Search

Aim To evaluate Save-A-Tooth ( SAT), EMT Toothsaver ( EMT) and Hank's Balanced Salt Solution ( HBSS) for their influence on the viability and proliferative capacity of human periodontal ligament fibroblasts (HPDLFs). Methodology Primary HPDLFs were seeded into 96-well cell culture plates and exposed to SAT, EMT, HBSS and water (negative control) for 0.5, 1, 3, 6, 12 and 24 h at room temperature (22 °C). After each exposure time, cell viability was measured through quantifying adenosine triphosphate ( ATP) using a luminescent dye. The proliferative capacity was also quantified using the PrestoBlue assay after 12 or 24 h storage in each medium. The data were analysed statistically by two-way anova and post hoc Least Significant Difference ( LSD) test ( P < 0.05). The morphology of the cells after 12 h storage was also investigated through live/dead viability/cytotoxicity kit together with fluorescence microscopy. Results There was no significant difference in cell viability amongst HBSS, SAT and EMT groups up to 6 h. SAT was effective in maintaining cell viability only up to 12 h and then became detrimental to HPDLF; after 24 h, the effectiveness of SAT in maintaining cell viability was similar to that of water ( P > 0.05). Amongst all the media, only EMT could maintain the proliferative capacity of HPDLFs significantly higher than the negative control, that is water ( P < 0.05) after 24 h storage. Conclusion EMT maintained the proliferative capacity of HPDLFs after 24 h storage. [ABSTRACT FROM AUTHOR]

In this study, we propose the "crystallization from supersaturated solution" method for producing drug-loaded polymeric micelles. This method involves the formation of solid drug-encapsulating crystals of a diblock copolymer through isothermal crystallization from a supersaturated solution of the copolymer in low molecular weight PEGs containing the drug, followed by dissolution of the crystals to obtain drug-loaded micelles. We fabricated and characterized micelles loaded with several model drugs (paclitaxel, rapamycin, and docetaxel) and their oligo(lactic acid) 8 -prodrugs using PEG 4kDa -b-PLA 2.2kDa as the micelle-forming copolymer and PEGs of varying molecular weights (200, 400, and 600 Da) as solvents. Our findings indicate that the molecular weight of the solvent PEG and the target drug loading significantly influence the physicochemical properties of the resulting micelles, including loading efficiency and particle size distribution. Micelles produced with PEG200 as the solvent exhibited the highest loading efficiency, followed by those made with PEG600 and PEG400 for all the drugs and prodrugs tested. Increasing the target drug loading enhanced both the loading efficiency and average particle size across all formulations. Furthermore, prodrug-loaded micelles showed higher loading efficiency and improved stability in aqueous solutions compared to their parent drug counterparts. Crystals encapsulating both parent drugs and prodrugs could be stored at room temperature for extended periods, producing micelles with no significant differences in loading efficiency and particle size distribution compared to freshly prepared micelles. Additionally, the crystals demonstrated a rapid dissolution rate, forming uniform micelles after just 5 s of hydration and agitation. Cytotoxicity studies against 4 T1 and MDA-MB-231 breast cancer cell lines revealed that the molecular weight of the PEG used as the solvent impacts the cytotoxicity of the resulting micelles, with those produced using PEG200 displaying the highest cytotoxicity, followed by PEG400 and PEG600. Overall, the crystallization from supersaturated solution method proves to be an effective platform for prolonged storage and rapid formation of stable, drug-loaded polymeric micelles. It has the potential to eliminate the need for freeze-drying in the formulation and storage of drug-loaded polymeric micelles. These findings highlight the method's potential for advancing drug delivery systems, particularly for the solubilization of hydrophobic drugs using micellar formulations., Competing Interests: Declaration of competing interest The authors disclose no conflict of interest., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Progressive forms of interstitial lung diseases, including idiopathic pulmonary fibrosis (IPF), are deadly disorders lacking non-invasive biomarkers for assessment of early disease activity, which presents a major obstacle in disease management. Excessive extracellular matrix (ECM) deposition is a hallmark of these disorders, with fibronectin being an abundant ECM glycoprotein that is highly upregulated in early fibrosis and serves as a scaffold for the deposition of other matrix proteins. Due to its role in active fibrosis, we are targeting fibronectin as a biomarker of early lung fibrosis disease activity via the PEGylated fibronectin-binding polypeptide (PEG-FUD). In this work, we demonstrate the binding of PEG-FUD to the fibrotic lung throughout the course of bleomycin-induced murine model of pulmonary fibrosis. We first analyzed the binding of radiolabeled PEG-FUD following direct incubation to precision cut lung slices from mice at different stages of experimental lung fibrosis. Then, we administered fluorescently labeled PEG-FUD subcutaneously to mice over the course of bleomycin-induced pulmonary fibrosis and assessed peptide uptake 24 h later through ex vivo tissue imaging. Using both methods, we found that peptide targeting to the fibrotic lung is increased during the fibrogenic phase of the single dose bleomycin lung fibrosis model (days 7 and 14 post-bleomycin). At these timepoints we found a correlative relationship between peptide uptake and fibrotic burden. These data suggest that PEG-FUD targets fibronectin associated with active fibrogenesis in this model, making it a promising candidate for a clinically translatable molecular imaging probe to non-invasively determine pulmonary fibrosis disease activity, enabling accelerated therapeutic decision-making., Competing Interests: Declarations. Ethics approval and consent to participate: Animal studies were approved by the University of Wisconsin-Madison IACUC (protocol numbers M005823 and M005532). No human subjects or tissues were used in this work. Competing interests: BRTJ, GSK, NS, and KB are inventors on an issued patent application (pub. no. US10828372B2; date of application granted: 10 November 2020) held by Wisconsin Alumni Research Foundation that covers the PEGylation of FN-binding peptides and their application in fibrosis. RH is a consultant to Archeus Technologies Inc. and MonoPar Therapeutics Inc. He is a member of the science advisory board of Archeus. None of these activities have any relation to imaging of pulmonary fibrosis. All other authors declare that they have no competing interests., (© 2025. The Author(s).)

