Silk Solution Research Articles

Rheological Study on Blend Solutions of Non-mulberry Silk Fibroin and Gelatin Biopolymers

In current research work, we have studied the blending effect of non-mulberry silk fibroin (10% weight/volume basis) and gelatin (20% weight/volume basis) in formic acid. Several blends as SF10G0, SF2G8, SF3G7, SF5G5 and SF0G10 have been made and their rheological behaviour was investigated. The blend solutions were subjected to a steady shear rheological study in the variety of range of shear rates, namely 0.01–500 sec-1 and the viscosities of blend solutions were noticed to decrease in comparison to pure silk solution. The frequency sweep was employed in dynamic rheological tests to determine complex viscosity of these solutions from range of angular sweep 0.1–100 rad/sec. The consistent shear-thinning behaviour was noted for all the blends. The difference in numerical values of shear and complex viscosities indicated disobedience of Cox-Merz rule. Such analysis can be utilised for tailoring the properties of solution prior to processing them to create a versatile range of materials.

Photocrosslinked Silk Fibroin Microgel Scaffolds for Biomedical Applications

AbstractSilk fibroin hydrogels are extensively explored for tissue engineering and regenerative medicine as an artificial extracellular matrix (ECM) that can support tissue growth. However, the nanometer pore size of hydrogels limits adequate cell, tissue, and vascular infiltration. Microgel scaffolds are an emerging class of microporous biomaterials formed by annealing small microscale hydrogels (microgels) into a 3D construct. In this work, silk microgels are generated using a microfluidic device that allows tuning of the microgel diameter (100–400 µm) and are stabilized via visible light‐initiated photo‐crosslinking of native tyrosine residues in silk. Microgels are then covalently annealed using silk solution as glue and the same cytocompatible visible light‐initiated crosslinking to form microgel scaffolds. Unlike the nano‐porosity of bulk photo‐crosslinked silk hydrogels, the microgel scaffolds have an average pore diameter of 29 ± 17 or 192 ± 81 µm depending on the microgel size, with enhanced mechanical properties compared to bulk hydrogels. This microporosity supports enhanced cell spreading and proliferation in vitro and increases scaffold remodeling in vivo, encouraging improved tissue infiltration and matrix deposition. The microgel size and material format also affect inflammatory responses in vivo. This work demonstrates that silk microgels and microgel scaffolds are promising candidates for tissue engineering and regenerative medicine applications.

Microfibre-Functionalised Silk Hydrogels.

Silk hydrogels have shown potential for tissue engineering applications, but several gaps and challenges, such as a restricted ability to form hydrogels with tuned mechanics and structural features, still limit their utilisation. Here, Bombyx mori and Antheraea mylitta (Tasar) silk microfibres were embedded within self-assembling B. mori silk hydrogels to modify the bulk hydrogel mechanical properties. This approach is particularly attractive because it creates structured silk hydrogels. First, B. mori and Tasar microfibres were prepared with lengths between 250 and 500 μm. Secondary structure analyses showed high beta-sheet contents of 61% and 63% for B. mori and Tasar microfibres, respectively. Mixing either microfibre type, at either 2% or 10% (w/v) concentrations, into 3% (w/v) silk solutions during the solution-gel transition increased the initial stiffness of the resulting silk hydrogels, with the 10% (w/v) addition giving a greater increase. Microfibre addition also altered hydrogel stress relaxation, with the fastest stress relaxation observed with a rank order of 2% (w/v) > 10% (w/v) > unmodified hydrogels for either fibre type, although B. mori fibres showed a greater effect. The resulting data sets are interesting because they suggest that the presence of microfibres provided potential 'flow points' within these hydrogels. Assessment of the biological responses by monitoring cell attachment onto these two-dimensional hydrogel substrates revealed greater numbers of human induced pluripotent stem cell-derived mesenchymal stem cells (iPSC-MSCs) attached to the hydrogels containing 10% (w/v) B. mori microfibres as well as 2% (w/v) and 10% (w/v) Tasar microfibres at 24 h after seeding. Cytoskeleton staining revealed a more elongated and stretched morphology for the cells growing on hydrogels containing Tasar microfibres. Overall, these findings illustrate that hydrogel stiffness, stress relaxation and the iPSC-MSC responses towards silk hydrogels can be tuned using microfibres.

