Polymer-induced Liquid-precursor Research Articles

Promotion effect of proanthocyanidin on dentin remineralization via the polymer induced liquid precursor process

Proanthocyanidin (PA) has demonstrated promise as a dental biomodifier for maintaining dentin collagen integrity, yet there is limited evidence regarding its efficacy in dentin repair. The aim of this study was to investigate the effect of PA on dentin remineralization through the polymer induced liquid precursor (PILP) process, as well as to assess the mechanical properties of the restored dentin. Demineralized dentin was treated with a PA-contained remineralization medium, resulting in the formation of PA-amorphous calcium phosphate (ACP) nanoparticles via the PILP process. The kinetics and microstructure of remineralized dentin were examined through the use of Fourier transform infrared spectroscopy(FTIR), attenuated total reflectance-FTIR, scanning electron microscopy, transmission electron microscopy. The results showed that the application of PA facilitated the process of dentin remineralization, achieving completion within 48 h, demonstrating a notable reduction in time required. Following remineralization, the mechanical properties of the dentin exhibited an elastic modulus of 15.89 ± 1.70 GPa and a hardness of 0.47 ± 0.08 GPa, which were similar to those of natural dentin. These findings suggest that combining PA with the PILP process can promote dentin remineralization and improve its mechanical properties, offering a promising new approach for dentin repair in clinical practice.

A Novel Biomineralized Collagen Liquid Crystal Hydrogel Possessing Bone-like Nanostructures by Complete In Vitro Fabrication.

The microstructure of bone consists of nano-hydroxyapatite (nano-HA) crystals aligned within the interspaces of collagen fibrils. To emulate this unique microstructure of bone, this work applied two biomimetic techniques to obtain bone-like microstructures in vitro, that is, combining the construction of collagen liquid crystal hydrogel (CLCH) with the application of a polymer-induced liquid precursor (PILP) mineralization process. Upon the elevation of pH, the collagen macromolecules within the collagen liquid crystal (CLC) were activated to self-assemble into CLCH, whose fibrils packed into a long and dense fiber bundle in high orientation, emulating the dense-packed matrix of bone. We demonstrated that the fibrillar mineralization of CLCH, leading to a bone-like nanostructured inorganic material part, can be achieved using the PILP crystallization process to pre-mineralize the dense collagen substrates of CLCH with CaCO3, immediately followed by the in situ mineral phase transformation of CaCO3 into weak-crystalline nano-HA. The combination of CLCH with the biomineralization process of PILP, together with the mineral phase transformation, achieved the in vitro simulation of the nanostructures of both the organic extracellular matrix (ECM) and inorganic ECM of bone. This design would constitute a novel idea for the design of three-dimension biomimetic bone-like material blocks for clinical needs.

Calcium phosphate nanoclusters modify periodontium remodeling and minimize orthodontic relapse.

Orthodontic relapse is one of the most prevalent concerns of orthodontic therapy. Relapse results in patients' teeth reverting towards their pretreatment positions, which increases the susceptibility to functional problems, dental disease, and substantially increases the financial burden for retreatment. This phenomenon is thought to be induced by rapid remodeling of the periodontal ligament (PDL) in the early stages and poor bone quality in the later stages. Current therapies, including fixed or removable retainers and fiberotomies, have limitations with patient compliance and invasiveness. Approaches using biocompatible biomaterials, such as calcium phosphate polymer-induced liquid precursors (PILP), is an ideal translational approach for minimizing orthodontic relapse. Here, post-orthodontic relapse is reduced after a single injection of high concentration PILP (HC-PILP) nanoclusters by altering PDL remodeling in the early stage of relapse and improving trabecular bone quality in the later phase. HC-PILP nanoclusters are achieved by using high molecular weight poly aspartic acid (PASP, 14 kDa) and poly acrylic acid (PAA, 450 kDa), which resulted in a stable solution of high calcium and phosphate concentrations without premature precipitation. In vitro results show that HC-PILP nanoclusters prevented collagen type-I mineralization, which is essential for the tooth-periodontal ligament (PDL)-bone interphase. In vivo experiments show that the PILP nanoclusters minimize relapse and improve the trabecular bone quality in the late stages of relapse. Interestingly, PILP nanoclusters also altered the remodeling of the PDL collagen during the early stages of relapse. Further in vitro experiments showed that PILP nanoclusters alter the fibrillogenesis of collagen type-I by impacting the protein secondary structure. These findings propose a novel approach for treating orthodontic relapse and provide additional insight into the PILP nanocluster's structure and properties on collagenous structure repair.

