Modulation of Biophysical Cues in Nature Inspired Patterning of Porous Silk Fibroin Scaffold for Replenishable Controlled Drug Delivery.

While a sticking plasteris enough for healing of most of the minor cuts they may get routinely, critical situations like surgical, gunshot, accidental or diabetic wounds;lacarations and other cutaneous deep cuts may require implants and simultaneous medications for healing. From the biophysical standpoint, an internal force-based physical surface stimulusis crucial for cellular sensing during wound repair. In this paper, the authors report the fabrication of a porous, biomimmetically patterned silk fibroin scaffold loaded with ampicillin, which exhibits controlled release of the drug along with possible replenishment of the same. In vitro swelling study reveals that the scaffolds with hierarchical surface patterns exhibit lower swelling and degradation than other types of scaffolds. The scaffolds, that show remarkable broad-spectrum antibacterial efficacy, exhibit Korsemeyer-Peppas model for the ampicillin release patterns due to the structural hydrophobicity imparted by the patterns. Four distinct cell-matrix adhesion regimes are investigated for the fibroblasts to eventually form cell sheets all over the hierarchical surface structures. 4',6-diamidino-2-phenylindole (DAPI) and Fluorescein Diacetate (FDA) fluorescent staining clearly demonstrate the superiority of patterned surface over its other variants. A comparative immunofluorescence study among collagen I, vinculin, and vimentin expressions substantiated the patterned surface to be superior to others.

Droplet Impact Dynamics on Biomimetic Replica of Yellow Rose Petals: Rebound to Micropinning Transition.

Rose petals exhibit a phenomenal wetting property of being sticky and superhydrophobic simultaneously. A recent study has shown that for short timescales, associated with drop impact phenomenon, lotus leaf and rose petal replicas exhibit similar wettability, thereby highlighting the difference between long and short time wettability. Also, short time wetting on rose petals of different colors remains completely unaddressed, as almost all existing study on wetting of rose petals have been performed with the classical red rose (Rosa chinensis). In this paper, we compare the drop impact studies on replicas of a yellow rose petal, with those on extensively studied red rose petal replicas and the lotus leaf over a wide range of Weber number (We), by varying the height of fall (h) from 10 to 375 mm. Our results reveal that over the replica of a yellow rose petal, the initial impact outcome varies from complete rebound to micro pinning and eventually complete pinning depending on the kinetic energy of the impacting drop, in contrast to that on red rose petal replica on which the droplet always pinned. Based on experimental finding, we present a comprehensive regime phase map of the post impact behavior of the drop on different surfaces as a function of impact height. We also present a simple scaling analysis to understand the combined effect of pattern height and periodicity on the critical h corresponding to wetting regime transition. Additionally, variation of maximum spreading diameter and spreading time with the h for the different surfaces is also discussed. The results highlight that the initial impact dynamics of a water drop over a topographically patterned substrate is a strong function of the topographical parameters and can be very different from the equilibrium wetting state.

Chitosan-collagen-fibrinogen uncrosslinked scaffolds possessing skin regeneration and vascularization potential.

Clinical success of regenerative medicine for treating deep-tissue skin injuries depends on the availability of skin grafts. Though bioengineered constructs are tested clinically, lack of neovascularization provide only superficial healing. Thus constructs, which promotes wound healing and supports vascularization has gained priority in tissue engineering. In this study, chitosan-collagen-fibrinogen (CCF) scaffold was fabricated using freeze-drying method without using any chemical crosslinkers. CCF scaffolds proved cytocompatibility and faster healing in in vitro scratch assay of primary human adult dermal fibroblasts cells with progressively increasing vascular endothelial growth factor-A and reducing vascular endothelial growth factor receptor 1 expressions. Skin regeneration evaluated on in vivo full thickness wound model confirmed faster remodeling with angiogenic signatures in CCF scaffold-implanted mice. Histopathological observations corroborated with stereo-zoom and SS-optical coherence tomography images of wound sites to prove the maturation of healing-bed, after 12 days of CCF implantation. Therefore, it is concluded that CCF scaffolds are promising for skin tissue regeneration and demonstrates pro-angiogenic potential.

Regenerative repair of full thickness skin wound assisted by dual crosslinking percolative gel casting maneuvered alginate hydrogel embedded with honey ghee blend resembles standard cutaneous properties.

