Furlani Franco, Malfatti Matilde Clarissa, Rondinella Alfredo, Campodoni Elisabetta, Sandri Monica, Fedrizzi Lorenzo, Tell Gianluca
Department of Medicine, University of Udine, Piazzale Kolbe 4, Udine, 33100, Italy.
Polytechnic Department of Engineering and Architecture, University of Udine, Via delle Scienze, 206, Udine, 33100, Italy.
J Biol Eng. 2024 Oct 25;18(1):60. doi: 10.1186/s13036-024-00458-9.
Hydroxyapatite (HA) is a bioceramic material widely used as a bone biomimetic substitute and can be synthesized by biomineralization, according to which HA nanoparticles are formed on a polymer template. Nevertheless, little is known about the effect of ion doping and biomineralization on cell metabolism, oxidative stress, and DNA damage. In the present contribution, we report on synthesizing and characterizing biomineralized chitosan as a polymer template with HA nanoparticles doped with magnesium (MgHA) and iron ions (FeHA). The physical-chemical and morphological characterization confirmed the successful synthesis of low crystalline ions-doped HA nanoparticles on the chitosan template, whereas the biochemical activity of the resulting nanoparticles towards human osteoblasts-like cells (MG63 and HOBIT) was investigated considering their effect on cell metabolism, proliferation, colony formation, redox status, and DNA damage extent. Data obtained suggest that particles enhance cell metabolism but partially limit cell proliferation. The redox status of cells was measured suggesting a slight increase in Reactive Oxygen Species production with chitosan biomineralized with iron-doped HA, whereas no effect with magnesium-doped HA and no effect of all formulations on the oxidation level of Peroxiredoxin. On the other hand, DNA damage was investigated by COMET assay, and expression and foci γH2AX. These latter tests indicated that HA-based nanoparticles promote DNA damage which is enhanced by chitosan thus suggesting that chitosan favors the nanoparticles' internalization by cells and modulates their biological activity. The potential DNA damage should be considered - and potentially exploited for instance in anticancer treatment - when HA-based particles are used to devise biomaterials.
羟基磷灰石(HA)是一种广泛用作骨仿生替代物的生物陶瓷材料,可通过生物矿化合成,在此过程中,HA纳米颗粒在聚合物模板上形成。然而,关于离子掺杂和生物矿化对细胞代谢、氧化应激和DNA损伤的影响,人们了解甚少。在本论文中,我们报道了合成并表征以生物矿化壳聚糖为聚合物模板、掺杂镁离子(MgHA)和铁离子(FeHA)的HA纳米颗粒。物理化学和形态学表征证实了在壳聚糖模板上成功合成了低结晶度的离子掺杂HA纳米颗粒,同时研究了所得纳米颗粒对人成骨样细胞(MG63和HOBIT)的生化活性,考虑了它们对细胞代谢、增殖、集落形成、氧化还原状态和DNA损伤程度的影响。获得的数据表明,颗粒增强了细胞代谢,但部分限制了细胞增殖。测量细胞的氧化还原状态表明,用铁掺杂HA生物矿化的壳聚糖使活性氧生成略有增加,而镁掺杂HA则无此影响,且所有制剂对过氧化物酶的氧化水平均无影响。另一方面,通过彗星试验以及γH2AX的表达和病灶来研究DNA损伤。后一项测试表明,基于HA的纳米颗粒会促进DNA损伤,而壳聚糖会增强这种损伤,因此表明壳聚糖有利于纳米颗粒被细胞内化并调节其生物活性。当使用基于HA的颗粒来设计生物材料时,应考虑潜在的DNA损伤——例如在抗癌治疗中可能会加以利用。