In Situ Construction of Morphologically Different Hydroxyapatite-Mineralized Structures on a Three-Dimensional Bionic Chitin Scaffold.

Slow healing at the tendon-bone interface is a prominent factor in the failure of tendon repair surgeries. The development of functional biomaterials with 3D gradient structures is urgently needed to improve tendon-bone integration. The crystalline form of hydroxyapatite (HAP) has a crucial impact on cell behavior, which directly influences protein adsorption, such as bone morphogenetic protein 2, the adhesion, proliferation, and osteogenic differentiation with cells. This work aimed to generate gradient mineral structures in situ by stabilizing calcium and phosphate ions using a polymer-induced liquid precursor process. To regulate the crystalline growth of HAP at the interface of β-chitin, this work made use of the surface properties of the organic matrix found in cuttlefish bone. These techniques allowed us to prepare an organic-inorganic composite gradient scaffold comprising plate-like HAP mineralized in situ on the surface of the scaffold and fibrous HAP in the scaffold's interior. Organic-inorganic composite gradient materials are anticipated for use in tendon-bone healing produced via the in situ construction of gradient-distributed HAP mineralization layers having varying crystalline morphologies on chitin scaffolds that possess a three-dimensional bionic structure.

Intrafibrillar mineralization of type I collagen with calcium carbonate and strontium carbonate induced by polyelectrolyte-cation complexes.

Calcium carbonate (CaCO3), possessing excellent biocompatibility, bioactivity, osteoconductivity and superior biodegradability, may serve as an alternative to hydroxyapatite (HAp), the natural inorganic component of bone and dentin. Intrafibrillar mineralization of collagen with CaCO3 was achieved through the polymer-induced liquid precursor (PILP) process for at least 2 days. This study aims to propose a novel pathway for rapid intrafibrillar mineralization with CaCO3 by sequential application of the carbonate-bicarbonate buffer and polyaspartic acid (pAsp)-Ca suspension. Fourier transform infrared (FTIR) spectroscopy, zeta potential measurements, atomic force microscopy/Kelvin probe force microscopy (AFM/KPFM), and three-dimensional stochastic optical reconstruction microscopy (3D STORM) demonstrated that the carbonate-bicarbonate buffer significantly decreased the surface potential of collagen and CO32-/HCO3- ions could attach to collagen fibrils via hydrogen bonds. The electropositive pAsp-Ca complexes and free Ca2+ ions are attracted to and interact with CO32-/HCO3- ions through electrostatic attractions to form amorphous calcium carbonate that crystallizes gradually. Moreover, like CaCO3, strontium carbonate (SrCO3) can deposit inside the collagen fibrils through this pathway. The CaCO3-mineralized collagen gels exhibited better biocompatibility and cell proliferation ability than SrCO3. This study provides a feasible strategy for rapid collagen mineralization with CaCO3 and SrCO3, as well as elucidating the tissue engineering of CaCO3-based biomineralized materials.

Fatigue and fracture of soft collagenous tissues mineralized in vitro

Biomineralization occurs in natural organisms, such as bones, teeth, shells and tendons. The formation mechanism and mechanical properties have been extensively studied for mineralized hard tissues, especially for bone and teeth. However, the research on the evolution of mechanical properties for mineralized soft tissues remains challenging due to the long period of mineralization. Here we use polymer-induced liquid-precursor (PILP) method to achieve accelerated in vitro mineralization of bovine pericardium with SrCO3. The modulus, tensile strength, toughness, and suture retention force of mineralized tissues have been characterized. After 30 days of mineralization, both the tensile strength and toughness decrease by two orders of magnitude. We carry out cyclic suture retention test to characterize the fatigue properties of mineralized tissues, and find that the cycle to failure of mineralized tissues is 10-100 times lower than that of unmineralized tissues. The degradation of mechanical properties results from the microstructure change of mineralized tissues, where collagen fibers change from curled to straight configuration. After mineralization, the interfibrous slippage is limited, and the fracture mode changes from fiber pullout to fiber break. Finite element analysis has been carried out to simulate the fracture and suture retention properties, and the simulation can well reproduce experimental results. The study may provide insights for tissue replacement and repairing.

