Upregulation Of Alkaline Phosphatase Research Articles

Oxygen vacancy healing boosts the piezoelectricity of bone scaffolds.

Although barium titanate (BaTiO3) presented tremendous potential in achieving self-powered stimulation to accelerate bone repair, pervasive oxygen vacancies restricted the full play of its piezoelectric performance. Herein, BaTiO3-GO nanoparticles were synthesized by the in situ growth of BaTiO3 on graphene oxide (GO), and subsequently introduced into poly-L-lactic acid (PLLA) powders to prepare PLLA/BaTiO3-GO scaffolds by laser additive manufacturing. During the synthesis process, CO and C-OH in GO would respectively undergo cleavage and dehydrogenation at high temperature to form negatively charged oxygen groups, which were expected to occupy positively charged oxygen vacancies in BaTiO3 and thereby inhibit the formation of oxygen vacancies. Moreover, GO could be partially reduced to reduced graphene oxide, which could act as a conductive phase to facilitate polarization charge transfer, thus further improving the piezoelectric performance. The results showed that the oxygen peak at the specific electron binding energy in O 1s declined from 54.4% to 14.6% and the Ti3+ peak that was positively correlated with oxygen vacancies apparently weakened for BaTiO3-GO, illustrating that the introduced GO significantly decreased the oxygen vacancy. As a consequence, the piezoelectric current of PLLA/BaTiO3-GO increased from 80 to 147.3 nA compared with that of PLLA/BaTiO3. The enhanced piezoelectric current effectively accelerated cell differentiation by upregulating alkaline phosphatase expression, calcium salt deposition and calcium influx. This work provides a novel insight for the design of self-powered stimulation scaffolds for bone regeneration.

Engineering three-dimensional bone macro-tissues by guided fusion of cell spheroids.

Bioassembly techniques for the application of scaffold-free tissue engineering approaches have evolved in recent years toward producing larger tissue equivalents that structurally and functionally mimic native tissues. This study aims to upscale a 3-dimensional bone in-vitro model through bioassembly of differentiated rat osteoblast (dROb) spheroids with the potential to develop and mature into a bone macrotissue. dROb spheroids in control and mineralization media at different seeding densities (1 × 104, 5 × 104, and 1 × 105 cells) were assessed for cell proliferation and viability by trypan blue staining, for necrotic core by hematoxylin and eosin staining, and for extracellular calcium by Alizarin red and Von Kossa staining. Then, a novel approach was developed to bioassemble dROb spheroids in pillar array supports using a customized bioassembly system. Pillar array supports were custom-designed and printed using Formlabs Clear Resin® by Formlabs Form2 printer. These supports were used as temporary frameworks for spheroid bioassembly until fusion occurred. Supports were then removed to allow scaffold-free growth and maturation of fused spheroids. Morphological and molecular analyses were performed to understand their structural and functional aspects. Spheroids of all seeding densities proliferated till day 14, and mineralization began with the cessation of proliferation. Necrotic core size increased over time with increased spheroid size. After the bioassembly of spheroids, the morphological assessment revealed the fusion of spheroids over time into a single macrotissue of more than 2.5mm in size with mineral formation. Molecular assessment at different time points revealed osteogenic maturation based on the presence of osteocalcin, downregulation of Runx2 (p < 0.001), and upregulated alkaline phosphatase (p < 0.01). With the novel bioassembly approach used here, 3D bone macrotissues were successfully fabricated which mimicked physiological osteogenesis both morphologically and molecularly. This biofabrication approach has potential applications in bone tissue engineering, contributing to research related to osteoporosis and other recurrent bone ailments.

