Blood, sweat, and tears: extraterrestrial regolith biocomposites with in vivo binders.

The proverbial phrase 'you can't get blood from a stone' is used to describe a task that is practically impossible regardless of how much force or effort is exerted. This phrase is well-suited to humanity's first crewed mission to Mars, which will likely be the most difficult and technologically challenging human endeavor ever undertaken. The high cost and significant time delay associated with delivering payloads to the Martian surface means that exploitation of resources in situ - including inorganic rock and dust (regolith), water deposits, and atmospheric gases - will be an important part of any crewed mission to the Red Planet. Yet there is one significant, but chronically overlooked, source of natural resources that will - by definition - also be available on any crewed mission to Mars: the crew themselves. In this work, we explore the use of human serum albumin (HSA) - a common protein obtained from blood plasma - as a binder for simulated Lunar and Martian regolith to produce so-called 'extraterrestrial regolith biocomposites (ERBs).' In essence, HSA produced by astronauts in vivo could be extracted on a semi-continuous basis and combined with Lunar or Martian regolith to 'get stone from blood', to rephrase the proverb. Employing a simple fabrication strategy, HSA-based ERBs were produced and displayed compressive strengths as high as 25.0 MPa. For comparison, standard concrete typically has a compressive strength ranging between 20 and 32 MPa. The incorporation of urea - which could be extracted from the urine, sweat, or tears of astronauts - could further increase the compressive strength by over 300% in some instances, with the best-performing formulation having an average compressive strength of 39.7 MPa. Furthermore, we demonstrate that HSA-ERBs have the potential to be 3D-printed, opening up an interesting potential avenue for extraterrestrial construction using human-derived feedstocks. The mechanism of adhesion was investigated and attributed to the dehydration-induced reorganization of the protein secondary structure into a densely hydrogen-bonded, supramolecular ß-sheet network - analogous to the cohesion mechanism of spider silk. For comparison, synthetic spider silk and bovine serum albumin (BSA) were also investigated as regolith binders - which could also feasibly be produced on a Martian colony with future advancements in biomanufacturing technology.

Non-covalent protein-based adhesives for transparent substrates-bovine serum albumin vs. recombinant spider silk.

Protein-based adhesives could have several advantages over petroleum-derived alternatives, including substantially lower toxicity, smaller environmental footprint, and renewable sourcing. Here, we report that non-covalently crosslinked bovine serum albumin and recombinant spider silk proteins have high adhesive strength on glass (8.53 and 6.28 MPa, respectively) and other transparent substrates. Moreover, the adhesives have high visible transparency and showed no apparent degradation over a period of several months. The mechanism of adhesion was investigated and primarily attributed to dehydration-induced reorganization of protein secondary structure, resulting in the supramolecular association of ß-sheets into a densely hydrogen-bonded network.

A comparative study of the effects of different bioactive fillers in PLGA matrix composites and their suitability as bone substitute materials: A thermo-mechanical and in vitro investigation.

Bone substitute composite materials with poly(L-lactide-co-glycolide) (PLGA) matrices and four different bioactive fillers: CaCO3, hydroxyapatite (HA), 45S5 Bioglass(®) (45S5 BG), and ICIE4 bioactive glass (a lower sodium glass than 45S5 BG) were produced via melt blending, extrusion and moulding. The viscoelastic, mechanical and thermal properties, and the molecular weight of the matrix were measured. Thermogravimetric analysis evaluated the effect of filler composition on the thermal degradation of the matrix. Bioactive glasses caused premature degradation of the matrix during processing, whereas CaCO3 or HA did not. All composites, except those with 45S5 BG, had similar mechanical strength and were stiffer than PLGA alone in compression, whilst all had a lower tensile strength. Dynamic mechanical analysis demonstrated an increased storage modulus (E') in the composites (other than the 45S5 BG filled PLGA). The effect of water uptake and early degradation was investigated by short-term in vitro aging in simulated body fluid, which indicated enhanced water uptake over the neat polymer; bioactive glass had the greatest water uptake, causing matrix plasticization. These results enable a direct comparison between bioactive filler type in poly(α-hydroxyester) composites, and have implications when selecting a composite material for eventual application in bone substitution.

Long-term in vitro degradation of PDLLA/bioglass bone scaffolds in acellular simulated body fluid.

The long-term (600days) in vitro degradation of highly porous poly(D,L-lactide) (PDLLA)/Bioglass-filled composite foams developed for bone tissue engineering scaffolds has been investigated in simulated body fluid (SBF). Foams of ∼93% porosity were produced by thermally induced phase separation (TIPS). The degradation profile for foams of neat PDLLA and the influence of Bioglass addition were comprehensively assessed in terms of changes in dimensional stability, pore morphology, weight loss, molecular weight and mechanical properties (dry and wet states). It is shown that the degradation process proceeded in several stages: (a) a quasi-stable stage, where water absorption and plasticization occurred together with weight loss due to Bioglass particle loss and dissolution, resulting in decreased wet mechanical properties; (b) a stage showing a slight increase in the wet mechanical properties and a moderate decrease in dimensions, with the properties remaining moderately constant until the onset of significant weight loss, whilst molecular weight continued to decrease; (c) an end stage of massive weight loss, disruption of the pore structure and the formation of blisters and embrittlement of the scaffold (evident on handling). The findings from this long-term in vitro degradation investigation underpin studies that have been and continue to be performed on highly porous poly(α-hydroxyesters) scaffolds filled with bioactive glasses for bone tissue engineering applications.