In this study, we propose a spatially patterned 3D-printed nanohydroxyapatite (nHA)/beta-tricalcium phosphate (β-TCP)/collagen composite scaffold incorporating human dental pulp-derived mesenchymal stem cells (hDP-MSCs) for bone regeneration in critical-sized defects. We investigated angiogenesis and osteogenesis in a rabbit critical-sized mandibular defect model treated with this engineered construct. The critical and synergistic role of collagen coating and incorporation of stem cells in the regeneration process was confirmed by including a cell-free uncoated 3D-printed nHA/β-TCP scaffold, a stem cell-loaded 3D-printed nHA/β-TCP scaffold, and a cell-free collagen-coated 3D-printed nHA/β-TCP scaffold in the experimental design, in addition to an empty defect. Posteuthanasia evaluations through X-ray analysis, histological assessments, immunohistochemistry staining, histomorphometry, and reverse transcription-polymerase chain reaction (RT-PCR) suggest the formation of substantial woven and lamellar bone in the cell-loaded collagen-coated 3D-printed nHA/β-TCP scaffolds. Histomorphometric analysis demonstrated a significant increase in osteoblasts, osteocytes, osteoclasts, bone area, and vascularization compared to that observed in the control group. Conversely, a significant decrease in fibroblasts/fibrocytes and connective tissue was observed in this group compared to that in the control group. RT-PCR indicated a significant upregulation in the expression of osteogenesis-related genes, including BMP2, ALPL, SOX9, Runx2, and SPP1. The findings suggest that the hDP-MSC-loaded 3D-printed nHA/β-TCP/collagen composite scaffold is promising for bone regeneration in critical-sized defects.

