Osteoblast Cell Growth Research Articles

Evolutionary Design of Self-Templated Supramolecular Fibrils Using M13 Bacteriophage for Tissue Engineering.

Biomaterials in nature form hierarchical structures and functions across various length scales through binding and assembly processes. Inspired by nature, we developed hierarchically organized tissue engineering materials through evolutionary screening and self-templating assembly. Leveraging the M13 bacteriophage (phage), we employed an evolutionary selection process against hydroxyapatite (HA) to isolate HA-binding phage (HAPh). The newly discovered phage exhibits a bimodal length, comprising 950 nm and 240 nm, where the synergistic effect of these dual lengths promotes the formation of supramolecular fibrils with periodic banded structures. The assembled HAPh fibrils show the capability of HA mineralization and the directional growth of osteoblast cells. When applied to a dentin surface, it induces the regeneration of dentin-like tissue structures, showcasing its potential applications as a scaffold in tissue engineering. The integration of evolutionary screening and self-templating assembly holds promise for the future development of hierarchically organized tissue engineering materials.

Investigating adhesion of primary human gingival fibroblasts and osteoblasts to orthodontic mini-implants by scanning electron microscopy

Miniscrews offer controlled anchorage and thus optimize tooth movement in orthodontic treatment. Nevertheless, failures such as soft tissue problems, instability due to loosening, partial osseointegration, or even device fracture can occur. While clinical technique can play a role in some of these problems, the manufacturer’s design and material choice influence how the implant interacts with the surrounding tissue. In some cases, the design and material may trigger unwanted bone and soft tissue responses. This in vitro study investigates how the implant surface affects cell adhesion and growth of human primary fibroblasts and osteoblasts on commercially available orthodontic TiAl6V4 miniscrews from three producers: tomas-pin SD N 08 (Dentaurum), OrthoEasy Pin (Forestadent), and Dual Top G2 (Promedia, Jeil Medical). Cell–implant interaction at the top, neck, and drilling part of the screws was assessed qualitatively by scanning electron microscopy. While both cell types adhered to and grew on all products, subtle differences in cell shape and spreading were detected, depending on the microstructure of the implant surface. This indicates that cell adhesion to implant surfaces can be controlled by manipulating the machining conditions.

Evaluation of Cardiospermum halicacabum on Bone Morphogenetic Protein-2 (BMP2) mRNA Expression in Osteoblast Cells.

Introduction Maintaining bone health is crucial for overall well-being, with osteoblasts playing a vital role in bone formation. Bone morphogenetic protein-2 (BMP2) is a key regulator, stimulating bone matrix synthesis and osteoblast differentiation. Recognizing BMP2's significance, there's growing interest in natural compounds, such as Cardiospermum halicacabum. This study explores Cardiospermum halicacabum's potential influence on BMP2 mRNA expression in osteoblast cells for insights into bone health modulation. Materials and methods This research utilized Cardiospermum halicacabum to explore its impact on MG-63 cells, a human osteoblast cell line. Osteoblast cells were cultured in Dulbecco's modified Eagle's medium (DMEM), supplemented with 10% heat-inactivated fetal bovine serum, and maintained at 37°C in a 5% CO2 and 95% air environment. Cell viability was evaluated by seeding osteoblast cells into 96-well plates and exposing them to different concentrations of Cardiospermum halicacabum (2.0 μg/ml and 20 μg/ml). The study observed both the promotion of osteoblast cell growth in MG-63 and morphological changes in thecells under an inverted light microscope at 10x magnification. Results were presented using one-way analysis of variance (ANOVA) conducted with IBM SPSS Statistics for Windows, Version 23 (Released 2015; IBM Corp., Armonk, New York, United States). Result The reverse transcription-polymerase chain (RT-PCR) results revealed an increased expression of BMP-2 mRNA fold change in comparison to the control group. A clear positive correlation was observed between the BMP-2 mRNA fold change and the notable increase in the concentration of Cardiospermum halicacabum. This investigation revealed a direct association of BMP-2 mRNA expressionwith the proliferation of osteoblast cells. Specifically, the BMP-2 mRNA fold change was recorded at 2.26±1.05 in Cardiospermum halicacabum at 2.0 μg/ml and 2.0 ± 0.84 at 20 μg/ml, with corresponding significances of 0.00, respectively. Conclusion Potential effects of Cardiospermum halicacabum on BMP-2 mRNA expression in osteoblast cellsand its role in bone health modulation revealed that Cardiospermum halicacabum may upregulate BMP-2 mRNA expression, suggesting its potential as a natural compound for enhancing bone formation. The observed positive correlation between Cardiospermum halicacabum concentration and BMP-2 mRNA fold change showed the significance of this botanical agent in promoting osteoblast cell proliferation. These results highlight the importance of further research to explore the applications of Cardiospermum halicacabum in managing bone disorders and improving overall bone health.