Alginate hydrogels containing different concentrations of magnesium-containing poly(lactic-co-glycolic acid) microspheres for bone tissue engineering

The easy loss of crosslinking ions in alginate can result in structural collapse and loss of its characteristics as a bone scaffold. A novel injectable tissue engineering scaffold containing poly(lactic-co-glycolic acid) (PLGA) microspheres and alginate was fabricated to improve alginate’s physiochemical and biological properties. MgCO3 and MgO were loaded at a 1:1 ratio into PLGA microspheres to form biodegradable PLGA microspheres containing magnesium (PMg). Subsequently, different concentrations of PMg were mixed into a Ca2+ suspension and employed as crosslinking agents for an alginate hydrogel. A pure Ca2+ suspension was used as the alginate crosslinking agent in the control group. The influence of PMg on the physiochemical properties of the injectable scaffolds, including the surface morphology, degradation rate, Mg2+ precipitation concentration, and the swelling rate, was investigated. MC3T3-E1 cells were seeded onto the hydrogels to evaluate the effect of the resultant alginate on osteoblastic attachment, proliferation, and differentiation. The physicochemical properties of the hydrogels, including morphology, degradation rate, and swelling ratio, were effectively tuned by PMg. Inductively coupled plasma-optical emission spectroscopy results showed that, in contrast to those in pure PMg, the magnesium ions (Mg2+) in alginate hydrogel containing PMg microspheres (Alg-PMg) were released in a dose-dependent and slow-releasing manner. Additionally, Alg-PMg with an appropriate concentration of PMg not only improved cell attachment and proliferation but also upregulated alkaline phosphatase activity, gene expression of osteogenic markers, and related growth factors. These findings indicate that PMg incorporation can regulate the physicochemical properties of alginate hydrogels. The resultant hydrogel promoted cell attachment, matrix mineralization, and bone regeneration. The hydrogel described in this study can be considered a promising injectable scaffold for bone tissue engineering.

Basal and inducible Osterix expression reflect equine mesenchymal progenitor cell osteogenic capacity.

Mesenchymal stem cells are characterized by their capacities for extensive proliferation through multiple passages and, classically, tri-lineage differentiation along osteogenic, chondrogenic and adipogenic lineages. This study was carried out to compare osteogenesis in equine bone marrow-, synovium- and adipose-derived cells, and to determine whether osteogenic capacity is reflected in the basal expression of the critical osteogenic transcription factors Runx2 and Osterix. Bone marrow, synovium and adipose tissue was collected from six healthy 2-year-old horses. Cells were isolated from these sources and expanded through two passages. Basal expression of Runx2 and Osterix was assessed in undifferentiated third passage cells, along with their response to osteogenic culture conditions. Bone marrow-derived cells had significantly higher basal expression of Osterix, but not Runx2. In osteogenic medium, bone-marrow cells rapidly developed dense, multicellular aggregates that stained strongly for mineral and alkaline phosphatase activity. Synovial and adipose cell cultures showed far less matrix mineralization. Bone marrow cells significantly up-regulated alkaline phosphatase mRNA expression and enzymatic activity at 7 and 14 days. Alkaline phosphatase expression and activity were increased in adipose cultures after 14 days, although these values were less than in bone marrow cultures. There was no change in alkaline phosphatase in synovial cultures. In osteogenic medium, bone marrow cultures increased both Runx2 and Osterix mRNA expression significantly at 7 and 14 days. Expression of both transcription factors did not change in synovial or adipose cultures. These results demonstrate that basal Osterix expression differs significantly in progenitor cells derived from different tissue sources and reflects the osteogenic potential of the cell populations.

Associations of urinary 1,3-butadiene metabolite with glucose homeostasis, prediabetes, and diabetes in the US general population: Role of alkaline phosphatase

The chemical – 1,3-butadiene (BD) is a volatile organic compound ubiquitous in the environment. However, the relationships and underlying mechanisms between BD exposure and glucose dyshomeostasis and diabetes in the general population remain unclear. We sought to explore the associations of BD exposure with glucose homeostasis, prediabetes, and diabetes, as well as the role of serum alkaline phosphatase (ALP) in these associations. This study included 5092 US general residents from the National Health and Nutrition Examination Survey with measurements of urinary BD metabolite (N-Acetyl-S-(3,4-dihydroxybutyl)-L-cysteine, DHBMA) and serum ALP. Glucose homeostasis was evaluated by fasting plasma glucose (FPG), fasting serum insulin (FINS), glycohemoglobin (HbA1c), and homeostasis model assessment of insulin resistance (HOMA-IR). HOMA-IR>2.6 was considered as insulin resistance (IR). Prediabetes and diabetes were determined according to the recommendations of the American Diabetes Association. The associations of DHBMA with glucose homeostasis, prediabetes, and diabetes were assessed by linear regression models and logistic regression models. The mediating role of ALP was evaluated by mediation analysis. We observed positive dose-response relationships of DHBMA level with glucose homeostasis indices and ALP levels, as well as with the risks of prediabetes and diabetes (all P < 0.05 and/or P for trend <0.05). Each 2-fold increase in DHBMA was associated with a 1.32%, 9.20%, 0.72%, and 10.64% increase in FPG, FINS, HbA1c, and HOMA-IR, respectively (all P < 0.05). And the corresponding odds ratios (ORs) and 95% confidence intervals (CIs) for IR, prediabetes, and diabetes were 1.36 (1.14, 1.61), 1.51 (1.26, 1.83), and 1.20 (0.90, 1.61), respectively. Furthermore, increased ALP significantly mediated 15.29%–41.12% of the associations of DHBMA with glucose dyshomeostasis and increased risks of prediabetes and diabetes. Our findings indicated that BD exposure was associated with glucose dyshomeostasis and increased risks of prediabetes and diabetes. The upregulation of ALP might play a significant role in these associations.