Modeling the 3-dimensional structure of the silkworm's spinning apparatus in silk production

The silk-spinning process of the silkworms transforms the liquid silk solution to a solid state under mild conditions, making it an attractive model for bioinspiration However, the precise mechanism behind silk expulsion remains largely unknown. Here we selected the silkworms as representative models to investigate the silk-spinning mechanism. We used serial block-face scanning electron microscopy (SBF-SEM) to reconstruct the three-dimensional structures of the spinnerets in silkworms at various stages and with different gene backgrounds. By comparing the musculature and duct deformation of these spinneret models during the spinning process, we were able to simulate the morphological changes of the spinneret. Based on the results, we proposed three essential factors for silkworm spinning: the pressure generated by the silk gland, the opening duct, and the pulling force generated by head movement. Understanding the silkworm spinning process provides insights into clarify the fluid-ejecting mechanism of a group of animals. Moreover, these findings are helpful to the development of biomimetic spinning device that mimics the push-and-pull dual-force system in silkworms. Statement of significanceThe silkworms’ spinning system produces fibers under mild conditions, making it an ideal candidate for bioinspiration. However, the mechanism of silk expulsion is unknown, and the three-dimensional structure of the spinneret is still uncertain. In this study, we reconstructed a detailed 3-dimensional model of the spinneret at near-nanometer resolution, and for the first time, we observed the changes that occur before and during the silk-spinning process. Our reconstructed models suggested that silkworms have the ability to control the spinning process by opening or closing the spinning duct. During the continuously spinning period, both the pressure generated by the silk gland and the pulling force resulting from head movement work in tandem to expel the silk solution. We believe that gaining a full understanding of the spinning process steps can advance our ability to spin synthetic fibers with properties comparable to those of native fibers by mimicking the natural spinning process.

Identification of Bioactive Solutions of Corn Silk (Zea mays L.) Extract and Biological Activity Test By Bioinformatics

Corn silk (Zea mays L.) is an agricultural by-product that is discarded as agricultural waste. Corn silk has secondary metabolite compounds that have the potential to become useful bioactive compounds. The purpose of this study is to recognize the bioactive potential of corn hair through compound identification and biological activity. The method used is preparation and extraction by maceration method followed by identification of compounds through LC-MS analysis method and Phytochemical test, bioinformatic PASS test based on data through PubChem, ChemSpider, and way2drug web. The results obtained from corn silk have a group of active compounds, including alkaloids, flavonoids, saponins, steroids and triterpenoids. Analysis of potential bioactivity received Acaciin compounds have high antioxidant bioactivity with a Pa value of 0.815, 2-arachidonoylglycerol has high potential antiinflammatory bioactivity with a Pa value of 0.736 and 1,2-di-O-methyl-4-[(2R)-2,4-dihydro butyramide shows moderate possible antibacterial activity with a Pa value of 0.643. This study concludes that corn silk has high bioactive potential as an antioxidant, anti-inflammatory and antibacterial.

Wild Silkworm Cocoon Waste Conversion into Tough Regenerated Silk Fibers by Solution Spinning.

Wild silkworm silk fibers have garnered attention owing to their softness, natural color, lightweight, and excellent mechanical properties. Because most wild silkworm cocoons obtained are pierced or dirty after the eclosion process, it is difficult to reel the long filament from the pierced cocoons to use as textile materials. Therefore, damaged wild silkworm cocoons are typically removed during the industrial process. Artificial silk spinning has been developed to transform domesticated silkworm silk solutions into regenerated silk fibers. However, regenerated fibers derived from wild silkworm silk have not been reported. Here, we produced regenerated silk fibers using a dry-wet spinning method using a dope solution derived from wild silkworm silk cocoon wastes. These regenerated silk fibers have thick and uniform diameters, unlike native silk fibers, contributing to their usefulness for sterilization and handling in medical applications. Moreover, they exhibited the same level of mechanical strength as their native counterparts. The molecular orientation and crystallinity of the regenerated silk fibers were adjustable by the drawing process, enabling the realization of their various tensile properties. This study promotes the utilization of unused protein resources to produce mechanically stable and tough silk-based fibers.

Instantaneous Formation of Silk Protein Aerosols and Fibers with a Portable Spray Device Under Ambient Conditions.