3D Poly (L-lactic acid) fibrous sponge with interconnected porous structure for bone tissue scaffold

Large bone defects, often resulting from trauma and disease, present significant clinical challenges. Electrospun fibrous scaffolds closely resembling the morphology and structure of natural ECM are highly interested in bone tissue engineering. However, the traditional electrospun fibrous scaffold has some limitations, including lacking interconnected macropores and behaving as a 2D scaffold. To address these challenges, a sponge-like electrospun poly(L-lactic acid) (PLLA)/polycaprolactone (PCL) fibrous scaffold has been developed by an innovative and convenient method (i.e., electrospinning, homogenization, progen leaching and shaping). The resulting scaffold exhibited a highly porous structure (overall porosity = 85.9 %) with interconnected, regular macropores, mimicking the natural extracellular matrix. Moreover, the incorporation of bioactive glass (BG) particles improved the hydrophilicity (water contact angle = 79.7°) and biocompatibility and promoted osteoblast cell growth. In-vitro 10-day experiment revealed that the scaffolds led to high cell viability. The increment of the proliferation rates was 195.4 % at day 7 and 281.6 % at day 10. More importantly, Saos-2 cells could grow, proliferate, and infiltrate into the scaffold. Therefore, this 3D PLLA/PCL with BG sponge holds great promise for bone defect repair in tissue engineering applications.

Scratch resistance and in-vitro biocompatibility of plasma-sprayed baghdadite coatings reinforced with carbon nanotubes

Hydroxyapatite (HA, Ca10(PO4)6(OH)2) coatings, commonly used for metallic implants, have limitations such as excessive brittleness, low fracture toughness, inadequate wear resistance, and slow osseointegration properties. In contrast, baghdadite (BAG, Ca3ZrSi2O9), a calcium zirconium silicate-based bioceramic, exhibits outstanding biological properties, to promote cell proliferation and differentiation of human osteoblasts along with adequate mechanical strength. In this study, plasma-sprayed HA, BAG, and carbon nanotubes (CNT, 1 wt%, and 2 wt%) reinforced BAG coatings are deposited on titanium (Ti) substrate to enhance its mechanical as well as biological properties. The microhardness values of CNT-reinforced BAG coatings increase due to a decrease in porosity and the retention of CNT inside the BAG matrix. Progressive loading scratch tests are performed, revealing a reduction in wear volume loss from 15.051 mm3 to 12.574 mm3 and scratch rate from 43.262 mm3N−1 m−1 to 36.172 mm3N−1 m−1 with the addition of 2 wt% CNT in BAG coatings. The scratch hardness test results demonstrated that introducing CNT into BAG coating significantly enhances its performance, showing a remarkable 38.7 % improvement. In-vitro cell culture studies indicate excellent cell adhesion, growth, and proliferation of MC3T3-E1 osteoblast cells on the surfaces of the CNT-reinforced BAG coatings.

A Porous Fluoride-Substituted Bovine-Derived Hydroxyapatite Scaffold Constructed for Applications in Bone Tissue Regeneration.