Deletion of the P2Y2 receptor aggravates internal elastic lamina calcification in chronic kidney disease mice through upregulation of alkaline phosphatase and lipocalin-2.

Calcification of the medial layer, inducing arterial stiffness, contributes significantly to cardiovascular mortality in patients with chronic kidney disease (CKD). Extracellular nucleotides block the mineralization of arteries by binding to purinergic receptors including the P2Y2 receptor. This study investigates whether deletion of the P2Y2 receptor influences the development of arterial media calcification in CKD mice. Animals were divided into: (i) wild type mice with normal renal function (control diet) (n= 8), (ii) P2Y2 R-/- mice with normal renal function (n= 8), (iii) wild type mice with CKD (n= 27), and (iv) P2Y2 R-/- mice with CKD (n= 22). To induce CKD, animals received an alternating (0.2-0.3%) adenine diet for 7 weeks. All CKD groups developed a similar degree of chronic renal failure as reflected by high serum creatinine and phosphorus levels. Also, the presence of CKD induced calcification in the heart and medial layer of the aortic wall. However, deletion of the P2Y2 receptor makes CKD mice more susceptible to the development of calcification in the heart and aorta (aortic calcium scores (median ± IQR), CKD-wild type: 0.34 ± 4.3mg calcium/g wet tissue and CKD-P2Y2 R-/- : 4.0± 13.2mg calcium/g wet tissue). As indicated by serum and aortic mRNA markers, this P2Y2 R-/- mediated increase in CKD-related arterial media calcification was associated with an elevation of calcification stimulators, including alkaline phosphatase and inflammatory molecules interleukin-6 and lipocalin 2. The P2Y2 receptor should be considered as an interesting therapeutic target for tackling CKD-related arterial media calcification.

Surface modifications to promote the osteoconductivity of ultra-high-molecular-weight-polyethylene fabrics for a novel biomimetic artificial disc prosthesis: An in vitro study.

A novel biomimetic artificial intervertebral disc (bioAID) for the cervical spine was developed, containing a hydrogel core representing the nucleus pulposus, an UHMWPE fiber jacket as annulus fibrosis, and titanium endplates with pins for mechanical fixation. Osseointegration of the UHMWPE fibers to adjacent bone structures is required to achieve proper biomimetic behavior and to provide long-term stability. Therefore, the aim of this study was to assess the osteoconductivity of several surface modifications of UHMWPE fabrics, 2D weft-knitted, using non-treated UHMWPE fibers (N), plasma treated UHMWPE fibers (PT), 10% hydroxy apatite (HA) loaded UHMWPE fibers (10%HA), plasma treated 10%HA UHMWPE fibers (PT-10%HA), 15%HA loaded UHMWPE fibers (15%HA) and plasma treated 15%HA UHMWPE fibers (PT-15%HA). Scanning electron microscopy (SEM) was used for surface characterization. Biological effects were assessed by evaluating initial cell attachment (SEM, DNA content), metabolic activity (PrestoBlue assay), proliferation, differentiation (alkaline phosphatase activity) and mineralization (energy dispersive x-ray, EDX analysis) using human bone marrow stromal cells. Plasma treated samples showed increased initial cell attachment, indicating the importance of hydrophilicity for cell attachment. However, incorporation only of HA or plasma treatment alone was not sufficient to result in upregulated alkaline phosphatase activity (ALP) activity. Combining HA loaded fibers with plasma treatment showed a combined effect, leading to increased cell attachment and upregulated ALP activity. Based on these results, combination of HA loaded UHMWPE fibers and plasma treatment provided the most promising fabric surface for facilitating bone ingrowth.