A variety of artificial silk spinning approaches have been attempted to mimic the natural spinning process found in silkworms and spiders, yet instantaneous silk fiber formation with hierarchical structure under physiological and ambient conditions without post-treatment procedures remains unaddressed. Here, we report a new strategy to fabricate silk protein-based aerosols and silk fibers instantaneously (< 1 s) in situ using a simple, portable, spray device, avoiding complicated and costly advanced manufacturing techniques. The key to success is the instantaneous conformational transition of silk fibroin from random coil to β-sheet right before spraying by mixing silk and polyethylene glycol (PEG) solutions in the spray device, allowing aerosols and silk fibers to be sprayed in situ, with further control achieved via the molecular weight of silk. The spinning process of the spray device is based on the use of green solvents, i.e., all steps of instant conformational transition of silk fibroin are carried out in aqueous conditions or with buffers at ambient conditions, in combination with shear and elongational flow caused by the hydraulic pressure generated in the spray container. The system supports a portable and user-friendly system that could be used for drug delivery carriers, wound coating materials and rapid silk fiber conformal coatings on surfaces.

Tuning the microstructure and mechanical properties of lyophilized silk scaffolds by pre-freezing treatment of silk hydrogel and silk solution

This work aims to understand how pre-freezing treatments (−20 °C, −80 °C or −196 °C (liquid nitrogen)) affect the microstructure, mechanical properties and secondary structure of silk scaffolds prepared from lyophilization of silk hydrogels and silk solutions. It is found that in comparison with silk solutions, silk hydrogels at the same silk fibroin concentrations produce scaffolds with more nanofibrous structures when they are pre-frozen at the different temperatures. Although pre-freezing with liquid nitrogen can produce nanofibrous scaffolds from either a silk solution (low concentration of 2%) or silk hydrogel (produced from 2 to 6% silk fibroin solutions), aligned macro-channels can be produced only from silk hydrogels. In addition, scaffolds obtained from silk hydrogels are dominated by β-sheets due to the crystallization process for gel network formation, while scaffolds prepared from silk solutions are largely amorphous. The findings of this work are important to tune the microstructure and mechanical properties of silk scaffolds.

Influence of lyophilization primary drying time and temperature on porous silk scaffold fabrication for biomedical applications.

Lyophilization of protein solutions, such as silk fibroin (silk), produces porous scaffolds useful for tissue engineering (TE). The impact of modifying lyophilization primary drying parameters on scaffold properties has not yet been explored previously. In this work, changes to primary drying duration and temperature were investigated using 3%, 6%, 9%, and 12% (w/v) silk solutions, via protocols labeled as Long Hold, Slow Ramp, and Standard. The 9% and 12% scaffolds were not successfully fabricated using the Standard protocol, while the Long Hold and Slow Ramp protocols resulted in scaffolds from all silk solution concentrations. Scaffolds fabricated using the Long Hold protocol had higher Young's moduli, smaller pore Feret diameters, and faster degradation. To investigate the utility of the different lyophilized scaffolds for in vitro cell culturing, the HepaRG liver cell line was cultured in the 3% to 12% scaffolds fabricated using the Long Hold protocol. The HepaRG cells grown in 3% scaffolds initially had greater lipid accumulation and metabolic activity than the other groups, although these differences were no longer apparent by Day 28. The deoxyribonucleic acid content of the HepaRG cells grown in 3% scaffold group was also initially significantly higher than the other groups. Significant differences in gene expression by 9% scaffolded HepaRG cells (CK19, HNFα) were seen on Day 14 while significant differences by 12% scaffolded HepaRG cells (ALB, APOA4) were seen on Day 28. Overall, modifying the primary drying parameters and silk concentration resulted in lyophilized scaffolds with tunable properties useful for TE applications.

Long-Term Preservation of SARS-CoV-2 RNA in Silk for Downstream RT-PCR Tests.

