Proliferation Of MC3T3-E1 Cells Research Articles

Investigations on microstructure and properties of five TiZr-based refractory multi-principal element alloys for clinical metallic biomaterials

The development of new materials with high strength, ductility, low modulus, wear resistance, and good biocompatibility remains a requirement in the field of clinical biomaterials. Based on the concept of high-entropy alloys, five TiZr-based refractory multi-principal element alloys (RMPEAs), i.e. MoTiZr, NbTiZr, MoTiVZr, NbTiVZr, and NbMoTiZr were designed and their microstructure, mechanical properties, elastic properties, and biocompatibility were assessed in this study. Results indicate that NbTiZr and NbTiVZr RMPEAs are single-phase BCC alloys, while the other alloys exhibit multiphase structures. In terms of mechanical behaviors, NbMoTiZr RMPEA has the highest hardness (496±15 HV) among the five alloys and also combines a high yield strength and good ductility, with values of 1275MPa and 21.66%, respectively. According to atomic calculations, the ideal shear strengths of the five alloys on the {110} plane are all lower than those on the {112} plane. Furthermore, NbMoTiZr RMPEA also exhibits a lower Young’s modulus than that of Ti6Al4V, which can reduce “stress shielding” effects. The in vitro biocompatibility evaluation revealed that NbMoTiZr RMPEA can support cell adhesion and proliferation of MC3T3-E1 cells and exhibits good biocompatibility comparable to Ti6Al4V. These results demonstrate the remarkable potential of NbMoTiZr RMPEA for orthopedic bioimplant applications.

Layer-by-layer self-assembly of PLL/CPP-ACP multilayer on SLA titanium surface: Enhancing osseointegration and antibacterial activity in vitro and in vivo

Implants are expected to possess both excellent osteointegration and antibacterial activity because poor osseointegration and infection are two major causes of titanium implant failure. In this study, we constructed layer-by-layer self-assembly films consisting of anionic casein phosphopeptides-amorphous calcium phosphate (CPP-ACP) and cationic poly (L-lysine) (PLL) on sandblasted and acid etched (SLA) titanium surfaces and evaluated their osseointegration and antibacterial performance in vitro and in vivo. The surface properties were examined, including microstructure, elemental composition, wettability, and Ca2+ ion release. The impact the surfaces had on the adhesion, proliferation and differentiation abilities of MC3T3-E1 cells were investigated, as well as the material’s antibacterial performance after exposure to the oral microorganisms such as Porphyromonas gingivalis (P. g) and Actinobacillus actinomycetemcomitans (A. a). For the in vivo studies, SLA and Ti (PLL/CA-3.0)10 implants were inserted into the extraction socket immediately after extracting the rabbit mandibular anterior teeth with or without exposure to mixed bacteria solution (P. g & A. a). Three rabbits in each group were sacrificed to collect samples at 2, 4, and 6 weeks of post-implantation, respectively. Radiographic and histomorphometry examinations were performed to evaluate the implant osseointegration. The modified titanium surfaces were successfully prepared and appeared as a compact nano-structure with high hydrophilicity. In particular, the Ti (PLL/CA-3.0)10 surface was able to continuously release Ca2+ ions. From the in vitro and in vivo studies, the modified titanium surfaces expressed enhanced osteogenic and antibacterial properties. Hence, the PLL/CPP-ACP multilayer coating on titanium surfaces was constructed via a layer-by-layer self-assembly technology, possibly improving the biofunctionalization of Ti-based dental implants.

Hai-Honghua medicinal liquor is a reliable remedy for fracture by promotion of osteogenic differentiation via activation of PI3K/Akt pathway

