骨再生研究中的应用

doi:10.3969/j.issn.1674-8115.2025.08.013

摘要/Abstract

摘要：

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。

关键词: 单细胞RNA测序, 骨再生, 颅颌面骨, 骨组织工程

Abstract:

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.

Key words: single-cell RNA sequencing, bone regeneration, craniofacial bones, bone tissue engineering

中图分类号:

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.

HUANG Zihan, HUANG Xinzhi. Application of single-cell RNA sequencing in bone regeneration[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2025, 45(8): 1053-1058.

图/表 1

参考文献 36

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Cell type	Key subpopulation	Location	Species	Feature	Reference
Immune cell	B cell	Hip callus	Human	Promote early-stage osteogenesis while inhibiting excessive bone formation in later stages	^[6]
	CD14⁺ dentritic cell	Hip callus	Human	Potential biomarkers of bone nonunion	^[8]
	M2 macrophage	Cranial periosteum	Swine	Secrete neuregulin-1 to recruit periosteal stromal cells	^[18]
Skeletal stem/progenitor cell	Inhba⁺progenitor cell	Femoral callus	Mouse	Promote SMAD2 phosphorylation, enhance progenitor cell proliferation and differentiation	^[10]
	CD168⁺skeletal stem cell	Femoral callus	Mouse	Osteogenic and chondrogenic differentiation	^[11]
	Ctsk⁺periosteal progenitor cell	Femoral callus	Mouse	Mechanosentive; chondrogenic differentiation	^[12]
	Sox9⁺progenitor cell	Cranial periosteum	Mouse	Osteogenic differentiation	^[19]
	Mkx⁺cell	Cranial suture	Mouse	Mechanosentive; osteogenic differentiation	^[20]
	Prrx1⁺cell	Mandibular callus	Canine	Mechanosentive; osteogenic differentiation	^[22]
	Krt14⁺Ctsk⁺cell	Maxillary sinus mucosa	Mouse	Co-expression of epithelial and mesenchymal characteristics	^[24]
	Ctsk⁺Ly6a⁺cell	Mandibular of bone marrow	Mouse	Osteogenic differentiation	^[28]
	Fat4⁺cell	Alveolar bone	Mouse	Osteogenic differentiation	^[29]
Skeletal muscle stem cell	Prrx1⁺Skeletal muscle stem cell	Femoral callus	Mouse	Promote fibrous tissue aggregation, leading to nonunion	^[13]

Cell type	Key subpopulation	Location	Species	Feature	Reference
Immune cell	B cell	Hip callus	Human	Promote early-stage osteogenesis while inhibiting excessive bone formation in later stages	^[6]
	CD14⁺ dentritic cell	Hip callus	Human	Potential biomarkers of bone nonunion	^[8]
	M2 macrophage	Cranial periosteum	Swine	Secrete neuregulin-1 to recruit periosteal stromal cells	^[18]
Skeletal stem/progenitor cell	Inhba⁺progenitor cell	Femoral callus	Mouse	Promote SMAD2 phosphorylation, enhance progenitor cell proliferation and differentiation	^[10]
	CD168⁺skeletal stem cell	Femoral callus	Mouse	Osteogenic and chondrogenic differentiation	^[11]
	Ctsk⁺periosteal progenitor cell	Femoral callus	Mouse	Mechanosentive; chondrogenic differentiation	^[12]
	Sox9⁺progenitor cell	Cranial periosteum	Mouse	Osteogenic differentiation	^[19]
	Mkx⁺cell	Cranial suture	Mouse	Mechanosentive; osteogenic differentiation	^[20]
	Prrx1⁺cell	Mandibular callus	Canine	Mechanosentive; osteogenic differentiation	^[22]
	Krt14⁺Ctsk⁺cell	Maxillary sinus mucosa	Mouse	Co-expression of epithelial and mesenchymal characteristics	^[24]
	Ctsk⁺Ly6a⁺cell	Mandibular of bone marrow	Mouse	Osteogenic differentiation	^[28]
	Fat4⁺cell	Alveolar bone	Mouse	Osteogenic differentiation	^[29]
Skeletal muscle stem cell	Prrx1⁺Skeletal muscle stem cell	Femoral callus	Mouse	Promote fibrous tissue aggregation, leading to nonunion	^[13]