Positive controls made of viral gene components are essential to validate the performance of diagnostic assays for pathogens like severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, most of them are target-specific, limiting their application spectrum when validating assays beyond their specified targets. The use of an inactivated whole-virus RNA reference standard could be ideal, but RNA is a labile molecule that needs cold chain storage and transportation to preserve its integrity and activity. The cold chain process stretches the already dwindling storage capacities, incurs huge costs, and limits the distribution of reference materials to low-resource settings. To circumvent these issues, we developed an inactivated whole-virus SARS-CoV-2 RNA reference standard and studied its stability in silk fibroin matrices, i.e., silk solution (SS) and silk film (SF). Compared to preservation in nuclease-free water (ddH2O) and SS, SF was more stable and could preserve the SARS-CoV-2 RNA reference standard at room temperature for over 21 weeks (∼6 months) as determined by reverse transcription polymerase chain reaction (RT-PCR). The preserved RNA reference standard in SF was able to assess the limits of detection of four commercial SARS-CoV-2 RT-PCR assays. In addition, SF is compatible with RT-PCR reactions and can be used to preserve a reaction-ready primer and probe mix for RT-PCR at ambient temperatures without affecting their activity. Taken together, these results offer extensive flexibility and a simpler mechanism of preserving RNA reference materials for a long time at ambient temperatures of ≥25 °C, with the possibility of eliminating cold chains during storage and transportation.

Toughening Wet-Spun Silk Fibers by Silk Nanofiber Templating.

Regenerated silk fibers typically fall short of silkworm cocoon fibers in mechanical properties due to reduced fiber crystal structure and alignment. One approach to address this has been to employ inorganic materials as reinforcing agents. The present study avoids the need for synthetic additives, demonstrating the first use of exfoliated silk nanofibers to control silk solution crystallization, resulting in all-silk pseudocomposite fibers with remarkable mechanical properties. Incorporating only 0.06wt% silk nanofibers led to a ≈44% increase in tensile strength (over 600MPa) and ≈33% increase in toughness (over 200kJkg-1 ) compared with fibers without silk nanofibers. These remarkable properties can be attributed to nanofiber crystal seeding in conjunction with fiber draw. The crystallinity nearly doubled from ≈17% for fiber spun from pure silk solution to ≈30% for the silk nanofiber reinforced sample. The latter fiber also shows a high degree of crystal orientation with a Herman's orientation factor of 0.93, a value which approaches that of natural degummed B. mori silk cocoon fiber (0.96). This study provides a strong foundation to guide the development of simple, eco-friendly methods to spin regenerated silk with excellent properties and a hierarchical structure that mimics natural silk.

Electrodeposition as a facile way for the preparation of piezoelectric ultrathin silk film–based flexible nanogenerators

This article demonstrates the use of the electrodeposition technique as a viable alternative for depositing uniform, ultra-smooth thin films of silk fibroin (SF). Herein, the electrodeposition of silk fibroin films from a homogeneous silk solution prepared by dissolving SF in formic acid (FA) solution is demonstrated. Ion electrodiffusion is used as the mechanism for the silk deposition. Optimizations were carried out to increase the β-sheet crystallinity, which in turn would enhance the inherent piezoelectric response of silk thin films, an exquisite property of silk that is seldom explored. The piezoelectric coefficient (d33) of electrodeposited silk thin film was experimentally figured out to be 8.39 pm/V. As a proof of concept, silk-based flexible nanogenerator was fabricated and tested successfully. Silk-based nanogenerator is able to produce a maximum open-circuit voltage of 1.02 V (peak–to-peak value) and short-circuit current of 0.8 mA at bending state. The device also exhibits good stability and reliability in terms of electrical outputs.

Bioformulation of Silk-Based Coating to Preserve and Deliver Rhizobium tropici to Phaseolus vulgaris Under Saline Environments.

Seed priming has been for a long time an efficient application method of biofertilizers and biocontrol agents. Due to the quick degradation of the priming agents, this technique has been limited to specific immediate uses. With the increase of awareness of the importance of sustainable use of biofertilizers, seed coating has presented a competitive advantage regarding its ability to adhere easily to the seed, preserve the inoculant, and decompose in the soil. This study compared primed Phaseolus vulgaris seeds with Rhizobium tropici and trehalose with coated seeds using a silk solution mixed with R. tropici and trehalose. We represented the effect of priming and seed coating on seed germination and the development of seedlings by evaluating physiological and morphological parameters under different salinity levels (0, 20, 50, and 75 mM). Results showed that germination and morphological parameters have been significantly enhanced by applying R. tropici and trehalose. Seedlings of coated seeds show higher root density than the freshly primed seeds and the control. The physiological response has been evaluated through the stomatal conductance, the chlorophyll content, and the total phenolic compounds. The stability of these physiological traits indicated the role of trehalose in the protection of the photosystems of the plant under low and medium salinity levels. R. tropici and trehalose helped the plant mitigate the negative impact of salt stress on all traits. These findings represent an essential contribution to our understanding of stress responses in coated and primed seeds. This knowledge is essential to the design of coating materials optimized for stressed environments. However, further progress in this area of research must anticipate the development of coatings adapted to different stresses using micro and macro elements, bacteria, and fungi with a significant focus on biopolymers for sustainable agriculture and soil microbiome preservation.