Ethnopharmacological relevanceHai-Honghua medicinal liquor (HHML), an external Chinese herbal formula preparation, is often applied to treat freshly closed tibia/fibular fractures, ankle fractures, and other bone-related disorders, but the related molecular mechanism is unclear. Aim of the studyTo evaluate the therapeutic effect of HHML in patients with tibial/fibular and ankle fractures, and to explore its related possible mechanism. Methods and materialsA total of 182 patients with tibia/fibular fractures and 183 patients with ankle fractures were enrolled in this study. A randomized, controlled, unblinded clinical trial was designed to evaluate the therapeutic effect of HHML on tibial/fibular and ankle fractures. The chemical compositions of HHML were analyzed by the HPLC-Q-Extractive MS/MS. Furthermore, a rat tibial fracture model was established to evaluate the therapeutic effects of HHML in promoting fracture healing, and the mouse embryonic osteoblasts cell line of MC3T3-E1 was further carried out to explore the mechanisms of HHML on osteoblast differentiation. ResultsIn the clinical evaluation, HHML treatment significantly shortened the time for pain and swelling in patients with tibial/fibular fractures (P < 0.01) and ankle fractures (P < 0.01), and the incidence of complications was significantly reduced as well. Subsequently, 116 constituents were identified from HHML via HPLC-Q-TOF-MS/MS analysis. In vivo, no obvious changes in weight were observed in HHML-treated rats. Moreover, the levels of bone formation markers (including osteocalcin (OCN), N-terminal propeptide of type I procollagen (PINP), alkaline phosphatase (ALP), calcium (Ca) and substance P) in rat serum were significantly increased in HHML-treated rats compared with model rats (P < 0.05). Micro-CT analysis showed bone mineral density (BMD), bone volume fraction (BV/TV), trabecular thickness (Tb.Th) of the HHML-treated rats were significantly increased (P < 0.05, vs. Model) while trabecular separation (Tb.Sp) and structure model index (SMI) values were significantly reduced (P < 0.05, vs. Model). Histological analysis showed that HHML treatment promoted the healing of fractures and cartilage repair, and increased the osteoblasts and collagen fibers. Furthermore, our results also revealed HHML could promote MC3T3-E1 cells proliferation and osteoblast differentiation via regulation of the runt-related transcription factor 2 (RUNX2), bone alkaline phosphatase (BALP), and OCN by activating phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) pathway, which confirmed by adding PI3K chemical inhibitor of LY294002. ConclusionHHML treatment is a reliable remedy for fractures in tibial and ankle by promotion of osteogenic differentiation via activation of PI3K/Akt pathway.

In-situ construction of bioactive nanostructures on Ti alloy surfaces through ultra-low concentration alkali treatment and low-temperature annealing

Osseointegration is crucial for the long-term stability of orthopedic implants. Nanosurfacing can impart osteoinductivity to the implant. However, the application potential of most available nanotechnologies in surface modification of implants is limited, and there is an urgent need to develop new effective nanosurfacing techniques with greater practicality. In this study, we for the first time, utilized an ultra-low concentration NaOH solution (0.55 mM) combined with low-temperature annealing to achieve solid-state dewetting (SSD) on Ti alloy surfaces, resulting in well-defined TiO2 nanoisland-like crystals. In vitro cell experiments demonstrated that the constructed nanostructures significantly affected the adhesion, proliferation, and osteogenic differentiation capability of MC3T3-E1 cells. The nanostructures formed at 550 °C exhibited the most pronounced enhancement of cell osteogenic differentiation capability. This method is simple, easy to operate, and scalable, holding significant application potential in the surface modification of Ti alloys.

Large artificial bone from 3D printed polycaprolactone/β-tricalcium phosphate (3D PCL/β-TCP) effectively promoting MC3T3-E1 cell adhesion, proliferation, and new bone formation

The use of 3D printing technology has advanced the bone tissue engineering, and constructing large artificial bones for repairing large-scale bone defects is highly significant. This study aimed to construct large artificial bones in a precise and controllable manner, focusing on repairing critical-sized bone defects. The researchers used 3D printing technology to synthesize 3D PCL/β-TCP, and then evaluated its ability to promote MC3T3-E1 cell adhesion, proliferation, and new bone formation through a series of characterizations. The results confirmed that 3D PCL/β-TCP, presented as a lattice structure similar to natural bone, could be used to prepare personalized artificial bone blocks based on the actual range of bone defects. The researchers also experimented with dipping 3D PCL/β-TCP bone blocks in different proportions of β-TCP and found that PT2 showed better mechanical and hydrophilic properties. Further experiments showed that PT2 presented excellent biocompatibility and outstanding capacity to promote MC3T3-E1 cell proliferation and adhesion, and that 3D PCL/β-TCP bone blocks possessed good osteogenic activity. Finally, it was suggested that PT2 exhibited the property of promoting new bone formation. These findings provide experimental data support for the repair of critical-sized bone defects.