Polymer-induced liquid precursors: How are they being used and what is the effect on dentin remineralization? A scoping review

To identify the effectiveness of polymer-induced liquid precursors (PILP) on dentin remineralization and the assessment approaches used to evaluate remineralization. The analyses were done into six stages: (1) Identifying the research question; (2) Identifying the studies; (3) Selecting relevant studies; (4) Plotting the data; (5) Reporting results; and (6) Risk of bias. The searches were carried out in the following databases: PubMed, Web of Science, and Scopus. A total of 247 articles were identified in the electronic database search. After applying the eligibility criteria, only 12 articles were included for data extraction. The outcomes of dentin remineralization patterns were plotted in association to PILP agent type and application mode. The intrafibrillar mineralization occurred in 78% of studies where polyaspartic acid (pAsp) was used, and 50% when the polyacrylic acid (PAA) was used. All the studies where PILP was used in a restorative material (resin composite, glass ionomer, and adhesive system)—extra and intrafibrillar mineralization pattern was found. The association of PILP agents with other materials with potential for mineralization showed beneficial results in remineralization, since they provide a medium to high concentration of calcium and phosphate. The pAsp showed better results for interfibrillar remineralization when compared to PAA. The association with Ca/P release materials or solution Ca P are essentials for mineralization via PILP agent. Further studies are needed to assess the effectiveness of remineralization through PILP agents because the level of evidence of the studies was low.

Biomimetic spatial control of Liesegang patterns of calcium phosphate in hydrogel employing sucrose and glycine

The organic-inorganic interactions are nature’s way of controlling the formation of biominerals. Of the biominerals, calcium phosphate (CaP) phases have been implicated in the structure of hard tissues such as teeth and bones. Hydroxyapatite (HA) is known to be the most stable and abundant CaP in hard tissues. Formation of HA takes place in a stepwise manner through amorphous precursors via a process called the PILP (polymer-induced liquid precursor). The present work aims to elaborate on the mechanism of HA formation via PILP using Liesegang band formation. Periodically precipitating (the Liesegang patterns) systems with varying concentrations of the precursors of CaP formation were employed to understand its crystallization in the presence of additives such as sucrose and glycine. The precipitate was characterized using various techniques such as image analysis, IR, SEM, and PXRD. The spatiotemporal analysis showed that the spacing coefficient (p) trend was pglycine>pcontrol>psucrose. It was revealed that the HA was the dominant CaP present in all the samples but with varying degrees of crystallinity. The systems containing sucrose have more crystallinity than that of glycine. In all the systems, the crystallinity went on decreasing from older rings formed at the top to newer rings formed at the bottom of the tubes, which implies the formation of amorphous liquid-like phase(s) as the intermediates. Based on the observations, a mechanism was proposed for the pre-and postponement of the bands formed in the Liesegang rings.

Synergistic intrafibrillar/extrafibrillar mineralization of collagen fibrils and scaffolds enhanced by introducing polyacrylamide to PILP for osteogenic differentiation

AbstractMineralization can help improve the mechanical properties and degradation rates of collagen scaffolds while ensuring good biocompatibility, thereby providing a suitable microenvironment for osteogenic differentiation. Intrafibrillar/extrafibrillar mineralization of collagen is promoted by polymer‐induced liquid precursors (PILP). In this study, polyacrylamide (PAM) was introduced to PILP which synergistically mineralized collagen scaffold intrafibrillarly and extrafibrillarly. PAM and polyacrylic acid (PAA) can stabilize the amorphous calcium phosphate (ACP), to form a PILP of PAM and PAA/ACP. As a control, another scaffold material was formed using the conventional mineralization method, that is, soaking collagen in a simulated body fluid. Collagen mineralization was characterized using SEM and TEM. After 7 days of mineralization with PILP, intrafibrillar crystallization of collagen was significantly higher than that in the control group, and the stiffness and modulus of this scaffold material significantly increased. Cellular experiment results indicated that the PILP‐mineralized collagen scaffold was biocompatible and promoted the osteogenic differentiation of MC3T3‐E1 pre‐osteoblasts. Biomimetic mineralization by PILP will assist the fabrication of mineralized collagen scaffold for bone repair applications.