Effect of Relative Humidity on the Electrospinning Performance of Regenerated Silk Solution.

Recently, the electrospun silk web has been intensively studied in terms of its biomedical applications, including tissue engineering scaffolds, due to its good biocompatibility, cytocompatibility, and biodegradability. In this study, the effect of relative humidity (RH) conditions on the morphology of electrospun silk fiber and the electrospinning production rate of silk solution was examined. In addition, the effect of RH on the molecular conformation of electrospun silk web was examined using Fourier transform infrared (FTIR) spectroscopy. As RH was increased, the maximum electrospinning rate of silk solution and fiber diameter of the resultant electrospun silk web were decreased. When RH was increased to 60%, some beads were observed, which showed that the electrospinnability of silk formic acid solution deteriorated with an increase in RH. The FTIR results showed that electrospun silk web was partially β-sheet crystallized and RH did not affect the molecular conformation of silk.

Mesoscale structure development reveals when a silkworm silk is spun

Silk fibre mechanical properties are attributed to the development of a multi-scale hierarchical structure during spinning. By careful ex vivo processing of a B. mori silkworm silk solution we arrest the spinning process, freezing-in mesoscale structures corresponding to three distinctive structure development stages; gelation, fibrilization and the consolidation phase identified in this work, a process highlighted by the emergence and extinction of ‘water pockets’. These transient water pockets are a manifestation of the interplay between protein dehydration, phase separation and nanofibril assembly, with their removal due to nanofibril coalescence during consolidation. We modeled and validated how post-draw improves mechanical properties and refines a silk’s hierarchical structure as a result of consolidation. These insights enable a better understanding of the sequence of events that occur during spinning, ultimately leading us to propose a robust definition of when a silkworm silk is actually ‘spun’.

Development of a mechanically matched silk scaffolded 3D clear cell renal cell carcinoma model

Development of a 3D, biomaterials-based model for clear cell renal cell carcinoma (ccRCC) would be advantageous for understanding disease progression in vitro. This study demonstrated the development of lyophilized silk scaffolds that mechanically match the experimentally determined Young's modulus for ex vivo ccRCC samples and normal kidney tissue. Scaffolds fabricated from silk solutions ranging from 3 to 12% (w/v) were evaluated through mechanical testing. Following mechanical characterization of ccRCC samples, it was demonstrated that 6% silk scaffolds mechanically matched ccRCC samples. No impact of pathological grade and stage on the calculated ccRCC modulus was observed and all tumors evaluated mechanically matched the 6% silk scaffold formulation. Stratifying tissue specimens based upon histological observations (e.g. evidence of high levels of collagen deposition) resulted in no significant differences between groups. To investigate the impact of a mechanically matched culturing environment on in vitro ccRCC disease characteristics a model ccRCC cell line, 786-O, was utilized. Scaffolded 786-O cells demonstrated increased lipid droplet accumulation, a hallmark of ccRCC, compared to standard two-dimensional (2D) culture conditions. Additionally, scaffolded 786-O cells demonstrated increased expression of genes associated with ccRCC aggressiveness (ex. VEGFA, TNF, and IL-6) or immune markers under investigation as therapeutic targets (ex. PDL1, CTLA4). Comparison with 786-O cells grown on non-mechanically matched scaffolds demonstrated that these improved ccRCC characteristics were driven by scaffold modulus. Overall, our findings support the use of silk scaffolds in replicating physiologic tumor behavior for clear cell renal cell carcinoma and provide a platform for investigating disease progression.

