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用于骨修复的纳米结构碳酸羟基磷灰石球体的细胞相容性和生物相容性

Cytocompatibility and biocompatibility of nanostructured carbonated hydroxyapatite spheres for bone repair.

作者信息

Calasans-Maia Mônica Diuana, Melo Bruno Raposo de, Alves Adriana Terezinha Neves Novellino, Resende Rodrigo Figueiredo de Brito, Louro Rafael Seabra, Sartoretto Suelen Cristina, Granjeiro José Mauro, Alves Gutemberg Gomes

机构信息

Departamento de Cirurgia Oral, Faculdade de Odontologia, Universidade Federal Fluminense, Niteroi, RJ, Brazil.

Faculdade de Odontologia, Universidade Federal Fluminense, Niteroi, RJ, Brazil.

出版信息

J Appl Oral Sci. 2015 Nov-Dec;23(6):599-608. doi: 10.1590/1678-775720150122.

Abstract

OBJECTIVE

The aim of this study was to investigate the in vitro and in vivo biological responses to nanostructured carbonated hydroxyapatite/calcium alginate (CHA) microspheres used for alveolar bone repair, compared to sintered hydroxyapatite (HA).

MATERIAL AND METHODS

The maxillary central incisors of 45 Wistar rats were extracted, and the dental sockets were filled with HA, CHA, and blood clot (control group) (n=5/period/group). After 7, 21 and 42 days, the samples of bone with the biomaterials were obtained for histological and histomorphometric analysis, and the plasma levels of RANKL and OPG were determined via immunoassay. Statistical analysis was performed by Two-Way ANOVA with post-hoc Tukey test at 95% level of significance.

RESULTS

The CHA and HA microspheres were cytocompatible with both human and murine cells on an in vitro assay. Histological analysis showed the time-dependent increase of newly formed bone in control group characterized by an intense osteoblast activity. In HA and CHA groups, the presence of a slight granulation reaction around the spheres was observed after seven days, which was reduced by the 42nd day. A considerable amount of newly formed bone was observed surrounding the CHA spheres and the biomaterials particles at 42-day time point compared with HA. Histomorphometric analysis showed a significant increase of newly formed bone in CHA group compared with HA after 21 and 42 days from surgery, moreover, CHA showed almost 2-fold greater biosorption than HA at 42 days (two-way ANOVA, p<0.05) indicating greater biosorption. An increase in the RANKL/OPG ratio was observed in the CHA group on the 7th day.

CONCLUSION

CHA spheres were osteoconductive and presented earlier biosorption, inducing early increases in the levels of proteins involved in resorption.

摘要

目的

本研究旨在调查与烧结羟基磷灰石(HA)相比,用于牙槽骨修复的纳米结构碳酸羟基磷灰石/海藻酸钙(CHA)微球的体外和体内生物学反应。

材料与方法

拔除45只Wistar大鼠的上颌中切牙,拔牙窝分别填充HA、CHA和血凝块(对照组)(每组/时期n = 5)。在7、21和42天后,获取含有生物材料的骨样本进行组织学和组织形态计量学分析,并通过免疫测定法测定血浆中RANKL和OPG的水平。采用双向方差分析和事后Tukey检验进行统计学分析,显著性水平为95%。

结果

在体外试验中,CHA和HA微球与人细胞和鼠细胞均具有细胞相容性。组织学分析显示,对照组新形成骨随时间增加,其特征为成骨细胞活性增强。在HA和CHA组中,7天后观察到微球周围有轻微的肉芽反应,到第42天时减少。与HA相比,在42天时间点观察到CHA微球和生物材料颗粒周围有大量新形成的骨。组织形态计量学分析显示,术后21天和42天,CHA组新形成骨比HA组显著增加,此外,在42天时CHA的生物吸附几乎是HA的2倍(双向方差分析,p<0.05),表明生物吸附性更强。在第7天,CHA组的RANKL/OPG比值增加。

结论

CHA微球具有骨传导性,且呈现出更早的生物吸附性,可诱导参与吸收的蛋白质水平早期升高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f5a/4716697/f50ba328d9f5/1678-7757-jaos-23-6-0599-gf01.jpg

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