Collagen Molecules Research Articles

The electron beam irradiation of collagen in the dry and gel states: The effect of the dose and water content from the primary to the quaternary levels

The aim of our study was to describe the impact of collagen in the gel and dry state to various doses of electron beam radiation (1, 10 and 25 kGy) which are using for food processing and sterilization. The changes in the chemical compositions (water, amino acids, lipids, glycosaminoglycans) were analyzed and the changes in the structure (triple-helix or β-sheet, the integrity of the collagen) were assessed. Subsequently, the impact of the applied doses on the mechanical properties, stability in the enzymatic environment, swelling and morphology were determined. The irradiated gels evinced enhanced degrees of cross-linking with only partial degradation. Nevertheless, an increase was observed in their stability manifested via a higher degree of resistance to the enzymatic environment, a reduction in swelling and, in terms of the mechanical behaviour, an approximation to the non-linear behavior of native tissues. In contrast, irradiation in the dry state exerted a somewhat negative impact on the observed properties and was manifested mainly via the scission of the collagen molecule and via a lower degree of stability in the aqueous and enzymatic environments. Neither the chemical composition nor the morphology was affected by irradiation.

Ultra-Rapid and Specific Gelation of Collagen Molecules for Transparent and Tough Gels by Transition Metal Complexation.

Collagen is the most abundant protein in the human body and one of the main components of stromal tissues in tumors which have a high elastic modulus of over 50kPa. Although collagen has been widely used as a cell culture scaffold for cancer cells, there have been limitations when attempting to fabricate a tough collagen gel with cells like a cancer stroma. Here, rapid gelation of a collagen solution within a few minutes by transition metal complexation is demonstrated. Type I collagen solution at neutral pH shows rapid gelation with a transparency of 81% and a high modulus of 1,781kPa by mixing with K2 PtCl4 solution within 3min. Other transition metal ions also show the same rapid gelation, but not basic metal ions. Interestingly, although type I to IV collagen molecules show rapid gelation, other extracellular matrices do not exhibit this phenomenon. Live imaging of colon cancer organoids in 3D culture indicates a collective migration property with modulating high elastic modulus, suggesting activation for metastasis progress. This technology will be useful as a new class of 3D culture for cells and organoids due to its facility for deep-live observation and mechanical stiffness adjustment.

Preparation and investigation of a novel antibacterial collagen-based material loaded with gentamicin following surface modification with citric acid for corneal tissue engineering

Corneal disease is an important clinical problem that affects millions of blind people and keratoplasty is currently the most successful treatment for corneal blindness. Unfortunately, there is a very high risk of bacterial infection during corneal transplantation. In this study, we proposed a novel synthetic collagen-based film for corneal therapy, and we effectively incorporated aminoglycoside gentamicin molecules onto the surface of the collagen film. We anticipate that this collagen-based substance will be antimicrobial and repair corneal tissue damage. Three steps were used to create this gentamicin-modified carboxylated collagen film, including: (i) Cross-link the collagen molecules with 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide and n-hydroxysuccinimide to create a collagen (Col) film. (ii) Citric acid was used to modify the Col film's surface in order to increase the number of carboxyl groups there (ColCA). (iii) Gentamicin molecules were grafted onto the surface of ColCA film by forming amide bonds (ColCA-GM). We discovered that this ColCA-GM film has good physicochemical properties and excellent biocompatibility. Furthermore, it was demonstrated that treating collagen films with citric acid significantly improved the antibacterial properties of ColCA-GM film. The outcomes point to a variety of potential applications for this novel film in corneal tissue engineering.