Hydroxyapatite is widely used in bone implantation because of its similar mineral composition to natural bone, allowing it to serve as a biocompatible osteoconductive support. A bovine-derived hydroxyapatite (BHA) scaffold was developed through an array of defatting and deproteinization procedures. The BHA scaffold was substituted with fluoride ions using a modified sol-gel method to produce a bovine-derived fluorapatite (BFA) scaffold. Fourier-transform infrared spectroscopy and X-ray diffraction analysis showed that fluoride ions were successfully substituted into the BHA lattice. According to energy dispersive X-ray analysis, the main inorganic phases contained calcium and phosphorus with a fluoride ratio of ~1-2 wt%. Scanning electron microscopy presented a natural microporous architecture for the BFA scaffold with pore sizes ranging from ~200-600 μm. The BHA scaffold was chemically stable and showed sustained degradation in simulated-body fluid. Young's modulus and yield strength were superior in the BFA scaffold to BHA. In vitro cell culture studies showed that the BFA was biocompatible, supporting the proliferative growth of Saos-2 osteoblast cells and exhibiting osteoinductive features. This unique technique of producing hydroxyapatite from bovine bone with the intent of producing high performance biomedically targeted materials could be used to improve bone repair.

Alginate-CaCO3 hybrid colloidal hydrogel with tunable physicochemical properties for cell growth

Biomaterials composed of food polysaccharides are of great interest for future biomedical applications due to their great biocompatibility, tunable mechanical properties, and complex architectural designs that play a crucial role in the modulation of cell adhesion and proliferation. In this work, a facile approach was designed to obtain novel 3D alginate-CaCO3 hybrid hydrogel particles in situ. Controlling the gel concentration from 3 to 20 mg·mL−1 allows us to control the alginate-CaCO3 hydrogel particles' size and density (size variation from 1.86 to 2.34 mm and density from 1.22 to 1.29 mg/mm3). This variable also has a considerable influence on the mineralization process resulting in CaCO3 particles with varied sizes and amounts within the hydrogel beads. The measurements of Young's modulus showed that the inclusion of CaCO3 particles into the alginate hydrogel improved its mechanical properties, and Young's modulus of these hybrid hydrogel particles had a linear relationship with alginate content and hydrogel particle size. Cell experiments indicated that alginate-CaCO3 hybrid hydrogel particles can support osteoblastic cell proliferation and growth. In particular, the amount of hydroxyapatite deposition on the cell membrane significantly increased after the treatment of cells with hybrid hydrogel particles, up to 20-fold. This work offers a strategy for constructing inorganic particle-doped polysaccharide hybrid hydrogel scaffolds that provide the potential to support cell growth.

In vitro and in vivo study of copper-doped bioactive glass for bone regeneration application

Bioactive glasses (BGs) are being explored in bone regeneration applications with great interest banking on their excellent bioactivity, angiogenesis, osteogenesis, and bone-bonding capacity. In this connection, the designing of metal-doped BG composite with enhanced biological properties is attracting the attention of the scientific community. With this motivation, the present study focuses on the synthesis of copper-doped 70S30C bioactive glass (CuBGs) using a simple precipitation method, wherein rice husk, eggshell, and copper sulfate were used as a precursor for silica, calcium, and copper. The objective of the study is to synthesize acid-free and binder-free CuBGs for bone regeneration application. The structural and textural properties of the CuBG samples were studied using XRD, FTIR, SEM-EDS, and BET techniques. In addition, in vitro bioactivity and degradation studies were performed in simulated body fluid (SBF). The antibacterial activity of the CuBGs shows an inhibitory effect against E. coli and S. aureus bacteria. The in vitro haemolysis study indicates haemo-compatibility and significantly improved angiogenesis after copper doping. The histology study of chick bone demonstrates the growth of osteoblast cells. Further, MG-63 cells were exposed to the CuBGs samples (concentrations up to 10 mg mL−1) to assess biocompatibility and it was observed that it did not exhibit any toxic effects. This is supported by an in vivo biocompatibility study indicating its non-toxic nature. The results reveal that CuBG plays an important role in enhancing bioactivity, antibacterial activity, and angiogenesis, and it is non-toxic making it a promising candidate for bone tissue engineering application.

