Antibacterial properties of marine algae incorporated polylactide acid membranes as an alternative to clinically applied different collagen membranes.

The reconstruction of bony defects in the alveolar crest poses challenges in dental practice. Guided tissue regeneration (GTR) and guided bone regeneration (GBR) procedures utilize barriers to promote bone regeneration and prevent epithelial growth. This study focuses on evaluating the antibacterial properties of marine algae-polylactic acid (PLA) composite membranes compared to commercially available collagen membranes. Marine algae (Corallina elongata, Galaxaura oblongata, Cystoseira compressa, Saragassum vulgare, and Stypopodium schimperi) were processed into powders and blended with PLA to fabricate composite membranes. Cytocompatibility assays using human periodontal ligament fibroblasts (n = 3) were performed to evaluate biocompatibility. Antibacterial effects were assessed through colony-forming units (CFU) and scanning electron microscopy (SEM) analysis of bacterial colonization on the membranes. The cytocompatibility assays demonstrated suitable biocompatibility of all marine algae-PLA composite membranes with human periodontal ligament fibroblasts. Antibacterial assessment revealed that Sargassum vulgare-PLA membranes exhibited the highest resistance to bacterial colonization, followed by Galaxaura oblongata-PLA and Cystoseira compressa-PLA membranes. SEM analysis confirmed these findings and revealed smooth surface textures for the marine algae-PLA membranes compared to the fibrous and porous structures of collagen membranes. Marine algae-PLA composite membranes show promising antibacterial properties and cytocompatibility for guided bone and tissue regeneration applications. Sargassum vulgare-PLA membranes demonstrated the highest resistance against bacterial colonization. These findings suggest that marine algae-PLA composite membranes could serve as effective biomaterials for infection control and tissue regeneration. Further in vivo validation and investigation of biodegradation properties are necessary to explore their clinical potential.

Platelet-Released Growth Factors Influence Wound Healing-Associated Genes in Human Keratinocytes and Ex Vivo Skin Explants.

Platelet-released growth factors (PRGFs) or other thrombocyte concentrate products, e.g., Platelet-Rich Fibrin (PRF), have become efficient tools of regenerative medicine in many medical disciplines. In the context of wound healing, it has been demonstrated that treatment of chronic or complicated wounds with PRGF or PRF improves wound healing in the majority of treated patients. Nevertheless, the underlying cellular and molecular mechanism are still poorly understood. Therefore, we aimed to analyze if PRGF-treatment of human keratinocytes caused the induction of genes encoding paracrine factors associated with successful wound healing. The investigated genes were Semaphorin 7A (SEMA7A), Angiopoietin-like 4 (ANGPLT4), Fibroblast Growth Factor-2 (FGF-2), Interleukin-32 (IL-32), the CC-chemokine-ligand 20 (CCL20), the matrix-metalloproteinase-2 (MMP-2), the chemokine C-X-C motif chemokine ligand 10 (CXCL10) and the subunit B of the Platelet-Derived Growth Factor (PDGFB). We observed a significant gene induction of SEMA7A, ANGPLT4, FGF-2, IL-32, MMP-2 and PDGFB in human keratinocytes after PRGF treatment. The CCL20- and CXCL10 gene expressions were significantly inhibited by PRGF therapy. Signal transduction analyses revealed that the PRGF-mediated gene induction of SEMA7A, ANGPLT4, IL-32 and MMP-2 in human keratinocytes was transduced via the IL-6 receptor pathway. In contrast, EGF receptor signaling was not involved in the PRGF-mediated gene expression of analyzed genes in human keratinocytes. Additionally, treatment of ex vivo skin explants with PRGF confirmed a significant gene induction of SEMA7A, ANGPLT4, MMP-2 and PDGFB. Taken together, these results describe a new mechanism that could be responsible for the beneficial wound healing properties of PRGF or related thrombocytes concentrate products such as PRF.

Organic Bone Matrix Component Type I and V Collagen Are Not Destructed in Bisphosphonate-Associated Osteonecrosis of the Jaws.