Structure, Stability and Binding Properties of Collagen-Binding Domains from Streptococcus mutans

Collagen-binding proteins (CBP), Cnm and Cbm, from Streptococcus mutans are involved in infective endocarditis caused by S. mutans because of their collagen-binding ability. In this study, we focused on the collagen-binding domain (CBD), which is responsible for the collagen-binding ability of CBP, and analyzed its structure, binding activity, and stability using CBD domain variants. The CBD consists of the N1 domain, linker, N2 domain, and latch (N1-N2~) as predicted from the amino acid sequences. The crystal structure of the Cnm/CBD was determined at a 1.81 Å resolution. N1_linker_N2 forms a ring structure that can enfold collagen molecules, and the latch interacts with N1 to form a ring clasp. N1 and N2 have similar immunoglobulin folds. The collagen-binding activities of Cbm/CBD and its domain variants were examined using ELISA. N1-N2~ bound to collagen with KD = 2.8 μM, and the latch-deleted variant (N1-N2) showed weaker binding (KD = 28 μM). The linker-deleted variant (N1N2~) and single-domain variants (N1 and N2) showed no binding activity, whereas the domain-swapped variant (N2-N1~) showed binding ability, indicating that the two N-domains and the linker are important for collagen binding. Thermal denaturation experiments showed that N1-N2 was slightly less stable than N1-N2~, and that N2 was more stable than N1. The results of this study provide a basis for the development of CBD inhibitors and applied research utilizing their collagen-binding ability.

Correlation between Mechanical Properties and Collagen Degeneration in Fibrous Tissue.

Fibrosis is a disease that causes abnormal accumulation of collagen and other extracellular matrix components. It can lead to organ failure and is responsible for one-third of all deaths worldwide. However, there is no cure for this disease, and the development of minimally invasive therapies is urgently needed. We have previously reported techniques for adjusting the shape and flexibility of fibrous tissue by traction while denaturing it with heat. However, studies comparing heat and traction on fibrous tissue are limited, so this paper examined that. Applying heat and traction to bovine Achilles tendon tissue has been shown to cause the denaturation of collagen molecules to accumulate in the tissue in response to these loads. Heat-induced collagen denaturation was nondirectional and omnidirectional, whereas mechanical stress-induced collagen denaturation was concentrated in the direction of traction. When both heat and traction were applied, collagen denaturation increased more than under a single load, indicating a synergistic effect.

Multifaced roles of desmoplastic reaction and fibrosis in pancreatic cancer progression: Current understanding and future directions.

Desmoplastic reaction is a fibrosis reaction that is characterized by a large amount of dense extracellular matrix (ECM) and dense fibrous stroma. Fibrotic stroma around the tumor has several different components, including myofibroblasts, collagen, and other ECM molecules. This stromal reaction is a natural response to the tissue injury process, and fibrosis formation is a key factor in pancreatic cancer development. The fibrotic stroma of pancreatic cancer is associated with tumor progression, metastasis, and poor prognosis. Reportedly, multiple processes are involved in fibrosis, which is largely associated with the upregulation of various cytokines, chemokines, matrix metalloproteinases, and other growth factors that promote tumor growth and metastasis. Fibrosis is also associated with immunosuppressive cell recruitment, such as regulatory T cells (Tregs) with suppressing function to antitumor immunity. Further, dense fibrosis restricts the flow of nutrients and oxygen to the tumor cells, which can contribute to drug resistance. Furthermore, the dense collagen matrix can act as a physical barrier to block the entry of drugs into the tumor, thereby further contributing to drug resistance. Thus, understanding the mechanism of desmoplastic reaction and fibrosis in pancreatic cancer will open an avenue to innovative medicine and improve the prognosis of patients suffering from this disease.

Interface‐Controlled Biomimetic Intrafibrillar Mineralization of Collagen: Effect of Ca2+/[PO4]3− Concentration Ratio