Short-chain fatty acids, acetate and propionate, directly upregulate osteoblastic differentiation

Short-chain fatty acids, including acetate, propionate, and butyrate are metabolites of dietary fibre produced by microbiota in the large intestine, have been proposed to contribute to effects on bone homeostasis. However, it is unclear whether they are used in osteoblasts and directly affect bone formation. We investigated whether short-chain fatty acids are absorbed in osteoblast cells and influence early osteoblastic differentiation using MC3T3-E1 cells. Acetate and propionate upregulated alkaline phosphatase activity, which is an osteoblast differentiation marker, and acetate upregulated alkaline phosphatase mRNA expression after treatment for 9 days, whereas butyrate did not in MC3T3-E1 cells. Butyrate was absorbed more rapidly and to a greater extent than acetate and propionate. These results indicate that short-chain fatty acids were used in osteoblastic cells, and particularly acetate and propionate directly upregulated differentiation in primary osteoblasts. Therefore, acetate and propionate might be useful for maintaining a positive balance of bone turnover.

Upregulation of Runt related transcription factor 1 (RUNX1) contributes to tendon–bone healing after anterior cruciate ligament reconstruction using bone mesenchymal stem cells

BackgroundAnterior cruciate ligament (ACL) injury could lead to functional impairment along with disabilities. ACL reconstruction often fails owing to the regeneration failure of tendon–bone interface. Herein, we aimed to investigate the effects of Runt related transcription factor 1 (RUNX1) on tendon–bone healing after ACL reconstruction using bone mesenchymal stem cells (BMSCs).MethodsBMSCs were isolated from the marrow cavity of rat femur, followed by the modification of RUNX1 with lentiviral system. Then, an ACL reconstruction model of rats was established with autografts.ResultsResults of flow cytometry exhibited positive-antigen CD44 and CD90, as well as negative-antigen CD34 and CD45 of the BMSCs. Then, we found that RUNX1-upregulated BMSCs elevated the decreased biomechanical strength of the tendon grafts after ACL reconstruction. Moreover, based on the histological observation, upregulation of RUNX1 was linked with better recovery around the bone tunnel, a tighter tendon–bone interface, and more collagen fibers compared to the group of BMSCs infected with LV-NC. Next, RUNX1-upregulated BMSCs promoted osteogenesis after ACL reconstruction, as evidenced by the mitigation of severe loss and erosion of the cartilage and bone in the tibial and femur area, as well as the increased number of osteoblasts identified by the upregulation of alkaline phosphatase, osteocalcin, and osteopontin in the tendon–bone interface.ConclusionElevated expression of RUNX1 contributed to tendon–bone healing after ACL reconstruction using BMSCs.

Endothelial cell-derived tetrahydrobiopterin prevents aortic valve calcification.

