Šebová Eva, Leal Filipa, Klusáček Rampichová Michala, Nirwan Viraj P, Fahmi Amir, Costa Pedro F, Filová Eva
Department of Tissue Engineering, Institute of Experimental Medicine, Czech Academy of Sciences, 14200 Prague, Czech Republic.
Biofabics Lda, Rua Alfredo Allen 455, 4200-135 Porto, Portugal.
Int J Mol Sci. 2025 Jun 4;26(11):5383. doi: 10.3390/ijms26115383.
This study investigates the impact of hydroxyapatite (HA) nanoparticles (NPs) on the cellular responses of poly(L-lactide-co-ε-caprolactone) (PLCL) scaffolds in bone tissue engineering applications. Three types of PLCL scaffolds were fabricated, varying in HANPs content. Saos-2 osteoblast-like cells (OBs) and THP-1-derived osteoclast-like cells (OCs) were co-cultured on the scaffolds, and cell proliferation was assessed using the MTS assay. The amount of double-stranded DNA (dsDNA) was quantified to evaluate cell proliferation. Expression levels of OBs and OCs markers were analyzed via quantitative polymerase chain reaction (qPCR) and the production of Collagen type I was visualized using confocal microscopy. Additionally, enzymatic activity of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP or ACP5) was measured to assess OB and OC function, respectively. Interestingly, despite the scaffold's structured character supporting the growth of the Saos-2 OBs and THP-1-derived OCs coculture, the incorporation of HANPs did not significantly enhance cellular responses compared to scaffolds without HANPs, except for collagen type I production. These findings suggest the need for further investigation into the potential benefits of HANPs in bone tissue engineering applications. Nevertheless, our study contributes valuable insights into optimizing biomaterial design for bone tissue regeneration, with implications for drug screening and material testing protocols.
本研究调查了羟基磷灰石(HA)纳米颗粒(NPs)对聚(L-丙交酯-共-ε-己内酯)(PLCL)支架在骨组织工程应用中细胞反应的影响。制备了三种类型的PLCL支架,其HA NPs含量不同。将Saos-2成骨样细胞(OBs)和THP-1衍生的破骨样细胞(OCs)在支架上共培养,并使用MTS法评估细胞增殖。对双链DNA(dsDNA)的量进行定量以评估细胞增殖。通过定量聚合酶链反应(qPCR)分析OBs和OCs标志物的表达水平,并使用共聚焦显微镜观察I型胶原蛋白的产生。此外,分别测量碱性磷酸酶(ALP)和抗酒石酸酸性磷酸酶(TRAP或ACP5)的酶活性以评估OB和OC的功能。有趣的是,尽管支架的结构特性支持Saos-2 OBs和THP-1衍生的OCs共培养的生长,但与不含HA NPs的支架相比,HA NPs的掺入并没有显著增强细胞反应,I型胶原蛋白产生除外。这些发现表明需要进一步研究HA NPs在骨组织工程应用中的潜在益处。然而,我们的研究为优化骨组织再生的生物材料设计提供了有价值的见解,对药物筛选和材料测试方案具有启示意义。