Effect of Silk Sericin Content on the Electrospun Silk Nanofibrous Membrane Property

Background: Silk sericin has a significant influence on the regenerated silk solution and silk-based materials property, while few reports were found to investigate this topic. Aim: The aim is to discuss the effect of silk sericin content on the electrospun silk nanofibrous membrane. Methods: Four degumming conditions (none degumming, boiling water degumming, 0.05 % Na2CO3 degumming, 0.5 % Na2CO3 degumming) were carried out for a systematic investigation in terms of (1) the silk sericin content after degumming; (2) the morphology of regenerated silk nanofibrous membrane was characterized by a Scanning Electron Microscope (SEM); and (3) structural properties of regenerated silk nanofibrous membrane by Fourier transform infrared (FTIR) spectroscopy, X-Ray Diffraction (XRD). Results and Conclusion: The results show that 0.5 % Na2CO3 degumming results in poor spinnability. The solutions derived from none degumming and boiling water degumming present high viscosity, leading to a hard silk nanofiber fabrication process. The silk nanofiber from the 0.05 % Na2CO3 degumming shows an easier fabrication process and better nanofiber morphology. These results will benefit the silk-based materials preparation, biomedical and separation application.

Spinning Regenerated Silk Fibers with Improved Toughness by Plasticizing with Low Molecular Weight Silk.

Low-molecular weight (LMW) silk was utilized as a LMW silk plasticizer for regenerated silk, generating weak physical crosslinks between high-molecular weight (HMW) silk chains in the amorphous regions of a mixed solution of HMW/LMW silk. The plasticization effect of LMW silk was investigated using mechanical testing, Raman spectroscopy, and wide-angle X-ray scattering (WAXS). Small amounts (10%) of LMW silk resulted in a 19.4% enhancement in fiber extensibility and 37.8% increase in toughness. The addition of the LMW silk facilitated the movement of HMW silk chains during drawing, resulting in an increase in molecular chain orientation when compared with silk spun from 100% HMW silk solution. The best regenerated silk fibers produced in this work had an orientation factor of 0.94 and crystallinity of 47.82%, close to the values of natural degummedBombyx mori silk fiber. The approach and mechanism elucidated here can facilitate artificial silk systems with enhanced properties.

The influence of metal ions on native silk rheology

Whilst flow is the basis for silk fibre formation, subtle changes in a silk feedstocks’ chemical environment may serve to increase both energetic efficiency and control hierarchical structure development during spinning. Despite the role of pH being largely understood, the influence of metal ions is not, only being inferred by correlative work and observations. Through a combination of rheology and microscopy, we provide a causative study of how the most abundant metal ions in the silk feedstock, Ca2+ and K+, affect its flow properties and structure. Our results show that Ca2+ ions increase viscosity and prevent molecular alignment and aggregation, providing ideal storage conditions for unspun silk. In contrast, the addition of K+ ions promotes molecular alignment and aggregation and therefore seems to transfer the silk feedstock into a spinning state which confirms recent ‘sticky reptation’ modelling hypotheses. Additionally, we characterised the influence of the ubiquitous kosmotropic agent Li+, used to prepare regenerated silk solutions, and find that it promotes molecular alignment and prevents aggregation which may permit a range of interesting artificial silk processing techniques to be developed. In summary, our results provide a clearer picture of how metal ions co-ordinate, control and thus contribute towards silk protein self-assembly which in turn can inspire structuring approaches in other biopolymer systems.

Microfluidic Silk Fibers with Aligned Hierarchical Microstructures.

The hierarchical structure of the ECM provides specific niches for tissues to regulate cell behavior, yet the challenge remains to design biomaterial systems for tissue regeneration to recreate such features in vitro. Here, we achieved this goal through the use of aligned hierarchical structures of native silk fibers, generated through the integration of "bottom-up" and "top-down" strategies to generate regenerated silk fibers with aligned nano- to micro-hierarchical structures. To achieve these designs, we assembled and dispersed silk nanofibers (SNF) in formic acid and spun them into fibers using bioinspired microfluidic chips with a geometry mimicking the native silk gland. The fibers generated using this device exhibited aligned hierarchical structure with fiber mechanical properties superior to fibers derived from more traditional spinning approaches with regenerated silk solutions. Besides the improved mechanical properties, Raman spectroscopic results indicated similarly aligned structures to native fibers and active control of cell proliferation, migration, and aggregate orientation. The results indicate the feasibility of developing bioactive silk fiber materials with hierarchical structures to facilitate utility in a range of cell and tissue regeneration scenarios.