Background and objectives: The investigation of the pathophysiology behind medication-related osteonecrosis (MRONJ) of the jaw mostly focuses on alterations in osteoclast and osteoblast cell activity, but changes in the organic and inorganic bone matrix have rarely been studied. The aim of this study was to investigate whether collagen, the main organic component of extracellular bone matrix, is destructed in osteonecrosis of the jaw secondary to antiresorptive medication. Material and methods: Bone samples of patients with MRONJ (n = 15, control group n = 3) were demineralized, and collagen fragments were separated from intact collagen pellets by ultrafiltration. The quantification of mature collagen cross-links hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) in pellets and ultrafiltrates was performed by high-performance liquid chromatography (HPLC). The detection of hydroxyproline (Hyp) was carried out using a spectrophotometric assay. In addition, collagen chains were analyzed by sodium dodecylsulfate-polyacrylamide gel (SDS-PAGE). Results: The results revealed significantly higher concentrations of HP, LP and Hyp in pellet samples. In addition, there were no significant differences between samples from MRONJ patients and those of the control group. These results were paralleled by SDS- PAGE. Conclusion: These findings suggest that MRONJ does not involve the destruction of type I and V collagen molecules, in contrast to previously reported destruction by osteoradionecrosis.

Platelet-Released Growth Factors Induce Genes Involved in Extracellular Matrix Formation in Human Fibroblasts.

Platelet concentrate products are increasingly used in many medical disciplines due to their regenerative properties. As they contain a variety of chemokines, cytokines, and growth factors, they are used to support the healing of chronic or complicated wounds. To date, underlying cellular mechanisms have been insufficiently investigated. Therefore, we analyzed the influence of Platelet-Released Growth Factors (PRGF) on human dermal fibroblasts. Whole transcriptome sequencing and gene ontology (GO) enrichment analysis of PRGF-treated fibroblasts revealed an induction of several genes involved in the formation of the extracellular matrix (ECM). Real-time PCR analyses of PRGF-treated fibroblasts and skin explants confirmed the induction of ECM-related genes, in particular transforming growth factor beta-induced protein (TGFBI), fibronectin 1 (FN1), matrix metalloproteinase-9 (MMP-9), transglutaminase 2 (TGM2), fermitin family member 1 (FERMT1), collagen type I alpha 1 (COL1A1), a disintegrin and metalloproteinase 19 (ADAM19), serpin family E member 1 (SERPINE1) and lysyl oxidase-like 3 (LOXL3). The induction of these genes was time-dependent and in part influenced by the epidermal growth factor receptor (EGFR). Moreover, PRGF induced migration and proliferation of the fibroblasts. Taken together, the observed effects of PRGF on human fibroblasts may contribute to the underlying mechanisms that support the beneficial wound-healing effects of thrombocyte concentrate products.

Silk Fiber-Reinforced Hyaluronic Acid-Based Hydrogel for Cartilage Tissue Engineering.

A continuing challenge in cartilage tissue engineering for cartilage regeneration is the creation of a suitable synthetic microenvironment for chondrocytes and tissue regeneration. The aim of this study was to develop a highly tunable hybrid scaffold based on a silk fibroin matrix (SM) and a hyaluronic acid (HA) hydrogel. Human articular chondrocytes were embedded in a porous 3-dimensional SM, before infiltration with tyramine modified HA hydrogel. Scaffolds were cultured in chondropermissive medium with and without TGF-ß1. Cell viability and cell distribution were assessed using CellTiter-Blue assay and Live/Dead staining. Chondrogenic marker expression was detected using qPCR. Biosynthesis of matrix compounds was analyzed by dimethylmethylene blue assay and immuno-histology. Differences in biomaterial stiffness and stress relaxation were characterized using a one-step unconfined compression test. Cell morphology was investigated by scanning electron microscopy. Hybrid scaffold revealed superior chondro-inductive and biomechanical properties compared to sole SM. The presence of HA and TGF-ß1 increased chondrogenic marker gene expression and matrix deposition. Hybrid scaffolds offer cytocompatible and highly tunable properties as cell-carrier systems, as well as favorable biomechanical properties.

