Electrospun L-Lysine/Amorphous Calcium Phosphate Loaded Core-Sheath Nanofibers for Managing Oral Biofilm Infections and Promoting Periodontal Tissue Repairment

Abstract

Introduction

Periodontitis, a chronic inflammatory disease prevalent worldwide, is primarily treated through GTR for tissue regeneration. The efficacy of GTR, however, remains uncertain due to potential infections and the intricate microenvironment of periodontal tissue. Herein, We developed a novel core-shell structure multifunctional membrane using a dual-drug-loaded coaxial electrospinning technique (Lys/ACP-CNF), contains L-lysine in the outer layer to aid in controlling biofilms after GTR regenerative surgery, and ACP in the inner layer to enhance osteogenic performance for accelerating alveolar bone repair.

Methods

The biocompatibility and cell adhesion were evaluated through CCK-8 and fluorescence imaging, respectively. The antibacterial activity was assessed using a plate counting assay. ALP, ARS, and RT-qPCR were used to examine osteogenic differentiation. Additionally, an in vivo experiment was conducted on a rat model with acute periodontal defect and infection. Micro-CT and histological analysis were utilized to analyze the in vivo alveolar bone regeneration.

Results

Structural and physicochemical characterization confirmed the successful construction of the core-shell fibrous structure. Additionally, the Lys/ACP-CNF showed strong antibacterial coaggregation effects and induced osteogenic differentiation of PDLSCs in vitro. The in vivo experiment confirmed that Lys/ACP-CNF promotes new bone formation.

Conclusion

Lys/ACP-CNF rapidly exhibited excellent antibacterial activity, protected PDLSCs from infection, and was conducive to osteogenesis, demonstrating its potential application for clinical periodontal GTR surgery.

Graphical Abstract

Keywords:

• anti-microbial therapy
• L-lysine
• electrospun membrane
• osteogenic
• periodontal tissue regeneration

Abbreviations

CNF, core-sheath nanofiber; Lys, L-lysine; GTR, Guided Tissue Regeneration; ACP, amorphous calcium phosphate; TCP, tricalcium phosphate; HA, hydroxyapatite; PDLSCs, periodontal ligament stem cells; Fn, F.nucleatum; Pg, P.gingivalis; PCL, Poly(ε-caprolactone); COL, TypeI collagen; PEG, Polyethylene Glycol; FTIR, Fourier Transform Infrared Spectrometer; XRD, X-ray diffraction; TEM, Transmission electron microscopy; HFLP, Hexafluoroisopropanol; PBS, phosphate-buffered saline; UV, Ultraviolet; SEM, Scanning electron microscope; TGA, Thermogravimetric analysis; CLSM, confocal laser scanning microscopy; BHI, brain-heart infusion; EPS, extracellular polysaccharide; CFU/mL, colony-forming units per milliliter; ARS, Alizarin Red S;ALP, Alkaline phosphatase; RT-qpcr, Reverse transcription quantitative polymerase chain reaction; SD rat, Sprague-Dawley rHFLPat; M1, the maxillary first molars; PDL, periodontal ligament; VOI, volume of interest; HU, Hounsfield Unit; BV/TV, the bone volume to tissue volume; Tb.Th, trabecular thickness; Tb. Sp, the ratio of trabecular separation; HE, Hematoxylin-eosin; WCA, water contact angle; OCN, osteocalcin; OPN, osteopontin; Runx2, runt related transcription factor 2; ALP, alkaline phosphatase; hagA, hemagglutinin A; hagB, hemagglutinin B; RagA, Ras-related GTP-binding protein.

Data Sharing Statement

The data, analytical methods and study materials that support the findings of this study can be made available on request to the corresponding authors who manage the information.

Ethics Approval and Consent to Participate

The experiments were approved by the Ethics Committee and the Animal Care and Use Committee of Nanjing Agricultural University (Approval Code: PZW2023039).

Acknowledgment

The authors expressed their gratitude to Xinyue Wang for providing grammar checking.

Author Contributions

Each author played a crucial role in the reported work, contributing to its conception, study design, execution, data acquisition, analysis, and interpretation. Additionally, they actively participated in drafting, revising, and critically reviewing the article. All authors provided final approval for the published version, reached consensus on the submitting journal, and collectively agree to be accountable for every aspect of the work.

Disclosure

The authors declare no conflicts of interest in this work.

Additional information

Funding

This work was supported by National Natural Science Foundation of China under Grant/Award Number 51972167, Postdoctoral research Foundation of Nanjing(2023) and Key projects of social development of Jiangsu Department of Science and Technology under Grant/Award Number BJ21006. The sponsors had no involvement in any of the stages from study design to submission of the paper for publication.