Assessment of antimicrobial microspheres as a prospective novel treatment targeted towards the repair of perianal fistulae.

BACKGROUND: None of the proposed materials tested for the management of perianal fistulae has proven to be a definitive treatment. AIM: To assess a new repair scaffold and drug delivery device conceived to target perianal fistula repair. METHODS: Poly(D,L-lactide-co-glycolide) porous microspheres containing either antibacterial silver-releasing degradable phosphate glass or metronidazole were prepared using thermally induced phase separation. RESULTS: Ion- and drug-release profiling of the microspheres revealed continued release of silver ions from microspheres filled with silver-doped phosphate glass and high encapsulation efficiency for metronidazole [78% and 82% for microspheres loaded with 2.5% and 1.3% (w/w), respectively]. Microbicidal activity was confirmed by growth inhibition of bacterial species (Staphylococcus aureus, Escherichia coli and Bacteroides fragilis), which characteristically dominate the colonization of perianal fistula tracts. Microspheres containing >3 mol% silver or metronidazole resulted in strong bacterial inhibition/kill against B. fragilis; the presence of one sphere containing >3 mol% silver had a potent inhibitory effect against all the microbes studied. Microspheres became rapidly integrated with host tissue following subcutaneous implantation into a rodent wound model. CONCLUSION: The study demonstrates a novel scaffold for guided tissue regeneration providing local release of antimicrobial agents sufficient to counter bacterial colonization and warrants further investigation.

Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering.

Biodegradable polymers and bioactive ceramics are being combined in a variety of composite materials for tissue engineering scaffolds. Materials and fabrication routes for three-dimensional (3D) scaffolds with interconnected high porosities suitable for bone tissue engineering are reviewed. Different polymer and ceramic compositions applied and their impact on biodegradability and bioactivity of the scaffolds are discussed, including in vitro and in vivo assessments. The mechanical properties of today's available porous scaffolds are analyzed in detail, revealing insufficient elastic stiffness and compressive strength compared to human bone. Further challenges in scaffold fabrication for tissue engineering such as biomolecules incorporation, surface functionalization and 3D scaffold characterization are discussed, giving possible solution strategies. Stem cell incorporation into scaffolds as a future trend is addressed shortly, highlighting the immense potential for creating next-generation synthetic/living composite biomaterials that feature high adaptiveness to the biological environment.

Bioactive and mechanically strong Bioglass-poly(D,L-lactic acid) composite coatings on surgical sutures.

New coating processes have been investigated for degradable (Vicryl) and nondegradable (Mersilk) sutures with the aim to develop Bioglass coated polymer fibers for wound healing and tissue engineering scaffold applications. First, the aqueous phase of a Bioglass particle slurry was replaced with a poly(D,L-lactic acid) (PDLLA) polymer dissolved in solvent dimethyle carbonate (DMC) to act as third phase. SEM observations indicated that this alteration significantly improved the homogeneity of the coatings. Second, a new coating strategy involving two steps was developed: the sutures were first coated with a Bioglass-PDLLA composite film followed by a second PDLLA coating. This two-step process of coating has addressed the problem of poor adherence of Bioglass particles on suture surfaces. The coated sutures were knotted to determine qualitatively the mechanical integrity of the coatings. The results indicated that adhesion strength of coatings obtained by the two-step method was remarkably enhanced. A comparative assessment of the bioactivity of one-step and two-step produced coatings was carried out in vitro using acellular simulated body fluid (SBF) for up to 28 days. Coatings produced by the two-step process were found to have similar bioactivity as the one-step produced coatings. The novel Bioglass/PDLLA/Vicryl and Bioglass/PDLLA/Mersilk composite sutures are promising bioactive materials for wound healing and tissue engineering applications.

Effect of iron on the surface, degradation and ion release properties of phosphate-based glass fibres.