In recent years, liquid-liquid phase separation (LLPS) has been recognized to act as a precursor to self-assembly in amphiphilic systems. In this study, we propose the use of entropy-driven LLPS to obtain a tunable precursor for polymeric micelle formation. In this new approach, an oligomer is utilized as a nonselective solvent for the block copolymer, allowing for the tuning of entropy and subsequent LLPS. A comprehensive model was developed using mean-field lattice theory to predict the conditions under which LLPS and micellization occur. The degree of polymerization of the solvent was found to have a significant impact on the phase behavior of the system, outweighing enthalpic contributions such as the interaction between the blocks of the copolymer and the solvent. Our model predicts that using a solvent with a degree of polymerization equal to or greater than 5 for a copolymer such as PEG 4kDa - b -PLA 2.2kDa will result in LLPS prior to complete micellization, regardless of the values of interaction parameters. It also suggests that phase-separated liquid and polymeric micelles can co-exist in such a mixture. We confirmed our model predictions using dynamic light scattering and phase microscopy when PEG200 was used as the solvent. Micellization for PEG 4kDa - b -PLA 2.2kDa /PEG200/water mixture occurred at 10-12% w/w water content, consistent with the model predictions. Furthermore, the LLPS-to-micelle transition was shown to be reversible by changing the temperature or water content, indicating that the phase-separated liquid may be in thermodynamic equilibrium with polymeric micelles.

Paclitaxel (PTX) is a frequently prescribed chemotherapy drug used to treat a wide variety of solid tumors. Oligo(lactic acid) 8 -PTX prodrug (o(LA) 8 -PTX) loaded poly(ethylene glycol)-b-poly(lactic acid) (PEG-b-PLA) micelles have higher loading, slower release and higher antitumor efficacy in murine tumor models over PTX-loaded PEG-b-PLA micelles. The goal of this work is to study plasma stability of o(LA) 8 -PTX-loaded PEG-b-PLA micelles and its pharmacokinetics after IV injection in rats. In rat plasma, o(LA) 8 -PTX prodrug is metabolized into o(LA) 1 -PTX and PTX. In human plasma, o(LA) 8 -PTX is metabolized more slowly into o(LA) 2 -PTX, o(LA) 1 -PTX, and PTX. After IV injection of 10 mg/kg PTX-equiv of o(LA) 8 -PTX prodrug loaded PEG-b-PLA micelles in Sprague-Dawley rats, metabolite abundance in plasma follows the order: o(LA) 1 -PTX > o(LA) 2 -PTX > o(LA) 4 -PTX > o(LA) 6 -PTX. Bile metabolite profiles of the o(LA) 8 -PTX prodrug is similar to plasma metabolite profiles. In comparison to equivalent doses of Abraxane®, plasma PTX exposure is two orders of magnitude higher for Abraxane® than PTX from o(LA) 8 -PTX prodrug loaded PEG-b-PLA micelles, and plasma o(LA) 1 -PTX exposure is fivefold higher than PTX from Abraxane®, demonstrating heightened plasma metabolite exposure for enhanced antitumor efficacy., (© 2023. The Author(s), under exclusive licence to American Association of Pharmaceutical Scientists.)

A new approach named PEG-assist is introduced for the production of drug-loaded polymeric micelles. The method is based on the use of PEG as the non-selective solvent for PEG-b-PLA in the fabrication procedure. Both hydration temperature and PEG molecular weight are shown to have a significant effect on the encapsulation efficiency of PTX in PEG 4kDa -b-PLA 2kDa micelles. The optimal procedure for fabrication includes the use of PEG 1kDa as the solvent at 60 °C, cooling the mixture to 40 °C, hydration at 40 °C, freezing at -80 °C and freeze-drying at -35 °C, 15 Pa. No significant difference (p > 0.05) in PTX encapsulation, average particle size and polydispersity index is observed between the samples before freeze-drying and after reconstitution of the freeze-dried cake. The prepared PTX formulations are stable at room temperature for at least 8 h. Scaling the batch size to 25× leads to no significant change (p > 0.05) in PTX encapsulation, average particle size and polydispersity index. PEG-assist method is applicable to other drugs such as 17-AAG, and copolymers of varied molecular weights. The use of no organic solvent, simplicity, cost-effectiveness, and efficiency makes PEG-assist a very promising approach for large scale production of drug-loaded polymeric micelles., Competing Interests: Declaration of Competing Interest The authors disclose no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Poly(3-caprolactone) (PCL)/β-tricalcium phosphate (β-TCP) composite scaffolds fabricated by three-dimensional (3D) printing are one of the common scaffolds for bone tissue regeneration. However, the main challenge of these 3D printed PCL/β-TCP scaffolds is the fact that many cells pass from porosities during in vitro cell seeding, leading to poor initial cell attachment. This study aimed to demonstrate the fabrication of a new collagen coating process for optimizing the hydrophilic property and cell-substrate interactions. This method may be used for coating collagen on any relevant biomedical constructs made of synthetic polymers to increase their biocompatibility and cell attachment., Materials and Methods: Porous composite scaffolds fabricated by 3D printing were coated with collagen by a novel method and compared to traditional methods. After plasma treatment, samples were inverted in a homogenized collagen solution, freeze-dried, stabilized by crosslinking, freeze-dried again, and fibrillated using a defined salt concentration. Samples were characterized by a 3D laser microscope, cytocompatibility assay, attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy, water absorption, protein absorption, and bioactivity assay., Results: Homogenized collagen at pH= 7 resulted in a very uniform layer on the surface of scaffolds with significantly higher cell proliferation (p < 0.05). Collagen-coated scaffolds showed significantly higher water absorption, protein absorption, and bioactivity compared to non-coated samples (p < 0.05)., Conclusion: The results demonstrate that both the pH and collagen structure influence the coating of scaffolds, while the concentrations used in this study do not have a significant difference in this aspect. The combination of homogenization and fibrillization makes scaffolds more biocompatible and desirable for bone tissue engineering., (Copyright © 2022 Elsevier B.V. All rights reserved.)