AbstractMineralized fibrils are important building blocks in bone tissue, formed by the hierarchical assembly of collagen molecules and crystalline hydroxyapatite (HAp). The mineralization pathway of HAp is reported as a nonclassical‐crystallization, but the nanoconfined crystallization in collagen fibrils remains poorly understood. The mechanism of intrafibrillar mineralization of collagen‐PDA fibrils in modified‐simulated body fluid (m‐SBF) solution is studied. Collagen‐amorphous calcium phosphate (ACP) fibrils are obtained by assembling collagen‐PDA fibrils with polyaspartic acid (pAsp) as a stabilizer. The ACP undergoes a phase transformation to HAp within the fibrils upon adjusting the phosphate concentration. It is found that the phase transformation of ACP to HAp in collagen fibrils can be accelerated with a 12 h incubation with 1/10 ratio of Ca2+ to [PO4]3−. A lower ratio of 1/1 and 1/5 results in a much slower phase transformation. This finding suggests that an elevated concentration of [PO4]3− is crucial for faster phase transformation. The relationship between the crystallization rate of HAp in the fibrils and the degree of mineralization is found to be linear in all cases, indicating an interface‐controlled process. This gives a better understanding of the mechanism of HAp mineralization in collagen fibrils, providing an effective approach to material design.

Cyanine dyes grafted onto collagen molecules for 3D printing of fluorescent biological scaffolds

In the photopolymerization 3D printing technology, preparing solid fluorescent biomaterials with fine structure is difficult due to the light barrier, aggregation quenching, and osmotic dissolution caused by adding fluorescent dyes, which severely limits their application in clinical practice. In this study, a bioactive fluorescence dye was prepared by coupling collagen type I and Cyanine IR-780 (Cyan) by dihydrolipoic acid (DHLA), in which 75.0% of primary amine of collagen was coupled with carboxyl of DHLA, and 48.4% of SH was coupled with Cyanine IR-780. The obtained collagen-based dye showed light stability under 405 nm LED light source and can be used in digital light processing-3D printing to fabricate hydrogel scaffolds containing bone collagen with fluorescent properties. The retained SH group in this macromolecular fluorescent dye can participate in the polymerization reaction in the photopolymerization process and improve the photopolymerization speed. With the formation of photopolymerized collagen polymer, the dye molecule shows fluorescence enhancement and antiaggregation caused quenching (anti-ACQ effect). The presented methods provide a foundation for the manufacturing of high-resolution solid 3D printed fluorescent structures, with direct relevance to biomedical optics and the broad adoption of fast manufacturing methods in fluorescence imaging.

Study on Interaction Mechanism between Neutral Salts and Collagen by Combining Experiments with Molecular Dynamics Simulation

The effect of salt on the collagen of hide/skin is of great significance in leather-making. However, the interaction between neutral salts and collagen has not been clear, since the microscopic interaction is hard to be observed directly from the macro level of hide/skin collagen. In this study, the collagen solutions in the typical neutral salts (NaCl, CaCl2, and Na2SO4) systems were used to explore the interaction mechanism between neutral salts and collagen via combining experiments with molecular dynamics (MD) simulation. The results of fluorescence measurements of pyrene, dynamic light scattering, atomic force microscopy, and isoelectric point suggested that the variation of the interaction between different neutral salts and collagen was accompanied with the changes in physicochemical properties of collagen. MD simulation further revealed more detailed information on the interaction mechanism between neutral salts and collagen at the molecular level. The computational results of non-bond energy of the collagen-salt model boxes indicated that the electrostatic interactions of different salts with collagen molecules had the order of CaCl2> Na2SO4> NaCl. The analyses of the visualized conformation and the radial distribution functions showed that CaCl2 with Ca2+ as contributing ion tended to form intramolecular salt bridges with collagen, while Na2SO4 with SO42-as contributing ion more likely formed salt bridges between collagen molecules in the shape of agglomerates. In contrast, NaCl with Cl-as contributing ion was scattered around the collagen models, and its effect on collagen was much smaller. The study elaborated the interaction mechanism of typical neutral salts and collagen to be helpful for further understanding and improving the use of neutral salts in many steps involved in leather production.

Strain-induced collagen denaturation is rate dependent in failure of cerebral arteries.

