| 摘要: | 多項研究表明，瓊脂糖由於具有良好的理化特性、生物相容性和熱可逆凝膠化行為，已被用作組織工程的生物材料。 然而很少有研究關注瓊脂糖與其他物質混合物的特性。本研究的目的是評估來自羧甲基纖維素 (CMC)-瓊脂糖混合物的三維 (3D) 多孔支架在生物體內骨再生的組織反應。羧甲基纖維素（CMC）/瓊脂糖多孔支架的生化特性、生物相容性和成骨分化潛能已在我們之前的研究中得到證實和論證。 然而，它們在生物體內的生物相容性、長期降解和生物學表現尚未得到研究和證實
此外，本實驗還評估了hDPSCs 細胞球體和 CMC/瓊脂糖支架組合在顱骨骨缺損模型中的骨癒合能力。製備樣本（Agarose 25/ CMC 75， Agarose 50/ CMC 50， Agarose 75/ CMC 25）並進行材料特徵分析。 再以 micro-CT、組織學、組織形態學分析評估 CMC/ Agarose支架在加入3D細胞球體後的體內骨形成或骨誘導成效。 體內實驗表明各組之間具有良好的血液相容性和生物相容性。在動物實驗中， CMC/Agarose 在配合加入 3D 細胞球體後的骨癒合促進效果，在8 週癒合後，micro-CT 結果顯示，相對於 CMC/Agarose (75/25)、CMC/Agarose (50/50)、CMC/Agarose (25/75)加入 3D 細胞球體，CMC/Agarose (75/25)與 CMC/Agarose (50/50)配合加入 3D 細胞球體後，有最佳的BV/TV (%)數值而且有統計學上的差異(p< 0.05)，此外，在 CMC/Agarose (75/25)與 CMC/Agarose (50/50)配合加入 3D 細胞球體的組別中，組織切片分析(H&E 與 Masson’s和IHC)進一步證實有更多的新生骨形成、更成熟的骨髓結構形成、更多生物材料的降解、更多新生血管、更多的骨細胞與更多的成骨細胞圍繞在生物材料周圍，這均證實CMC/Agarose (50/50)在加入 3D 細胞球體後，能夠誘導骨癒合以及血管生成。 總體而言，我們的研究結果證實， CMC/Agarose(50/50)結合 3D 細胞球體具備良好生物相容性和骨癒合能力，未來相當有潛能應用於骨組織工程應用以及成為骨替代生物材料。
Multiple studies have indicated that agarose, due to its favorable physicochemical properties, biocompatibility, and thermoreversible gelation behavior, has been used as a biomaterial in tissue engineering. However, there has been limited research on the characteristics of agarose when mixed with other substances.The purpose of this study is to evaluate the tissue response of three-dimensional (3D) porous scaffolds made from a mixture of carboxymethyl cellulose (CMC) and agarose in bone regeneration within a biological organism. The biochemical properties, biocompatibility, and osteogenic differentiation potential of CMC/agarose porous scaffolds have been previously validated in our research. However, their in vivo biocompatibility, long-term degradation, and biological performance have not been thoroughly studied and confirmed.
Additionally, the study assesses the bone-healing capacity of hDPSCs cell spheres combined with CMC/agarose scaffolds in a cranial bone defect model. Samples (Agarose 25/CMC 75, Agarose 50/CMC 50, Agarose 75/CMC 25) were prepared, and material characteristic analyses were conducted. Subsequently, micro-CT, histology, tissue morphology, and immunofluorescence staining (IHC) were employed to evaluate the in vivo bone formation or osteoinductive effectiveness of CMC/agarose scaffolds after the addition of 3D cell spheres.
In vivo experiments indicated good blood compatibility and biocompatibility among the groups. In the animal experiments, the CMC/agarose scaffolds, when combined with the addition of 3D cell spheres, exhibited enhanced bone healing promotion. After 8 weeks of healing, micro-CT results demonstrated that, compared to CMC/agarose (75/25), CMC/agarose (50/50), and CMC/agarose (25/75) with added 3D cell spheres, CMC/agarose (75/25) and CMC/agarose (50/50) had the optimal BV/TV (%) values with statistical significance (p < 0.05). Furthermore, in the groups where CMC/agarose (75/25) and CMC/agarose (50/50) were combined with 3D cell spheres, histological slice analysis (H&E and Masson) further confirmed more new bone formation, mature bone marrow structure, greater material degradation, increased neovascularization, more osteoblasts, and more osteocytes surrounding the biomaterial. These findings collectively support that CMC/agarose (50/50), when combined with 3D cell spheres, can induce bone healing and vascular regeneration.
In conclusion, our research results confirm that the combination of CMC/agarose (50/50) with 3D cell spheres possesses excellent biocompatibility and bone healing capabilities. They hold significant potential for applications in bone tissue engineering and as substitutes for bone biomaterials in the future. |