A modular design composed of 3D-printed polycaprolactone (PCL) as the load-bearing module, and dual porosity gelatin foam as the bio-reactive module, was developed and characterized in this study. Surface treatment of the PCL module through aminolysis-aldehyde process was found to yield a stronger interface bonding compared to NaOH hydrolysis, and therefore was used in the fabrication procedure. The modular scaffold was shown to significantly improve the mechanical properties of the gelatin foam. Both compressive modulus and ultimate strength was found to increase over 10 times when the modular design was employed. The bio-reactive module i.e., gelatin foam, presented a dual porosity network of 100-300 μm primary and <10 μm secondary pores. SEM images revealed excellent attachment of DPSCs to the bio-reactive module.

Objectives: The aim of this work was to combine engineered hard and soft tissue, adopting a new method for interfacial adhesion of osteo-mucosal construct. We hypothesized that the chemical procedure involved in this method not only adheres the components, but also improves the cell growth inside them., Methods: 3D-printed functionally-graded porous hard-tissue scaffolds were characterized, functionalized by aminolysis and tyrosinase, and accommodated by human osteoblast cells. Introducing amino groups through aminolysis and inducing dopaquinones by tyrosinase can take part in the Michael additions to cause the adhesion. Subsequently, fully-differentiated engineered oral mucosa was formed directly on the surface of hard tissue. Constructs were assessed in term of morphology, structure, chemical composition, histology, and cytocompatibility. Interfacial adhesion was compared to a control group prepared by using a biological glue for the attachment of the soft and hard tissues., Results: The data confirmed higher proliferation of osteoblast cells via aminolysis and improved osteoblast cells distribution and differentiation by incorporation of tyrosinase in collagen. There was evidence of multilayered, stratified epithelium on the osteo-mucosal model with viable fibroblasts and osteoblasts within the lamina propria and bone tissue layers. Our method of adhesion resulted in cohesive debonding within the engineered soft tissue; while in the control group, adhesive debonding and complete separation of the oral mucosa from the hard tissue was observed. Although the shear strength of the osteo-mucosal model (157.6 kDa ± 25.1) was slightly higher than that of the control group (149.4 kDa ± 23.1), there was no statistically significant difference between them (p > 0.05). However, the advantage of our in situ adhesion approach is the absence of a barrier like glue which can disrupt direct cellular communications between tissues., Significance: This study provides a novel method of directly combining tissue-engineered human bone with oral mucosa, which has the potential to improve cell-ingrowth and tissue integration. This engineered tissue construct, after further optimization, can be used clinically as a graft material in various oral surgeries and can also be employed as an in vitro model to investigate many aspects of oral diseases and examine dental materials and oral health care products as a replacement of in vivo models., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Pressure-assisted coating (PAC) is introduced to coat 3D-printed polymeric scaffolds with β-tricalcium phosphate (β-TCP) for tissue engineering applications. The method consists of four steps: infiltration of ceramic particles into the porous structure of the polymeric scaffold, dehydration of the slurry, compaction of ceramic particles around the scaffold, and heat treatment. The optimal coating is obtained at an infiltration speed of 400 mm/min followed by complete dehydration, compaction under ca. 8 MPa pressure, and subsequent heat treatment at 65 °C. The outcome is a uniformly coated scaffold with no deformation or structural defects, as confirmed by micro-CT analysis and laser and scanning electron microscopy. Scaffolds coated using the PAC method present superior interface bonding strength compared to those coated with a biomimetic approach. The contact angle decreased from 75.2 ± 1.4° for the uncoated scaffold to 39.6 ± 9.6° for the PAC specimen. PAC also increased the surface roughness from 0.66 ± 0.08 to 6.89 ± 0.26 μm and doubled the number of attached cells on the 3 rd day of culture. The described method is applicable to different structures, object sizes, pore sizes, and shapes. For instance, in-depth coating of a 10 mm × 10 mm ( D × H ) cone with a 58 ± 4 μm-thick layer of β-TCP can be achieved using PAC. The method can be used to coat other polymers, such as poly(lactic- co -glycolic acid) (PLGA). Successful coating of β-TCP on 3D-printed PLGA scaffolds is also presented as a proof of concept.