Morinda Officinalis-derived extracellular vesicle-like particles: Anti-osteoporosis effect by regulating MAPK signaling pathway

BackgroundPostmenopausal osteoporosis (PMOP) is a systemic bone disease characterized by low bone mass and microstructural damage. Morinda Officinalis (MO) contains various components with anti-PMOP activities. Morinda Officinalis-derived extracellular vesicle-like particles (MOEVLPs) are new active components isolated from MO, and no relevant studies have investigated their anti-osteoporosis effect and mechanism. PurposeTo investigate the alleviating effect of MOEVLPs on PMOP and the underlying mechanism. MethodsDifferential centrifugation and ultracentrifugation were used to isolate MOEVLPs from MO. Transmission electron microscopy (TEM), flow nano analyzer, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), agarose gel electrophoresis, and thin-layer chromatography were employed to characterize MOEVLPs. PMOP mouse models were utilized to examine the anti-PMOP effect of MOEVLPs. H&E and immunohistochemical staining were used for drug safety and osteogenic effect assessment. Mouse embryo osteoblast precursor cells (MC3T3-E1) were used in vitro experiments. CCK-8 kit, alizarin red staining, proteomic, bioinformatic analyses, and western blot were used to explore the mechanism of MOEVLPs. ResultsIn this study, MOEVLPs from MO were successfully isolated and characterized. Animal experiments demonstrated that MOEVLPs exhibited specific femur targeting, were non-toxic to the heart, liver, spleen, lung, kidney, and aorta, and possessed anti-PMOP properties. The ability of MOEVLPs to strengthen bone formation was better than that of alendronate. In vitro experiments, results revealed that MOEVLPs did not significantly enhance osteogenic differentiation in MC3T3-E1 cells. Instead, MOEVLPs promoted the proliferation of MC3T3-E1 cells. Proteomic and bioinformatic analyses suggested that the proliferative effect of MOEVLPs was closely associated with the mitogen-activated protein kinase (MAPK) signaling pathway, particularly the altered expression of cAMP response element-binding protein (CREB) and ribosomal S6 kinase 1 (RSK1). Western blot results further confirmed these findings. ConclusionOur studies successfully isolated high-quality MOEVLPs and demonstrated that MOEVLPs can alleviate PMOP by promoting osteoblast proliferation through the MAPK pathway. MOEVLPs have the potential to become a novel and natural anti-PMOP drug.

Unraveling the effects of nano-grained microstructure on mechanical behavior, corrosion, and in vitro biocompatibility of 316L austenitic stainless steel

Nanograins are often beneficial in achieving better cytocompatibility for metallic implants. In this work, we have achieved grains from the micro to nano range in 316L austenitic stainless steel by a combination of cryo-rolling (50%, 70%, and 90% thickness reduction) followed by annealing (750–1000 °C for 2–20 min). The combined effect of grain refinement along with texture on its mechanical, corrosion, and in vitro cellular behavior was thoroughly investigated. Microstructural observations revealed nanograins formation (grain size = 144 nm) only for the 90CRA sample, whereas grain refinement in the micron range was observed for 0-70CRA samples. Electron backscatter diffraction (EBSD) studies showed fully austenitic structures for all the annealed samples while low Kernel Average Misorientation (KAM) values indicated strain-free microstructures. Orientation distribution function (ODF) maps showed the presence of strong γ-fiber (<111>||ND) and weak α-fiber (<110>||ND) in the 90CRA sample. The grain refinement led to an increase in the yield strength and ultimate tensile strength from 470.6 ± 16.21 MPa and 858.35 ± 31.82 MPa (0CRA) to 1662.06 ± 58.44 MPa and 2112.3 ± 51.62 MPa (90CRA), respectively. The potentiodynamic polarization study revealed that the corrosion rate of the 90CRA sample was about eight times less than that of the 0CRA sample. This was further corroborated by Mott-Schottky analysis, which showed a lowering of both acceptor and donor densities with the reduction in grain size. The evolution of a strong <111> texture during cryo-rolling contributed towards enhanced corrosion resistance of nano-grained 316L. Furthermore, grain refinement improved wettability, which in turn led to better cell proliferation and differentiation. At all culture days, MC3T3-E1 cell proliferation and differentiation were considerably higher for nano-grained 316L micron-sized counterparts. Thus, grain refinement to the nano range is an effective technique for the property enhancement of biomedical implants.

Modulation of the microRNA-6089/E2F transcription factor2 axis by querceting: implications for osteoblast viability, proliferation, migration, and osteogenic differentiation in fracture healing.

