Osteogenic Function Research Articles

Akt phosphorylates and regulates the osteogenic activity of Osterix

Osterix (Osx), a zinc-finger transcription factor is required for osteoblast differentiation and new bone formation during embryonic development. Akt is a member of the serine/threonine-specific protein kinase and plays important roles in osteoblast differentiation. The function of Osterix can be also modulated by post-translational modification. But, the precise molecular signaling mechanisms between Osterix and Akt are not known. In this study, we investigated the potential regulation of Osterix function by Akt in osteoblast differentiation. We found that Akt phosphorylates Osterix and that Akt activation increases protein stability, osteogenic activity and transcriptional activity of Osterix. We also found that BMP-2 increases the protein level of Osterix in an Akt activity-dependent manner. These results suggest that Akt activity enhances the osteogenic function of Osterix, at least in part, through protein stabilization and that BMP-2 regulates the osteogenic function of Osterix, at least in part, through Akt.

Cell and gene therapy for bone repair

Improving bone repair remains an important and challenging issue. Therapeutic approaches to amplify osteogenic cell pool or function include cell and gene therapies. We identified genes that promote human mesenchymal cell differentiation and bone formation. Targeting these or other genes may improve the efficacy of cell therapy for bone repair.

Protein kinase A phosphorylates and regulates the osteogenic activity of Dlx5

Dlx5 transcription factor plays important roles in osteoblast differentiation and its transcription is regulated by many osteogenic signals including BMP-2. Recent studies suggest that the function of Dlx5 is also regulated post-translationally by protein kinases such as p38 and CaMKII. Protein kinase A (PKA) is involved in several steps of osteoblast differentiation and its activity has been shown necessary, yet not sufficient, for BMP-induced osteoblast differentiation. PKA is a ubiquitous cellular kinase that phosphorylates serine and threonine residues(s) of target proteins. In this study, we investigated the potential regulation of Dlx5 function by PKA in osteoblast differentiation. We found that PKA phosphorylates Dlx5 and that PKA activation increases the protein stability, osteogenic activity and transcriptional activity of Dlx5. We also found that BMP-2 increases the protein level of Dlx5 in a PKA activity-dependent manner. These results suggest that PKA activity enhances the osteogenic function of Dlx5, at least in part, through protein stabilization and that BMP-2 regulates the osteogenic function of Dlx5, at least in part, through PKA.

Influence of sterilization methods on cell behavior and functionality of osteoblasts cultured on TiO 2 nanotubes

We investigated the adhesion, proliferation and osteogenic functionality of osteoblasts cultured on titanium dioxide (TiO 2) nanotubes in response to different sterilization methods (dry autoclaving vs. wet autoclaving). We prepared various sizes (30–100 nm diameter) of TiO 2 nanotubes on titanium substrates by anodization, sterilized nanotubes by different conditions, and seeded osteoblast cells onto the nanotube surfaces with two different cell seeding densities (10,000 vs. 50,000 cells/well in 12-culture well). The result of this study indicates that the adhesion, proliferation and alkaline phosphatase activity of osteoblasts cultured on only the larger 70 and 100 nm TiO 2 nanotube arrays were dramatically changed by the different sterilization conditions at a low cell seeding density. However, with a higher cell seeding density (50,000 cells/well in 12-cell culture well), the results revealed no significant difference among altered nanotube geometry, 30–100 nm diameters, nor sterilization methods. Next, it was revealed that the nanofeatures of proteins adhered on nanotubular TiO 2 morphology are altered by the sterilization method. It was determined that this protein adhesion effect, in combination with the cell density of osteoblasts seeded onto such TiO 2 nanotube surfaces, has profound effects on cell behavior. This study clearly shows that these are some of the important in vitro culture factors that need to be taken into consideration, as well as TiO 2 nanotube diameters which play an important role in the improvement of cell behavior and functionality.

Transcriptional control mechanisms associated with the nucleotide receptor P2X7, a critical regulator of immunologic, osteogenic, and neurologic functions.

The nucleotide receptor P2X(7) is an attractive therapeutic target and potential biomarker for multiple inflammatory and neurologic disorders, and it is expressed in several immune, osteogenic, and neurologic cell types. Aside from its role in the nervous system, it is activated by ATP released at sites of tissue damage, inflammation, and infection. Ligand binding to P2X(7) stimulates many cell responses, including calcium fluxes, MAPK activation, inflammatory mediator release, and apoptosis. Much work has centered on P2X(7) action in cell death and mediator processing (e.g., pro-interleukin-1 cleavage by the inflammasome), but the contribution of P2X(7) to transcriptional regulation is less well defined. This review will focus on the growing evidence for the importance of nucleotide-mediated gene expression, highlight several animal models, human genetic, and clinical studies that support P2X(7) as a therapeutic target, and discuss the latest developments in anti-P2X(7) clinical trials.