Hydroxyapatite coated titanium with curcumin and epigallocatechin gallate for orthopedic and dental applications

Titanium and its alloy are clinically used as an implant material for load-bearing applications to treat bone defects. However, the lack of biological interaction between bone tissue and implant and the risk of infection are still critical challenges in clinical orthopedics. In the current work, we have developed a novel approach by first 1) modifying the implant surface using hydroxyapatite (HA) coating to enhance bioactivity and 2) integrating curcumin and epigallocatechin gallate (EGCG) in the coating that would induce chemopreventive and osteogenic potential and impart antibacterial properties to the implant. The study shows that curcumin and EGCG exhibit controlled and sustained release profiles in acidic and physiological environments. Curcumin and EGCG also show in vitro cytotoxicity toward osteosarcoma cells after 11 days, and the dual system shows a ~94 % reduction in bacterial growth, indicating their in vitro chemopreventive potential and antibacterial efficacy. The release of both curcumin and EGCG was found to be compatible with osteoblast cells and further promotes their growth. It shows a 3-fold enhancement in cellular viability in the dual drug-loaded implant compared to the untreated samples. These findings suggest that multifunctional HA-coated Ti6Al4V implants integrated with curcumin and EGCG could be a promising strategy for osteosarcoma inhibition and osteoblast cell growth while preventing infection.

Enhanced knee joint treatment using a hybrid hyaluronic acid-alginate filler reinforced with hydroxyapatite-titanium nanoparticles for sports-related injuries

Sports injuries, particularly bone injuries, have become increasingly prevalent in recent years. As athletes push themselves harder and become more competitive, the risk of injury rises. Additionally, the aging population has contributed to the increased prevalence of bone injuries, including those related to the knee joint. As a result, significant research efforts have focused on developing new techniques for knee joint repair. In this study, a novel hyaluronic acid-Alginate filler composition containing hydroxyapatite-titanium nanoparticles (HA/Ti-NPs) was prepared through freeze-drying. The crystalline properties of the porous scaffolds were analyzed using X-ray diffraction (XRD), while scanning electron microscope (SEM) images were used to visualize the scaffold structure. The morphological investigation demonstrated that adding 15 wt% Ti-NP to the scaffold reduces percentages of porosity to about 76.5 ± 5, while increasing the elastic modulus. Furthermore, the presence of nanoparticles decreased the rate of degradation, increased crystallinity, reduced pore size, and promoted calcium release, creating a favorable environment for bone tissue growth. The scaffold's degradation pH is close to neutral, which is ideal for bone tissue treatment. Adding 15 wt% Ti-NP reduces the contact angle from 120° to 93°. The composite scaffolds promote the proliferation and growth of osteoblast cells, and after 48 h, degraded material improved cell viability, growth, and proliferation. The three-component hyaluronic acid-Alginate/HA scaffolds with 15 wt% Ti-NP exhibited suitable morphological, mechanical, physical, and chemical properties for bone applications, as well as high potential for joint bonding. These porous scaffolds have the potential to be used as bone fillers of sport injuries and further research is needed to confirm their efficacy.

An investigation into the cytotoxic effects of microbubbles and their constituents on osteosarcoma and bone marrow stromal cells