Quercetin is widely distributed in plants as a flavonol compound with multiple biological activities. It has been found that quercetin can regulate bone homeostasis through multiple pathways and targets. This study investigated the role and specific molecular mechanisms of quercetin in regulating osteoblast viability, proliferation, migration and osteogenic differentiation. A mouse model of traumatic fracture was established and then 100 mg/kg quercetin corn oil suspension was gavaged at the same time every day for 28 days. miR-6089 and E2F transcription factor 2 (E2F2) expression levels in mice were measured. Fracture healing in mice was observed. MC3T3-E1 cells were transfected with plasmids targeting miR-6089 and E2F2, and cell viability, proliferation, migration, apoptosis, and osteogenic differentiation were determined. The targeting relationship between miR-6089 and E2F2 was verified. In vivo experiments showed that quercetin significantly increased osteocalcin (OCN) expression (P<0.05) and promoted fracture healing in traumatic fracture (TF) mice. miR-6089 expression was down-regulated (P<0.05) and E2F2 expression was up-regulated (P<0.05) in TF mice. Quercetin promoted miR-6089 expression and inhibited E2F2 expression (both P<0.05). In vitro results showed that quercetin promoted miR-6089 expression and inhibited E2F2 expression in a dose-dependent manner (both P<0.05). Quercetin dose-dependently promoted MC3T3-E1 cell viability, proliferation, migration, and osteogenic differentiation, and inhibited MC3T3-E1 cell apoptosis (all P<0.05). Up-regulating miR-6089 further promoted MC3T3-E1 cell viability, proliferation, migration and osteogenic differentiation, and inhibited MC3T3-E1 cell apoptosis (all P<0.05). miR-6089 targeted and regulated E2F2 expression. Up-regulating E2F2 attenuated the promoting effect of up-regulated miR-6089 on MC3T3-E1 cell viability, proliferation, migration, osteogenic differentiation, and inhibition of apoptosis (all P<0.05). We conclude that quercetin enhances osteoblast viability, proliferation, migration, and osteogenic differentiation by modulating the miR-6089/E2F2 axis, thereby promoting fracture healing.

Preparation, Characterization, and Bioactivities of Cobalt, Strontium and Fluorine Co-Doped Oxide Films on Titanium Surface for Clinical Application

Titanium and titanium alloys are receiving widespread attention due to their excellent comprehensive mechanical properties, corrosion resistances, and biocompatibilities. However, titanium metal itself is biologically inert in physiological environments, and after implantation, it is surrounded by a layer of encapsulating fibrous membrane, making it difficult to form solid bonds with the tissue. Plasma electrolytic oxidation is a new technology used to prepare bioactive porous ceramic membranes on the surfaces of titanium and titanium alloys. It has application prospects for biomimetic surface modifications of titanium alloys. In this study, a cobalt, strontium and fluorine codoped oxide film (TAM-CSF) was prepared on a titanium surface via plasma electrolytic oxidation. The surface characteristics of the film were evaluated with field emission scanning electron microscopy, energy spectrum analyses, atomic force microscopy, profilometry and X-ray photoelectron spectroscopy. Additionally, the corrosion performance of the material was evaluated with an electrochemical workstation. The biocompatibility and bioactivity of the film were tested with in vitro cell experiments. The results showed that the TAM-CSF on the titanium surface had a porous morphology, and the CSF was uniformly doped on the film surface. TAM-CSF improved the surface roughness of the titanium. This film exhibited good biocompatibility and promoted the extension and proliferation of MC3T3-E1 cells. It was possible to prepare TAM-CSF on titanium surfaces via plasma electrolytic oxidation. The film exhibited a good morphology and good biological activity and has clinical application prospects.

Anti-osteoporosis effect of Mori Follum- Portulaca oleracea L.- Lycium barbarum L. extract based on network pharmacology and experimental validation and its utilization in functional yogurt

In China, Mori Follum, Portulaca oleracea L., and Lycium barbarum L. are traditional Chinese medicine and food used in treating osteoporosis. However, the active compounds, targets, and specific molecular mechanisms of the triangular herb composed of "Mori Follum, Portulaca oleracea L., and Lycium barbarum L." (MPL) have not been fully elucidated. In this study, the chemical components of MPL were first identified by LC-MS. Next, network pharmacology was used to identify the core targets and key pathways of MPL in anti-osteoporosis. And molecular docking simulations were conducted. The prediction results were validated by in vitro experiments using MC3T3-E1 cells. This study screened 22 active compounds from MPL, with a total of 59 targets involved in the treatment of osteoporosis. IL6, TNF, PTGS2, AKT1, ESR1, TP53, HIF1A, PPARG, PRKACA are core targets, and PI3K/AKT is a key pathway. Molecular docking showed that Wongnin, Hirsutine, and Cryptotanshinone strongly bind with AKT1 and ESR1. MPL could promoted MC3T3-E1 cell proliferation and differentiation and regulated the expression of key proteins in the PI3K/AKT/Runx2 signaling pathway. Further, MPL was used as a prebiotic-assisted fermentation to obtain functional yogurt with osteogenic activity. Moreover, MPL1 could improve the fermentation rate, exhibiting better ideal physicochemical properties and sensory quality of yogurt. This study provides a foundation for developing MPL as an anti-osteoporosis functional food.