Regenerative Potential of Platelet Concentrate Lysate in Mechanically Injured Cartilage and Matrix-Associated Chondrocyte Implantation In Vitro.

Adjuvant therapy in autologous chondrocyte implantation (ACI) can control the post-traumatic environment and guide graft maturation to support cartilage repair. To investigate both aspects, we examined potential chondro-regenerative effects of lysed platelet concentrate (PC) and supplementary interleukin 10 (IL-10) on mechanically injured cartilage and on clinically used ACI scaffolds. ACI remnants and human cartilage explants, which were applied to an uniaxial unconfined compression as injury model, were treated with human IL-10 and/or PC from thrombocyte concentrates. We analyzed nuclear blebbing/TUNEL, sGAG content, immunohistochemistry, and the expression of COL1A1, COL2A1, COL10A1, SOX9, and ACAN. Post-injuriously, PC was associated with less cell death, increased COL2A1 expression, and decreased COL10A1 expression and, interestingly, the combination with Il-10 or Il-10 alone had no additional effects, except on COL10A1, which was most effectively decreased by the combination of PC and Il-10. The expression of COL2A1 or SOX9 was statistically not modulated by these substances. In contrast, in chondrocytes in ACI grafts the combination of PC and IL-10 had the most pronounced effects on all parameters except ACAN. Thus, using adjuvants such as PC and IL-10, preferably in combination, is a promising strategy for enhancing repair and graft maturation of autologous transplanted chondrocytes after cartilage injury.

In Vitro Comparison of 2 Clinically Applied Biomaterials for Autologous Chondrocyte Implantation: Injectable Hydrogel Versus Collagen Scaffold.

OBJECTIVE: In autologous chondrocyte implantation (ACI), there is no consensus about used bioscaffolds. The aim of this study was to perform an in vitro comparative analysis of 2 clinically applied biomaterials for cartilage lesion treatment. DESIGN: Monolayer expanded human chondrocytes (n = 6) were embedded in a collagen scaffold (CS) and a hyaluronic acid-based hydrogel (HA). Cells were cultured in chondropermissive medium supplemented with and without interleukin-10 (IL-10) and bone morphogenetic protein-2 (BMP-2). Gene expression of chondrogenic markers (COL1A1, COL2A1, COL10A1, ACAN, SOX9) was detected via quantitative real-time-polymerase chain reaction (RT-qPCR). Biosynthesis of matrix compounds, cell viability, morphology as well as migration from surrounding native bovine cartilage into cell-free scaffolds were analyzed histologically. Adhesion of the material to adjacent cartilage was investigated by a custom-made push-out test. RESULTS: The shift of COL1/2 ratio toward COL2A1 was more pronounced in HA, and cells displayed a more spherical morphology compared with CS. BMP-2 and IL-10 significantly increased COL2A1, SOX9, and ACAN expression, which was paralleled by enhanced staining of glycosaminoglycans (GAGs) and type 2 collagen in histological sections of CS and HA. COL10A1 was not significantly expressed in HA and CS. Better interfacial integration and enhanced cell invasion was observed in CS. Push-out tests using CS showed higher bonding strength to native cartilage. CONCLUSION: HA-based hydrogel revealed a more chondrocyte-like phenotype but only allowed limited cell invasion, whereas CS were advantageous in terms of cellular invasion and interfacial adhesion. These differences may be clinically relevant when treating cartilaginous or osteochondral defects.

Robot-assisted scaffold implantation and two-stage flap raising of the greater omentum for reconstruction of the facial skeleton: Description of a novel technique.

BACKGROUND: Bone regeneration in the greater omentum is a promising strategy in facial skeleton reconstruction. This feasibility-study aims to perform robot-assisted scaffold implantation and second-stage flap raising. METHODS: Firstly, scaffolds were implanted into the greater omentum using the da Vinci Xi surgical system or conventional technique in five miniature pigs, respectively. After 3 months a free flap including the gastrocolic vascular pedicle was harvested and procedures were evaluated. RESULTS: The average operation time was 59.2 min for open surgery and 73.8 min for robot-assisted surgery. The average incision length of open surgery was 11.6 cm. Both techniques led to successful scaffold implantation without any complication. In all cases the scaffolds were integrated as intended and resulted in bone formation. CONCLUSIONS: Current study demonstrated that the implantation of scaffolds into the greater omentum and flap harvesting using da Vinci Xi surgical system is a promising minimally-invasive approach in regenerative surgery.