A novel thieno[2,3-d]pyrimidine derivative inhibiting vascular endothelial growth factor receptor-2: A story of computer-aided drug discovery.

Following the pharmacophoric features of vascular endothelial growth factor receptor 2 (VEGFR-2) inhibitors, a novel thieno[2,3-d]pyrimidine derivative has been designed and its activity against VEGFR-2 has been demonstrated by molecular docking studies that showed an accurate binding mode and an excellent binding energy. Furthermore, the recorded binding was confirmed by a series of molecular dynamics simulation studies, which also revealed precise energetic, conformational, and dynamic changes. Additionally, molecular mechanics with generalized Born and surface area solvation and polymer-induced liquid precursors studies were conducted and verified the results of the MD simulations. Next, in silico absorption, distribution, metabolism, excretion, and toxicity studies have also been conducted to examine the general drug-like nature of the designed candidate. According to the previous results, the thieno[2,3-d]pyrimidine derivative was synthesized. Fascinatingly, it inhibited VEGFR-2 (IC50 = 68.13 nM) and demonstrated strong inhibitory activity toward human liver (HepG2), and prostate (PC3) cell lines with IC50 values of 6.60 and 11.25 µM, respectively. As well, it was safe and showed a high selectivity index against normal cell lines (WI-38). Finally, the thieno[2,3-d]pyrimidine derivative arrested the growth of the HepG2 cells at the G2/M phase inducing both early and late apoptosis. These results were further confirmed through the ability of the thieno[2,3-d]pyrimidine derivative to induce significant changes in the apoptotic genes levels of caspase-3, caspase-9, Bcl-2 associated X-protein, and B-cell lymphoma 2.

Influence of Crosslinking Methods on Biomimetically Mineralized Collagen Matrices for Bone-like Biomaterials.

To assist in bone defect repair, ideal bone regeneration scaffolds should exhibit good osteoconductivity and osteoinductivity, but for load-bearing applications, they should also have mechanical properties that emulate those of native bone. The use of biomimetic processing methods for the mineralization of collagen fibrils has resulted in interpenetrating composites that mimic the nanostructure of native bone; however, closely matching the mechanical properties of bone on a larger scale is something that is still yet to be achieved. In this study, four different collagen crosslinking methods (EDC-NHS, quercetin, methacrylated collagen, and riboflavin) are compared and combined with biomimetic mineralization via the polymer-induced liquid-precursor (PILP) process, to obtain bone-like collagen-hydroxyapatite composites. Densified fibrillar collagen scaffolds were fabricated, crosslinked, and biomimetically mineralized using the PILP process, and the effect of each crosslinking method on the degree of mineralization, tensile strength, and modulus of the mineralized scaffolds were analyzed and compared. Improved modulus and tensile strength values were obtained using EDC-NHS and riboflavin crosslinking methods, while quercetin and methacrylated collagen resulted in little to no increase in mechanical properties. Decreased mineral contents appear to be necessary for retaining tensile strength, suggesting that mineral content should be kept below a percolation threshold to optimize properties of these interpenetrating nanocomposites. This work supports the premise that a combination of collagen crosslinking and biomimetic mineralization methods may provide solutions for fabricating robust bone-like composites on a larger scale.

Crosslinking Improve Demineralized Dentin Performance and Synergistically Promote Biomimetic Mineralization by CaP_PILP.

