Dental-tissue-derived stem cells have been used for tissue engineering owing to their ease of isolation and efficacy in in vitro and in vivo proliferation and differentiation. Nanohydroxyapatite/chitosan/gelatin (nHA/CG) three-dimensional porous scaffolds are promising for bone tissue engineering, especially jaw bone regeneration, because of their structural and functional similarity to natural bone. In our previous study, the efficiency of scaffolds with stem cell complexes in osteogenesis was confirmed in vivo in immunocompromised mice. However, studies on the bone regeneration efficiency of stem cell-seeded nHA/CG scaffolds using large animal jaw bone defect models have not been conducted. This study evaluated the bone regeneration potential of the nHA/CG scaffolds with transplanted human periodontal ligament stem cells (hPDLSCs) in critical-sized jaw bone defects in minipigs. The hPDLSCs isolated from periodontal ligaments of discarded teeth (postorthodontic purposes) were seeded onto the nHA/CG scaffolds. The scaffold was successfully synthesized according to our previous studies. Forty-eight critical-sized jaw bone defects were created in 12 minipigs. The defects were randomly assigned to one of three groups [scaffolds with seeded hPDLSCs (hPDLSCs/nHA/CG), only scaffold (nHA/CG), and a negative control group, ie, no cells and scaffolds implanted into defects] to investigate jaw bone regeneration. The bone regeneration capacities of the three groups were assessed for up to 12 weeks. The results showed that the hPDLSCs adhered well to the nHA/CG scaffold in vitro, and the cell-nHA/CG composites significantly increased new bone formation and generated large bones with normal architectures and vascularization in vivo compared to the nHA/CG and control groups. Immunohistochemistry staining showed that runt-related transcription factor 2 (Runx2) was highly expressed in the bone marrow formed in the hPDLSCs/nHA/CG group. This study provides strong evidence for future clinical applications of the nHA/CG scaffolds transplanted with hPDLSCs to regenerate the bone in large jaw bone defects.

中文翻译：

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移植纳米羟基磷灰石/壳聚糖/明胶3D多孔支架的人牙周膜干细胞促进猪颌骨再生

牙科组织来源的干细胞因其易于分离和在体外和体内增殖和分化中的功效而被用于组织工程。纳米羟基磷灰石/壳聚糖/明胶（nHA/CG）三维多孔支架由于其结构和功能与天然骨相似，因此有望用于骨组织工程，尤其是颌骨再生。在我们之前的研究中，在免疫功能低下的小鼠体内证实了具有干细胞复合物的支架在成骨中的效率。然而，尚未使用大型动物颌骨缺损模型对接种干细胞的 nHA/CG 支架的骨再生效率进行研究。本研究评估了移植人牙周膜干细胞 (hPDLSC) 的 nHA/CG 支架在小型猪的临界尺寸颌骨缺损中的骨再生潜力。从废弃牙齿（正畸目的）的牙周韧带中分离的 hPDLSCs 被接种到 nHA/CG 支架上。根据我们之前的研究成功合成了支架。在 12 头小型猪中产生了 48 个临界尺寸的颌骨缺损。将缺损随机分配到三组 [接种 hPDLSCs 的支架 (hPDLSCs/nHA/CG)、仅支架 (nHA/CG) 和阴性对照组中的一组，即没有细胞和支架植入缺损] 以研究下颌骨再生。对三组的骨再生能力进行了长达 12 周的评估。结果表明，hPDLSCs 在体外很好地粘附到 nHA/CG 支架上，与 nHA/CG 和对照组相比，细胞-nHA/CG 复合物显着增加了新骨形成并在体内生成了具有正常结构和血管化的大骨. 免疫组化染色显示runt相关转录因子2（Runx2）在hPDLSCs/nHA/CG组形成的骨髓中高表达。这项研究为移植 hPDLSCs 的 nHA/CG 支架的未来临床应用提供了强有力的证据，以在大颌骨缺损中再生骨骼。免疫组化染色显示runt相关转录因子2（Runx2）在hPDLSCs/nHA/CG组形成的骨髓中高表达。这项研究为移植 hPDLSCs 的 nHA/CG 支架的未来临床应用提供了强有力的证据，以在大颌骨缺损中再生骨骼。免疫组化染色显示runt相关转录因子2（Runx2）在hPDLSCs/nHA/CG组形成的骨髓中高表达。这项研究为移植 hPDLSCs 的 nHA/CG 支架的未来临床应用提供了强有力的证据，以在大颌骨缺损中再生骨骼。