Quantifying Osteogenic Cell Degradation of Silk Biomaterials

The degradation of silk protein films by human mesenchymal stem cells (hMSCs), osteoblasts and osteoclasts, cells involved in osteogenic functions in normal and diseased bone, was assessed in vitro. The involvement of specific matrix metalloproteinases (MMPs) and integrin signaling in the degradation process was determined. Scanning electron microscopy (SEM) and atomic force microscopy (AFM) were used to quantitatively compare degradation by the different cell types using surface patterned silk films. Osteoblasts and osteoclasts demonstrated significant degradation of the silk films in vitro in comparison to the hMSCs and the film controls without cells. The osteoclasts degraded the silk films the most and also generated the highest level of MMPs 1 and 2. The osteoblasts upregulated integrins α5 and β1, while the osteoclasts upregulated integrins α2 and β1. There was significant contrast in responses on the silk matrices between osteogenic cells versus undifferentiated hMSCs to illustrate in vitro the role of cell type on matrix remodeling. These are important issues in matching biomaterial matrix features and studies in vitro to remodeling in vivo, in both normal and disease tissue systems. Cell populations and niche factors impact tissue regeneration, wound healing, physiological state, and the ability to better understand the role of different cell types is critical to overall regenerative outcomes.

Activation of the Transcription Factor FosB/Activating Protein-1 (AP-1) Is a Prominent Downstream Signal of the Extracellular Nucleotide Receptor P2RX7 in Monocytic and Osteoblastic Cells*

Activation of the ionotropic P2RX7 nucleotide receptor by extracellular ATP has been implicated in modulating inflammatory disease progression. Continuous exposure of P2RX7 to ligand can result in apoptosis in many cell types, including monocytic cells, whereas transient activation of P2RX7 is linked to inflammatory mediator production and the promotion of cell growth. Given the rapid hydrolysis of ATP in the circulation and interstitial space, transient activation of P2RX7 appears critically important for its action, yet its effects on gene expression are unclear. The present study demonstrates that short-term stimulation of human and mouse monocytic cells as well as mouse osteoblasts with P2RX7 agonists substantially induces the expression of several activating protein-1 (AP-1) members, particularly FosB. The potent activation of FosB after P2RX7 stimulation is especially noteworthy considering that little is known concerning the role of FosB in immunological regulation. Interestingly, the magnitude of FosB activation induced by P2RX7 stimulation appears greater than that observed with other known inducers of FosB expression. In addition, we have identified a previously unrecognized role for FosB in osteoblasts with respect to nucleotide-induced expression of cyclooxygenase-2 (COX-2), which is the rate-limiting enzyme in prostaglandin biosynthesis from arachidonic acid and is critical for osteoblastic differentiation and immune behavior. The present studies are the first to link P2RX7 action to FosB/AP-1 regulation in multiple cell types, including a role in nucleotide-induced COX-2 expression, and support a role for FosB in the control of immune and osteogenic function by P2RX7.

Red yeast rice stimulates osteoblast proliferation and increases alkaline phosphatase activity in MC3T3-E1 cells

Red yeast (Monascus purpureus) is used as a traditional hypocholesterolemic dietary food component in Asia due to its bioactive component, lovastatin. Recently, new evidence suggesting that the statins in red yeast enhance bone formation has been reported, but more research is still needed in order to support these claims of osteogenic effects. Therefore, in this study, we hypothesized that red yeast rice (in which red yeast is fermented) can improve osteogenic function through osteoblast cell proliferation and differentiation. We studied the effect of methanol extract of red yeast rice powder (RYRP) on osteoblast proliferation and differentiation by measuring mitochondrial enzyme activity and bone marker alkaline phosphatase (ALP) activity, respectively. Osteoblast-like MC3T3-E1 cells were cultured in various concentrations of RYRP methanol extract (0.001-1 mg/mL) during the osteoblast differentiation period (1, 5, 10, and 15 days). As measured by 3-[4,5-dimethylthiazol-2-y]-2,5-diphenyltetrazolium bromide assay, RYRP extracts stimulated cell proliferation during a 24-hour period, compared to cooked white rice powder extract. The most pronounced effect was observed at the concentration range between 0.075 and 0.1 mg/mL. This RYRP stimulatory effect for cell proliferation was observed during the whole osteogenic period. Cellular (synthesized) ALP activity was increased at a RYRP extract concentration of 0.075 mg/mL during 15 days of culture, but the medium (secreted) ALP activity did not show any significant change. This cellular ALP activity stimulation by RYRP extract was confirmed by the staining of ALP activity on cell matrix layers for matrix calcification. The results imply that RYRP extract may increase osteogenic effect by stimulating cell proliferation and ALP activity in osteoblastic cells.

