Preparation of functional polyhydroxyalkanoate microspheres and their antibacterial activity and osteogenic effect evaluation.
10.7507/1002-1892.202303136
- Author:
Jianfei WU
1
;
Binglong WANG
1
;
Yu LIU
1
;
Daixu WEI
1
Author Information
1. Zigong Affiliated Hospital of Southwest Medical University, Zigong Psychiatric Research Center, Zigong Institute of Brain Science, Zigong Sichuan, 643020, P. R. China.
- Publication Type:Journal Article
- Keywords:
Bone tissue engineering;
bone morphogenetic protein 2;
human β-defensin 3;
microsphere;
polyhydroxyalkanoate;
scaffold material
- MeSH:
Humans;
Osteogenesis;
Polyhydroxyalkanoates;
Microspheres;
Alkaline Phosphatase;
Anti-Bacterial Agents/pharmacology*;
Coloring Agents;
Escherichia coli
- From:
Chinese Journal of Reparative and Reconstructive Surgery
2023;37(8):929-936
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct polyhydroxyalkanoate (PHA) microspheres loaded with bone morphogenetic protein 2 (BMP-2) and human β-defensin 3 (HBD3), and evaluate the antibacterial activity of microspheres and the effect of promoting osteogenic differentiation, aiming to provide a new option of material for bone tissue engineering.
METHODS:The soybean lecithin (SL)-BMP-2 and SL-HBD3 were prepared by SL-mediated introduction of growth factors into polyesters technology, and the functional microsphere (f-PMS) containing BMP-2 and HBD3 were prepared by microfluidic technology, while pure microsphere (p-PMS) was prepared by the same method as the control. The morphology of microspheres was observed by scanning electron microscopy and the water absorption was detected; the release curves of BMP-2 and HBD3 in f-PMS were detected by ELISA kit. The antibacterial effect of microspheres in Staphylococcus aureus and Escherichia coli was tested with the LIVE/DEADTM BacLightTM bacterial staining kit; the biocompatibility of microspheres was tested using Transwell and cell counting kit 8 (CCK-8). The effect of microspheres on osteogenic differentiation was determined by collagen type Ⅰ (COL-1) immunofluorescence staining and alkaline phosphatase (ALP) concentration.
RESULTS:In this experiment, the f-PMS and p-PMS were successfully constructed. Morphological characteristics showed that p-PMS surface was rough and distributed with micropores of 1-3 μm, while f-PMS surface was smooth and existed white granular material. There was no significant difference in water absorption between the two groups (P>0.05). The release curves of BMP-2 and HBD3 in the f-PMS and p-PMS were basically the same, showing both early sudden release and late slow release. The antibacterial activity of f-PMS was significantly higher than that of p-PMS in the test that against Staphylococcus aureus and Escherichia coli (P<0.05), but there was no significant difference in biocompatibility between the two groups (P>0.05). The results of osteogenic differentiation of human BMSCs showed that the fluorescence intensity of osteogenic specific protein COL-1 of f-PMS was significantly higher than that in p-PMS, and the activity of ALP in f-PMS was also significantly higher than that in p-PMS (P<0.05).
CONCLUSION:The p-PHA have good antibacterial activity and biocompatibility, and can effectively promote the osteogenic differentiation of human BMSCs, which is expected to be applied to bone tissue engineering in the future.
