Development and performance evaluation of a novel elastic bacterial nanocellulose/polyurethane small caliber artificial blood vessels.

There is an increasing demand for small-diameter blood vessels. Currently, there is no clinically available small-diameter artificial vessel. Bacterial nanocellulose (BNC) has vast potential for applications in artificial blood vessels due to its good biocompatibility. At the same time, medical polyurethane (PU) is a highly elastic polymer material widely used in artificial blood vessels. This study reports a composite small-diameter BNC/PU conduit using a non-solvent-induced phase separation method with the highly hydrophilic BNC tube as the skeleton and the hydrophobic polycarbonate PU as the filling material. The results revealed that the compliance and mechanical matching of BNC/PU tubes were higher than BNC tubes; the axial/radial mechanical strength, burst pressure, and suture strength were significantly improved; the blood compatibility and cell compatibility were also excellent. The molecular and subcutaneous embedding tests showed that the composite tubes had lighter inflammatory reactions. The results of the animal substitution experiments showed that the BNC/PU tubes kept blood flow unobstructed without tissue proliferation after implantation in rats for 9 months. Thus, the BNC/PU small-diameter vascular prosthesis had the potential for long-term patency and acted as an ideal material for small-diameter vessels.

Production of novel elastic bacterial nanocellulose/polyvinyl alcohol conduits via mercerization and phase separation for small-caliber vascular grafts application.

Size and properties of tubular bacterial nanocellulose (BNC) can be regulated by controllable mercerization with thinner tube walls, better mechanical properties, and improved biocompatibility. Although mercerized BNC (MBNC) conduits have considerable potential as small-caliber vascular grafts (<6 mm), poor suture retention and lack of compliance that cannot match natural blood vessels increase the difficulty of surgery and limit potential clinical application. Polyvinyl alcohol (PVA) is a kind of hydrophilic polymer with good biocompatibility and elasticity, which can precipitate in alkaline solutions. In this study, novel elastic mercerized BNC/PVA conduits (MBP) are manufactured combining mercerization of BNC tubes with precipitation and phase separation of PVA with thinner tube wall, improved suture retention, better elasticity, good hemocompatibility and great cytocompatibility. The MBP obtained with 12.5 % PVA is selected for transplantation in a rat abdominal aorta model. For 32 weeks, normal blood flow is observed using Doppler sonographic inspection, which demonstrates long-term patency. Immunofluorescence staining results also indicate the formation of endothelium and smooth muscle layers. The results indicate the introduction of PVA, and its phase separation into mercerization of tubular BNC can endow MBP conduits with better compliance and suture retention, making it a promising candidate for blood vessel replacement.

Vascular cells responses to controlled surface structure and properties of bacterial nanocellulose artificial blood vessel after mercerization.

Therapeutic benefits of small caliber artificial blood vessels to cure cardio and cerebrovascular diseases are mainly limited by their low patency during long-term transplantation. Bacterial nanocellulose (BNC), as a natural polysaccharide mainly synthesized by a bacterium Komagataeibatacter xylinus, has shown great potential in small-caliber vascular graft applications due to its shape controllability, and furthermore its physical surface structure can be adjusted with different treatments. However, influences of physical surface structure and properties of BNC conduits on behaviors of vascular cells have not been investigated. In this work, mercerized BNC conduits (MBNC) with different surface roughness and stiffness were constructed by controlled alkali (NaOH) treatment. The changes of surface structures and properties significantly affected the behaviors of vascular cells and gene expression; meanwhile, the cell seeding density also affected the cell responses. After mercerization with NaOH concentration > 10 %, it was observed that the increased stiffness of MBNC decreased several functional gene expressions of human vascular endothelial cells, and the pathological transformation of smooth muscle cells was inhibited. This study demonstrates physical surface structure of MBNC conduits will critically regulate functions and behaviors of vascular cells and it also provides important designing parameters to improve the long-term patency of BNC-based conduits.

Functionalization of Aminoalkylsilane-Grafted Bacterial Nanocellulose with ZnO-NPs-Doped Pullulan Electrospun Nanofibers for Multifunctional Wound Dressing.

High moisture permeability, excellent mechanical properties in a wet state, high water-holding capability, and high exudate absorption make bacterial nanocellulose (BNC) a favorable candidate for biomedical device production, especially wound dressings. The lack of antibacterial activity and healing-promoting ability are the main drawbacks that limit its wide application. Pullulan (Pul) is a nontoxic polymer that can promote wound healing. Zinc oxide nanoparticles (ZnO-NPs) are well-known as a safe antibacterial agent. In this study, aminoalkylsilane was chemically grafted on a BNC membrane (A-g-BNC) and used as a bridge to combine BNC with Pul-ZnO-NPs hybrid electrospun nanofibers. FTIR results confirmed the successful production of A-g-BNC/Pul-ZnO. The obtained dressing demonstrated blood clotting performance better than that of BNC. The dressing showed an ability to release ZnO, and its antibacterial activity was up to 5 log values higher than that of BNC. The cytotoxicity of the dressing toward L929 fibroblast cells clearly showed safety due to the proliferation of fibroblast cells. The animal test in a rat model indicated faster healing and re-epithelialization, small blood vessel formation, and collagen synthesis in the wounds covered by A-g-BNC/Pul-ZnO. The new functional dressing, fabricated with a cost-effective and easy method, not only showed excellent antibacterial activity but could also accelerate wound healing.