BackgroundUltrasound-responsive microbubbles offer a means of achieving minimally invasive, localised drug delivery in applications including regenerative medicine. To facilitate their use, however, it is important to determine any cytotoxic effects they or their constituents may have. The aim of this study was to test the hypothesis that phospholipid-shelled microbubbles are non-toxic to human bone-derived cells at biologically-relevant concentrations. MethodsMicrobubbles were fabricated using combinations of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dibehenoyl-sn-glycero-3-phosphocholine (DBPC), polyoxyethylene(40) stearate (PEG40S) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene-glycol)-2000] (DSPE-PEG2000). Microbubble size and concentration were measured as a function of time and temperature by optical microscopy. Effects on MG63 osteosarcoma and human bone marrow stromal cells (BMSCs) were measured for up to 72 h by assay for viability, metabolic activity and proliferation. ResultsDBPC:DSPE-PEG2000 microbubbles were significantly more stable than DSPC:PEG40S microbubbles under all conditions tested. Serum-containing medium had no detrimental effect on microbubble stability, but storage at 37 °C compared to at 4 °C reduced stability for both preparations, with almost complete dissolution of microbubbles at times ≥24 h. DSPC:PEG40S microbubbles had greater inhibitory effects on cell metabolism and growth than DBPC:DSPE-PEG2000 microbubbles, with PEG40S found to be the principle inhibitory component. These effects were only evident at high microbubble concentrations (≥20% (v/v)) or with prolonged culture (≥24 h). Increasing cell-microbubble contact by inversion culture in a custom-built device had no inhibitory effect on metabolism. ConclusionsThese data indicate that, over a broad range of concentrations and incubation times, DBPC:DSPE-PEG2000 and DSPC:PEG40S microbubbles have little effect on osteoblastic cell viability and growth, and that PEG40S is the principle inhibitory component in the formulations investigated.

Guar-Based Injectable Hydrogel for Drug Delivery and In Vitro Bone Cell Growth.

Injectable hydrogels offer numerous advantages in various areas, which include tissue engineering and drug delivery because of their unique properties such as tunability, excellent carrier properties, and biocompatibility. These hydrogels can be administered with minimal invasiveness. In this study, we synthesized an injectable hydrogel by rehydrating lyophilized mixtures of guar adamantane (Guar-ADI) and poly-β-cyclodextrin (p-βCD) in a solution of phosphate-buffered saline (PBS) maintained at pH 7.4. The hydrogel was formed via host-guest interaction between modified guar (Guar-ADI), obtained by reacting guar gum with 1-adamantyl isocyanate (ADI) and p-βCD. Comprehensive characterization of all synthesized materials, including the hydrogel, was performed using nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and rheology. The in vitro drug release study demonstrated the hydrogel's efficacy in controlled drug delivery, exemplified by the release of bovine serum albumin (BSA) and anastrozole, both of which followed first-order kinetics. Furthermore, the hydrogel displayed excellent biocompatibility and served as an ideal scaffold for promoting the growth of mouse osteoblastic MC3T3 cells as evidenced by the in vitro biocompatibility study.

Structural, cell culture, antibacterial and in vitro bioactivity analysis of highly-ordered Al2O3 nanotube coating with decorated Ag2O nanoparticles on Ti-6Al-4V

The alumina nanotubular arrays (Al2O3 NTs) are a promising candidate for application in bone and dental implants for their better mechanical and chemical behavior in the human body compared to titanium. Nevertheless, the biocompatibility and antibacterial activity of Al2O3 NTs still need further improvement to meet the requirements of safe implants. In this research, Al2O3 NTs coating was proposed on Ti-6Al-4 V via magnetron sputtering pure aluminum followed by an anodization and sealing process, then heat treated for 1 h at 450 °C. After that, Ag2O nanoparticles (NPs) were decorated on the nanotubular walls under controlled conditions using magnetron sputtering. The findings demonstrated that the proposed Al2O3 NTs-Ag2O NPs promoted in-vitro bioactivity alongside excellent antibacterial activity against E.coli and S.aureus bacteria within 4 h and 6 h, respectively. Additionally, the cell-culture showed healthy growth of the human osteoblast cells (HOb) on the Al2O3 NTs-Ag2O NPs. The new design of coating also shows that the total cumulative Ag+released after 14 days is significantly low which is safe and provides the long-term antibacterial activity. It is illustrated that Al2O3 NTs-Ag2O NPs have promising biomedical properties to enhance the biofunctionality of biomedical-grade Ti-6Al-4 V implants.