Tetrahydrobiopterin (BH4) is a critical determinant of the biological function of endothelial nitric oxide synthase. The present study was to investigate the role of valvular endothelial cell (VEC)-derived BH4 in aortic valve calcification. Plasma and aortic valve BH4 concentrations and the BH4:BH2 ratio were significantly lower in calcific aortic valve disease patients than in controls. There was a significant decrease of the two key enzymes of BH4 biosynthesis, guanosine 5'-triphosphate cyclohydrolase I (GCH1) and dihydrofolate reductase (DHFR), in calcified aortic valves compared with the normal ones. Endothelial cell-specific deficiency of Gch1 in Apoe-/- (Apoe-/-Gch1fl/flTie2Cre) mice showed a marked increase in transvalvular peak jet velocity, calcium deposition, runt-related transcription factor 2 (Runx2), dihydroethidium (DHE), and 3-nitrotyrosine (3-NT) levels in aortic valve leaflets compared with Apoe-/-Gch1fl/fl mice after a 24-week western diet (WD) challenge. Oxidized LDL (ox-LDL) induced osteoblastic differentiation of valvular interstitial cells (VICs) co-cultured with either si-GCH1- or si-DHFR-transfected VECs, while the effects could be abolished by BH4 supplementation. Deficiency of BH4 in VECs caused peroxynitrite formation increase and 3-NT protein increase under ox-LDL stimulation in VICs. SIN-1, the peroxynitrite generator, significantly up-regulated alkaline phosphatase (ALP) and Runx2 expression in VICs via tyrosine nitration of dynamin-related protein 1 (DRP1) at Y628. Finally, folic acid (FA) significantly attenuated aortic valve calcification in WD-fed Apoe-/- mice through increasing DHFR and salvaging BH4 biosynthesis. The reduction in endothelial-dependent BH4 levels promoted peroxynitrite formation, which subsequently resulted in DRP1 tyrosine nitration and osteoblastic differentiation of VICs, thereby leading to aortic valve calcification. Supplementation of FA in diet attenuated hypercholesterolaemia-induced aortic valve calcification by salvaging BH4 bioavailability.

Magnesium Modified β-Tricalcium Phosphate Induces Cell Osteogenic Differentiation In Vitro and Bone Regeneration In Vivo.

In vitro, in vivo, and clinical studies have shown how the physicochemical and biological properties of β-tricalcium phosphate (β-TCP) work in bone regeneration. This study aimed to improve the properties of β-TCP by achieving optimum surface and bulk β-TCP chemical/physical properties through the hydrothermal addition of magnesium (Mg) and to later establish the biocompatibility of β-TCP/Mg for bone grafting and tissue engineering treatments. Multiple in vitro and in vivo analyses were used to complete β-TCP/Mg physicochemical and biological characterization. The addition of MgO brought about a modest rise in the number of β-TCP surface particles, indicating improvements in alkaline phosphatase (ALP) activity on day 21 (p < 0.05) and in the WST-1assay on all days (p < 0.05), with a corresponding increase in the upregulation of ALP and bone sialoprotein. SEM analyses stated that the surfaces of the β-TCP particles were not altered after the addition of Mg. Micro-CT and histomorphometric analysis from rabbit calvaria critical defects resulted in β-TCP/Mg managing to reform more new bone than the control defects and β-TCP control at 2, 6, and 8 weeks (* p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001). The hydrothermal addition of MgO to the β-TCP surfaces ameliorated its biocompatibility without altering its surface roughness resulting from the elemental composition while enhancing cell viability and proliferation, inducing more bone regeneration by osteoconduction in vivo and osteoblastic differentiation in vitro.

Pro-inflammatory mediators released by activated monocytes promote aortic valve fibrocalcific activity

BackgroundCalcific aortic valve disease (CAVD) is the most prevalent heart valve disorder in the elderly. Valvular fibrocalcification is a characteristic pathological change. In diseased valves, monocyte accumulation is evident, and aortic valve interstitial cells (AVICs) display greater fibrogenic and osteogenic activities. However, the impact of activated monocytes on valular fibrocalcification remains unclear. We tested the hypothesis that pro-inflammatory mediators from activated monocytes elevate AVIC fibrogenic and osteogenic activities.Methods and resultsPicro-sirius red staining and Alizarin red staining revealed collagen and calcium depositions in cultured human AVICs exposed to conditioned media derived from Pam3CSK4-stimulated monocytes (Pam3 CM). Pam3 CM up-regulated alkaline phosphatase (ALP), an osteogenic biomarker, and extracellular matrix proteins collagen I and matrix metalloproteinase-2 (MMP-2). ELISA analysis identified high levels of RANTES and TNF-α in Pam3 CM. Neutralizing RANTES in the Pam3 CM reduced its effect on collagen I and MMP-2 production in AVICs while neutralizing TNF-α attenuated the effect on AVIC ALP production. In addition, Pam3 CM induced NF-κB and JNK activation. While JNK mediated the effect of Pam3 CM on collagen I and MMP-2 production, NF-κB was critical for the effect of Pam3 CM on ALP production in AVICs.ConclusionsThis study demonstrates that activated monocytes elevate the fibrogenic and osteogenic activities in human AVICs through a paracrine mechanism. TNF-α and RANTES mediate the pro-fibrogenic effect of activated monocytes on AVICs through activation of JNK, and TNF-α also activates NF-κB to elevate AVIC osteogenic activity. The results suggest that infiltrated monocytes elevate AVIC fibrocalcific activity to promote CAVD progression.