Phosphate-based glass fibres (PGF) have the unique characteristic of being completely soluble in an aqueous environment, releasing bioactive and biocompatible ions. They have been proposed as tissue engineering scaffolds for craniofacial skeletal muscle regeneration, where myoblasts are seeded directly onto the fibres. Studies have shown that these cells have a preference in their initial attachment to fibres of certain composition and size, which in turn control the rate of degradation. This study investigated the relationship between the surface properties, degradation properties and ion release (cationic and anionic species) by altering the chemical composition of the PGF. Iron oxide (Fe2O3) was incorporated into glasses containing P2O5 (50 mol%), CaO (30 mol%) and Na2O (20 mol%). Six glass compositions with Fe2O3 ranging from 0 to 5 mol% by replacing the equivalent Na2O mol% were investigated. Contact angle measurements showed that polar interactions occurring on the glass surfaces diminished with increasing Fe2O3 content. This behaviour was reflected in the estimated surface energies of the glasses, where the overall surface energy decreased with increasing Fe2O3 content due to the decrease in polar or acid/base component. The incorporation of up to 5 mol% Fe2O3 into PGF resulted in a significant reduction in the degradation rate (by two orders of magnitude), which can be related to the formation of more hydration resistant P-O-Fe bonds. However, the degradation rate increased with decreasing fibre diameter (comparing average diameters of 31.6 +/- 6.5 microm versus 13.1 +/- 1.3 microm) for a given mass of fibre, and this is related to the surface area to volume ratio. Taken together the results suggest that fibres with the larger diameters and containing 3-5 mol% Fe2O3 could initially be a more durable scaffold than ones with 1 or 2 mol% Fe2O3 for initial cell attachment.

Mechanical properties of highly porous PDLLA/Bioglass composite foams as scaffolds for bone tissue engineering.

This study developed highly porous degradable composites as potential scaffolds for bone tissue engineering. These scaffolds consisted of poly-D,L-lactic acid filled with 2 and 15 vol.% of 45S5 Bioglass particles and were produced via thermally induced solid-liquid phase separation and subsequent solvent sublimation. The scaffolds had a bimodal and anisotropic pore structure, with tubular macro-pores of approximately 100 microm in diameter, and with interconnected micro-pores of approximately 10-50 microm in diameter. Quasi-static and thermal dynamic mechanical analysis carried out in compression along with thermogravimetric analysis was used to investigate the effect of Bioglass on the properties of the foams. Quasi-static compression testing demonstrated mechanical anisotropy concomitant with the direction of the macro-pores. An analytical modelling approach was applied, which demonstrated that the presence of Bioglass did not significantly alter the porous architecture of these foams and reflected the mechanical anisotropy which was congruent with the scanning electron microscopy investigation. This study found that the Ishai-Cohen and Gibson-Ashby models can be combined to predict the compressive modulus of the composite foams. The modulus and density of these complex foams are related by a power-law function with an exponent between 2 and 3.

Development and characterisation of silver-doped bioactive glass-coated sutures for tissue engineering and wound healing applications.

A novel silver-doped bioactive glass powder (AgBG) was used to coat resorbable Vicryl (polyglactin 910) and non-resorbable Mersilk surgical sutures, thereby imparting bioactive, antimicrobial and bactericidal properties to the sutures. Stable and homogeneous coatings on the surface of the sutures were achieved using an optimised aqueous slurry-dipping technique. Dynamic mechanical analysis (DMA) was used to investigate the viscoelastic parameters of storage modulus and tandelta and thermal transitions of the as-received and composite (coated) sutures. The results generally showed that the bioactive glass coating did not affect the dynamic mechanical and thermal properties of the sutures. The in vitro bioactivity of the sutures was tested by immersion in simulated body fluid (SBF). After only 3 days of immersion in SBF, bonelike hydroxyapatite formed on the coated suture surfaces, indicating their enhanced bioactive behaviour. Resorbable sutures with bioactive coatings as fabricated here, in conjunction with 3-D textile technology, may provide attractive materials for producing 3-D scaffolds with controlled porosities for tissue engineering applications. The bactericidal properties imparted by the Ag-containing glass coating open also new opportunities for use of the composite sutures in wound healing and body wall repair.

In vitro evaluation of novel bioactive composites based on Bioglass-filled polylactide foams for bone tissue engineering scaffolds.

Highly porous poly(DL-lactic acid) (PDLLA) foams and Bioglass-filled PDLLA composite foams were characterized and evaluated in vitro as bone tissue engineering scaffolds. The hypothesis was that the combination of PDLLA with Bioglass in a porous structure would result in a bioresorbable and bioactive composite, capable of supporting osteoblast adhesion, spreading and viability. Composite and unfilled foams were incubated in simulated body fluid (SBF) at 37 degrees C to study the in vitro degradation of the polymer and to detect hydroxyapatite (HA) formation, which is a measure of the materials' in vitro bioactivity. HA was detected on all the composite samples after incubation in SBF for just 3 days. After 28 days immersion the foams filled with 40 wt % Bioglass developed a continuous layer of HA. The formation of HA for the 5 wt % Bioglass-filled foams was localized to the Bioglass particles. Cell culture studies using a commercially available (ECACC) human osteosarcoma cell line (MG-63) were conducted to assess the biocompatibility of the foams and cell attachment to the porous substrates. The osteoblast cell infiltration study showed that the cells were able to migrate through the porous network and colonize the deeper regions within the foam, indicating that the composition of the foams and the pore structures are able to support osteoblast attachment, spreading, and viability. Rapid formation of HA on the composites and the attachment of MG-63 cells within the porous network of the composite foams confirms the high in vitro bioactivity and biocompatibility of these materials and their potential to be used as scaffolds in bone tissue engineering and repair.