to explore the effects of optimal crosslinking (chemical treatment) on demineralized dentin matrix and the possible synergism with calcium phosphate polymer-induced liquid precursor (CaP-PILP) bionic remineralization (physical treatment), and offer benefit to the clinic of resin-dentin bonding and dentin hypersensitivity. demineralized dentin was treated with glutaraldehyde (GA), carbodiimide (EDC), and procyanidin (PA) for crosslinking, followed by CaP-PILP biomimetic remineralization. The morphology, surface mechanical and physio-chemical properties, and enzymatic resistance were evaluated regardless of the modification. the collagen fibers appeared morphologically complete with higher surface microhardness and characteristic peaks of amide I-III bands were visible after GA, PA, and EDC crosslinking. Collagen collapse and dissolution were seen in untreated demineralized dentin with enzyme attack, while the collagen fiber structure remained intact in GA- and PA-treated specimens. The lamellar mineral phase was visible at 2 days and the dentin tubules were almost completely enclosed at 4-6 days after PA crosslinking and mineralization. However, demineralized collagen fibers and open tubules were still visible between the dentinal tubules on day 8 in the control group. the structure integrity, enzyme resistance, and mechanical properties of the collagen fiber network could be significantly preserved by GA and PA crosslinking than EDC and no treatment. While, strongest synergistic effects were observed in PA on bionic remineralization by CaP-PILP, and further significantly improve the quality and shorten the duration of mineralization. These findings would be beneficial for dental clinical practice of resin-dentin bonding and dentin hypersensitivity.

Optimizing the biodegradability and osteogenesis of biogenic collagen membrane via fluoride-modified polymer-induced liquid precursor process

ABSTRACT Biogenic collagen membranes (BCM) have been widely used in guided bone regeneration (GBR) owing to their biodegradability during tissue integration. However, their relatively high degradation rate and lack of pro-osteogenic properties limit their clinical outcomes. It is of great importance to endow BCM with tailored degradation as well as pro-osteogenic properties. In this study, a fluoride-modified polymer-induced liquid precursor (PILP) based biomineralization strategy was used to convert the collagen membrane from an organic phase to an apatite-based inorganic phase, thus achieving enhanced anti-degradation performance as well as osteogenesis. As a result, three phases of collagen membranes were prepared. The original BCM in the organic phase induced the mildest inflammatory response and was mostly degraded after 4 weeks. The organic-inorganic mixture phase of the collagen membrane evoked a prominent inflammatory response owing to the fluoride-containing amorphous calcium phosphate (F-ACP) nanoparticles, resulting in active angiogenesis and fibrous encapsulation, whereas the inorganic phase induced a mild inflammatory response and degraded the least owing to the transition of F-ACP particles into calcium phosphate with high crystallinity. Effective control of ACP is key to building novel apatite-based barrier membranes. The current results may pave the way for the development of advanced apatite-based membranes with enhanced barrier performances.

Towards resorbable 3D-printed scaffolds for craniofacial bone regeneration.

This review will briefly examine the development of 3D-printed scaffolds for craniofacial bone regeneration. We will, in particular, highlight our work using Poly(L-lactic acid) (PLLA) and collagen-based bio-inks. This paper is a narrative review of the materials used for scaffold fabrication by 3D printing. We have also reviewed two types of scaffolds that we designed and fabricated. Poly(L-lactic acid) (PLLA) scaffolds were printed using fused deposition modelling technology. Collagen-based scaffolds were printed using a bioprinting technique. These scaffolds were tested for their physical properties and biocompatibility. Work in the emerging field of 3D-printed scaffolds for bone repair is briefly reviewed. Our work provides an example of PLLA scaffolds that were successfully 3D-printed with optimal porosity, pore size and fibre thickness. The compressive modulus was similar to, or better than, the trabecular bone of the mandible. PLLA scaffolds generated an electric potential upon cyclic/repeated loading. The crystallinity was reduced during the 3D printing. The hydrolytic degradation was relatively slow. Osteoblast-like cells did not attach to uncoated scaffolds but attached well and proliferated after coating the scaffold with fibrinogen. Collagen-based bio-ink scaffolds were also printed successfully. Osteoclast-like cells adhered, differentiated, and survived well on the scaffold. Efforts are underway to identify means to improve the structural stability of the collagen-based scaffolds, perhaps through mineralization by the polymer-induced liquid precursor process. 3D-printing technology is promising for constructing next-generation bone regeneration scaffolds. We describe our efforts to test PLLA and collagen scaffolds produced by 3D printing. The 3D-printed PLLA scaffolds showed promising properties akin to natural bone. Collagen scaffolds need further work to improve structural integrity. Ideally, such biological scaffolds will be mineralized to produce true bone biomimetics. These scaffolds warrant further investigation for bone regeneration.