Corrosion behavior, antibacterial ability, and osteogenic activity of Zn-incorporated Ni-Ti-O nanopore layers on NiTi alloy

• Zn-incorporated nanopores layers with different lengths were prepared on the NiTi substrate through anodization and hydrothermal treatment. • The loaded amounts and release kinetics of Zn ions was controlled by the Ni-Ti-O nanopores. • Zn incorporation enhanced the corrosion resistance. • Zn-incorporated nanopores layers enhanced the antibacterial property and exhibited long-lasting antibacterial ability. • Zn incorporation promoted the generation of alkaline phosphatase products and significantly promotesd type I collagen synthesis and extracellular matrix mineralization. Development of bone fixation devices with excellent corrosion resistance, antibacterial ability, and osteogenic activity is critical for promoting fracture healing. In this study, Zn-incorporated nanopore (NP) layers were prepared on the NiTi alloy through anodization and hydrothermal treatment. Results show that Zn can be evenly incorporated into the NP layers in the form of ZnTiO 3 . The Zn-incorporated samples exhibit good corrosion resistance and significantly reduce Ni 2+ release. Meanwhile, the samples can continuously release Zn 2+ , which is responsible for excellent long-term antibacterial ability. Furthermore, the synergetic effect of Zn 2+ release and nanoporous structure of the NP layers endues the NiTi alloy excellent osteogenic activity, as verified by upregulated alkaline phosphatase activity, secretion of type I collagen, and extracellular matrix mineralization. Therefore, Zn-incorporated Ni-Ti-O NP layers have great potential as biomedical coatings of NiTi-based implant materials.

Micro-arc oxidation-assisted sol-gel preparation of calcium metaphosphate coatings on magnesium alloys for bone repair

Calcium phosphate coating is an attractive surface modification strategy for magnesium alloys, since it can increase their corrosion resistance and endow them with osteogenic function simultaneously. Herein, a calcium metaphosphate (CMP) coating was fabricated on magnesium alloy by using sol-gel approach assisted with micro-arc oxidation pre-treatment. Scanning electron microscopy showed that the micro-pores and cracks in micro-arc oxidation inner layer generated during the pre-treatment process were sealed by the grainy sol-gel outer layer. Energy dispersive spectrometry and X-ray diffraction results demonstrated the identity of the coating as CMP. The cross-cut test showed that the adhesion of CMP coating was strong. Applying bare magnesium alloy substrate as a control, the CMP coating surface was rougher and more hydrophilic. The potentiodynamic polarization test demonstrated that the corrosion resistance was significantly improved by using CMP coating. Hydrogen evolution in immersion test further confirmed that the degradation rate was decelerated within 14 days. Moreover, CMP coating facilitated the adhesion speed, spreading area, and focal adhesion formation of bone marrow stem cells. The number of cells in the active proliferating state and proliferated cells present on the CMP coating also increased. Additionally, CMP coating upregulated alkaline phosphatase activity and osteogenic gene expression in cells. In summary, the micro-arc oxidation assisted sol-gel CMP coatings increased the corrosion resistance and promoted the interfacial cell behavior for magnesium alloy implants, which might inform the further development of surface modifications on magnesium alloys for bone related applications.

Antibacterial and osteogenic activities of clindamycin-releasing mesoporous silica/carboxymethyl chitosan composite hydrogels.