Elucidating the role of the GC/GR/GLUT1 axis in steroid-induced osteonecrosis of the femoral head: A proteomic approach

BackgroundSteroid-induced osteonecrosis of the femoral head (SONFH) is a prevalent and incapacitating condition that affects the hip joint. Unfortunately, early diagnostic and treatment measures are limited. MethodsOur study employed Tandem Mass Tag (TMT) labeling mass spectrometry (MS)-based quantitative proteome to compare the proteins of femoral head tissues in patients with SONFH with those of patients who sustained femoral neck fracture (FNF). We investigated the level and effects of glucose transporter member 1 (GLUT1) in SONFH patients and MC3T3-E1 cells and examined the function and molecular mechanism of GLUT1 in the context of SONFH using in vivo and in vitro approaches. ResultsThe SONFH group exhibited significant changes in protein expression levels compared to the fracture group. Specifically, we observed the up-regulation of 86 proteins and the down-regulation of 138 proteins in the SONFH group. Among the differentially expressed proteins, GLUT1 was down-regulated and associated with glucose metabolic processes in the SONFH group. Further analysis using Parallel Reaction Monitoring (PRM), WB, and PCR confirmed that the protein was significantly down-regulated in both femoral head tissue samples from SONFH patients and dexamethasone-treated MC3T3-E1 cells. Moreover, overexpression of GLUT1 effectively reduced glucocorticoid (GC)-induced apoptosis and the suppression of osteoblast proliferation and osteogenic differentiation in MC3T3-E1 cells, as well as GC-induced femoral head destruction in GC-induced ONFH rat models. Additionally, our research demonstrated that GC down-regulated GLUT1 transcription via glucocorticoid receptors in MC3T3-E1 cells. ConclusionsGLUT1 was down-regulated in patients with SONFH; furthermore, down-regulated GLUT1 promoted apoptosis and inhibited osteoblast ossification in dexamethasone-induced MC3T3-E1 cells and contributed to GC-induced femoral head destruction in a SONFH rat model. Glucocorticoids inhibited the transcriptional activity of GLUT1, leading to a reduction in the amount and activity of GLUT1 in the cells and ultimately promoting apoptosis and inhibiting osteoblast ossification via the GC/GR/GLUT1 axis in SONFH.

Preclinical Evaluation of Bioactive Small Intestinal Submucosa-PMMA Bone Cement for Vertebral Augmentation.

In vertebroplasty and kyphoplasty, bioinert poly(methyl methacrylate) (PMMA) bone cement is a conventional filler employed for quick stabilization of osteoporotic vertebral compression fractures (OVCFs). However, because of the poor osteointegration, excessive stiffness, and high curing temperature of PMMA, the implant loosens, the adjacent vertebrae refracture, and thermal necrosis of the surrounding tissue occurs frequently. This investigation addressed these issues by incorporating the small intestinal submucosa (SIS) into PMMA (SIS-PMMA). In vitro analyses revealed that this new SIS-PMMA bone cement had improved porous structure, as well as reduced compressive modulus and polymerization temperature compared with the original PMMA. Furthermore, the handling properties of SIS-PMMA bone cement were not significantly different from PMMA. The in vitro effect of PMMA and SIS-PMMA was investigated on MC3T3-E1 cells via the Transwell insert model to mimic the clinical condition or directly by culturing cells on the bone cement samples. The results indicated that SIS addition substantially enhanced the proliferation and osteogenic differentiation of MC3T3-E1 cells. Additionally, the bone cement's biomechanical properties were also assessed in a decalcified goat vertebrae model with a compression fracture, which indicated the SIS-PMMA had markedly increased compressive strength than PMMA. Furthermore, it was proved that the novel bone cement had good biosafety and efficacy based on the International Standards and guidelines. After 12 weeks of implantation, SIS-PMMA indicated significantly more osteointegration and new bone formation ability than PMMA. In addition, vertebral bodies with cement were also extracted for the uniaxial compression test, and it was revealed that compared with the PMMA-implanted vertebrae, the SIS-PMMA-implanted vertebrae had greatly enhanced maximum strength. Overall, these findings indicate the potential of SIS to induce efficient fixation between the modified cement surface and the host bone, thereby providing evidence that the SIS-PMMA bone cement is a promising filler for clinical vertebral augmentation.