Implantation of air-dried bacterial nanocellulose conduits in a small-caliber vascular prosthesis rabbit model.

There are no small-caliber (<6 mm) vascular prostheses so far commercially available around the globe. Bacterial nanocellulose (BNC) is considered a promising material for small-caliber artificial blood vessel applications. Although BNC hydrogel-like (BNC-Gel) materials possess a 3D network structure, facilitating nutrient exchange when used as vascular prostheses, they are difficult to suture during surgery due to their softness. Furthermore, a water content greater than 99% prevents the material from convenient methods of preservation and transport. Air-drying the BNC (BNC-Dry) would solve these problems. The comparative morphology, mechanical properties, hemocompatibility, and cytocompatibility of the BNC-Gel and BNC-Dry conduits of 3 mm in diameter were recorded in the present study, the results indicating that the mechanical properties, hemocompatibility, and cytocompatibility of BNC-Dry conduits were superior to conduits of BNC-Gel. Forty-six days after replacement of the carotid artery in New Zealand white rabbits, the BNC-Dry conduits remained patent. Composite blood vessels composed of cellulose and autologous tissue were harvested for immunohistochemistry and immunofluorescence staining. Sections demonstrated that the outer walls of the conduits were wrapped with autologous tissue. Contractile smooth muscle cells (SMCs) were observed on the outer surface of the conduit, similar to that observed in natural blood vessels. BNC-Dry conduits exhibited excellent performance and possessed properties convenient for surgical applications as small-diameter blood vessels.

Improved Performance of Bacterial Nanocellulose Conduits by the Introduction of Silk Fibroin Nanoparticles and Heparin for Small-Caliber Vascular Graft Applications.

Bacterial nanocellulose (BNC) is a promising material for small-caliber artificial blood vessels, although promoting its anticoagulant properties with more rapid endothelialization would improve long-term patency. Silk fibroin nanoparticles (SFNP) were introduced into the luminal wall surface of BNC conduits both with and without heparin (Hep) through pressurization followed by fixation. Hep was introduced in two ways: (1) embedded within SF nanoparticles to form SF-HepNPs for construction of the BNC-SF-HepNP conduit and (2) chemically grafted onto BNC and BNC-SFNP to form BNC-Hep and BNC-SFNP-Hep conduits. Fourier transform infrared spectroscopy confirmed the formation of SF-HepNPs, although they did not incorporate into the fibrillar network due to their large size. Hep was successfully grafted onto BNC and BNC-SFNP, verified by toluidine blue staining. The hemocompatibility and cytocompatibility of the five samples (BNC, BNC-SFNP, BNC-SF-HepNP, BNC-Hep, and BNC-SFNP-Hep conduits) were compared in vitro. The heparinized BNC-Hep and BNC-SFNP-Hep conduits improved the anticoagulant properties, and BNC-SFNP-Hep promoted human umbilical vein endothelial cell proliferation but also controlled excessive human arterial smooth muscle cell proliferation, assisting rapid endothelialization and improving lumen patency. No significant inflammatory reaction or material degradation was observed after subcutaneous implantation for 4 weeks. Autogenous tissues were observed around the conduits, and cells infiltrated into the edges of all samples, the BNC-SFNP conduit causing the deepest infiltration, providing an appropriate microenvironment for angiogenesis when used in small-caliber blood vessel applications. Few inflammatory cells were found around the BNC-Hep and BNC-SFNP-Hep conduits. Thus, the anticoagulant properties of the BNC-SFNP-Hep conduit and its stimulation of endothelialization suggest that it has great potential in clinical applications as a small-caliber artificial blood vessel.

Bacterial Nanocellulose-Enhanced Alginate Double-Network Hydrogels Cross-Linked with Six Metal Cations for Antibacterial Wound Dressing.

Alginate (Alg) and bacterial nanocellulose (BNC) have exhibited great potential in biomedical applications, especially wound dressing. Non-toxicity and a moisture-maintaining nature are common features making them favorable for functional dressing fabrication. BNC is a natural biopolymer that promotes major advances to the current and future biomedical materials, especially in a flat or tubular membrane form with excellent mechanical strength at hydrated state. The main drawback limiting wide applications of both BNC and Alg is the lack of antibacterial activity, furthermore, the inherent poor mechanical property of Alg leads to the requirement of a secondary dressing in clinical treatment. To fabricate composite dressings with antibacterial activity and better mechanical properties, sodium alginate was efficiently incorporated into the BNC matrix using a time-saving vacuum suction method followed by cross-linking through immersion in separate solutions of six cations (manganese, cobalt, copper, zinc, silver, and cerium). The results showed the fabricated composites had not only pH-responsive antibacterial activities but also improved mechanical properties, which are capable of acting as smart dressings. All composites showed non-toxicity toward fibroblast cells. Rat model evaluation showed the skin wounds covered by the dressings healed faster than by BNC.