NVP-BEZ235, a dual pan class I PI3 kinase and mTOR inhibitor, promotes osteogenic differentiation in human mesenchymal stromal cells

Osteoblasts are bone-forming cells derived from mesenchymal stromal cells (MSCs) that reside within the bone marrow. In response to a variety of factors, MSCs proliferate and differentiate into mature, functional osteoblasts. Several studies have shown previously that suppression of the PI3K and mTOR signaling pathways in these cells strongly promotes osteogenic differentiation, which suggests that inhibitors of these pathways may be useful as anabolic bone agents. In this study we examined the effect of BEZ235, a newly developed dual PI3K and mTOR inhibitor currently in phase I-II clinical trials for advanced solid tumors, on osteogenic differentiation and function using primary MSC cultures. Under osteoinductive conditions, BEZ235 strongly promotes osteogenic differentiation, as evidenced by an increase in mineralized matrix production, an upregulation of genes involved in osteogenesis, including bone morphogenetic proteins (BMP2, -4, and -6) and transforming growth factor β1 (TGF-β1) superfamily members (TGFB1, TGFB2, and INHBE), and increased activation of SMAD signaling molecules. In addition, BEZ235 enhances de novo bone formation in calvarial organotypic cultures. Using pharmacologic inhibitors to delineate mechanism, our studies reveal that suppression of mTOR and, to a much lesser extent PI3K p110α, mediates the osteogenic effects of BEZ235. As confirmation, shRNA-mediated knockdown of mTOR enhances osteogenic differentiation and function in SAOS-2 osteoblast-like cells. Taken together, our findings suggest that BEZ235 may be useful in treating PI3K/mTOR-dependent tumors associated with bone loss, such as the hematologic malignancy multiple myeloma.

Bone Morphogenetic Proteins and its Receptors; Therapeutic Targets in Cancer Progression and Bone Metastasis?

Breast and prostate cancer are osteotropic cancers, i.e., carcinomas that have a special predilection to form bone metastases. At postmortem examination, approximately 70% of patients dying of these cancers have evidence of metastatic bone disease. Bone Morphogenetic Proteins (BMPs) were first identified by their ability to induce ectopic bone formation in vivo. Since prostate cancer cells express several BMPs, BMPs have been implicated in the osteoblastic phenotype of bone metastases. In addition to their osteogenic function, BMPs turned out to be multifunctional proteins regulating cell growth, differentiation, migration, and apoptosis in various target cells, including breast and prostate cancer cells. Especially in the last decade, studies have focused on the role of several BMPs in osteotropic cancers. In this review, the role of BMPs, particularly that of BMP7, in breast and prostate cancer will be discussed.

A Helix‐Loop‐Helix Motif within a Genomic DNaseI Footprint of the Runx2 Gene Promoter is Essential for Basal Bone Tissue‐Specific Transcription

Transcriptional mechanisms controlling Runx2 levels are critical for its osteogenic function during osteoblast differentiation. Here we examine the chromatin organization of the genomic Runx2 locus in intact cells. Chromatin immuno‐precipitations were performed with antibodies of modified histones and PCR probes spanning 120 kbp surrounding its bone‐specific (P1) promoter. The results show that a ~10 kbp genomic segment around the P1 promoter interacts with highly acetylated histones H3 and H4 reflecting an open chromatin conformation. Within this acetylated domain, we find a single major DNaseI hypersensitive (DHS) region that resolves into a cluster of smaller sites (−0.4 ~0.0 kbp) and coincides with the promoter segment that controls basal Runx2 transcription in osteoblasts. Genomic DNase I footprinting indicates that the DHS region contains several protein/DNA interaction sites (e.g., at AP1, HLH/E‐box and Runx motifs). HLH/E‐box occupancy of the DHS region persists in all mesenchymal cell types examined. Occupancy of the AP1 binding site occurs only during S phase when Runx2 expression is minimal. Point‐mutation of the HLH/E box causes a dramatic reduction of the P1 promoter activity. We propose that protein/DNA interactions at the AP1 and HLH/E box motifs may control Runx2 transcription when osteoblasts transit into a quiescent and differentiated state. Supported by NIH grant AR039588.