Conventional treatment of jaw bone infection is often ineffective at controlling bacterial infection and enhancing bone regeneration. Biodegradable composite hydrogels comprised of carboxymethyl chitosan (CMCS) and clindamycin (CDM)-loaded mesoporous silica nanoparticles (MCM-41), possessing dual antibacterial activity and osteogenic potency, were developed in the present study. CDM was successfully loaded into both untreated and plasma-treated MCM-41 nanoparticles, denoted as (p)-MCM-41, followed by the incorporation of each of CDM-loaded (p)-MCM-41 into CMCS. The resulting CDM-loaded composite hydrogels, (p)-MCM-41-CDM-CMCS, demonstrated slow degradation rates (about 70% remaining weight after 14-day immersion), while the CDM-free composite hydrogel entirely disintegrated after 4-day immersion. The plasma treatment was found to improve drug loading capacity and slow down initial drug burst effect. The prolonged releases of CDM from both (p)-MCM-41-CDM-CMCS retained their antibacterial effect against Streptococcus sanguinis for at least 14 days in vitro. In vitro assessment of osteogenic activity showed that the CDM-incorporated composite hydrogel was cytocompatible to human mesenchymal stem cells (hMSCs) and induced hMSC mineralization via p38-dependent upregulated alkaline phosphatase activity. In conclusion, novel (p)-MCM-41-CDM-CMCS hydrogels with combined controlled release of CDM and osteogenic potency were successfully developed for the first time, suggesting their potential clinical benefit for treatment of intraoral bone infection.

Bone Piezoelectricity-Mimicking Nanocomposite Membranes Enhance Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells by Amplifying Cell Adhesion and Actin Cytoskeleton.

Ferroelectric biomaterials have been widely investigated and demonstrated to enhance osteogenesis by simulating the inherent electrical properties of bone tissues. Nevertheless, the underlying biological processes are still not wellunderstood. Hence, this study investigated the underlying biological processes by which bone piezoelectricity-mimicking barium titanate/poly(vinylidene fluoride-trifluoroethylene) nanocomposite membranes (BTO nanocomposite membranes) promote osteogenesis of Bone Marrow Mesenchymal Stem Cells (BMSCs). Ourresults revealed that the piezoelectric coefficient (d33) of nanocomposite membranes aftercontrolled corona poling was similar to that of native bone, and exhibited highly-stable piezoelectrical properties and concentrated surface electrical potential. These nanocomposite membranes significantly enhanced the adhesion and spreading of BMSCs, which was manifested as increased number and area of mature focal adhesions. Furthermore, the nanocomposite membranes significantly promoted the expression of integrin receptors genes (α1, α5 andβ3), which in turn enhanced osteogenesis of BMSCs, as manifested by upregulated Alkaline Phosphatase (ALP) and Bone Morphogenetic Protein 2 (BMP2) expression levels. Further investigations found that the Focal Adhesion Kinase (FAK)-Extracellular Signal-Regulated Kinase1/2 (ERK 1/2) signaling axis may be involved in the biological process of polarized nanocomposite membrane-induced osteogenesis. This study thus provides useful insights for betterunderstanding of the biological processes by which piezoelectric or ferroelectric biomaterials promote osteogenesis.

Correction to: Improving exposure of anodically ordered Ni–Ti–O and corrosion resistance and biological properties of NiTi alloys by substrate electropolishing

Although Ni–Ti–O nanopores (NPs) can be fabricated by anodization of mechanically polished NiTi alloys, the top disordered layer is difficult to remove thus hindering the functionality of the Ni–Ti–O NPs. In this work, an electropolishing (EP) pretreatment was performed on the NiTi substrate prior to anodization to thoroughly expose the NPs. Our results show that the EP pretreatment for 5 min perfectly removes the top disordered layer on the Ni–Ti–O NPs to expose the underlying NPs and consequently, the corrosion resistance and antibacterial ability are enhanced. The exposed NPs can elongate bone marrow mesenchymal stem cells, which may be responsible for the upregulated alkaline phosphatase activity, secretion of Type I collagen, and extracellular matrix mineralization. These results suggest that EP is a desirable pretreatment before anodization of the NiTi alloys because the irregular surface layer on the Ni–Ti–O NPs can be removed to enhance the corrosion resistance and biological functions.