Poly(ethylene glycol)-block-poly(D,L-lactic acid) (PEG-b-PLA) and poly(ethylene glycol)-block-poly(ε-caprolactone) (PEG-b-PCL) form nano-assemblies, including micelles and nanoparticles, that increase the water solubility of anticancer drugs for injection. PEG-b-PLA and PEG-b-PCL are less toxic than commonly used organic solvents or solubilizers for injection, such as Cremophor EL® in Taxol®. Formulating paclitaxel in PEG-b-PLA micelles, as Genexol-PM®, permits dose escalation over Taxol®, enhancing antitumor efficacy in breast, lung and ovarian cancers. To expand the repertoire of anticancer drugs for injection, acyl and oligo(lactic acid) ester prodrugs have been synthesized for PEG-b-PLA and PEG-b-PCL nano-assemblies, compatibility, and novel nanomedicines for injection. Notably, acyl and oligo(lactic acid) taxane prodrugs delivered by PEG-b-PLA and PEG-b-PCL nano-assemblies display heightened plasma exposure, reduction in biodistribution into major organs and enhanced tumor exposure in murine tumor models, versus parent anticancer drugs in conventional formulations. As a result, acyl and oligo(lactic acid) ester prodrugs are less toxic and induce durable antitumor responses. In summary, acyl and oligo(lactic acid) ester prodrugs widen the range of anticancer drugs that can be tested safely and effectively by using PEG-b-PLA and PEG-b-PCL nano-assemblies, and they display superior anticancer efficacy over parent anticancer drugs, which are often approved products. Oligo(lactic acid) ester taxane prodrugs are in pre-clinical development as novel drug combinations and immunotherapy combinations for cancer therapy., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Statement of Problem: Soft-tissue attachment to different surfaces may play a pivotal role in the long-term success of dental implants. However, studies on the issue, especially on newer materials, are sparse., Purpose: The purpose of this in vitro study was to evaluate the viability and adhesion of human gingival fibroblasts (HGFs) on different implant abutment materials with specific surface modifications., Material and Methods: One hundred and fifty specimens in 6 experimental groups were evaluated: smooth-machined titanium alloy (Ti), laser-modified titanium (TiL), smooth-machined polyetheretherketone (PEEK) (P), laser-modified PEEK (PL), plasma-treated PEEK (PP), laser- and plasma-treated PEEK (PLP). Machined Ti was considered as the control group. Surface roughness (S a ), water contact angle (WCA), and X-ray photoelectron spectroscopy (XPS) were measured. HGF attachment and proliferation were observed at 1, 3, and 7 days after cell seeding. Comparison of the means among the groups was performed with 1-way analysis of variance (ANOVA) with post hoc comparison using the Tukey test (α=.05)., Results: S a values of the laser modified groups were significantly higher than those of the nonmodified (smooth-machined) groups (P<.001). WCAs were significantly different among PEEK groups, and plasma-sprayed groups had the lowest WCAs. XPS analysis of both Ti and PEEK groups showed laser treatment did not have any significant effect on the surface composition of the PEEK as the same bonds with similar ratio/fraction were detected in the spectrum of the modified specimens. Scanning electron microscopy (SEM) revealed more functionally oriented HGF cells on the laser-grooved surfaces. On the first, third, and seventh day of proliferation, the titanium groups showed no significant differences (P>.05). On the first and third days of proliferation, the plasma sprayed groups (PP, PLP) showed significantly greater proliferation than all experimental groups (P<.001). On the seventh day of proliferation, statistically significant differences were observed between all PEEK groups and between all PEEK groups and the Ti group (P<.001), with the exception of the PL and P groups and the PLP and Ti groups (P>.05)., Conclusions: Laser-modified titanium and PEEK surfaces led to guided gingival fibroblast attachment. Plasma treatment of PEEK surfaces increased the wettability of this polymer and improved proliferation of HGF., (Copyright © 2020 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.)