While soft tissues are commonly damaged by mechanical loading, the manifestation of this damage at the microstructural level is not fully understood. Specifically, while rate-induced stiffening has been previously observed in cerebral arteries, associated changes in microstructural damage patterns following high-rate loading are largely undefined. In this study, we stretched porcine middle cerebral arteries to failure at 0.01 and >150 s-1, both axially and circumferentially, followed by probing for denatured tropocollagen using collagen hybridizing peptide (CHP). We found that collagen fibrils aligned with the loading direction experienced less denaturation following failure tests at high than low rates. Others have demonstrated similar rate dependence in tropocollagen denaturation during soft tissue failure, but this is the first study to quantify this behavior using CHP and to report it for cerebral arteries. These findings may have significant implications for traumatic brain injury and intracranial balloon angioplasty. We additionally observed possible tropocollagen denaturation in vessel layers primarily composed of fibrils transversely aligned to the loading axis. To our knowledge, this is the first observation of collagen denaturation due to transverse loading, but further research is needed to confirm this finding. STATEMENT OF SIGNIFICANCE: Previous work shows that collagen hybridizing peptide (CHP) can be used to identify collagen molecule unfolding and denaturation in mechanically overloaded soft tissues, including the cerebral arteries. But experiments have not explored collagen damage at rates relevant to traumatic brain injury. In this work, we quantified collagen damage in cerebral arteries stretched to failure at both high and low rates. We found that the collagen molecule is less damaged at high than at low rates, suggesting that damage mechanisms of either the collagen molecule or other elements of the collagen superstructure are rate dependent. This work implies that arteries failed at high rates, such as in traumatic brain injury, will have different molecular-level damage patterns than arteries failed at low rates. Consequently, improved understanding of damage characteristics may be expanded in the future to better inform clinically relevant cases of collagen damage such as angioplasty and injury healing.

Bone beads enveloped with vascular endothelial cells for bone regenerative medicine.

The transplantation of pre-vascularized bone grafts is a promising strategy to improve the efficacy of engraftment and bone regeneration. We propose a hydrogel microbead-based approach for preparing vascularized and high-density tissue grafts. Mesenchymal stem cell-encapsulated collagen microgels (2µL), termed bone beads, were prepared through spontaneous constriction, which improved the density of the mesenchymal stem cells and collagen molecules by more than 15-fold from the initial day of culture. Constriction was attributed to cell-attractive forces and involved better osteogenic differentiation of mesenchymal stem cells than that of spheroids. This approach was scalable, and ∼2000 bone beads were prepared semi-automatically using a liquid dispenser and spinner flask. The mechanical stimuli in the spinner flask further improved the osteogenic differentiation of the mesenchymal stem cells in the bone beads compared with that in static culture. Vascular endothelial cells readily attach to and cover the surface of bone beads. The in vitro assembly of the endothelial cell-enveloped bone beads resulted in microchannel formation in the interspaces between the bone beads. Significant effects of endothelialization on in vivo bone regeneration were shown in rats with cranial bone defects. The use of endothelialized bone beads may be a scalable and robust approach for treating large bone defects. STATEMENT OF SIGNIFICANCE: A unique aspect of this study is that the hMSC-encapsulated collagen microgels were prepared through spontaneous constriction, leading to the enrichment of collagen and cell density. This constriction resulted in favorable microenvironments for the osteogenic differentiation of hMSCs, which is superior to conventional spheroid culture. The microgel beads were then enveloped with vascular endothelial cells and assembled to fabricate a tissue graft with vasculature in the interspaces among the beads. The significant effects of endothelialization on in vivo bone regeneration were clearly demonstrated in rats with cranial bone defects. We believe that microgel beads covered with vascular endothelial cells provide a promising approach for engineering better tissue grafts for bone-regenerative medicine.

Exploring the Interplay between Polyphenols and Lysyl Oxidase Enzymes for Maintaining Extracellular Matrix Homeostasis.