Tailorable mechanical and degradation properties of KCl-reticulated and BDDE-crosslinked PCL/chitosan/κ-carrageenan electrospun fibers for biomedical applications: Effect of the crosslinking-reticulation synergy

The development of intricated and interconnected porous mats is desired for many applications in biomedicine and other relevant fields. The mats that comprise the use of natural, bioactive, and biodegradable polymers are the focus of current research activities. In the present work, crosslinked fibers with improved characteristics were produced by incorporating 1,4-butanediol diglycidyl ether (BDDE) into a polymer formulation containing polycaprolactone (PCL), chitosan (CS), and κappa-carrageenan (κ-C). A slight variation of formic acid (FA)/acetic acid (AA) ratio used as a solvent system, significantly affected the characteristics of the produced fiber mats. Both polysaccharides and BDDE played a major role in tailoring mechanical properties when fibrous scaffolds were reticulated under KCl-mediated basic conditions for determined periods of time at 50 °C. In vitro biological assessment of the electrospun fiber mats revealed proliferation of MC3T3-E1 cells when incubated for 1 and 7 days. After staining the cells with 4′,6-diamidino-2-phenylindole (DAPI)/rhodamine phalloidin an autofluorescence response was observed by fluorescence microscopy in the scaffold manufactured using a solvent with higher FA/AA ratio due to the formation of microfibers. The results demonstrated the potential of the BDDE-crosslinked PCL/CS/κ-C electrospun fibers as promising materials for biomedical applications that may include soft and bone tissue regeneration.

Sol-gel derived bioactive glasses containing boron and strontium: Bioactivity, biocompatibility, and antibacterial properties

This study was aimed at the biological characterization of bioactive glasses in the systems of (60-x) SiO2–36CaO–4P2O5–xB2O3, and (60-x) SiO2–31CaO–4P2O5–5SrO-xB2O3 (x = 0, 1, 5, 10 and 15 mol%) synthesized through sol-gel technique. The influence of boron and strontium addition on in vivo bioactivity, antibacterial efficiency, and cell viability was studied. The morphological and structural assessments indicated that the bioactive glasses with chemical compositions of 50SiO2–31CaO–4P2O5–5SrO-10B2O3 (BG-5S10B), and 45SiO2–31CaO–4P2O5–5SrO-15B2O3 (BG-5S15B) possessed the highest bioactivity, owing to faster ion release and superior ability to induce apatite precipitation. The findings of the cell viability assay highlighted significant improvements in the proliferation and differentiation of osteoblastic MC3T3-E1 cells through the incorporation of strontium and boron (up to 10 %). Notably, the BG-5S10B sample exhibited exceptional biocompatibility and substantial antibacterial effectiveness on both E. coli and S. aureus. A combination of favorable biological properties makes BG-5S10B a highly promising option for use in bone tissue engineering applications.

Lysophosphatidic Acid/Polydopamine-Modified nHA Composite Scaffolds for Enhanced Osteogenesis via Upregulating the Wnt/Beta-Catenin Pathway.

Guided bone regeneration (GBR) technology has been widely used for the regeneration of periodontal bone defects. However, the limited mechanical properties and bone regeneration potential of the currently available GBR membranes often limit their repair effectiveness. In this paper, serum-derived growth factor lysophosphatidic acid (LPA) nanoparticles and dopamine-decorative nanohydroxyapatite (pDA/nHA) particles were double-loaded into polylactic-glycolic acid/polycaprolactone (PLGA/PCL) scaffolds as an organic/inorganic biphase delivery system, namely, PP-pDA/nHA-LPA scaffolds. Physicochemical properties and osteogenic ability in vitro and in vivo were performed. Scanning electron microscopy and mechanical tests showed that the PP-pDA/nHA-LPA scaffolds had a 3D bionic scaffold structure with improved mechanical properties. In vitro cell experiments demonstrated that the PP-pDA/nHA-LPA scaffolds could significantly enhance the attachment, proliferation, osteogenic differentiation, and mineralization of MC3T3-E1 cells. In vivo, the PP-pDA/nHA-LPA scaffolds exhibited great cytocompatibility and cell recruitment ability in 2- and 4-week subcutaneous implantation experiments and significantly promoted bone regeneration in the periodontal defect scaffold implantation experiment. Moreover, LPA-loaded scaffolds were confirmed to enhance osteogenic activities by upregulating the expression of β-catenin and further activating the Wnt/β-catenin pathway. These results demonstrate that the biphase PP-pDA/nHA-LPA delivery system is a promising material for the GBR.

