Department of Family Oral Health, Faculty of Dentistry, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia.
PeerJ. 2022 May 2;10:e13356. doi: 10.7717/peerj.13356. eCollection 2022.
The selection of appropriate scaffold plays an important role in ensuring the success of bone regeneration. The use of scaffolds with different materials and their effect on the osteogenic performance of cells is not well studied and this can affect the selection of suitable scaffolds for transplantation. Hence, this study aimed to investigate the comparative ability of two different synthetic scaffolds, mainly hydroxyapatite (HA) and polycaprolactone (PCL) scaffolds in promoting and bone regeneration.
cell viability, morphology, and alkaline phosphatase (ALP) activity of MC3T3-E1 cells on HA and PCL scaffolds were determined in comparison to the accepted model outlined for two-dimensional systems. An study involving the transplantation of MC3T3-E1 cells with scaffolds into an artificial bone defect of 4 mm length and 1.5 mm depth in the rat's left maxilla was conducted. Three-dimensional analysis using micro-computed tomography (micro-CT), hematoxylin and eosin (H&E), and immunohistochemistry analyses evaluation were performed after six weeks of transplantation.
MC3T3-E1 cells on the HA scaffold showed the highest cell viability. The cell viability on both scaffolds decreased after 14 days of culture, which reflects the dominant occurrence of osteoblast differentiation. An early sign of osteoblast differentiation can be detected on the PCL scaffold. However, cells on the HA scaffold showed more prominent results with intense mineralized nodules and significantly ( < 0.05) high levels of ALP activity with prolonged osteoblast induction. Micro-CT and H&E analyses confirmed the results with bone formation were significantly ( < 0.05) greater in HA scaffold and was supported by IHC analysis which confirmed stronger expression of osteogenic markers ALP and osteocalcin.
Different scaffold materials of HA and PCL might have influenced the bone regeneration ability of MC3T3-E1. Regardless, and bone regeneration was better in the HA scaffold which indicates its great potential for application in bone regeneration.
选择合适的支架对于确保骨再生的成功至关重要。使用不同材料的支架及其对细胞成骨性能的影响尚未得到充分研究,这可能会影响适合移植的支架的选择。因此,本研究旨在比较两种不同的合成支架(主要是羟基磷灰石(HA)和聚己内酯(PCL)支架)在促进骨再生方面的能力。
通过比较二维系统概述的公认模型,确定 MC3T3-E1 细胞在 HA 和 PCL 支架上的细胞活力、形态和碱性磷酸酶(ALP)活性。在大鼠左侧上颌骨 4 毫米长、1.5 毫米深的人工骨缺损中进行了将 MC3T3-E1 细胞与支架一起移植的动物实验。在移植 6 周后,通过微计算机断层扫描(micro-CT)、苏木精和伊红(H&E)和免疫组织化学分析进行三维分析。
HA 支架上的 MC3T3-E1 细胞显示出最高的细胞活力。培养 14 天后,两种支架上的细胞活力均下降,这反映了成骨细胞分化的主导发生。在 PCL 支架上可以检测到成骨细胞分化的早期迹象。然而,HA 支架上的细胞表现出更明显的结果,有强烈的矿化结节,并且随着成骨细胞诱导时间的延长,ALP 活性显著升高(<0.05)。micro-CT 和 H&E 分析证实了骨形成的结果,HA 支架上的骨形成明显(<0.05)更多,并通过免疫组织化学分析得到证实,该分析证实了更强的成骨标志物 ALP 和骨钙素的表达。
HA 和 PCL 等不同的支架材料可能影响了 MC3T3-E1 的骨再生能力。无论如何,HA 支架在和骨再生方面表现更好,这表明其在骨再生中的应用潜力巨大。