Abstract 3828: Herbal extracts from lycii radicis corex and achyranthes japonica prevent multiple myeloma progression

Abstract Multiple Myeloma (MM) is a plasma cell malignancy that, despite advances in treatment, remains incurable. In over 90% of MM patients, aberrant bone remodeling occurs and results in osteolytic lesions. Korean traditional medicine has a long-standing interest in healthy foods to enhance the immune system, energy-boost, and Yin-and-Yang balance. Traditional medicine has used herbal extracts (HE) from natural plants with fewer side effects and long-term treatment tolerance. In earlier studies, lycii radices cortex (LRC) and achyranthes japonica (AJ) containing HE have demonstrated the ability to enhance cell growth and mineralization of osteoblast cells while also inhibiting osteoclast differentiation. The current study investigated the effects of LRC+AJ containing HE on murine 5TGM1 MM-bearing mice.This HE significantly increased the mouse survivals of both Treatment before Transplant (TbT) and Treatment after Transplant (TaT) groups with median survivals of undetermined and 52.5 days, respectively, while MM control had a median survival of 42 days. The Mantel-Cox test found that TbT and TaT mice were significantly different from the control group (P=0.0014 and 0.0182, respectively). In histomorphometric analysis, the spines were scanned by micro-CT and revealed that TbT and TaT groups had significantly increased bone volume over total volume (BV/TV) than control. To see if HE affects cell growth of myeloma cells and osteoblast cells, we further investigated the HE on preosteoblast cell line MC3T3-E1, mouse MM cell line 5TGM1-Luc, and human MM cell line U-266. Cells were treated with HE, and viability was assessed at 48 and 96 hours post-treatment. Remarkably, LRC+AJ containing HE increased MC3T3-E1 cell growth while it decreased 5TGM1 and U-266 cell viability. In addition, we found the DAP3G in HE used in this study.Our results demonstrated that LRC+AJ HE prevents MM and promotes bone formation in 5TGM1 engrafted NSG mice. We also found that LRC+AJ HE suppresses myeloma cell growth and enhances osteoblast cell survival. Although it is yet to be defined, a correlation of osteoblast activity and myeloma cell inhibition suggests a possible connection of these events to prevent MM progression. Citation Format: Syed Hassan Mehdi, Sun-OK Lee, Dam Huh, Donghoon Yoon. Herbal extracts from lycii radicis corex and achyranthes japonica prevent multiple myeloma progression. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3828.

Effects of Electronic Cigarette E-Liquids on the Adhesion and Growth of Osteoblast Cells

Electronic cigarettes (E-cigarettes) were designed to replace traditional cigarettes to decrease the harmful effects of smoking. The liquid in e-cigarettes contains different chemicals, such as Propylene Glycol (PG) and Vegetable Glycerin (VG), and may contain nicotine. To be more attractive, e-liquids are supplemented with flavorings. Because vaped e-liquids are shown to be damaging to oral tissues, similar effects could occur in the oral cavity with non-vaped e-liquids. We evaluated the effects of non-vaped e-liquid constituents on osteoblast behavior. For this purpose, human osteoblast cells (MG-63) were exposed to e-liquid containing 70% PG -30% VG with and without tobacco flavor and with or without nicotine at 12, 18 mg/mL, or no nicotine content. The e-liquids were used at various concentrations (0, 1, and 5%). To evaluate the effect of the e-liquids on osteoblast morphology and adhesion, optical microscope observations and viable cell counting using trypan blue exclusion were used. Cell growth was also analyzed using the Methyl Thiazol Tetrazolium (MTT) assay, while cytotoxicity was tested by measuring Lactate Dehydrogenase (LDH) levels after cell exposure for 24 h to the e-liquids. Our results show that e-liquids induced significant morphological changes evidenced by round cell forms, with no contact between cells. Adhesion was significantly reduced, particularly in the presence of nicotine. E-liquids at a concentration of 5% significantly reduced osteoblast growth. This effect was observed with both flavored and non-flavored e-liquid with or without nicotine. The decreased osteoblast adhesion and growth after exposure to e-liquid was confirmed by increased levels of LDH. Overall results indicate the potentially harmful effects of e-liquid non-vaped chemicals on bone cells, which could lead to the impairment of bone regeneration and tissue remodeling processes.