Bovine serum albumin-modified 3D printed alginate dialdehyde-gelatin scaffolds incorporating polydopamine/SiO2-CaO nanoparticles for bone regeneration

Three-dimensional (3D) printing allows precise manufacturing of bone scaffolds for patient-specific applications and is one of the most recently developed and implemented technologies. In this study, bilayer and multimaterial alginate dialdehyde-gelatin (ADA-GEL) scaffolds incorporating polydopamine (PDA)/SiO2-CaO nanoparticle complexes were 3D printed using a pneumatic extrusion-based 3D printing technology and further modified on the surface with bovine serum albumin (BSA) for application in bone regeneration. The morphology, chemistry, and in vitro bioactivity of PDA/SiO2-CaO nanoparticle complexes were characterized (n = 3) and compared with those of mesoporous SiO2-CaO nanoparticles. Successful deposition of the PDA layer on the surface of the SiO2-CaO nanoparticles allowed better dispersion in a liquid medium and showed enhanced bioactivity. Rheological studies (n = 3) of ADA-GEL inks consisting of PDA/SiO2-CaO nanoparticle complexes showed results that may indicate better injectability and printability behavior compared to ADA-GEL inks incorporating unmodified nanoparticles. Microscopic observations of 3D printed scaffolds revealed that PDA/SiO2-CaO nanoparticle complexes introduced additional topography onto the surface of 3D printed scaffolds. Additionally, the modified scaffolds were mechanically stable and elastic, closely mimicking the properties of natural bone. Furthermore, protein-coated bilayer scaffolds displayed controllable absorption and biodegradation, enhanced bioactivity, MC3T3-E1 cell adhesion, proliferation, and higher alkaline phosphatase (ALP) activity (n = 3) compared to unmodified scaffolds. Consequently, the present results confirm that ADA-GEL scaffolds incorporating PDA/SiO2-CaO nanoparticle complexes modified with BSA offer a promising approach for bone regeneration applications.

Molecular mechanism of resveratrol promoting differentiation of preosteoblastic MC3T3-E1 cells based on network pharmacology and experimental validation

The purpose of this study was to investigate the mechanism by which resveratrol (Res) inhibits apoptosis and promotes proliferation and differentiation of pre-osteoblastic MC3T3-E1 cells, laying the groundwork for the treatment of osteoporosis (OP). The TCMSP database was used to find the gene targets for Res. The GeneCards database acquire the gene targets for OP. After discovering the potential target genes, GO, KEGG, and Reactome enrichment analysis were conducted. Verifying the major proteins involved in apoptosis can bind to Res using molecular docking. CCK8 measured the proliferative activity of mouse pre-osteoblasts in every group following Res intervention. Alkaline phosphatase staining (ALP) and alizarin red staining to measure the ability of osteogenic differentiation. RT-qPCR to determine the expression levels of Runx2 and OPG genes for osteogenic differentiation ability of cells. Western blot to measure the degree of apoptosis-related protein activity in each group following Res intervention. The biological processes investigated for GO of Res therapeutic OP involved in cytokine-mediated signaling pathway, negative regulation of apoptotic process, Aging, extrinsic apoptotic signaling pathway in absence of ligand, according to potential therapeutic target enrichment study. Apoptosis, FoxO signaling pathway, and TNF signaling pathway are the primary KEGG signaling pathways. Recactome pathways are primarily engaged in Programmed Cell Death, Apoptosis, Intrinsic Apoptotic Pathway, and Caspase activation via extrinsic apoptotic signaling pathways. This research established a new approach for Res treatment of OP by demonstrating how Res controls the apoptosis-related proteins TNF, IL6, and CASP3 to suppress osteoblast death and increase osteoclastogenesis.