Surface Modified \u03b2-Tricalcium phosphate enhanced stem cell osteogenic differentiation in vitro and bone regeneration in vivo

A major number of studies have demonstrated Beta-tricalcium phosphate (β-TCP) biocompatibility, bioactivity, and osteoconductivity characteristics in bone regeneration. The aim of this research was to enhance β-TCP's biocompatibility, and evaluate its physicochemical properties by argon glow discharge plasma (GDP) plasma surface treatment without modifying its surface. Treated β-TCP was analyzed by scanning electron microscopy (SEM), energy-dispersive spectrometry, X-ray photoelectron spectroscopy (XPS), X-ray diffraction analysis, and Fourier transform infrared spectroscopy characterization. To evaluate treated β-TCP biocompatibility and osteoblastic differentiation, water-soluble tetrazolium salts-1 (WST-1), immunofluorescence, alkaline phosphatase (ALP) assay, and quantitative real-time polymerase chain reaction (QPCR) were done using human mesenchymal stem cells (hMSCs). The results indicated a slight enhancement of the β-TCP by GDP sputtering, which resulted in a higher Ca/P ratio (2.05) than the control. Furthermore, when compared with control β-TCP, we observed an improvement of WST-1 on all days (p < 0.05) as well as of ALP activity (day 7, p < 0.05), with up-regulation of ALP, osteocalcin, and Osteoprotegerin osteogenic genes in cells cultured with the treated β-TCP. XPS and SEM results indicated that treated β-TCP’s surface was not modified. In vivo, micro-computed tomography and histomorphometric analysis indicated that the β-TCP test managed to regenerate more new bone than the untreated β-TCP and control defects at 8 weeks (p < 0.05). Argon GDP treatment is a viable method for removing macro and micro particles of < 7 μm in size from β-TCP bigger particles surfaces and therefore improving its biocompatibility with slight surface roughness modification, enhancing hMSCs proliferation, osteoblastic differentiation, and stimulating more new bone formation.

Scleraxis upregulated by transforming growth factor-β1 signaling inhibits tension-induced osteoblast differentiation of priodontal ligament cells via ephrin A2.

During tooth movement in orthodontic treatment, bone formation and resorption occur on the tension and compression sides of the alveolar bone, respectively. Although the bone formation activity increases in the periodontal ligament (PDL) on the tension side, the PDL itself is not ossified and maintains its homeostasis, indicating that there are negative regulators of bone formation in the PDL. Our previous report suggested that scleraxis (Scx) has an inhibitory effect on ossification of the PDL on the tension side through the suppression of calcified extracellular matrix formation. However, the molecular biological mechanisms of Scx-modulated inhibition of ossification in the tensioned PDL are not fully understood. The aim of the present study is to clarify the inhibitory role of Scx in osteoblast differentiation of PDL cells and its underlying mechanism. Our in vivo experiment using a mouse experimental tooth movement model showed that Scx expression was increased during early response of the PDL to tensile force. Scx knockdown upregulated expression of alkaline phosphatase, an early osteoblast differentiation marker, in the tensile force-loaded PDL cells in vitro. Transforming growth factor (TGF)-β1-Smad3 signaling in the PDL was activated by tensile force and inhibitors of TGF-β receptor and Smad3 suppressed the tensile force-induced Scx expression in PDL cells. Tensile force induced ephrin A2 (Efna2) expression in the PDL and Efna2 knockdown upregulated alkaline phosphatase expression in PDL cells under tensile force loading. Scx knockdown eliminated the tensile force-induced Efna2 expression in PDL cells. These findings suggest that the TGF-β1-Scx-Efna2 axis is a novel molecular mechanism that negatively regulates the tensile force-induced osteoblast differentiation of PDL cells.

Improving exposure of anodically ordered Ni–Ti–O and corrosion resistance and biological properties of NiTi alloys by substrate electropolishing

Although Ni–Ti–O nanopores (NPs) can be fabricated by anodization of mechanically polished NiTi alloys, the top disordered layer is difficult to remove thus hindering the functionality of the Ni–Ti–O NPs. In this work, an electropolishing (EP) pretreatment was performed on the NiTi substrate prior to anodization to thoroughly expose the NPs. Our results show that the EP pretreatment for 5 min perfectly removes the top disordered layer on the Ni–Ti–O NPs to expose the underlying NPs and consequently, the corrosion resistance and antibacterial ability are enhanced. The exposed NPs can elongate bone marrow mesenchymal stem cells, which may be responsible for the upregulated alkaline phosphatase activity, secretion of Type I collagen, and extracellular matrix mineralization. These results suggest that EP is a desirable pretreatment before anodization of the NiTi alloys because the irregular surface layer on the Ni–Ti–O NPs can be removed to enhance the corrosion resistance and biological functions.