Runx2 regulates diverse gene expression programs to (de‐) sensitize the osteogenic and/or mitogenic responses of osteoblast progenitors and osteosarcoma cells

The osteogenic and cell cycle regulatory functions of Runt‐related transcription factor 2 (Runx2) are perturbed in osteosarcoma cells. We investigated which target genes of Runx2 control proliferation of normal osteoblasts and osteosarcoma cells by Affymetrix expression profiling and ChIP‐on‐chip analysis using NimbleGen arrays. Target gene analysis shows that Runx2 modulates normal osteoblast proliferation by regulating distinct programs of genes involved in G protein‐coupled receptor signaling, sterol/steroid metabolism, the fibroblast growth factor/proteoglycan signaling axis, RNA helicases and heat shock proteins. Strikingly, our data reveal that the gene expression programs controlled by Runx2 in osteoprogenitors are fundamentally different from those in osteosarcomas. Thus cancer‐related perturbations in normal osteoblast growth and differentiation switch the biological function of Runx2 from a growth suppressor to a putative oncoprotein that supports the tumorigenic and/or metastatic potential of osteosarcoma cells.

Phagocytosis of Apoptotic Cells Modulates Mesenchymal Stem Cells Osteogenic Differentiation to Enhance IL-17 and RANKL Expression on CD4+ T Cells

Osteogenic differentiation of mesenchymal stem cells (MSC) is important to homeostatic bone remodeling. Infiltration of mesenchymal progenitor cells to inflamed joints has been reported in collagen-induced arthritis murine model and in patients with rheumatoid arthritis (RA). Therapeutic application of MSC in RA has been suggested and under investigation. However, the underlying mechanisms on what triggers the migration of MSC from bone marrow (BM) to inflamed joints and how MSC acts in the joints remains elusive. As hemopoietic stem cells and MSC act reciprocally and excessive apoptotic cells (AC) are observed in the BM of patients with RA, we hypothesize that AC may alter MSC osteogenic differentiation resulting in bone erosion in RA. In this study, we demonstrated for the first time that MSC were able to phagocytose AC and this phagocytosis enhanced MSC osteogenic differentiation. AC-treated MSC under osteogenic differentiation expressed CXC-chemokine receptor (CXCR)-4 and CXCR5, which might enable them to migrate toward the inflamed joints. In addition, AC-treated MSC secreted interleukin (IL)-8, monocyte chemoattractant protein-1, and RANTES, which might induce chemotaxis of CD4+ T cells to the inflamed joints. Interestingly, by coculturing AC-treated MSC under osteogenic differentiation with CD4+ T cells, T helper (Th) 17 cells development was significantly enhanced and these Th17 cells promoted osteoclasts formation and bone resorption. Furthermore, the induction of Th17 cells was dependent on increased IL-6 production from major histocompatibility complex class II-expressing AC-treated MSC under osteogenic differentiation. This data provide a novel insight on the role of AC in modulating MSC osteogenic differentiation and function in inflammatory bone diseases.

Vitreous cryopreservation of tissue engineered bone composed of bone marrow mesenchymal stem cells and partially demineralized bone matrix

Cryopreservation of tissue engineered products by maintaining their structure and function is a prerequisite for large-scale clinical applications. In this study, we examined the feasibility of cryopreservation of tissue engineered bone (TEB) composed of osteo-induced canine bone marrow mesenchymal stem cells (cBMSCs) and partially demineralized bone matrix (pDBM) scaffold by vitrification. A novel vitreous solution named as VS442 containing 40% dimethyl-sulfoxide (DMSO), 40% EuroCollins (EC) solution and 20% basic culture medium (BCM) was developed. After being cultured in vitro for 8 days, cell/scaffold complex in VS442 was subjected to vitreous preservation for 7 days and 3 months, respectively. Cell viability, proliferation and osteogenic differentiation of cBMSCs in TEB after vitreous cryopreservation were examined with parallel comparisons being made with those cryopreserved in VS55 vitreous solution. Compared with that cryopreserved in VS55, cell viability and subsequent proliferative ability of TEB in VS442 after being rewarmed were significantly higher as detected by live/dead staining and DNA assay. The level of alkaline phosphatase (ALP) expression and osteocalcin (OCN) deposition in VS442 preserved TEB was also higher than those in the VS55 group since 3 days post-rewarm. Both cell viability and osteogenic capability of the VS55 group were found to be declined to a negligible level within 15 days post-rewarm. Furthermore, it was observed that extending the preservation of TEB in VS442 to 3 months did not render any significant effect on its survival and osteogenic potential. Thus, the newly developed VS442 vitreous solution was demonstrated to be more efficient in maintaining cellular viability and osteogenic function for vitreous cryopreservation of TEB over VS55.