Collagen, the most abundant structural protein found in mammals, plays a vital role as a constituent of the extracellular matrix (ECM) that surrounds cells. Collagen fibrils are strengthened through the formation of covalent cross-links, which involve complex enzymatic and non-enzymatic reactions. Lysyl oxidase (LOX) is responsible for catalyzing the oxidative deamination of lysine and hydroxylysine residues, resulting in the production of aldehydes, allysine, and hydroxyallysine. These intermediates undergo spontaneous condensation reactions, leading to the formation of immature cross-links, which are the initial step in the development of mature covalent cross-links. Additionally, non-enzymatic glycation contributes to the formation of abnormal cross-linking in collagen fibrils. During glycation, specific lysine and arginine residues in collagen are modified by reducing sugars, leading to the creation of Advanced Glycation End-products (AGEs). These AGEs have been associated with changes in the mechanical properties of collagen fibers. Interestingly, various studies have reported that plant polyphenols possess amine oxidase-like activity and can act as potent inhibitors of protein glycation. This review article focuses on compiling the literature describing polyphenols with amine oxidase-like activity and antiglycation properties. Specifically, we explore the molecular mechanisms by which specific flavonoids impact or protect the normal collagen cross-linking process. Furthermore, we discuss how these dual activities can be harnessed to generate properly cross-linked collagen molecules, thereby promoting the stabilization of highly organized collagen fibrils.

Multifilament Collagen Fiber Bundles with Tendon-like Structure and Mechanical Performance.

Collagen multifilament bundles comprised of thousands of monofilaments are prepared by multipin contact drawing of an entangled polymer solution consisting of collagen and poly(ethylene oxide) (PEO). The multifilament bundles are hydrated in graded concentrations of PEO and phosphate buffered saline (PBS) to promote assembly of collagen fibrils within each monofilament while preserving the structure of the multifilament bundle. Multiscale structural characterization reveals that the hydrated multifilament bundle contains properly folded collagen molecules packed in collagen fibrils containing microfibrils, staggered by exactly one-sixth of the microfibril D-band spacing to produce a periodicity of 11nm. Sequence analysis predicts that in this structure, phenylalanine residues are close enough within and between microfibrils to become ultraviolet C (UVC) crosslinked. In agreement with this analysis, the ultimate tensile strength (UTS) and Young's modulus of the hydrated collagen multifilament bundles crosslinked by UVC radiation increase nonlinearly with total UVC energy to reach values in the range of native tendons without damage to the collagen molecules. This fabrication method recapitulates the structure of a tendon across multiple length scales and offers tunability in tensile properties using only collagen molecules and no other chemical additives in addition to PEO, which is almost entirely removed during the hydration process.

Advanced Glycation End Products as a Potential Target for Restructuring the Ovarian Cancer Microenvironment: A Pilot Study.

Ovarian cancer is the sixth leading cause of cancer-related death in women, and both occurrence and mortality are increased in women over the age of 60. There are documented age-related changes in the ovarian cancer microenvironment that have been shown to create a permissive metastatic niche, including the formation of advanced glycation end products, or AGEs, that form crosslinks between collagen molecules. Small molecules that disrupt AGEs, known as AGE breakers, have been examined in other diseases, but their efficacy in ovarian cancer has not been evaluated. The goal of this pilot study is to target age-related changes in the tumor microenvironment with the long-term aim of improving response to therapy in older patients. Here, we show that AGE breakers have the potential to change the omental collagen structure and modulate the peritoneal immune landscape, suggesting a potential use for AGE breakers in the treatment of ovarian cancer.

Mechanochemistry of Collagen.