Hope is that tissue engineering will provide a solution to meet the growing needs for bone substitutes. Among the potential solutions, three-dimensional (3D) printing is a promising method to fabricate functional bone substitutes especially for treatment of complex and critical-sized bone defects. Despite its encouraging achievements, 3D printing of bone substitutes still faces serious challenges including mechanical strength, shape complexity, optimization of pore parameters, and vascularization. The newer approach, that is, 3D bioprinting, is also confronted with challenges, which have prevented the realization of the dream of fabricating functional patient-specific bone substitutes. This article reviews the major challenges toward 3D printing and bioprinting of bone substitutes and recent studies addressing them. Potential solutions for each challenge and future directions are also provided. Impact Statement This review provides a current overview of the challenges in 3D (bio)printing of bone substitutes and summarizes the potential solutions.

Purpose: In this work, the effects of sodium chloride (NaCl) on gene expression of planktonic Streptococcus mutans cells are investigated. Also assessed are the effects of NaCl on zeta potential of sound and demineralized dentin., Methods: The relative level of glucosyltransferase B ( gtfB ), gtfC and gtfD transcription of S. mutans in the presence of NaCl was evaluated by quantitative polymerase chain reaction (qPCR). The osmolality of varying salt (NaCl) concentrations and their influence on the zeta potential of sound and demineralized dentin was investigated as well., Results: NaCl significantly reduced the expression of gtfB and C genes in planktonic S. mutans ; whereas, gtf D gene expression significantly increased in the presence of NaCl (P < 0.05). NaCl at concentrations of 37.5 mg/ml reduced zeta potential of demineralized dentin, while no significant decrease of zeta potential was found when sound dentin was exposed to this concentration., Conclusion: NaCl reduces the expression of some gtfs in S. mutans and increases negative potential charge of demineralized dentin.

3D dual porosity protein-based scaffolds have been developed using the combination of foaming and freeze-drying. The suggested approach leads to the production of large, highly porous scaffolds with negligible shrinkage and deformation compared to the conventional freeze-drying method. Scanning electron microscopy, standard histological processing and mercury intrusion porosimetry confirmed the formation of a dual network in the form of big primary pores (243 ± 14 µm) embracing smaller secondary pores (42 ± 3 µm) opened onto their surface, resembling a vascular network. High interconnectivity of the pores, confirmed by micro-CT, is shown to improve diffusion kinetics and support a relatively uniform distribution of isolated human dental pulp stem cells within the scaffold compared to conventional scaffolds. Dual network scaffolds indicate more than three times as high cell proliferation capability as conventional scaffolds in 14 days.