Bioactive Composite Methacrylated Gellan Gum for 3D-Printed Bone Tissue-Engineered Scaffolds

Gellan gum (GG) was chemically modified with methacrylic moieties to produce a photocrosslinkable biomaterial ink, hereinafter called methacrylated GG (GGMA), with improved physico-chemical properties, mechanical behavior and stability under physiological conditions. Afterwards, GGMA was functionalized by incorporating two different bioactive compounds, a naturally derived eumelanin extracted from the black soldier fly (BSF-Eumel), or hydroxyapatite nanoparticles (HAp), synthesized by the sol-gel method. Different ink formulations based on GGMA (2 and 4% (w/v)), BSF-Eumel, at a selected concentration (0.3125 mg/mL), or HAp (10 and 30% wHAp/wGGMA) were developed and processed by three-dimensional (3D) printing. All the functionalized GGMA-based ink formulations allowed obtaining 3D-printed GGMA-based scaffolds with a well-organized structure. For both bioactive signals, the scaffolds with the highest GGMA concentration (4% (w/v)) and the highest percentage of infill (45%) showed the best performances in terms of morphological and mechanical properties. Indeed, these scaffolds showed a good structural integrity over 28 days. Given the presence of negatively charged groups along the eumelanin backbone, scaffolds consisting of GGMA/BSF-Eumel demonstrated a higher stability. From a mechanical point of view, GGMA/BSF-Eumel scaffolds exhibited values of storage modulus similar to those of GGMA ones, while the inclusion of HAp at 30% (wHAp/wGGMA) led to a storage modulus of 32.5 kPa, 3.5-fold greater than neat GGMA. In vitro studies proved the capability of the bioactivated 3D-printed scaffolds to support 7F2 osteoblast cell growth and differentiation. BSF-Eumel and HAp triggered a different time-dependent physiological response in the osteoblasts. Specifically, while the ink with BSF-Eumel acted as a stimulus towards cell proliferation, reaching the highest value at 14 days, a higher expression of alkaline phosphatase activity was detected for scaffolds consisting of GGMA and HAp. The overall findings demonstrated the possible use of these biomaterial inks for 3D-printed bone tissue-engineered scaffolds.

Effectiveness of Marine Bioactive Compound Fucoidan in Stimulating Osteoblast Cells Formation: A Systematic Review

BACKGROUND: Bone loss in the oral cavity can occur in various situations, including periodontitis-related issues and complications after tooth extraction. A damaged or lost alveolar bone can be restored through a tissue engineering approach. Fucoidan, a marine biopolymer derived from brown algae, is one biomaterial that aids bone regeneration since it contains biomaterials that can generate new osteoblast cells. AIM: The purpose of this review is to determine whether fucoidan can be employed to promote osteoblast cell growth during bone repair. METHODS: The search strategy was performed in PubMed; Elsevier (Scopus); ScienceDirect; Libgen, and Google Scholar. In addition, a manual hand searching was performed to locate and identify additional studies. RESULTS: Based on the reviewed articles, it has been discovered that five met the inclusion criteria and found that the marine bioactive compound fucoidan can significantly increase the expression of ALP activity. CONCLUSION: Fucoidan is considered to have biological properties, including antithrombotic, anticoagulant, and antioxidant. It also serves as a phenotypic marker during the early stages of osteoblastic differentiation.

The effect of Cissus quadrangularis Salisb. extract on maturation of rat mesenchymal stem cells.