The collagen molecular family is the result of nearly one billion years of biological evolution. It is a unique family of proteins, the majority of which provide general mechanical support to biological tissues. Fibril forming collagens are the most abundant collagens in vertebrate animals and are generally found in positions that resist tensile loading. In animals, cells produce fibril-forming collagen molecules that self-assemble into larger structures known as collagen fibrils. Collagen fibrils are the fundamental, continuous, load-bearing elements in connective tissues, but are often further aggregated into larger load-bearing structures, fascicles in tendon, lamellae in cornea and in intervertebral disk. We know that failure to form fibrillar collagen is embryonic lethal, and excessive collagen formation/growth (fibrosis) or uncontrolled enzymatic remodeling (type II collagen: osteoarthritis) is pathological. Collagen is thus critical to vertebrate viability and instrumental in maintaining efficient mechanical structures. However, despite decades of research, our understanding of collagen matrix formation is not complete, and we know still less about the detailed mechanisms that drive collagen remodeling, growth, and pathology. In this perspective, we examine the known role of mechanical force on the formation and development of collagenous structure. We then discuss a mechanochemical mechanism that has the potential to unify our understanding of collagenous tissue assembly dynamics, which preferentially deposits and grows collagen fibrils directly in the path of mechanical force, where the energetics should be dissuasive and where collagen fibrils are most required. We term this mechanism: Mechanochemical force-structure causality. STATEMENT OF SIGNIFICANCE: : Our mechanochemical-force structure causality postulate suggests that collagen molecules are components of mechanochemically-sensitive and dynamically-responsive fibrils. Collagen molecules assemble preferentially in the path of applied strain, can be grown in place by mechanical extension, and are retained in the path of force through strain-stabilization. The mechanisms that drive this behavior operate at the level of the molecules themselves and are encoded into the structure of the biomaterial. The concept might change our understanding of structure formation, enhance our ability to treat injuries, and accelerate the development of therapeutics to prevent pathologies such as fibrosis. We suggest that collagen is a mechanochemically responsive dynamic element designed to provide a substantial "material assist" in the construction of adaptive carriers of mechanical signals.

Mechanics of isolated individual collagen fibrils.

Collagen fibrils are the fundamental structural elements in vertebrate animals and compose a structural framework that provides mechanical support to load-bearing tissues. Understanding how these fibrils initially form and mechanically function has been the focus of a myriad of detailed investigations over the last few decades. From these studies a great amount of knowledge has been acquired as well as a number of new questions to consider. In this review, we examine the current state of our knowledge of the mechanical properties of extant fibrils. We emphasize on the mechanical response and related deformation of collagen fibrils upon tension, which is the predominant load imposed in most collagen-rich tissues. We also illuminate the gaps in knowledge originating from the intriguing results that the field is still trying to interpret. STATEMENT OF SIGNIFICANCE: Collagen is the result of millions of years of biological evolution and is a unique family of proteins, the majority of which provide mechanical support to biological tissues. Cells produce collagen molecules that self-assemble into larger structures, known as collagen fibrils. As simple as they appear under an optical microscope, collagen fibrils display a complex ultrastructural architecture tuned to the external forces that are imposed upon them. Even more complex is the way collagen fibrils deform under loading, and the nature of the mechanisms that drive their formation in the first place. Here, we present a cogent synthesis of the state-of-knowledge of collagen fibril mechanics. We focus on the information we have from in vitro experiments on individual, isolated from tissues, collagen fibrils and the knowledge available from in silico tests.

The Role of Integrin Receptor's α and β Subunits of Mouse Mesenchymal Stem Cells on the Interaction of Marine-Derived Blacktip Reef Shark (Carcharhinus melanopterus) Skin Collagen.

Marine collagen (MC) has recently attracted more attention in tissue engineering as a biomaterial substitute due to its significant role in cellular signaling mechanisms, especially in mesenchymal stem cells (MSCs). However, the actual signaling mechanism of MC in MSC growth, which is highly influenced by their molecular pattern, is poorly understood. Hence, we investigated the integrin receptors (α1β1, α2β1, α10β1, and α11β1) binding mechanism and proliferation of MCs (blacktip reef shark collagen (BSC) and blue shark collagen (SC)) compared to bovine collagen (BC) on MSCs behavior through functionalized collagen molecule probing for the first time. The results showed that BSC and SC had higher proliferation rates and accelerated scratch wound healing by increasing migratory rates of MSCs. Cell adhesion and spreading results demonstrated that MC had a better capacity to anchor MSCs and maintain cell morphology than controls. Living cell observations showed that BSC was gradually assembled by cells into the ECM network within 24 h. Interestingly, qRT-PCR and ELISA revealed that the proliferative effect of MC was triggered by interacting with specific integrin receptors such as α2β1, α10β1, and α11β1 of MSCs. Accordingly, BSC accelerated MSCs' growth, adhesion, shape, and spreading by interacting with specific integrin subunits (α2 and β1) and thereby triggering further signaling cascade mechanisms.