Background: The aim of this study was to evaluate the surgical handling and clinical applicability of a specific 3D-printed membrane design fabricated using a gelatin, elastin and sodium hyaluronate blend for conjunctival reconstruction and compare it with amniotic membrane (AM), which is normally used in such surgeries., Methods: 3D printing technique was employed to fabricate the membrane based on gradient design. Prior to printing, rheometry was employed to optimize the ink composition. The printed membranes were then fully characterized in terms of physical and mechanical properties. In vitro viability, proliferation and adhesion of human limbal epithelial cells were assessed using MTT assay and scanning electron microscopy (SEM), respectively. Prior to in vivo experiment, surgical handling of each membrane was evaluated by three surgeons. In vivo evaluation was conducted through implanting the gelatin-based membranes and AM on induced conjunctival defects in rabbits (n = 8). Clinical observations, including epithelialization, inflammation severity, scar tissue formation and presence of granulation tissue, were recorded from day 1 through day 28. Histological examination was performed on all enucleated eyes on day 28. In addition to H&E staining, specific stains including Periodic Acid Schiff staining, Masson's Trichrome staining and immuno-histochemical staining for α-SMA were further used to assess goblet cell proliferation, healed sub-epithelial stroma and scar tissue formation and the presence of myofibroblasts, respectively., Results: Among all the examined compositions, a blend of 8% w/v gelatin, 2% w/v elastin and 0.5% w/v sodium hyaluronate was found to be appropriate for printing. The printed membranes had favorable optical characteristics (colorless and transparent), and the surgical handling was significantly easier compared to AM. Epithelial cells cultivated on the membranes indicated suitable viability and proliferation, and SEM images presented appropriate cell adhesion on the surface of the membranes. Clinical observations suggested similar epithelialization time (approximately 3 weeks) for both the membrane and AM grafted eyes but significantly lower levels of clinical inflammation in the membrane group from day 1 through day 28 (p = 0.01), which is a key advantage of using the printed membranes over the AM. Histological examination showed similar qualities in the healed epithelium in terms of cell morphology and cell layers. However, twice the density of goblet cells per 100 cells was observed in the gelatin-based membrane grafted group. Remnant of the degraded implant was seen in only 3 of the membranes, but in 7 of the AM grafted eyes. Inflammation and granulomatous reaction was significantly higher in sections containing the AM compared to membrane (p < 0.01 and p = 0.01, respectively). α-SMA staining was more evident, but not significantly different from the gelatin-based membrane, for the AM group (p = 0.25)., Conclusion: The designed gelatin-based membrane offers the necessary physical and mechanical characteristics needed for successful ocular surface/conjunctival defect construction and may be considered a promising alternative to AM due to a more predictable degradation pattern, higher goblet cell density on the healed epithelium, less inflammation and reduced scar tissue formation., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Recent advances in three-dimensional printing technology have led to a rapid expansion of its applications in tissue engineering. The present study was designed to develop and characterize an in vitro multi-layered human alveolar bone, based on a 3D printed scaffold, combined with tissue engineered oral mucosal model. The objective was to incorporate oral squamous cell carcinoma (OSCC) cell line spheroids to the 3D model at different anatomical levels to represent different stages of oral cancer. Histological evaluation of the 3D tissue model revealed a tri-layered structure consisting of distinct epithelial, connective tissue, and bone layers; replicating normal oral tissue architecture. The mucosal part showed a well-differentiated stratified oral squamous epithelium similar to that of the native tissue counterpart, as demonstrated by immunohistochemistry for cytokeratin 13 and 14. Histological assessment of the cancerous models demonstrated OSCC spheroids at three depths including supra-epithelial level, sub-epithelial level, and deep in the connective tissue-bone interface. The 3D tissue engineered composite model closely simulated the native oral hard and soft tissues and has the potential to be used as a valuable in vitro model for the investigation of bone invasion of oral cancer and for the evaluation of novel diagnostic or therapeutic approaches to manage OSCC in the future., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Three-dimensional (3D) printing is currently being intensely studied for a diverse set of applications, including the development of bioengineered tissues, as well as the production of functional biomedical materials and devices for dental and orthopedic applications. The aim of this study was to develop and characterize a 3D-printed hybrid construct that can be potentially suitable for guided tissue regeneration (GTR). For this purpose, the rheology analyses have been performed on different bioinks and a specific solution comprising 8% gelatin, 2% elastin and 0.5% sodium hyaluronate has been selected as the most suitable composition for printing a structured membrane for GTR application. Each membrane is composed of 6 layers with strand angles from the first layer to the last layer of 45, 135, 0, 90, 0 and 90°. Confirmed by 3D Laser Measuring imaging, the membrane has small pores on one side and large pores on the other to be able to accommodate different cells like osteoblasts, fibroblasts and keratinocytes on different sides. The ultimate cross-linked product is a 150μm thick flexible and bendable membrane with easy surgical handling. Static and dynamic mechanical testing revealed static tensile modules of 1.95±0.55MPa and a dynamic tensile storage modulus of 314±50kPa. Through seeding the membranes with fibroblast and keratinocyte cells, the results of in vitro tests, including histological analysis, tissue viability examinations and DAPI staining, indicated that the membrane has desirable in vitro biocompatibility. The membrane has demonstrated the barrier function of a GTR membrane by thorough separation of the oral epithelial layer from the underlying tissues. In conclusion, we have characterized a biocompatible and bio-resorbable 3D-printed structured gelatin/elastin/sodium hyaluronate membrane with optimal biostability, mechanical strength and surgical handling characteristics in terms of suturability for potential application in GTR procedures., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Objective: A systematic characterization of hybrid scaffolds, fabricated based on combinatorial additive manufacturing technique and freeze-drying method, is presented as a new platform for osteoblastic differentiation of dental pulp cells (DPCs)., Methods: The scaffolds were consisted of a collagenous matrix embedded in a 3D-printed beta-tricalcium phosphate (β-TCP) as the mineral phase. The developed construct design was intended to achieve mechanical robustness owing to 3D-printed β-TCP scaffold, and biologically active 3D cell culture matrix pertaining to the Collagen extracellular matrix. The β-TCP precursor formulations were investigated for their flow-ability at various temperatures, which optimized for fabrication of 3D printed scaffolds with interconnected porosity. The hybrid constructs were characterized by 3D laser scanning microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and compressive strength testing., Results: The in vitro characterization of scaffolds revealed that the hybrid β-TCP/Collagen constructs offer superior DPCs proliferation and alkaline phosphatase (ALP) activity compared to the 3D-printed β-TCP scaffold over three weeks. Moreover, it was found that the incorporation of TCP into the Collagen matrix improves the ALP activity., Significance: The presented results converge to suggest the developed 3D-printed β-TCP/Collagen hybrid constructs as a new platform for osteoblastic differentiation of DPCs for craniomaxillofacial bone regeneration., (Copyright © 2017. Published by Elsevier Ltd.)