We report synergism in scarless cutaneous wound repair by alginate hydrogel (HGSAG) embedded with an optimized blend of characterized Jamun honey and characterized indigenously prepared ghee. Thorough screening and characterization of honey and ghee are carried out followed by obtaining a novel dual crosslinking percolative gel casting fabrication method to come up with HGSAG showing superior chemical stability, and mechanical strength (Nanoindentation study; lowest stiffness: 0.71 ± 0.19 µN/nm), and surface morphology (SEM; highest roughness: 0.13 ± 0.04 µm) to other variants. In vitro swelling study and degradation behavior study show intermediate swelling (swelling index: 0.59 ± 0.008 in 98 h) and required restricted degradation (PBS: 73.38 ± 0.55%, DMEM: 83.48 ± 0.69% in 10 days) for HGSAG which is necessary for providing nutrients to cells and in vivo therapeutic efficacy. We observe the remarkable antibacterial efficacy of HGSAG against Staphylococcus mutans and Escherichia coli. This particular substrate also shows decent 3T3 fibroblasts viability, cell-cell communication followed by cell-matrix interaction, and proliferation compared to other variants. Molecular gene expression studies by quantitative RT-PCR technique reveal strong upregulation of collagen I, CD26, and TGF-ß3 while downregulation in the case of TGF-ß1 which eventually substantiates scarless wound healing potential of HGSAG. Wound closure kinetics is most rapidly and successfully underpinned by HGSAG while compared to other alternatives including marketed healing patches. Regular close monitoring using histopathological studies and real-time imaging by Swept-Source Optical Coherence Tomography of in vivo wound model treated with HGSAG come up with the fascinating result of scarless healing (HGSAG treated epithelial thickness: 62.96 ± 0.67 µm, unwounded akin epithelial thickness: 62.56 ± 0.34 µm) within 12 days of wounding. Thus, the work highlights modified and stabilized alginate hydrogel embedded with honey and ghee blend as a potential scarless full-thickness cutaneous wound healing bio-scaffold.

Amyloids Formed by Nonaromatic Amino Acid Methionine and Its Cross with Phenylalanine Significantly Affects Phospholipid Vesicle Membrane: An Insight into Hypermethioninemia Disorder.

The incorrect metabolic breakdown of the nonaromatic amino acid methionine (Met) leads to the disorder called hypermethioninemia via an unknown mechanism. To understand the molecular level pathogenesis of this disorder, we prepared a DMPC lipid membrane, the mimicking setup of the cell membrane, and explored the effect of the millimolar level of Met on it. We found that Met forms toxic fibrillar aggregates that disrupt the rigidity of the membrane bilayer, and increases the dynamic response of water molecules surrounding the membrane as well as the heterogeneity of the membrane. Such aggregates strongly deform red blood cells. This opens the requirement to consider therapeutic antagonists either to resist or to inhibit the toxic amyloid aggregates against hypermethioninemia. Moreover, such disrupting effect on membrane bilayer and cytotoxicity along with deformation effect on RBC by the cross amyloids of Met and Phenylalanine (Phe) was found to be most virulent. This exclusive observation of the enhanced virulent effect of the cross amyloids is expected to be an informative asset to explain the coexistence of two amyloid disorders.

Cross-sectional visual comparison of remineralization efficacy of various agents on early smooth surface caries of primary teeth with swept source optical coherence tomography.

PURPOSE: Sweptsource optical coherence tomography (SS-OCT) permits cross-sectional observation of surface/subsurface characteristics of enamel including early carious lesions (ECL) or remineralization non-invasively.This study aimed to visually compare the cross-sectional remineralizing efficacy of various agents on ICDAS-II scores-1&2 by using SS-OCT and histology. METHODS: Baseline SS-OCT (grey-scale/false-colour) and histology was performed on the randomly selected two samples with scores-1&2. Four remineralizing agents [fluoride-varnish (FV), CPP-ACP, nanohydroxy-paste (NHP) and silver-diamine-fluoride (SDF)]were evaluated for 2-or 6-weeks post-remineralization using SS-OCT and histology. RESULTS: Score-1&2 baseline SS-OCT images showed a linear-shaped demineralization with dentino-enamel junction (DEJ) visible; and bowl-shaped demineralization with DEJ invisible respectively. Remineralizing agents were assessed on the basis of their ability to remineralize the surface, subsurface and made visualize the DEJ in score-2. SS-OCT showed an outer growth layer in post-remineralization score-1, 2-weeks samples with FV and NHP. All the agents showed progressive subsurface remineralization in 6 weeks. Active lesions showed rapid uptake of minerals on surface. Subsurface mineralization in pigmented score-2 matched sound enamel with NHP and SDF. Surface remineralization was comparable in FV and SDF followed by NHP. SDF demonstrated deeper subsurface remineralization followed by NHP and CPP-ACP. CONCLUSION: SS-OCT images correlated to histology. SS-OCT could monitor surface/subsurface in-situde/remineralization activity non-invasively.