2D magnesium phosphate resorbable coating to enhance cell adhesion on titanium surfaces

Titanium and its alloys are essential metals for orthopedic implant manufacturing due to their exceptional mechanical properties and biocompatibility, used extensively for treating various orthopedic conditions. However, Titanium (Ti) implants have a disadvantage due to lack of bioactivity, potentially affecting osseointegration and osteoconductive capabilities, and may take several months to integrate with bone tissue. In this work, we prepared a layer of 2D magnesium phosphate (MgPi) coating on the surface of titanium surfaces via the spin-coating technique. Various techniques were used to study the phase composition of the coatings, including FTIR, Raman spectroscopy, NMR, and XRD analysis. Morphology and chemical analysis were performed using Atomic force microscopy and SEM/EDX. Nano-scratch test and water contact angle measurements were used to measure adhesion strength and wettability. In addition, in vitro cell assays were used to assess cell adhesion and viability to determine how the MC3T3-E1 osteoblast-like cells reacted to the different treated Ti substrates. AFM results showed that the surface roughness became lower after coatings. MgPi-coated samples showed higher hydrophilicity, protein adsorption, and cell viability than uncoated samples. The nano-scratch test showed that the MgPi coating showed better adherence to chemically and thermally treated samples compared to untreated samples. The deposited MgPi coating has good adhesion to the Ti-substrates. Most significantly, compared to uncoated control (Ti) (p < 0.005) and chemically treated coated samples CT-MgPi (p < 0.005), MC3T3-E1 cell proliferation was significantly increased on thermochemical coated surfaces. These findings point to resorbable two-dimensional MgPi coatings as a potential candidate for promoting Ti implant osseointegration.

Alginate-PVDF piezoelectric hydrogel containing calcium copper titanate- hydroxyapatite as a self-powered scaffold for bone tissue engineering and energy harvesting

We have developed a piezoelectric hydrogel composite comprising cross-linked polyvinylidene fluoride-sodium alginate (PVDF-SA) and calcium copper titanate nanowires (CCTO) as the piezoelectric filler, and hydroxyapatite nanoparticles (HA) as the mineral filler. A freeze-drying method was used to prepare piezoelectric hydrogel scaffold. The PVDF/SA-CCTOs-HA hydrogel composite with 0.5% CCTO and 0.4% HA content demonstrated significantly improved piezoelectric performance. Specifically, it exhibited the highest piezoelectric performance among all samples, with VOC of 7 V and ISC of 3.5 μA. These values were 5.5 and 4.9 times higher, than those of the pure PVDF-SA hydrogel scaffold. The rheology studies explored the viscoelastic properties of hydrogel and found that the hydrogel's solid characteristics led to a higher elastic modulus in comparison to the viscous modulus. The composite exhibited higher compressive and tensile strengths at 8.2 and 0.8 MPa, respectively, compared to the pure hydrogel, which has strengths of 6.3 and 0.4 MPa. In vitro studies revealed the good cytocompatibility, cell proliferation of MC3T3-E1 cells on piezoelectric composite. The PVDF-SA/CCTO-HA demonstrated significant antibacterial properties, offering further advantages for its practical applications. The hydrogel composite can identify various human activities, such as wrist and elbow movements, finger pressing and releasing, and heel pressure. Furthermore, it demonstrated its energy storage capability by charging a 1 μF capacitor. This work offers a practical approach to fabricate hydrogel scaffolds possessing favorable piezoelectric characteristics, biocompatibility, robust mechanical properties, and enhanced proliferation, thereby enabling their extensive utilization in bone tissue engineering applications, human movement detection, and energy harvesting.

Scallop mantle-derived exosomes promote osteoblast formation and alleviate osteoporosis in mice

Osteoporosis, the most prevalent metabolic bone disease, has emerged as a serious public health concern. Safe and effective therapeutic interventions for osteoporosis are urgently needed. Here, we demonstrated that scallop mantle-derived exosomes (SMEs) promoted osteogenesis and prevented osteoporosis in ovariectomy-induced osteoporotic mice. Specifically, we successfully isolated SMEs with typical exosome morphology and desirable particle size. SMEs were found to contain abundant functional lipids, metabolites, and osteogenic-related proteins. Cell experiments revealed that SMEs could be internalized by osteoblastic MC3T3-E1 cells and promoted the proliferation, differentiation, and mineralization of MC3T3-E1 cells. In addition, SMEs survived the hostile gastrointestinal conditions and accumulated in the bone of mice after oral administration. Further mice investigations demonstrated that orally administered SMEs efficiently alleviated the bone loss, and improved the bone microstructure and biochemical properties in an ovariectomy-induced osteoporosis mouse model. Overall, these results reveal the osteogenesis effect of dietary SMEs, suggesting the potential of SMEs as an alternative or adjunct therapeutic agent in preventing or ameliorating osteoporosis.