The aim of this research was to develop chitosan/gelatin/keratin composite containing hydrocortisone sodium succinate as a buccal mucoadhesive patch to treat desquamative gingivitis, which was fabricated through an environmental friendly process. Mucoadhesive films increase the advantage of higher efficiency and drug localization in the affected region. In this research, mucoadhesive films, for the release of hydrocortisone sodium succinate, were prepared using different ratios of chitosan, gelatin and keratin. In the first step, chitosan and gelatin proportions were optimized after evaluating the mechanical properties, swelling capacity, water uptake, stability, and biodegradation of the films. Then, keratin was added at different percentages to the optimum composite of chitosan and gelatin together with the drug. The results of surface pH showed that none of the samples were harmful to the buccal cavity. FTIR analysis confirmed the influence of keratin on the structure of the composite. The presence of a higher amount of keratin in the composite films resulted in high mechanical, mucoadhesive properties and stability, low water uptake and biodegradation in phosphate buffer saline (pH = 7.4) containing 10 4  U/ml lysozyme. The release profile of the films ascertained that keratin is a rate controller in the release of the hydrocortisone sodium succinate. Finally, chitosan/gelatin/keratin composite containing hydrocortisone sodium succinate can be employed in dental applications.

A solvent-free microsphere sintering technique was developed to fabricate scaffolds with pore size gradient for tissue engineering applications. Poly(D,L-Lactide) microspheres were fabricated through an emulsification method where TiO 2 nanoparticles were employed both as particulate emulsifier in the preparation procedure and as surface modification agent to improve bioactivity of the scaffolds. A fine-tunable pore size gradient was achieved with a pore volume of 30±2.6%. SEM, EDX, XRD and FTIR analyses all confirmed the formation of bone-like apatite at the 14 th day of immersion in Simulated Body Fluid (SBF) implying the ability of our scaffolds to bond to living bone tissue. In vitro examination of the scaffolds showed progressive activity of the osteoblasts on the scaffold with evidence of increase in its mineral content. The bioactive scaffold developed in this study has the potential to be used as a suitable biomaterial for bone tissue engineering and hard tissue regeneration., (Copyright © 2016 Elsevier Inc. All rights reserved.)