Orally Administered Lactoferrin Preserves Bone Mass and Microarchitecture in Ovariectomized Rats

Lactoferrin (LF) is reported to stimulate osteoblast proliferation and inhibit osteoclast activity in bone cell culture. However, the effect of oral LF on bone in osteoporosis needs to be explored. Three-month-old female Sprague-Dawley rats (n = 70) were assigned to the following groups: sham-operated, ovariectomized (OVX) untreated, OVX + bovine serum albumin (BSA; 85 mg/kg body weight), OVX + LF (0.85 mg/kg, 8.5 mg/kg, and 85 mg/kg body weight), and OVX + 17β-estradiol (E2; 10 μg/kg body weight). After 3 mo of treatment, E2 completely prevented the OVX-induced bone loss. OVX rats treated with LF were protected against the OVX-induced reduction of bone volume, trabecular number, and thickness, and the elevation of trabecular separation was prevented. LF also increased bone mineral density and increased the parameters of mechanical strength at 8.5- and 85-mg/kg doses. Greater bone formation and reduced bone resorption, as assessed by biochemical markers of bone remodeling, occurred in rats administered LF. LF at 8.5- and 85-mg/kg concentrations caused a significant decrease in serum calcium, but this reduction did not occur in rats fed 0.85 mg/kg LF. In addition, serum tumor necrosis factor-α and interleukin-6 production were suppressed and serum calcitonin was elevated significantly in LF-fed rats at all 3 doses. These findings indicated that oral LF not only preserved bone mass but also improved bone microarchitecture. The absorption of LF peptides and their effects on bone cells could to some extent account for the osteogenic function of oral LF.

Treatment of early osteonecrosis of femoral head by core decompression and autograft of mesenchymal stem cell cultured in vitro under arthroscopy: experiment with rabbits

To observe the therapeutic effect of core decompression and autograft of mesenchymal stem cells (MSCs) cultured in vitro under arthroscopy in treatment of early osteonecrosis of femoral head. Thirty-six rabbits were made into models of early osteonecrosis of femoral head of the right leg, and then randomly divided into 3 equal groups: Group A group as control group, Group B group undergoing simple core decompression under arthroscopy by clearing out of necrotic bone, and Group C undergoing core decompression and implantation of MSCs. Four and 8 weeks later 6 rabbits from each group underwent roentgenography and then were killed with their femoral heads taken out to undergo histological examination. Four weeks later new bone was evident in the implantation field of Group C while fibrous tissues were evident in the femoral head of Group B. Pathological examination showed regular shape and normal density of femoral head in Group C and irregular shape and low density of femoral head in Group B. X-ray photograph 8 weeks after treatment showed that the femoral head articular surface of Group A collapsed, uneven density and defect of spongy bone were seen in Group B, and the shape of femoral head of Group C was regular without collapse and the original defect had been repaired. With strong osteogenic function, MSCs improve the repair of ischemic necrosis of femoral head and can be used in treatment of early osteonecrosis of femoral head via core decompression and autograft of MSCs cultured in vitro under arthroscopy.

(−)-Epigallocatechin gallate inhibition of osteoclastic differentiation via NF-κB

People who regularly drink tea have been found to have a higher bone mineral density (BMD) and to be at less risk of hip fractures than those who do not drink it. Green tea catechins such as (−)-epigallocatechin gallate (EGCG) have been reported to increase osteogenic functioning in mesenchymal stem cells. However, its effect on osteoclastogenesis remains unclear. In this study, we investigated the effect of EGCG on RANKL-activation osteoclastogenesis and NF-κB in RAW 264.7, a murine preosteoclast cell line. EGCG (10–100 μM) significantly suppressed the RANKL-induced differentiation of osteoclasts and the formation of pits in murine RAW 264.7 cells and bone marrow macrophages (BMMs). EGCG appeared to target osteoclastic differentiation at an early stage but had no cytotoxic effect on osteoclast precursors. In addition, it significantly inhibited RANKL-induced NF-κB transcriptional activity and nuclear translocation. We conclude that EGCG inhibits osteoclastogenesis through its activation of NF-κB.