Characterization of the effect of chromium salts on tropocollagen molecules and molecular aggregates

Though the capability of chromium treatment to improve the stability and mechanical properties of collagen fibrils is well-known, the influence of different chromium salts on collagen molecules (tropocollagen) is not well characterized. In this study, the effect of Cr3+ treatment on the conformation and hydrodynamic properties of collagen was studied using atomic force microscopy (AFM) and dynamic light scattering (DLS). Statistical analysis of contours of adsorbed tropocollagen molecules using the two-dimensional worm-like chain model revealed a reduction of the persistence length (i.e., the increase of flexibility) from ≈72 nm in water to ≈56–57 nm in chromium (III) salt solutions. DLS studies demonstrated an increase of the hydrodynamic radius from ≈140 nm in water to ≈190 nm in chromium (III) salt solutions, which is associated with protein aggregation. The kinetics of collagen aggregation was shown to be ionic strength dependent. Collagen molecules treated with three different chromium (III) salts demonstrated similar properties such as flexibility, aggregation kinetics, and susceptibility to enzymatic cleavage. The observed effects are explained by a model that considers the formation of chromium-associated intra- and intermolecular crosslinks. The obtained results provide novel insights into the effect of chromium salts on the conformation and properties of tropocollagen molecules.

3D Tortuosity and Diffusion Characterization in the Human Mineralized Collagen Fibril Using a Random Walk Model.

Bone tissue is mainly composed at the nanoscale of apatite minerals, collagen molecules and water that form the mineralized collagen fibril (MCF). In this work, we developed a 3D random walk model to investigate the influence of bone nanostructure on water diffusion. We computed 1000 random walk trajectories of water molecules within the MCF geometric model. An important parameter to analyse transport behaviour in porous media is tortuosity, computed as the ratio between the effective path length and the straight-line distance between initial and final points. The diffusion coefficient is determined from the linear fit of the mean squared displacement of water molecules as a function of time. To achieve more insight into the diffusion phenomenon within MCF, we estimated the tortuosity and diffusivity at different quotes in the longitudinal direction of the model. Tortuosity is characterized by increasing values in the longitudinal direction. As expected, the diffusion coefficient decreases as tortuosity increases. Diffusivity outcomes confirm the findings achieved by experimental investigations. The computational model provides insights into the relation between the MCF structure and mass transport behaviour that may contribute to the improvement of bone-mimicking scaffolds.

Exploration of 2D and 2.5D Conformational Designs Applied on Epoxide/Collagen-Based Integrative Biointerfaces with Device/Tissue Heterogeneous Affinity.

Collagen and multifunctional epoxides, which are respectively the common constituents of natural and polymer interfaces, were combined to fabricate integrative biointerfaces with device/tissue heterogeneous affinity. Further, the traditional 2D and advanced 2.5D conformational designs were achieved on collagen-based biointerfaces. The 2D conformational biointerfaces were formed by the self-entanglement of collagen molecules based on extensive hydrogen bonds among molecules, and the lamellar structures of 2D conformational biointerfaces could act as barriers to protect both biointerfaces and substrates from enzymes and corrosion. The unique stacking structures of 2.5D conformational biointerfaces were formed by cross-linking microaggregates that were established and connected by epoxy cross-linking bonds and provided the extra 0.5D degree of freedom on structure design and functional specialization through artificially manipulating the constituents and density of microaggregates. Besides, the intersecting channels among microaggregates gave 2.5D biointerfaces diffusion behaviors, which further brought good wettability and biodegradability. The integrative biointerfaces behaved well on cell viability and enhanced the cell adhesion strength in vitro, which could be attributed to the collaborations of collagen and epoxy groups. The subcutaneous implant model in rats was utilized to investigate soft tissue response, and the results demonstrated that the tissues around implantation areas healed well and without calcification or infection. The coating of integrative biointerfaces alleviated the fibrosis around implantation areas, and the inflammatory responses and foreign body reactions were improved.