Biomimetic Mineralization of Fibrillar Collagen with Strontium-doped Hydroxyapatite

Fibrillar collagen structures mineralized with hydroxyapatite using the polymer-induced liquid precursor (PILP) process have been explored as synthetic models for studying biomineralization of human hard tissues and have also been applied in the fabrication of scaffolds for hard tissue regeneration. Strontium has important biological functions in bone and has been used as a therapeutic agent for treating diseases that result in bone defects, such as osteoporosis. Here, we developed a strategy to mineralize collagen with Sr-doped hydroxyapatite (HA) using the PILP process. Doping with Sr altered the crystal lattice of HA and inhibited the degree of mineralization in a concentration-dependent manner, but did not affect the unique formation of intrafibrillar minerals using the PILP. The Sr-doped HA nanocrystals were aligned in the [001] direction but did not recapitulate the parallel alignment of the c-axis of pure Ca HA in relation to the collagen fiber long axis. The mimicry of doping Sr in PILP-mineralized collagen can help understand the doping of Sr in natural hard tissues and during treatment. The fibrillary mineralized collagen with Sr-doped HA will be explored in future work as biomimetic and bioactive scaffolds for regeneration of bone and tooth dentin.

Calcium Phosphate Delivery Systems for Regeneration and Biomineralization of Mineralized Tissues of the Craniofacial Complex.

Calcium phosphate (CaP)-based materials have been extensively used for mineralized tissues in the craniofacial complex. Owing to their excellent biocompatibility, biodegradability, and inherent osteoconductive nature, their use as delivery systems for drugs and bioactive factors has several advantages. Of the three mineralized tissues in the craniofacial complex (bone, dentin, and enamel), only bone and dentin have some regenerative properties that can diminish due to disease and severe injuries. Therefore, targeting these regenerative tissues with CaP delivery systems carrying relevant drugs, morphogenic factors, and ions is imperative to improve tissue health in the mineralized tissue engineering field. In this review, the use of CaP-based microparticles, nanoparticles, and polymer-induced liquid precursor (PILPs) amorphous CaP nanodroplets for delivery to craniofacial bone and dentin are discussed. The use of these various form factors to obtain either a high local concentration of cargo at the macroscale and/or to deliver cargos precisely to nanoscale structures is also described. Finally, perspectives on the field using these CaP materials and next steps for the future delivery to the craniofacial complex are presented.

A variable mineralization time and solution concentration intervene in the microstructure of biomimetic mineralized collagen and potential osteogenic microenvironment.

The absence of a conducive bone formation microenvironment between fractured ends poses a significant challenge in repairing large bone defects. A promising solution is to construct a bone formation microenvironment that mimics natural bone tissue. Biomimetic mineralized collagen possesses a chemical composition and microstructure highly similar to the natural bone matrix, making it an ideal biomimetic bone substitute material. The microstructure of biomimetic mineralized collagen is influenced by various factors, and its biomineralization and microstructure, in turn, affect its physicochemical properties and biological activity. We aimed to utilize mineralization time and solution concentration as variables and employed the polymer-induced liquid precursor strategy to fabricate mineralized collagen with diverse microstructures, to shed light on how mineralization parameters impact the material microstructure and physicochemical properties. We also investigated the influence of microstructure and physicochemical properties on cell biocompatibility and the bone-forming microenvironment. Through comprehensive characterization, we examined the physical and chemical properties of I-EMC under various mineralization conditions and assessed the in vitro and in vivo biocompatibility and osteogenic performance. By investigating the relationship between mineralization parameters, material physicochemical properties, and osteogenic performance, we revealed how microstructures influence cellular behaviors like biocompatibility and osteogenic microenvironment. Encouragingly, mineralization solutions with varying concentrations, stabilized by polyacrylic acid, successfully produced intrafibrillar and extrafibrillar mineralized collagen. Compared to non-mineralized collagen, all mineralized samples demonstrated improved bone-forming performance. Notably, samples prepared with a 1× mineralization solution exhibited relatively smooth surfaces with even mineralization. Extending the mineralization time enhanced the degree of mineralization and osteogenic performance. Conversely, samples prepared with a 2× mineralization solution had rough surfaces with large calcium phosphate particles, indicating non-uniform mineralization. Overall, our research advances the potential for commercial production of mineralized collagen protein products, characterized by dual biomimetic properties, and their application in treating various types of bone defects.