Degenerative diseases, such as osteoporosis, could be treated by stem cells. The aim of this study was to identify the gene expression of bone marrow mesenchymal stem cells (BM-MSC) derived from Sprague Dawley rats and to assess the effect of Cissus quadrangularis Salisb. extract on their maturation into bone cells. The BM-MSC were divided into three groups: (a) BM-MSCs + osteoblast cell growth basal medium as the positive control; (b) BM-MSCs + Dulbecco's modified eagle's medium (DMEM) + 0.3 mg/mL methanol extract of C. quadrangularis as methanol group; and (c) BM-MSC + DMEM + 0.3 mg/mL ethyl acetate extract of C. quadrangularis as ethyl acetate group. A relative quantification approach using was used to analyze the expression of the alp (alkaline phosphatase) gene, with the beta-actin gene was used to normalize the expression of the alp gene. The intra-assay variation was calculated to validate the RT-qPCR data. Our study found that the intra-assay variation value was acceptable, with most of the coefficients of variability (CV) value <5. Ethyl acetate solvent outperformed methanol solvent in extracting the active compound C. quadrangularis. In the ethyl acetate extract group, the expression of the alp gene increased three times compared to the positive control. In methanol extract group, the expression of alp gene was lower six times compared to positive control. This study suggests that C. quadrangularis extracts using ethyl acetate could induce the maturation of BM-MSCs. However, further studies are warrant to confirm this effect using different indicators.

Synergistic effect of nanostructure and calcium ions on improving the bioactivity of titanium implants.

Surface structure and composition play essential roles in the osseointegration of titanium implants. In the present study, a nanoscale surface structure incorporated with calcium ions was fabricated on a titanium surface by hydrothermal treatment. The characteristics of the surfaces were analysed, and the bioactivity of the samples was evaluated in vitro and in vivo. nm-Ti and nm/Ca-Ti surfaces were significantly more hydrophilic than control-Ti surfaces. nm/Ca-Ti samples showed much faster bone-like apatite precipitation in simulated body fluid than the other samples. The results of MC3T3-E1 cell tests demonstrated that both nm-Ti and nm/Ca-Ti surfaces accelerated cell adhesion and proliferation. The highest level of osteogenesis-related genes (Runx2, bone morphogenetic protein-2, osteopontin and osteocalcin) were observed in nm/Ca-Ti samples, followed by nm-Ti samples. Alizarin red staining experiment showed that the amount of extracellular matrix mineralized nodules in nm/Ca-Ti group was significantly higher than others. In animal experiments using SD rats, nm/Ca-Ti showed the highest value of new bone formation at two and four weeks. The present study suggested that the nanostructure and calcium ions showed synergetic effects on accelerating bone-like apatite precipitation and osteoblast cell growth and differentiation. Animal experiment further indicated that such surface could promote early osteogenesis.

A novel flexible, conductive, and three-dimensional reduced graphene oxide/polyurethane scaffold for cell attachment and bone regeneration

Development of conductive and three-dimensional scaffolds with elastic properties and shape-recovery capability for bone regeneration within irregular bone cavities has been challenging. Polyurethanes (PUs) are intrinsically elastic polymers; however, their mechanical performance, biocompatibility, and functionality need to be improved for application as implants and biomedical devices. Herein, application of a novel flexible, conductive, and three-dimensional polyurethane scaffold (3DPU), fabricated through coating a commercial PU foam with graphene oxide (GO) and its subsequent reduction with ascorbic acid, described as 3DrGO/PU, is investigated for bone regeneration. The 3DrGO/PU scaffold supported the growth and proliferation of mouse osteoblast cells (MG-63) with strong mineralization and cell attachment. It is likely that the electrically conductive macro-porous 3DrGO/PU scaffold, provides bioactivity and promote nucleation and growth of hydroxyapatite (HA) in the simulated body fluid. Experiments assessing in vivo bone formation in rat calvarial skull defects provided clear evidence for efficacy of the 3D scaffold for treatment of irregular bone defects. The results of this study are promising, as they present easy production of a cost-effective green fabricated scaffold. It also offers a potential for cell attachment and capture, to be used in future tissue engineering and even biosensing applications.