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富含血小板纤维蛋白 (PRF) 修饰的纳米羟基磷灰石/壳聚糖/明胶/海藻酸钠支架增加了人牙髓干细胞 (DPSC) 和由 DPSC 分化而来的成骨细胞的黏附性和活力。

Platelet-rich fibrin (PRF) modified nano-hydroxyapatite/chitosan/gelatin/alginate scaffolds increase adhesion and viability of human dental pulp stem cells (DPSC) and osteoblasts derived from DPSC.

机构信息

Dental Research Center, School of Dentistry, Pontificia Universidad Javeriana, Bogotá, Colombia.

Dental Research Center, School of Dentistry, Pontificia Universidad Javeriana, Bogotá, Colombia.

出版信息

Int J Biol Macromol. 2024 Jul;273(Pt 1):133064. doi: 10.1016/j.ijbiomac.2024.133064. Epub 2024 Jun 10.

Abstract

Bone tissue regeneration strategies have incorporated the use of natural polymers, such as hydroxyapatite (nHA), chitosan (CH), gelatin (GEL), or alginate (ALG). Additionally, platelet concentrates, such as platelet-rich fibrin (PRF) have been suggested to improve scaffold biocompatibility. This study aimed to develop scaffolds composed of nHA, GEL, and CH, with or without ALG and lyophilized PRF, to evaluate the scaffold's properties, growth factor release, and dental pulp stem cells (DPSC), and osteoblast (OB) derived from DPSC viability. Four scaffold variations were synthesized and lyophilized. Then, degradation, swelling profiles, and morphological analysis were performed. Furthermore, PDGF-BB and FGF-B growth factors release were quantified by ELISA, and cytotoxicity and cell viability were evaluated. The swelling and degradation profiles were similar in all scaffolds, with pore sizes ranging between 100 and 250 μm. FGF-B and PDGF-BB release was evidenced after 24 h of scaffold immersion in cell culture medium. DPSC and OB-DPSC viability was notably increased in PRF-supplemented scaffolds. The nHA-CH-GEL-PRF scaffold demonstrated optimal physical-biological characteristics for stimulating DPSC and OB-DPSC cell viability. These results suggest lyophilized PRF improves scaffold biocompatibility for bone tissue regeneration purposes.

摘要

骨组织再生策略已经纳入了天然聚合物的使用,如羟基磷灰石(nHA)、壳聚糖(CH)、明胶(GEL)或藻酸盐(ALG)。此外,血小板浓缩物,如富含血小板的纤维蛋白(PRF),被建议改善支架的生物相容性。本研究旨在开发由 nHA、GEL 和 CH 组成的支架,有或没有 ALG 和冻干 PRF,以评估支架的特性、生长因子释放以及来源于牙髓干细胞(DPSC)的成骨细胞(OB)的活力。合成并冻干了四种支架变体。然后,进行降解、溶胀曲线和形态分析。此外,通过 ELISA 定量测定 PDGF-BB 和 FGF-B 生长因子的释放,并评估细胞毒性和细胞活力。所有支架的溶胀和降解曲线相似,孔径在 100 到 250 μm 之间。在支架浸入细胞培养基 24 小时后,观察到 FGF-B 和 PDGF-BB 的释放。PRF 补充支架显著增加了 DPSC 和 OB-DPSC 的活力。nHA-CH-GEL-PRF 支架表现出最佳的物理生物学特性,可刺激 DPSC 和 OB-DPSC 的细胞活力。这些结果表明,冻干 PRF 可提高支架的生物相容性,从而促进骨组织再生。

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