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An Insight into Nano Silver Fluoride-Coated Silk Fibroin Bioinspired Membrane Properties for Guided Tissue Regeneration.

ABSTRACT: The current work focuses on the development of a novel electrospun silk fibroin (SF) nonwoven mat as a GTR membrane with antibacterial, biomineralization and biocompatible properties. The γ-poly glutamic acid (γ-PGA)-capped nano silver fluoride (NSF) and silver diamine fluoride (SDF) were first synthesized, which were dip-coated onto electrospun silk fibroin mats (NSF-SF and SDF-SF). UV-Vis spectroscopy and TEM depicted the formation of silver nanoparticles. NSF-SF and SDF-SF demonstrated antibacterial properties (against Porphyromonas gingivalis) with 3.1 and 6.7 folds higher relative to SF, respectively. Post-mineralization in simulated body fluid, the NSF-SF effectively promoted apatite precipitation (Ca/P ~1.67), while the SDF-SF depicted deposition of silver nanoparticles, assessed by SEM-EDS. According to the FTIR-ATR deconvolution analysis, NSF-SF portrayed ~75% estimated hydroxyapatite crystallinity index (CI), whereas pure SF and SDF-SF demonstrated ~60%. The biocompatibility of NSF-SF was ~82% when compared to the control, while SDF-coated samples revealed in vitro cytotoxicity, further needing in vivo studies for a definite conclusion. Furthermore, the NSF-SF revealed the highest tensile strength of 0.32 N/mm and 1.76% elongation at break. Therefore, it is substantiated that the novel bioactive and antibacterial NSF-SF membranes can serve as a potential candidate, shedding light on further in-depth analysis for GTR applications.

SUBMITTER: Pandey A

PROVIDER: S-EPMC8401509 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The dissolution of Bombyx mori silk fibroin (SF) films in formic acid (FA) for the preparation of electrospinning dopes is widely exploited to produce electrospun SF scaffolds. The SILKBridge® nerve conduit is an example of medical device having in its wall structure an electrospun component produced from an FA spinning dope. Though highly volatile, residual FA remains trapped into the bulk of the SF nanofibers. The purpose of this work is to investigate the type and strength of the interaction between FA and SF in electrospun mats, to quantify its amount and to evaluate its possible toxicological impact on human health. The presence of residual FA in SF mats was detected by FTIR and Raman spectroscopy (new carbonyl peak at about 1,725 cm-1) and by solid state NMR, which revealed a new carbonyl signal at about 164.3 ppm, attributed to FA by isotopic 13C substitution. Changes occurred also in the spectral ranges of hydroxylated amino acids (Ser and Thr), demonstrating that FA interacted with SF by forming formyl esters. The total amount of FA was determined by HS-GC/MS analysis and accounted for 247 ± 20 μmol/g. The greatest part was present as formyl ester, a small part (about 3%) as free FA. Approximately 17% of the 1,500 μmol/g of hydroxy amino acids (Ser and Thr) theoretically available were involved in the formation of formyl esters. Treatment with alkali (Na2CO3) succeeded to remove the greatest part of FA, but not all. Alkali-treated electrospun SF mats underwent morphological, physical, and mechanical changes. The average diameter of the fibers increased from about 440 nm to about 480 nm, the mat shrunk, became stiffer (the modulus increased from about 5.5 MPa to about 7 MPa), and lost elasticity (the strain decreased from about 1 mm/mm to about 0.8 mm/mm). Biocompatibility studies with human adult dermal fibroblasts did not show significant difference in cell proliferation (313 ± 18 and 309 ± 23 cells/mm2 for untreated and alkali-treated SF mat, respectively) and metabolic activity. An in-depth evaluation of the possible toxicological impact of residual FA was made using the SILKBridge® nerve conduit as case study, following the provisions of the ISO 10993-1 standard. The Potential Patient Daily Intake, calculated from the total amount of FA determined by HS-GC/MS, was 2.4 mg/day and the Tolerable Exposure level was set to 35.4 mg/day. This allowed to obtain a value of the Margin of Safety of 15, indicating that the amount of FA left on SF mats after electrospinning does not raise concerns for human health.

Project description:Tissue engineering has become a new approach for repairing bony defects. Highly porous osteoconductive scaffolds perform the important role for the success of bone regeneration. By biomimetic strategy, apatite-coated porous biomaterial based on silk fibroin scaffolds (SS) might provide an enhanced osteogenic environment for bone-related outcomes. To assess the effects of apatite-coated silk fibroin (mSS) biomaterials for bone healing as a tissue engineered bony scaffold, we explored a tissue engineered bony graft using mSS seeded with osteogenically induced autologous bone marrow stromal cells (bMSCs) to repair inferior mandibular border defects in a canine model. The results were compared with those treated with bMSCs/SS constructs, mSS alone, SS alone, autologous mandibular grafts and untreated blank defects. According to radiographic and histological examination, new bone formation was observed from 4 weeks post-operation, and the defect site was completely repaired after 12 months for the bMSCs/mSS group. In the bMSCs/SS group, new bone formation was observed with more residual silk scaffold remaining at the center of the defect compared with the bMSCs/mSS group. The engineered bone with bMSCs/mSS achieved satisfactory bone mineral densities (BMD) at 12 months post-operation close to those of normal mandible (p>0.05). The quantities of newly formed bone area for the bMSCs/mSS group was higher than the bMSCs/SS group (p<0.01), but no significant differences were found when compared with the autograft group (p>0.05). In contrast, bony defects remained in the center with undegraded silk fibroin scaffold and fibrous connective tissue, and new bone only formed at the periphery in the groups treated with mSS or SS alone. The results suggested that apatite-coated silk fibroin scaffolds combined with bMSCs could be successfully used to repair mandibular critical size border defects and the premineralization of these porous silk fibroin protein scaffolds provided an increased osteoconductive environment for bMSCs to regenerate sufficient new bone tissue.

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An Insight into Nano Silver Fluoride-Coated Silk Fibroin Bioinspired Membrane Properties for Guided Tissue Regeneration.

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