Human bone derived cell culture on PLGA flat sheet membranes of different lactide:glycolide ratio

Providing a scaffold that can supply nutrients on a large scale (several cubic centimeters) is the key to successfully regenerating vascularized tissue: biodegradable membranes are a promising new scaffold suited to this purpose. Poly(lactic-co-glycolic-acid) (PLGA) flat sheet membranes of different lactide:glycolide ratios, prepared by phase inversion using 1-methyl-2-pyrrolidinone (NMP) as the solvent and water as the nonsolvent, were compared by assessing attachment, proliferation and osteogenic function of human bone derived cells (HBDC). Three different lactide:glycolide ratios, 50:50, 75:25, and 100:0, were compared to tissue culture polystyrene (TCPS). For attachment, 50:50 and 75:25 had similar numbers to TCPS but 100:0 had significantly fewer cells than TCPS. 50:50 and 75:25 had significantly lower HBDC numbers after 7 days but 100:0 had similar numbers compared to TCPS. For proliferation the cell number on the membranes were similar to each other. After 3 weeks, osteoblastic function of the HBDC, shown by mineralization and alkaline phosphatase activity, was present but was significantly lower compared to the TCPS control but similar when the membranes were compared. PLGA membranes fabricated from a range of ratios support HBDC culture so the optimum scaffold composition can be selected based on other factors, such as degradation rate.

Altered fracture callus formation in chondromodulin-I deficient mice

Chondromodulin-I (Chm-I) is a glycoprotein that stimulates the growth of chondrocytes and inhibits angiogenesis in vitro. Mice lacking the Chm1 gene show abnormal bone metabolism and pathological angiogenesis in cardiac valves in the mature stage although they develop normally without aberrations in endochondral bone formation during embryogenesis or in cartilage development during growth. These findings indicate that Chm-I is critical under conditions of stress such as bone repair through endochondral ossification of a fracture callus. We carried out the present study to examine the expression and role of Chm-I in bone repair using a stabilized tibial fracture model, and compared fracture healing in Chm1 knockout ( Chm1 −/−) mice with that in wild-type mice. Chm-I mRNA and protein localized in the external cartilaginous callus in the reparative phase of fracture healing. Radiological examination showed a delayed union in Chm1 −/− mice although the fracture site was covered with both external and internal calluses. Chm1 null mutation reduced external cartilaginous callus formation as judged by marked decrease of type X collagen alpha 1 (Col10a1) expression and the total amount of cartilage matrix. Interestingly, the majority of chondrocytes in the periosteal callus failed to differentiate into mature chondrocytes in Chm1 −/− mice, while the hypertrophic maturation of chondrocytes between the cortices was not affected. These results suggest that Chm-I is involved in hypertrophic maturation of periosteal chondrocytes. Although a direct effect of Chm-I on bones is still unclear, bony callus formation was increased while external cartilaginous callus decreased in Chm1 −/− mice. We conclude that in the absence of Chm1, predominant primary bone healing occurs due to an indirect effect induced by reduction of cartilaginous callus rather than to a direct effect on osteogenic function, resulting in a delayed union.

In Vitro Investigation of Orthopedic Titanium-Coated and Brushite-Coated Surfaces Using Human Osteoblasts in the Presence of Gentamycin

Anti-infective coatings have been developed to protect the surfaces of cementless implants from bacterial colonization that is known to be a prerequisite for device-related infection. The aim of this study is to investigate the effect of brushite-coated arthroplasty surfaces on human osteoblasts and to evaluate the impact of concomitant exposure to gentamycin. We cultured human osteoblasts (hFOB 1.19) on brushite-coated and uncoated titanium alloy in the presence of gentamycin and analyzed cell function and vitality. Our results show that brushite-coated titanium alloy surfaces supported the function of osteoblasts and the expression of extracellular matrix even in the presence of highly dosed gentamycin. Brushite-coated titanium alloy surfaces supported osteogenic function, indicating that this coating could enhance implant osteointegration in vivo. Concomitant exposure to gentamycin slightly decreased osteoblastic activity in vitro, suggesting that there might also be negative effects in vivo. However, in vivo studies are necessary to validate these in vitro findings.