Preparation and properties of asymmetric porous antimicrobial nZnO/P34HB composite guided bone regeneration membrane

Chang LIU; Wei LIU; Jingmei LUO; Li HAN; Chao ZHANG; Wanrong TANG

doi:10.16016/j.2097-0927.202404134

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Preparation and properties of asymmetric porous antimicrobial nZnO/P34HB composite guided bone regeneration membrane

Biomedical Engineering | 更新时间：2026-01-09

- Preparation and properties of asymmetric porous antimicrobial nZnO/P34HB composite guided bone regeneration membrane
- Journal of Army Medical University Vol. 46, Issue 24, Pages: 2745-2754(2024)
- 作者机构：
  
  637000 四川南充，川北医学院附属医院口腔科
- 作者简介：
- 基金信息：
  
  the Scientific Research Initiation Foundation for Doctors of North Sichuan Medical College(CBY22-QDA11)
- DOI：10.16016/j.2097-0927.202404134
  CLC： R318.08;R782.13;R944.9
- Received：28 April 2024，
  
  Revised：2024-10-11，
  
  Published：30 December 2024
- 稿件说明：
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Chang LIU, Wei LIU, Jingmei LUO, et al. Preparation and properties of asymmetric porous antimicrobial nZnO/P34HB composite guided bone regeneration membrane[J]. Journal of Army Medical University, 2024, 46(24): 2745-2754.
DOI：

Chang LIU, Wei LIU, Jingmei LUO, et al. Preparation and properties of asymmetric porous antimicrobial nZnO/P34HB composite guided bone regeneration membrane[J]. Journal of Army Medical University, 2024, 46(24): 2745-2754. DOI： 10.16016/j.2097-0927.202404134.

摘要

目的

制备不对称多孔抗菌纳米氧化锌(zinc oxide nanoparticle

nZnO)/聚3-羟基丁酸酯4-羟基丁酸酯[Poly (3-hydroxybutyrate-co-4-hydroxy butyrate) copolymer

P34HB]复合引导骨再生(guided bone regeneration

GBR)膜

并检测其成骨性能。

方法

本研究采用超声分散技术、溶剂置换法和冷冻干燥技术制备负载nZnO的P34HB复合GBR膜(实验分为空白对照组和0%、0.5%、1.0%、1.5% nZnO/P34HB材料组)

采用扫描电子显微镜(scanning electron microscope

SEM)、能谱仪(energy dispersive spectrometer

EDS)等明确nZnO的负载并观察其结构; CCK-8检测细胞增殖能力; 划痕实验检测细胞迁移能力; 蛋白吸附实验检测GBR膜的蛋白吸附能力; 酶解实验检测GBR膜的体外降解; 成骨实验检测碱性磷酸酶(alkaline phosphatase