Investigation of Topographical Alterations in Titanium-Zirconium-Alloy Implant Threads following Er:YAG Irradiation.

The aim of the current experimental study was to comparatively assess the surface alterations in titanium and titanium-zirconium alloy implants in terms of thread pitch topography after irradiation with an Er:YAG laser, which is recommended in the literature for its sterilizing effect in the treatment of contaminated implant surfaces. Roxolid® and SLA® (Sand-blasted, Large-grit, Acid-etched) implants from Straumann® company with the same macro topography were investigated. The surface treatment was carried out using a wavelength of 2940 nm, 60 s irradiation time, a frequency of 10 Hz, and energies between 120 mJ and 250 mJ. The alterations were quantitatively analyzed by conducting roughness analysis via white light interferometry and qualitatively using SEM images. Roxolid® could particularly maintain its surface topography at a level of 160 mJ. At an energy level of 250 mJ, the surface properties of the pitch could be significantly altered for the first time. Compared to the Standard Plus dental implants studied, no distinct removal of the material from the surface was detected. The alloy properties of Roxolid® confirm the manufacturer's statement in terms of stability and could offer advantages in peri-implantitis management if decontamination has been selected. However, as a part of a respective strategy, smoothening of a Roxolid® implant surface requires a significantly higher energy level compared to SLA-Standard® dental implants.

Learning Curve and Comparison of Dynamic Implant Placement Accuracy Using a Navigation System in Young Professionals.

The aim of the current study was to evaluate the learning curve and accuracy of implant placement by young professionals using a dynamic computer-assisted surgical system for dental implant placement. Ten students tried to place eight implants with a dynamic surgical system in predefined positions on two consecutive weekends, resulting in 160 implant placements in total. Postoperatively, the positions of the implants were scanned with an intraoral scanner and compared for deviations at the entry point, the apex, as well as angular deviations to the master model. The mean values of all measurements improved; statistical significance was found for the changes in the angle as well as for the position of the implants to the apex (p < 0.001). Furthermore, the young professionals indicated subjective improvement in handling the dynamic surgery system. Navigated surgical dental implant placement can be learned quickly and can support young professionals in everyday clinical practice, especially in difficult anatomic situations.

Role of myeloid early endothelial progenitor cells in bone formation and osteoclast differentiation in tissue construct based on hydroxyapatite poly(ester-urethane) scaffolds.

Engineering of a vascularized bone construct is a highly challenging task which needs to take into account the impact of different components on the bone regeneration process. Bone repair influencing factors in such constructs range from the material properties and scaffold design, to the interaction of different cell types contributing to bone formation and remodeling or neovascularization, respectively. In this context, early endothelial progenitor cells (EPC), mononuclear cells isolated from the peripheral blood, express the endothelial marker CD31 but also a series of myeloid markers and have been shown to support the formation of vessel-like structures. These cells are also characterized by a highly adaptable phenotype influenced by other cells creating an instructive niche. The present study was designed to investigate the impact of EPC on bone formation or remodeling using a co-culture system of outgrowth endothelial cells, mature endothelial cells isolated from the peripheral blood cell cultures, and mesenchymal stem cells grown on hydroxyapatite poly(ester-urethane) scaffolds. The formation of vessel-like structures in these constructs was shown by CLSM and immunohistochemistry and further evaluated by real time RT-PCR. Osteogenic differentiation in these constructs was investigated by von Kossa, Alizarin Red, and real time PCR. Data indicated that osteogenic differentiation occurred within the constructs after 14 days of culture but without a direct influence by EPC in this process. Finally, although we observed a series of osteoclast related makers in the constructs when EPC were included, no indications for an increased osteoclast-like activity, which might lead to increased bone resorption, were observed. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1922-1932, 2016.