Subcutaneous implantation of tooth germ stem cells over the masseter muscle in mice: An in vivo pilot study

IF 3.4 3区环境科学与生态学 Q3 CELL & TISSUE ENGINEERING

Regenerative Therapy Pub Date : 2025-02-11 DOI:10.1016/j.reth.2025.01.021

Nosrat Nourbakhsh , Ghazaleh Baniebrahimi , Sahar Talebi , Ardeshir Talebi , Mohammad Hossein Nasr Esfahani , Bijan Movahedian , Maziar Manshayi , Navid Naghdi , Fatemeh Ejeian , Elahe Masaeli , Seyed Ali Mosaddad

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引用次数: 0

Abstract

Objectives

This study aimed to explore the potential of tooth germ stem cells for regenerating tooth-like structures by subcutaneously implanting first molar tooth germ stem cells over the masseter muscle in mice.

Methods

Five pairs of house mice, Mus musculus, were selected for mating. At gestational day 14 (E14), the fetuses were extracted, and the first molar tooth germ at the cap stage was isolated. Tooth germ stem cells were prepared into a suspension and seeded onto scaffolds, which were then implanted subcutaneously over the masseter muscle in male mice. The control group (n = 5 male mice) received acellular scaffolds implanted at the same site. After 20 days, the regenerated tissues were resected and analyzed histologically using hematoxylin and eosin (H & E) staining, Masson's trichrome staining, and immunohistochemical (IHC) staining for cytokeratin (CK) and vimentin markers.

Results

H & E staining showed the formation of integrated oval structures at the implant site in all samples. Masson's trichrome staining identified dispersed accumulations of cellular mineralized matrix within the connective tissue. IHC staining was positive for vimentin, confirming the mesenchymal origin of the loose tissue at the center, indicating future dental pulp development. Positive CK staining indicated the ectodermal origin of dense peripheral tissues, suggesting the future formation of inner enamel epithelium. The combined immunohistochemical results for vimentin and CK confirmed the ecto-mesenchymal origin of the regenerated tissue, which resembled a late bell-stage tooth germ observed around gestational days 17.5–18 and showed early indications of dentin formation (D0).

Conclusion

The study indicates that tooth germ stem cells may have the potential to produce dense, tooth-like structures when implanted subcutaneously in mice. These findings provide preliminary insights into the possible applications of tooth germ stem cells in regenerative dental tissue engineering.

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牙胚干细胞在小鼠咬肌上的皮下植入：一项体内初步研究

目的通过在小鼠咬肌上皮下植入第一磨牙胚干细胞，探讨牙胚干细胞再生牙样结构的潜力。方法选择家鼠小家鼠5对进行交配。妊娠第14天（E14）取出胎体，分离帽期第一磨牙胚。研究人员将牙胚干细胞制备成悬浮液，并将其植入支架上，然后将支架植入雄性小鼠的咬肌皮下。对照组（雄性小鼠5只）在同一部位植入脱细胞支架。20d后，切除再生组织，用苏木精和伊红(H &；细胞角蛋白（CK）和波形蛋白标记物的染色、马松三色染色和免疫组化（IHC）染色。ResultsH,E染色显示植入部位形成完整的椭圆形结构。马松三色染色鉴定结缔组织内分散堆积的细胞矿化基质。免疫组化染色显示vimentin阳性，证实了中间松散组织的间质起源，预示着未来牙髓的发育。CK染色阳性提示致密外周组织起源于外胚层，提示将来会形成内釉质上皮。vimentin和CK的联合免疫组化结果证实了再生组织的外间质起源，类似于妊娠17.5-18天左右观察到的钟状晚期牙胚，并显示出牙本质形成的早期迹象（D0）。结论牙胚干细胞在小鼠体内皮下移植后，有可能产生致密的牙样结构。这些发现为牙胚干细胞在再生牙组织工程中的应用提供了初步的见解。

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来源期刊

Regenerative Therapy Engineering-Biomedical Engineering

CiteScore

6.00

自引率

2.30%

发文量

106

审稿时长

49 days

期刊介绍： Regenerative Therapy is the official peer-reviewed online journal of the Japanese Society for Regenerative Medicine. Regenerative Therapy is a multidisciplinary journal that publishes original articles and reviews of basic research, clinical translation, industrial development, and regulatory issues focusing on stem cell biology, tissue engineering, and regenerative medicine.