ALP)活性; 茜素红染色观察钙化结节; 体外抑菌实验检测GBR膜的抑菌能力。

结果

本研究成功制备了负载0%、0.5%、1.0%、1.5% nZnO的4组nZnO/P34HB复合GBR膜

SEM结果显示

该复合膜一面疏松多孔

另一面致密; EDS表明复合膜上出现锌元素的掺入。细胞

增殖实验结果显示

0.5%、1.0% nZnO/P34HB组促进MC3T3-E1增殖且增殖数量均高于空白对照组、0%、1.5% nZnO/P34HB组(

0.05);细胞划痕实验结果显示

在含有0.5% nZnO/P34HB组的浸提液培养基中MC3T3-E1明显越过划痕边缘进行迁移

且48 h时的0.5% nZnO/P34HB组细胞迁移率高于其他3个实验材料组(

0.05);蛋白吸附实验结果显示

4组实验材料组光密度值差异无统计学意义

表明nZnO的加入不会降低材料本身的蛋白吸附能力; 酶解实验结果显示

4组实验材料组的质量均随着酶解时间增加而持续降低

28 d后仍保持较低的降解率; ALP活性检测结果显示

0.5% nZnO/P34HB组ALP染色更深且活性高于其他3个实验材料组(

0.01)。茜素红染色结果显示

0.5% nZnO/P34HB组与其他3个实验材料组比较

观察到较多的典型红色深染钙化结节

0%、0.5% nZnO/P34HB组的细胞矿化程度在4组中更高(

0.01)。体外抑菌实验结果显示

0.5% nZnO/P34HB组平板上的菌落数量显著少于空白对照组和0% nZnO/P34HB组(

0.01)。

结论

本研究成功制备了负载nZnO的P34HB复合GBR膜

筛选出的0.5% nZnO/P34HB组具有不对称多孔结构

能够促进细胞增殖

利于细胞成骨

同时具有抑菌能力。

Abstract

Objective

To develop an asymmetric porous guided bone regeneration (GBR) membrane made from antibacterial zinc oxide nanoparticle (nZnO)/poly (3-hydroxybutyrate-co-4-hydroxy butyrate) copolymer (P34HB)

and detect its osteogenic performance.

Methods

Ultrasonic dispersion technique

solvent displacement method and freeze-drying technique were used to prepare P34HB composite GBR membrane loaded with nZnO at different doses (0%

0.5%

1.0% and 1.5%).Scanning electron microscopy (SEM) and energy spectrometry (EDS) were conducted to determine the load of nZnO and observe the structure of the prepared membrane.Then MC3T3-E1 cells were implanted in a 24-well culture plate with the prepared GBR membrane pre-laid in the well.Thus the experiment included blank control group (no membrane) and groups with membranes composing 0%

0.5%

1.0% and 1.5% nZnO

respectively.CCK-8 and cell scratch assays were used to detect cell proliferation capacity and migration ability.The protein adsorption capacity of GBR membrane was detected by protein adsorption experiment.The

in vitro

degradation of GBR membrane was tested; the act

ivity of alkaline phosphatase (ALP) was detected with chemical test.And calcified nodules were observed with alizzarin red staining; the antibacterial ability of GBR membranes was tested with

in vitro

antibacterial experiments.

Results

Four groups of nZnO/P34HB composite GBR membranes with 0%

0.5%

1.0% and 1.5% nZnO were successfully prepared.SEM results showed that the composite membrane was porous on one side and dense on the other side.EDS indicated the incorporation of zinc elements in the composite membrane.The results of cell proliferation assay showed that 0.5% and 1.0% nZnO/P34HB group promoted the proliferation of MC3T3-E1 cells

when compared with the blank control group and 0% and 1.5% nZnO/P34HB groups (

0.05).The results of cell scratch assay showed that MC3T3-E1 cells in the 0.5% nZnO/P34HB group migrated significantly over the scratch edge

with the cell mobility higher than that of the other 3 membrane groups in 48 h after culture (

0.01).Protein adsorption experiment displayed there was no significant difference in optical density of the 4 group of membrane

indicating that the addition of nZnO had no effect on the protein adsorption capacity of the material itself.The results of enzymatic experiment showed that though the quality of the 4 membranes was continuously decreased with elapse of time

the degradation rate was still relatively lower after 28 d of treatment.ALP staining and alizarin red staining displayed that the 0.5% nZnO/P34HB group obtained the strongest ALP activity and the largest amount of calcified nodules when compared with the other 3 groups (

0.01).Higher mineralization was observed in the 0% and 0.5% nZnO/P 34 HB groups than the other 2 groups (

0.01).The results of

in vitro

antibacterial experiments showed that the number of colonies in the 0.5% nZnO/P 34 HB group was significantly less than that in the blank control

group and the 0% nZnO/P34HB group (

0.01).

Conclusion

Our prepared P34HB composite GBR membrane loaded with nZnO is in asymmetric porous structure

and can promote cell proliferation

facilitate cell osteogenesis and have bacteriostatic ability.

关键词

Keywords

references

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