纳米颗粒增强的手性超分子水凝胶成骨性能

doi:10.19894/j.issn.1000-0518.210487

摘要/Abstract

摘要：

手性超分子水凝胶能够仿生细胞外手性微环境，在组织工程中具有特殊的意义，但其强度和稳定性较低，仍然面临着巨大的挑战。本文将无机羟基磷灰石纳米颗粒（HAP）引入到苯丙氨酸衍生物手性超分子水凝胶（LPF）中以改善其力学性能和生物功能。圆二色光谱和扫描电子显微镜结果显示，HAP掺入后LPF组装手性发生反转。与纯LPF水凝胶相比，杂化水凝胶具有优异的力学性能和长期稳定性。LPF储能模量G'值仅为452 Pa，在24 h内迅速崩塌；而LPF/HAP水凝胶的G'提高到3 353 Pa，保持凝胶态时间超过3个月。HAP纳米粒子表面上丰富的Ca²⁺离子与LPF羧基形成的配位作用是水凝胶强化的关键因素。此外，HAP加入还提高了对细胞的成骨刺激，表现为LPF/HAP基质上成骨细胞碱性磷酸酶（ALP）活性的增加，这不仅因为羟基磷灰石的骨诱导能力，还归因于HAP使纳米纤维的刚度和稳定性明显改善。通过无机纳米颗粒的力学增强和功能化，手性超分子水凝胶在骨组织工程中具有广阔的应用前景。

关键词: 超分子水凝胶, 无机纳米颗粒, 杂化, 力学稳定性, 成骨, 手性

Abstract:

Chiral supramolecular hydrogels that can mimic the chiral extracellular microenvironment are of special significance in tissue engineering， however， it still remains a great challenge for their low long lifetime stability. Herein， we introduce inorganic hydroxyapatite nanoparticles （HAP） to the well-established phenylalanine derivative chiral supramolecular hydrogel （LPF）， aiming to improve its mechanical performance as well as biofunctions. Morphological studies revealed an inversion of the chirality of LPF after HAP incorporation. Rheological experiments confirmed the long-term stability and superior mechanical property of the hybrid hydrogel over the neat hydrogel， where LPF/HAP hydrogel exhibited an improved elastic modulus of 3353 Pa， standing over three months， and LPF hydrogel collapsed soon within 24 hours with a much lower storage modulus （G'） value of 452 Pa. The abundant coordination interactions supplied by Ca²⁺ ions on the surface of HAP nanoparticles crosslinking with the carboxyl group are considered to be the crucial factors of the mechanical strengthening. Moreover， the HAP incorporation led to an enhanced stimulation of the osteogenesis of the osteoblastic cells， demonstrating by the increasement of the alkaline phosphatase （ALP） activity on LPF/HAP substrate， which ascribed not only to the osteoinductive ability of hydroxyapatite， but also to the improved stiffness and stability induced by HAP. The findings indicate that the chiral supramolecular hydrogel is promising in bone tissue engineering via inorganic nanoparticle strengthening and functionalization.

Key words: Supramolecular hydrogel, Inorganic nanoparticle, Hybrid, Mechanical stability, Osteogenesis, Chirality.

中图分类号:

O629.7

赵常利, 秦明高, 窦晓秋, 冯传良. 纳米颗粒增强的手性超分子水凝胶成骨性能[J]. 应用化学, 2022, 39(1): 177-187.

ZHAO Chang-Li, QIN Ming-Gao, DOU Xiao-Qiu, FENG Chuan-Liang. High Mechanical Stability and Osteogenesis of Chiral Supramolecular Hydrogel Induced by Inorganic Nanoparticles[J]. Chinese Journal of Applied Chemistry, 2022, 39(1): 177-187.

图/表 6

图示1 HAP掺入苯丙氨酸衍生物超分子水凝胶（LPF）对力学性能和成骨细胞分化行为影响的示意图

图1 凝胶LPF （A）、LC （B）、LN （C）、LT（D）和LH（E）的SEM图及CD光谱（F）

Fig.1 SEM images of LPF （A）， LC （B）， LN （C）， LT （D）， LH （E） xerogels； CD spectra of the hydrogels indicate signal inversion of LH around 260 nm compared to LPF （F）

图2 水凝胶形成后（A）、静置24 h的形态（B）及水凝胶样品的流变性能（频率扫描）（C）

Fig.2 Photograph images of the hydrogels right after hydrogel formation （A） 24 hours after hydrogel formation（B）， and the rheological measurements of hydrogel samples for frequency sweep （C）

图3 水凝胶的ATR-IR光谱

Fig.3 ATR-IR spectra of the hydrogels

图4 （A）荧光显微镜观察显示在不同基质上培养1 d和3 d后的活死细胞（比例尺=100 μm）；（B）根据活死染色图像计算细胞密度；（C）在不同基质上培养1、3和5 d后CCK-8吸光度结果；（D）ALP活性（*表示P < 0.05）

Fig.4 （A） Fluorescence microscopy images of the live and dead MC3T3-E1 cells after culture for 1 day and 3 days on different substrates（Scale bar： 100 μm）；（B） Calculated cell density according to the live-dead staining images；（C） Optical density results of the CCK-8 test after incubation for 1， 3 and 5 days on different substrates；（D） ALP activity （* denotes P < 0.05）

图5 MC3T3-E1细胞在LPF、LC、LN、LT和LH表面培养1 d后的荧光显微镜图像。细胞核呈蓝色，细胞骨架呈红色（比例尺=40 μm）

Fig.5 Fluorescence microscopy images of MC3T3-E1 cells after 1 day cultured on LPF， LC， LN， LT and LH substrates. Cell nucleus was stained in blue； cytoskeleton was stained in red （Scale bar = 40 μm）

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