Improved Mesenchymal Stem Cell Proliferation, Differentiation, Epithelial Transition, and Restrained Senescence on Hierarchically Patterned Porous Honey Silk Fibroin Scaffolds.

We report a significant improvement of adipose-derived mesenchymal stem cells' (ADMSCs) biocompatibility and proliferation on hierarchically patterned porous honey-incorporated silk fibroin scaffolds fabricated using a combination of soft lithography and freeze-drying techniques. Parametric variations show enhanced surface roughness, swelling, and degradation rate with good pore interconnectivity, porosity, and mechanical strength for soft-lithographically fabricated biomimetic microdome arrays on the 2% honey silk fibroin scaffold (PHSF2) as compared to its other variants, which eventually made PHSF2 more comparable to the native environment required for stem cell adhesion and proliferation. PHSF2 also exhibits sustained honey release with remarkable antibacterial efficacy against methicillin-resistant Staphylococcus aureus (MRSA). Honey incorporation (biochemical cue) influences microdome structural features, that is, biophysical cues (height, width, and periodicity), which further allows ADMSCs pseudopods (filopodia) to grasp the microdomes for efficient cell-cell communication and cell-matrix interaction and regulates ADMSCs behavior by altering their cytoskeletal rearrangement and thereby increases the cellular spreading area and cell sheet formation. The synergistic effect of biochemical (honey) and biophysical (patterns) cues on ADMSCs studied by the nitro blue tetrazolium assay and DCFDA fluorescence spectroscopy reveals limited free radical generation within cells. Molecular expression studies show a decrease in p53 and p21 expressions validating ADMSCs senescence inhibition, which is further correlated with a decrease in cellular senescence-associated ß galactosidase activity. We also show that an increase in CDH1 and CK19 molecular expressions along with an increase in SOX9, RUNX2, and PPARγ molecular expressions supported by PHSF2 justify the substrate's efficacy of underpinning mesenchymal to epithelial transition and multilineage trans-differentiation. This work highlights the fabrication of a naturally healing nutraceutical (honey)-embedded patterned porous stand-alone tool with the potential to be used as smart stem cells delivering regenerative healing implant.

Dual cross-linked honey coupled 3D antimicrobial alginate hydrogels for cutaneous wound healing.

We report potentiation of healing efficacy of alginate by value addition at its structural level. Dual crosslinked (ionically and covalently) sodium alginate hydrogel coupled with honey (HSAG) brings about an intermediate stiffness in the fabric, confers consistent swelling property and limits erratic degradation of the polymer which ultimately provides conducive milieu to cellular growth and proliferation. In this work honey concentrations in HSAGs are varied from 2% to 10%. FTIR, XRD and nanoindentation studies on the HSAGs exhibited physicochemical integrity. In vitro degradation study provided the crucial finding on 4% HSAG having controlled degradation rate up to 12 days with a weight loss of 87.36 ± 1.14%. This particular substrate also has an ordered crystalline surface morphology with decent cellular viability (HaCaT and 3T3) and antimicrobial potential against Methicillin Resistant Staphylococcus aureus (MRSA) and Escherichia coli. The in vivo wound contraction kinetics on murine models (4% HSAG treated wound contraction: 94.56 ± 0.1%) has been monitored by both invasive (histopathology) and noninvasive (Swept Source Optical Coherence Tomography) imaging and upon corroborating them it evidenced that 4% HSAG treated wound closure achieved epithelial thickness resembling to that of unwounded skin. Thus, the work highlights structurally modified alginate hydrogel embedded with honey as a potential antimicrobial healing agent.