CA1 induced dental follicle stem cells co-culture with dental pulp stem cells and loaded three-dimensional printed PCL/β-TCP scaffold: a novel strategy for alveolar cleft bone regeneration

IF 4.2 3区 医学 Q2 ENGINEERING, BIOMEDICAL
Hao-Ran Zhao, Jin-Ze Zhao, Ning Zhao, Ling-Fa Xue, Yao-Xiang Xu, Jin Yue, Wen-Lin Xiao
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引用次数: 0

Abstract

Background

Bone tissue engineering for alveolar clefts is in the early stages of development, and more research is needed to determine the optimal cell types, growth factors and delivery methods for the therapy.

Methods

We co-cultured Carbonic anhydrase 1 (CA1) induced dental follicle stem cells (DFSCs) with dental pulp stem cells (DPSCs). In vitro, the Lentivirus vector overexpressing CA1 (LV-CA1) gene was constructed, transfected into DFSCs, and co-cultured with DPSCs indirectly. Osteoblast biomarkers in differentiated DFSCs were detected using quantitative real-time polymerase chain reaction and Western blotting. In vivo, establish a rat alveolar cleft model, transplanted stem cell Polycaprolactone/β-tricalcium phosphate (PCL/β-TCP) three-dimensional printed composite scaffold and samples were collected at 4 and 8 weeks postoperatively. The osteogenic effect was evaluated through micro computed tomography and histomorphometric analysis.

Results

In vitro, the activity of DFSCs in the LV-CA1+Co-culture group was increased, and the mRNA and protein expressions of CA1, Alkaline phosphatase (ALP), Bone morphogenetic proteins 2 (BMP2), and Runt-related transcription factor 2 (RUNX2) were amplified to varying degrees (P < 0.05). In vivo, micro-CT displayed at 4 and 8 weeks postoperatively, the LV-CA1+Co-culture group had a considerably higher percentage of new bone development (39.1% and 56.9%) (P < 0.05) than the other two groups. Histomorphometric analysis displayed the LV-CA1+Co-culture group had more newly formed bone trabeculae and immature collagen.

Conclusion

A strategy based on a novel osteogenic gene CA1 and dental-derived mesenchymal stem cells co-culture is applied to the alveolar cleft, providing a novel idea for the application of bone tissue engineering in alveolar cleft bone grafting.

Graphical Abstract

CA1诱导牙滤泡干细胞与牙髓干细胞共培养及负载三维打印PCL/β-TCP支架:牙槽裂骨再生的新策略
骨组织工程治疗肺泡裂尚处于早期发展阶段,需要更多的研究来确定治疗的最佳细胞类型、生长因子和递送方式。方法采用碳酸酐酶1 (CA1)诱导的牙滤泡干细胞(DFSCs)与牙髓干细胞(DPSCs)共培养。体外构建过表达CA1 (LV-CA1)基因的慢病毒载体,转染到DFSCs中,与DPSCs间接共培养。采用实时定量聚合酶链反应和Western blotting检测分化的DFSCs成骨细胞生物标志物。在体内,建立大鼠牙槽裂模型,移植干细胞聚己内酯/β-磷酸三钙(PCL/β-TCP)三维打印复合支架,并于术后4周和8周采集标本。通过显微计算机断层扫描和组织形态学分析评估成骨效果。结果在体外,LV-CA1+共培养组DFSCs活性升高,CA1、碱性磷酸酶(ALP)、骨形态发生蛋白2 (BMP2)、runt相关转录因子2 (RUNX2) mRNA和蛋白表达均不同程度扩增(P < 0.05)。在体内,术后4周和8周的micro-CT显示,LV-CA1+共培养组的新生骨成形率(39.1%和56.9%)明显高于其他两组(P < 0.05)。组织形态学分析显示LV-CA1+共培养组新生骨小梁和未成熟胶原较多。结论基于新型成骨基因CA1与牙源性间充质干细胞共培养的策略可应用于牙槽骨裂的修复,为骨组织工程技术在牙槽骨裂骨移植中的应用提供了新的思路。图形抽象
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来源期刊
Journal of Materials Science: Materials in Medicine
Journal of Materials Science: Materials in Medicine 工程技术-材料科学:生物材料
CiteScore
8.00
自引率
0.00%
发文量
73
审稿时长
3.5 months
期刊介绍: The Journal of Materials Science: Materials in Medicine publishes refereed papers providing significant progress in the application of biomaterials and tissue engineering constructs as medical or dental implants, prostheses and devices. Coverage spans a wide range of topics from basic science to clinical applications, around the theme of materials in medicine and dentistry. The central element is the development of synthetic and natural materials used in orthopaedic, maxillofacial, cardiovascular, neurological, ophthalmic and dental applications. Special biomedical topics include biomaterial synthesis and characterisation, biocompatibility studies, nanomedicine, tissue engineering constructs and cell substrates, regenerative medicine, computer modelling and other advanced experimental methodologies.
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