Facile fabrication of biomimetic silicified gelatin scaffolds for angiogenesis and bone regeneration by a bioinspired polymer-induced liquid precursor

Bioinspired materials with dual capabilities of pro-osteogenesis and pro-angiogenesis have attracted increasing scientific interest in the field of orthopedics. Herein, inspired by the mineralization process of natural bone, a biomimetic strategy is presented to fabricate highly bioactive hybrid nanocomposite scaffolds via impregnation of Si-containing polymer-induced liquid precursor (PILP) into biocompatible porous gelatin scaffolds followed by in situ free radial polymerization. The well-designed biohybrid scaffold exhibited not only a micro/nano fiber-like porous architecture (pore size: 100–200 μm; porosity: 80–90 %), high specific surface area (≈20 m2/g), vastly improved mechanical performance, tunable swelling and degradation behaviors but also a sustained release profile of bioactive Si ions up to 21 days. In vitro experiments revealed the excellent biological performance of the resulting biohybrid scaffold, including good biomineralization capacity and biocompatibility, establishing a favorable microenvironment for facilitating cell adhesion, proliferation, migration, osteogenic differentiation, and vascularization. In rat calvarial critical-size defect models, the newly developed biohybrid scaffold could potentially trigger a chain of biological events: stimulating the polarization of M2 macrophages, recruiting endogenous stem cells and endothelial cells at the injury site to enable a suitable regenerative microenvironment for accelerating coupled osteogenesis and angiogenesis, and eventually promoting vascularized bone regeneration.

Assessment of Optimal Conditions for Marine Invertebrate Cell-Mediated Mineralization of Organic Matrices.

Cellular strategies and regulation of their crystallization mechanisms are essential to the formation of biominerals, and harnessing these strategies will be important for the future creation of novel non-native biominerals that recapitulate the impressive properties biominerals possess. Harnessing these biosynthetic strategies requires an understanding of the interplay between insoluble organic matrices, mineral precursors, and soluble organic and inorganic additives. Our long-range goal is to use a sea anemone model system (Nematostella vectensis) to examine the role of intrinsically disordered proteins (IDPs) found in native biomineral systems. Here, we study how ambient temperatures (25–37 °C) and seawater solution compositions (varying NaCl and Mg ratios) will affect the infiltration of organic matrices with calcium carbonate mineral precursors generated through a polymer-induced liquid-precursor (PILP) process. Fibrillar collagen matrices were used to assess whether solution conditions were suitable for intrafibrillar mineralization, and SEM with EDS was used to analyze mineral infiltration. Conditions of temperatures 30 °C and above and with low Mg:Ca ratios were determined to be suitable conditions for calcium carbonate infiltration. The information obtained from these observations may be useful for the manipulation and study of cellular secreted IDPs in our quest to create novel biosynthetic materials.

The Role of Process-Directing Agents on Enamel Lesion Remineralization: Fluoride Boosters.

The aim of this study was to investigate the effects of two process-directing agents (polyaspartic acid and osteopontin) used in a polymer-induced liquid-precursor (PILP) process on the remineralization of bacteria-induced enamel demineralization. Enamel demineralization lesions (depths of about 180–200 µm) were created and exposed to Streptococcus mutans, cultured with a 10% sucrose solution for 21 days, and remineralized using a PILP process (pH = 7.4, 14 days) with a calcium phosphate solution containing either polyaspartic acid or osteopontin in the presence or absence of fluoride (0.5 ppm). The specimens were examined under scanning electron microscopy. The fluoride was successfully incorporated into the PILP remineralization process for both polyaspartic acid and osteopontin. When the fluoride was added to the PILP remineralization solution, there was more uniform remineralization throughout the lesion than with either polyaspartic acid or osteopontin alone. However, in the absence of these process-directing agents, fluoride alone showed less remineralization with the formation of a predominantly surface-only layer. The PILP remineralization process relies on the ability of process-directing agents to stabilize calcium phosphate ions and holds promise for enamel lesion remineralization, and these agents, in the presence of fluoride, seem to play an important role as a booster or supplement in the continuation of remineralization by reducing the mineral gains at the surface layer.